HK1226719A1 - Isochromene derivatives as phosphoinositide 3-kinases inhibitors - Google Patents
Isochromene derivatives as phosphoinositide 3-kinases inhibitors Download PDFInfo
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- HK1226719A1 HK1226719A1 HK16114804.9A HK16114804A HK1226719A1 HK 1226719 A1 HK1226719 A1 HK 1226719A1 HK 16114804 A HK16114804 A HK 16114804A HK 1226719 A1 HK1226719 A1 HK 1226719A1
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Description
Technical Field
The present invention relates to compounds that inhibit inositol phosphate 3-kinase (PI 3K, infra); in particular, the present invention relates to compounds which are isochromene (isochromene) derivatives, to processes for preparing such compounds, to pharmaceutical compositions containing them and to their therapeutic use.
More specifically, the compounds of the invention are inhibitors of the activity or function of class I PI3K, and more specifically they are inhibitors of the activity or function of the PI3K α, PI3K β, PI3K and/or PI3K γ isoforms of class I PI 3K. Accordingly, the compounds of the present invention may be used to treat a number of disorders associated with the PI3K enzyme mechanism, such as respiratory diseases including asthma, Chronic Obstructive Pulmonary Disease (COPD) and cough; allergic diseases including allergic rhinitis and atopic dermatitis; autoimmune diseases, including rheumatoid arthritis and multiple sclerosis; inflammatory diseases, including inflammatory bowel disease; cardiovascular diseases including thrombosis and atherosclerosis; hematological malignancies; cystic fibrosis; neurodegenerative diseases; pancreatitis; multiple organ failure; renal disease; platelet aggregation; cancer; sperm motility; organ transplantation and particularly transplant rejection; transplant rejection; lung injury; and pain, including pain associated with rheumatoid arthritis or osteoarthritis, back pain, systemic inflammatory pain, postherpetic neuralgia (postherpetic neuralgia), diabetic neuropathy, inflammatory neuropathic pain, trigeminal neuralgia, and central pain.
Background
In biochemistry, kinases are a type of enzyme that transfers phosphate groups from high energy donor molecules, such as ATP, to specific substrates, a process known as phosphorylation. In particular, the PI3K enzyme is a lipase kinase that phosphorylates Phosphoinositols (PI) on the 3' -hydroxyl group of the inositol ring (Panayotou et al Trends Cell Biol 2: 358-60 (1992)). It is well known that PI located on the plasma membrane may act as a second messenger in the signaling cascade via docking proteins containing pleckstrin-homology (PH), FYVE, PX and other phospholipid-binding domains (Vanhaaesebroeck B et al, Annu. Rev. biochem 70, 535-602, 2001; Katso R et al, Annu. Rev. cell Dev. biol.17, 615-675, 2001).
Thus, PI can function as a second messenger in many cellular processes, including signal transduction, membrane trafficking and transport regulation, cytoskeletal organization, cell survival and death, and many additional functions.
PI can bind to the lipid bilayer of the cell membrane through two fatty acids linked to the cytosolic inositol ring via glycerophosphate linkers. The PI inositol ring can be phosphorylated by the PI3K enzyme, resulting in the regulation of cell growth, survival and proliferation. For this reason, PI3K phosphorylation by the enzyme PI is one of the most relevant signal transduction events associated with mammalian cell surface receptor activation (Cantley LC, Science 296, 1655-7, 2002; Vanhaaesebroeck B et al, Annu. Rev. biochem 70, 535-602, 2001).
PI3K enzymes have been classified into 3 classes based on sequence homology, structure, binding partners, activation patterns, and substrate preference: class I PI3K, class II PI3K, and class III PI3K (Vanhaaesebroeck B et al, exp. cell Res.253(1), 239-54, 1999; and Leslie NR et al, chem. Rev.101(8), 2365-80, 2001).
Class I PI3K converts phosphoinositide- (4,5) -diphosphate (PI (4,5) P2) to phosphoinositide- (3,4,5) -triphosphate (PI (3,4,5) P3), which functions as a second messenger. The signaling cascade for activation due to increased intracellular levels of PI (3,4,5) P3 is negatively regulated by the action of 5 '-specific and 3' -specific phosphatases (Vanhaaesebroeck B et al, Trends biochem. Sci.22(7), 267-72, 1997; Katso R et al, Annu. Rev. cell Dev. biol.17, 615-75, 2001; and Toker A, cell. mol. Life Sci.59(5), 761-79, 2002).
Class II PI3K enzymes are the most recently identified type of PI3K and their exact function remains unclear.
Class III PI3K enzymes consist of single family members that are structurally related to class I PI3K enzymes and are apparently important in endocytosis and vesicle transport. However, there is some evidence that class III PI3K may be involved in immune cell processes such as phagocytosis and Toll-like receptor (TLR) signaling.
Class I PI3K enzymes may be further classified into class IA and class IB based on their activation mechanism.
In more detail, class IA PI3K enzymes comprise 3 closely related isoforms: PI3K α, PI3K β and PI3K, whereas class IB contains only the PI3K γ isoform. These enzymes are composed of a group of 4 types called p 110: heterodimers consisting of catalytic subunits of alpha (α), beta (β), delta () and gamma (γ) isoforms, which are related in composition to regulatory subunits. The first 2 p110 isoforms (α and β) are ubiquitously expressed and involved in cell differentiation and proliferation. Therefore, PI3K α and PI3K β enzymes are widely studied as targets for the development of new chemotherapeutic agents.
In addition, p110 and p110 γ isoforms are predominantly expressed in leukocytes and are important in immune response activation, such as leukocyte migration, B and T cell activation, and mast cell degranulation. Therefore, PI3K and PI3K γ isoforms are strongly associated with inflammatory respiratory diseases.
Recently, inhibitor derivatives of the PI3K enzyme known in the art can generally inhibit the isoforms (α, β, and γ isoforms), and they can contribute to the individual effects of the specific isoforms in various diseases.
Thus, specific activity assays for class IA inhibitors of one specific PI3K α, PI3K β, PI3K and PI3K γ isoform have been developed to identify suitable properties for treating disorders associated with the PI3K enzyme mechanism. Such disorders may include, for example, respiratory diseases selected from idiopathic chronic cough, cough-variant asthma, cough associated with breast tumor or lung cancer, viral or post-viral cough, Upper Airway Cough Syndrome (UACS), or post-nasal drip cough; or cough associated with acid and non-acid gastroesophageal reflux disease, asthma, chronic bronchitis, Chronic Obstructive Pulmonary Disease (COPD), interstitial lung disease, Idiopathic Pulmonary Fibrosis (IPF), congestive heart disease, sarcoidosis, infection (e.g., pertussis), viral infection, including viral respiratory infection and viral respiratory disease exacerbation; non-viral respiratory infections including aspergillosis and leishmaniasis; allergic diseases including allergic rhinitis and atopic dermatitis; autoimmune diseases, including rheumatoid arthritis and multiple sclerosis; inflammatory diseases, including inflammatory bowel disease; cardiovascular diseases including thrombosis and atherosclerosis; hematological malignancies; neurodegenerative diseases; pancreatitis; multiple organ failure; renal disease; platelet aggregation; cancer; sperm motility; graft rejection; transplant rejection; lung injury; and pain, including pain associated with rheumatoid arthritis or osteoarthritis, back pain, systemic inflammatory pain, post-herpetic neuralgia, diabetic neuropathy, inflammatory neuropathic pain (trauma), trigeminal neuralgia, and central pain.
Given the number of pathological responses mediated by the PI3K enzyme, there is a continuing need for PI3K enzyme inhibitors that can be used to treat a number of disorders. The present invention therefore relates to novel compounds which are inhibitors of the PI3K α, PI3K β, PI3K and PI3K γ isoforms of the class I PI3K enzyme, which may generally have desirable therapeutic properties for the reasons set out above.
In particular, the compounds of the invention may have selectivity for the isoform of PI3K enzyme or for gamma and isoform of PI3K enzyme far beyond other isoforms of the same enzyme.
Brief description of the invention
The invention relates to compounds of formula (I),
r, R therein1、R2、R3、R4、R5CY, Z, m, n and p are as reported in the detailed description of the invention below, said compounds acting as inhibitors of phosphoinositide 3-kinase; the present invention relates to processes for their preparation, pharmaceutical compositions comprising them alone or in combination with one or more active ingredients in admixture with one or more pharmaceutically acceptable carriers.
The invention also provides suitable devices for delivering pharmaceutical compositions of the compounds of the invention.
In one aspect, the invention provides the use of a compound of the invention in the manufacture of a medicament.
In another aspect, the invention provides the use of a compound of the invention for the preparation of a medicament for the prevention and/or treatment of any disease characterized by hyperactivity of phosphoinositide-3-kinase (PI3K) and/or in which inhibition of PI3K activity is desired, and in particular by selective inhibition or gamma enzyme isoforms exceeding the alpha and beta isoforms.
The present invention also provides a method for the prevention and/or treatment of any disease for which PI3K enzyme inhibition is desired, comprising administering to a patient in need of such treatment a therapeutically effective amount of a compound of the present invention.
In particular, the compounds of the present invention may be administered alone or in combination with additional active ingredients in order to prevent and/or treat respiratory diseases characterized by inflammatory airway obstruction, e.g., cough, asthma, COPD and IPF.
It will be apparent to those skilled in the art that the compounds of the present invention may also be represented by the following general formula (I')
R, R therein1、R2、R3、R4、R5CY, Z, m, n and p are as reported in the detailed description of the invention, the bondsIs a double bond, and the M group is an arylene-diyl group, in particular a 1, 2-phenylene-diyl group.
Alternative values for the M group may be 3,4-, 2,3-, or 4, 5-thiophenediyl, 4, 5-thiazolediyl; saturated or partially saturated analogs thereof, and the like.
Compounds having alternative values for such M groups are, for example, 5- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-7H-thieno [2,3-c ] pyran-7-one; 6- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -7-phenyl-4H-thieno [3,2-c ] pyran-4-one; 6- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -7-phenyl-4H-pyrano [3,4-d ] thiazol-4-one.
In the description of the synthetic routes used hereinafter to prepare the compounds of the invention (schemes 1-9), the M group may be represented by formula (I ") above, where M refers to a 1, 2-phenylene-diyl group.
Detailed Description
The present invention relates to a class of compounds which act as phosphoinositide 3 kinase (PI3K) inhibitors.
The compounds inhibit the activity or function of class I PI3K, and more specifically, they are inhibitors derivatives of the activity or function of PI3K α, PI3K β, PI3K γ, and/or PI3K isoforms of class I PI 3K.
The present invention relates to compounds of formula (I):
wherein:
each R, when present, is independently selected from:
- OR6；
- SR6；
- S(O)q-R8；
- NR10R11；
-a halogen;
- (C1-C6) An alkyl group;
- (C1-C6) A haloalkyl group;
- (C3-C7) A cycloalkyl group;
- (C5-C7) A cycloalkenyl group;
- (C2-C6) An alkenyl group;
- (C2-C6) An alkynyl group;
-substituted or unsubstituted aryl; and
-substituted or unsubstituted heteroaryl;
R1and R2Are both H or combine to form an oxo group (═ O);
R3and R4The same or different, at each occurrence is independently selected from:
- H；
- (C1-C6) An alkyl group; and
- (C1-C6) A haloalkyl group;
R5selected from:
- H；
- OR7；
- SR7；
- S(O)q-R9；
-a halogen;
- NR12R13；
- CN；
- C(O)NR12R13；
- COOR14；
- (C1-C6) An alkyl group;
- (C1-C6) A haloalkyl group;
- (C1-C6) A hydroxyalkyl group;
- (C1-C6) An aminoalkyl group;
- (C3-C7) A cycloalkyl group;
aryl (C)1-C3) An alkyl group;
- (C5-C7) A cycloalkenyl group;
- (C2-C6) An alkenyl group;
- (C2-C6) An alkynyl group;
- (C2-C6) An aminoalkynyl group;
-substituted or unsubstituted (C)3-C6) A heterocycloalkyl group;
-substituted or unsubstituted aryl; and
-substituted or unsubstituted heteroaryl;
R6、R7and R14The same or different, at each occurrence is independently selected from:
- H；
- (C1-C6) An alkyl group;
- (C1-C6) A haloalkyl group;
- (C1-C6) A hydroxyalkyl group;
- (C1-C6) An aminoalkyl group;
aryl (C)1-C6) An alkyl group;
- (C1-C6) An alkanoyl group;
-an arylcarbonyl group; and
aryl (C)2-C4) An alkanoyl group;
R8and R9The same or different, at each occurrence is independently selected from
- (C1-C6) An alkyl group;
- (C1-C6) A haloalkyl group;
- (C1-C6) A hydroxyalkyl group;
- (C1-C6) An aminoalkyl group;
-substituted or unsubstituted aryl;
-substituted or unsubstituted heteroaryl; and
- NR12R13；
R10、R11、R12and R13The same or different, at each occurrence, is independently selected from H, (C)1-C6) Aminoalkyl radical, (C)1-C6) Hydroxyalkyl and (C)1-C6) An alkyl group; or R10And R 11Or R12And R13May form, together with the nitrogen atom to which they are attached, a 5-to 6-membered heterocyclic group;
z, when present, is an atom or group independently selected each time from O, NH, C (O), NHC (O), C (O) NH, S (O) and S (O)2；
m is 0 or 1;
n is 1 or 2;
p is 0 or an integer from 1 to 3;
q is 1 or 2;
cy is selected from:
-substituted or unsubstituted (C)3-C6) A heterocycloalkyl group;
-substituted or unsubstituted aryl; and
-substituted or unsubstituted heteroaryl;
or a pharmaceutically acceptable salt thereof.
The term "pharmaceutically acceptable salt" as used herein refers to derivatives of the compounds of formula (I) wherein the parent compound is suitably modified by conversion of any free acid or basic group, if present, into the corresponding addition salt with any base or acid which would normally be expected to be pharmaceutically acceptable.
Suitable examples of such salts may thus include inorganic or organic acid addition salts of basic residues such as amino groups as well as inorganic or organic base addition salts of acidic residues such as carboxyl groups.
The cations of the inorganic bases which may suitably be used to prepare salts within the scope of the present invention comprise ions of alkali or alkaline earth metals, such as potassium, sodium, calcium or magnesium.
Those obtained by reacting a main compound functioning as a base with an inorganic acid or an organic acid to form a salt include, for example, hydrochloride, hydrobromide, sulfate, phosphate, methanesulfonate, camphorsulfonate, acetate, oxalate, maleate, fumarate, succinate and citrate.
Definition of
The term "halogen atom" as used herein includes fluorine, chlorine, bromine and iodine, preferably chlorine or fluorine.
Term "(C)1-Cx) Alkyl ", wherein x is an integer greater than 1, refers to a straight or branched chain alkyl group wherein the number of carbon atoms of the composition is in the range of 1-x. Particularly preferred alkyl groups are methyl, ethyl, n-propyl, isopropyl and tert-butyl.
Expression "(C)1-Cx) Haloalkyl "means as defined above" (C)1-Cx) An "alkyl" group in which one or more hydrogen atoms are replaced by one or more halogen atoms, which may be the same or different from each other.
Said (C)1-Cx) Examples of haloalkyl groups thus may include halogenated, polyhalogenated and perhalogenated alkyl groups, such as trifluoromethyl or difluoromethyl groups.
By analogy, the term "(C)1-Cx) Hydroxyalkyl "or" (C)1-Cx) Aminoalkyl "means as defined above" (C)1-Cx) An alkyl "group in which one or more hydrogen atoms are replaced by one or more hydroxyl (OH) or amino groups, respectively.
In this specification, unless otherwise provided, the definition of aminoalkyl embraces one or more (NR)10R11) A substituted alkyl group.
With reference to the substituents R as defined above10、R11、R12And R13In this context, it may be further explained when R is 10And R11Or R12And R13When taken together with the nitrogen atom to which they are attached to form a 5-to 6-membered heterocyclic group, at least one other ring carbon atom on the heterocyclic group may be replaced by at least one heteroatom or hetero-group (e.g., N, NH, S, or O) or may carry an-oxo (═ O) substituent group. The heterocyclic group may further be optionally substituted at available points in the ring, i.e. at carbon atoms or at available heteroatoms or hetero-groups for substitution. Examples of such heterocyclic groups are thus 1-pyrrolidinyl, 1-piperidinyl, 1-piperazinyl, 4-morpholinyl, piperazin-4-yl-2-one, 4-methylpiperazin-1-yl.
Term "(C)3-Cy) Cycloalkyl "wherein y is an integer greater than 3, refers to a saturated cyclic hydrocarbon group containing 3-y ring carbon atoms. Non-limiting examples include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cycloheptyl.
The term "aryl (C)1-Cx) Alkyl "refers to an aryl ring attached to a straight or branched alkyl group with a number of constituent carbon atoms in the range of 1-x, such as phenylmethyl, phenylethyl or phenylpropyl.
Expression of derivation "(C)3-Cz) Heterocycloalkyl "means a saturated or partially unsaturated monocyclic ring (C)3-Cz) Cycloalkyl groups, wherein z is an integer greater than 3, wherein at least one ring carbon atom is replaced by at least one heteroatom or hetero-group (e.g., N, NH, S, or O). (C) 3-Cz) Non-limiting examples of heterocycloalkyl groups are represented by the following: pyrrolidinyl, imidazolidinyl, thiazolidinyl, piperazinyl, piperidinyl, morpholinyl, thiomorpholinyl, dihydro-or tetrahydro-pyridinyl, tetrahydropyranyl, pyranyl, 2H-or 4H-pyranyl, dihydro-or tetrahydrofuranyl, 1, 3-dioxolan-2-yl groups, and the like. As defined above (C)3-Cz) The heterocycloalkyl group may optionally be further on available points in the ring, i.e. carbon atoms, or may be available as a substituted heteroatom or hetero-groupIs substituted. For example, a tetrahydro-pyridinyl group, when further substituted, may be substituted on the-NH group, such as in the following examples: 1-benzyl-1, 2,3, 6-tetrahydropyridin-4-yl, 1- (cyclopropylmethyl) -1,2,3, 6-tetrahydropyridin-4-yl, 1-acetyl-1, 2,3, 6-tetrahydropyridin-4-yl, 1- (pyridin-4-ylmethyl) -1,2,3, 6-tetrahydropyridin-4-yl.
Term "(C)2-Cx) Alkenyl "refers to a straight or branched, conjugated or unconjugated carbon chain having one or more double bonds in either the cis or trans configuration, where the number of atoms is 2-x.
By analogy, the term "(C)5-Cy) Cycloalkenyl ", wherein y is an integer greater than 5, refers to cyclic hydrocarbon groups containing from 5 to y ring carbon atoms and one or two double bonds, wherein cycloalkenyl may be further optionally substituted with one or more groups such as amino groups.
Term "(C)2-Cx) Alkynyl "refers to a straight or branched carbon chain having one or more triple bonds, wherein the number of atoms is 2-x.
By analogy, the term "(C)2-Cx) Aminoalkynyl "means as defined above" (C)2-Cx) Alkynyl "groups wherein one or more hydrogen atoms are replaced by one or more amino groups, and wherein the amino groups may further optionally be replaced by one or more (C)1-C6) Alkyl groups.
The expression "aryl" refers to a monocyclic, bicyclic or tricyclic ring system having 5 to 20, preferably 5 to 15 ring atoms, wherein at least one ring is aromatic.
The expression "heteroaryl" refers to a monocyclic, bicyclic or tricyclic ring system having 5 to 20, preferably 5 to 15 ring atoms, wherein at least one ring is aromatic and wherein at least one ring atom is a heteroatom or a heteroaromatic group (e.g. N, NH, S or O).
Examples of suitable aryl or heteroaryl monocyclic systems include, for example, phenyl, thienyl (also referred to herein as thiophenyl or thiophenyl), pyrrolyl, pyrazolyl, imidazolyl, isoxazolyl, oxazolyl, isothiazolyl, thiazolyl, pyridyl, pyrimidinyl, pyrazinyl, furanyl groups, and the like.
Examples of suitable aryl or heteroaryl bicyclic ring systems include naphthyl, biphenylene, purinyl, pteridinyl, pyrazolopyrimidinyl, benzotriazolyl, quinolinyl, isoquinolinyl, indolyl, isoindolyl, benzothienyl, benzodioxinyl, dihydrobenzodioxinyl, indenyl, dihydro-indenyl, dihydrobenzodiazepine, benzoxazinyl, and the like.
Examples of suitable aryl or heteroaryl tricyclic ring systems include fluorenyl groups and benzofused derivatives of the heteroaryl bicyclic ring systems exemplified above.
Term "(C)1-Cx) Alkanoyl "refers to an alkylcarbonyl group (e.g., (C)1-Cx) Alkyl (CO) where x is an integer greater than 1), where the radical "alkyl" has the meaning defined above. Non-limiting examples include acetyl, propionyl, butyryl.
The expression "arylcarbonyl" refers to an aryl- (CO) -group, wherein the group "aryl" has the meaning defined above. A non-limiting example is represented by benzoyl.
The term "aryl (C)2-Cx) Alkanoyl "refers to aryl (C)2-Cx) Alkylcarbonyl groups, wherein x is an integer greater than 2, wherein aryl and alkyl have the meaning defined above. Non-limiting examples are represented by phenylacetyl, phenylpropionyl or phenylbutyryl groups;
by analogy, the expression "heteroaryl (C)1-Cx) Alkyl "and" (C)3-Cy) Cycloalkyl (C)1-Cx) Alkyl "means a radical substituted by one or more heteroaryl or (C) as defined above3-Cy) Cycloalkyl group substituted "(C)1-Cx) Alkyl radical”。
Aryl radical (C)1-C6) Examples of alkyl groups include, for example, the phenylmethyl group referred to herein as benzyl. Heteroaryl (C)1-C6) Examples of alkyl groups include, for example, pyridin-4-ylmethyl. (C) 3-C7) Cycloalkyl (C)1-C6) Examples of alkyl groups include, for example, cyclopropylmethyl.
The expression "ring system" as used herein refers to a monocyclic or bicyclic ring system, which may be saturated, partially saturated or unsaturated, such as aryl, (C)3-C7) Cycloalkyl group, (C)3-C6) Heterocycloalkyl or heteroaryl.
When R is present, as will be apparent to those skilled in the art3And R4In the meantime, the compounds of formula (I) may comprise at least one stereocentre, i.e. represented by the carbon atom bearing the asterisk (—) In (IA), and may thus exist as optical stereoisomers.
If the compounds of the invention have at least one stereocenter, they can thus be present as enantiomers. If the compounds of the invention have two or more stereogenic centers, they may also exist as diastereomers. It is to be understood that all such single enantiomers, diastereomers and mixtures thereof in any proportion are intended to be encompassed by the scope of the present invention. The absolute configuration (R) or (S) of the carbon (. lambda.) is assigned based on the Cahn-Ingold-Prelog nomenclature based on the preference of the groups.
Atropisomers are stereoisomers resulting from the hindered rotation about a single bond, where the steric tension barrier to addressing is high enough to enable the isolation of conformers (Bringmann G et al, Angew. Chemie intermediates Ed.44(34), 5384-5427, 2005. doi: 10.1002/anie.200462661).
Oki is defined as a conformational isomer that interconverts at a half-life greater than 1000 seconds at a given temperature (Oki M, Topics in Stereochemistry 14, 1-82, 1983).
Atropisomers differ from other chiral compounds in that in many cases they may be in thermal equilibrium, while other forms of chiral isomerization are often only possible chemically.
It is possible to separate atropisomers by chiral resolution methods such as selective crystallization. In atropiso-enantioselective or atropiselective syntheses, one atropisomer is formed with loss of the other. The enantioselective synthesis can be carried out by using a chiral auxiliary, such as a Corey Bakshi Shibata (CBS) catalyst, an asymmetric catalyst derived from proline; or by a process based on thermodynamic equilibrium when the isomerization reaction favors one atropisomer over another.
Racemic forms of the compounds of formula (I) and individual atropisomers (substantially free of their corresponding enantiomers) and stereoisomerically enriched atropisomer mixtures are included within the scope of the present invention.
In a preferred embodiment, the present invention relates to a compound of formula (IA) as defined above, wherein n ═ 1, R 3Has the same meaning as above except for H, R4Is H and the absolute configuration of the chiral carbon (×) is (R).
In another embodiment, the preferred configuration of carbon (×) is (S).
In a preferred embodiment, the compounds of formula (I) as described in the present invention are present as a mixture of diastereomers.
A first group of preferred compounds are compounds of formula (I) wherein:
R1and R2Are both H or combine to form an oxo group (═ O);
R3is selected fromH and (C)1-C6) An alkyl group;
R4is H;
R5is selected from H; halogen; OR (OR)7(ii) a Aryl radical (C)1-C3) An alkyl group; (C)5-C7) Cycloalkenyl group, (C)2-C6) Alkynyl, (C)2-C6) Aminoalkynyl, substituted or unsubstituted (C)3-C6) A heterocycloalkyl group; substituted or unsubstituted aryl and substituted or unsubstituted heteroaryl;
R、R7m, n, p, Z and CY are as defined above.
A more preferred group of compounds are compounds of formula (I) wherein:
p is 0 or 1;
r is absent or selected from halogen and (C)1-C6) An alkyl group;
R1and R2Are both H or combine to form an oxo group (═ O);
R3selected from H, methyl, ethyl and propyl;
R4is H;
R5is selected from H; fluorine; bromine; a phenyl group; a phenyl methyl group; 2-, 3-or 4-pyridyl; 5-thiazolyl; 2-,3-, 4-or 5-thienyl, 1H-pyrazol-4-yl, 2-,4-, 5-or 6-pyrimidinyl, cyclohexenyl, prop-1-ynyl, 1,2,3, 6-tetrahydropyridin-4-yl, 1,2,5, 6-tetrahydropyridin-3-yl, 8-azabicyclo [3.2.1 ]Oct-2-en-3-yl and 3, 6-dihydro-2H-pyran-4-yl, optionally substituted with one or more groups selected from halogen, (C)1-C6) Alkyl, OR7、-S(O)q-R9、-C(O)NR10R11、COOR14、(C1-C6) Hydroxyalkyl, substituted or unsubstituted (C)1-C6) Aminoalkyl radical, (C)1-C6) Alkanoyl, substituted or unsubstituted (C)3-C6) Heterocycloalkyl, aryl (C)1-C6) Alkyl, heteroaryl (C)1-C6) Alkyl, (C)3-C7) Cycloalkyl (C)1-C6) Alkyl and NR10R11；
R7、R9、R10、R11、R14M, n, q, Z and CY are as defined above.
An even more preferred group of compounds are compounds of formula (I) wherein:
p is 0;
r is absent;
R1and R2Are both H or combine to form an oxo group (═ O);
R3selected from H, methyl, ethyl and propyl;
R4is H;
R5is selected from phenyl; a phenyl methyl group; 2-, 3-or 4-pyridyl; 5-thiazolyl, 2-,3-, 4-or 5-thienyl, 1,2,3, 6-tetrahydropyridin-4-yl and 3, 6-dihydro-2H-pyran-4-yl, optionally substituted with one or more groups selected from fluoro, bromo, methyl, methoxy, amino, dimethylamino, 4-morpholinosulfonyl, 4- (2-morpholinoethoxy), 4-morpholinomethyl and 4-piperazinylmethyl; piperidin-1-ylmethyl, 4-methylpiperazin-1-carbonyl, (2- (dimethylamino) ethyl) -carbonyl, acetyl, phenylmethyl, phenylmethoxy-carbonyl, 4,5, 5-tetramethyl-1, 3-dioxolan-2-yl, pyrrolidin-1-ylmethyl, bis (2-hydroxyethyl) aminomethyl, hydroxymethyl, dimethylaminomethyl, (dimethylamino) propyl, 4- (2-hydroxyethyl) piperazin-1-yl) methyl, piperazin-2-on-1-ylmethyl, cyclopropylmethyl, hydroxycarbonyl, pyridin-4-ylmethyl;
m, n, Z and CY are as defined above.
A second group of preferred compounds are compounds of formula (I) wherein:
R1and R2Are both H or combine to form an oxo group (═ O);
R3selected from H, methyl, ethyl and propyl;
R4is H;
R5is selected from H; halogen; OR (OR)7(ii) a Aryl radical (C)1-C3) An alkyl group; substituted or unsubstituted (C)3-C6) A heterocycloalkyl group; substituted or unsubstituted aryl and substituted or unsubstituted heteroaryl;
z, when present, is an atom or group independently selected each time from O, NH, C (O), NHC (O), C (O) NH, S (O) and S (O)2；
CY is heteroaryl selected from 7H-purin-7-yl; 9H-purin-9-yl; 9H-purin-6-yl; 1H-pyrazolo [3,4-d]Pyrimidin-1-yl; 1H-pyrazolo [3,4-d]Pyrimidin-4-yl; 2H-pyrazolo [3,4-d]Pyrimidin-2-yl; 2-,4-, 5-or 6-pyrimidinyl; and 2-pyrazinyl, pyrrolo [2,3-d ]]Pyrimidin-7-yl, pyrazolo [1,5-a ]]Pyrimidin-3-yl, pyrido [3,2-d]Pyrimidin-4-yl, pyrido [2,3-d ]]Pyrimidin-8-yl-5-one, thieno [3,2-d]Pyrimidin-4-yl, thieno [2,3-d ]]Pyrimidin-4-yl all of which are optionally substituted with one or more groups selected from halogen, (C)1-C6) Alkyl, (C)1-C6) Haloalkyl, CN, NR10R11Optionally substituted aryl and optionally substituted heteroaryl selected from the group consisting of phenyl, 1H-indazol-4-yl, 1H-indazol-5-yl, 1H-indazol-6-yl, 2-,3-,4-,5-, 6-pyridyl, 1H-1H-pyrazol-3-yl, pyrazol-4-yl, pyrazol-5-yl, pyrazin-2-yl, pyrimidin-5-yl, pyridazin-4-yl and 2-,4-, 5-thiazolyl;
R、R7、R10、R11M, n, p are as defined above.
A second group of more preferred compounds are compounds of formula (I) wherein:
R1and R2Combine to form an oxo group (═ O);
R3selected from H, methyl or ethyl;
R4is H;
R5is selected from H; halogen selected from fluorine and bromine; aryl which is phenyl; aryl being phenylmethyl (C)1-C3) An alkyl group; selected from 2-, 3-or 4-pyridyl; 5-thiazolyl, 2-,3-, 4-or 5-thienyl heteroaryl; (C) selected from 1,2,3, 6-tetrahydropyridin-4-yl and 3, 6-dihydro-2H-pyran-4-yl3-C6) Heterocycloalkyl and 4-cyclohexenyl, all of which are optionally substituted with one or more groups selected from fluoro, bromo, methyl, methoxy, dimethylamino, morpholinosulfonyl, morpholinoethoxy, morpholinomethyl, and piperazinylmethyl; 4-methylpiperazin-1-carbonyl, 4- (2-hydroxyethyl) piperazin-1-yl-methyl, piperazin-2-on-1-yl-methyl, pyridin-4-ylmethyl;
z, when present, is an atom or group independently selected each time from O, NH, C (O), NHC (O), C (O) NH, S (O) and S (O)2；
CY is heteroaryl selected from 7H-purin-7-yl; 9H-purin-9-yl; 9H-purin-6-yl; 1H-pyrazolo [3,4-d]Pyrimidin-1-yl; 1H-pyrazolo [3,4-d]Pyrimidin-4-yl; 2H-pyrazolo [3,4-d ]Pyrimidin-2-yl; and 2-,4-, 5-or 6-pyrimidinyl; 2-pyrazinyl, all of which are optionally substituted with one or more groups selected from Cl, Br, F, I, methyl, trifluoromethyl, CN; NH (NH)2；NH-CH3；N(CH3)2(ii) a 3-methyl-1H-indazol-5-yl, 1H-indazol-4-yl; 3-fluoro-5-hydroxyphenyl; 1- (3-fluoro-4-hydroxyphenyl); 6-,5-, 4-hydroxypyridin-3-yl, 6-, 5-methoxypyridin-3-yl, 5-aminopyridin-3-yl, 5-fluoropyridin-3-yl, 5-fluoro-6-hydroxypyridin-3-yl 6- (methylsulfonyl) pyridin-3-yl, 5-hydroxy-6-methylpyridin-3-yl, 6-,5- (hydroxymethyl) pyridin-3-yl, 2-aminothiazol-5-yl; 2- (acetylamino) - (thiazol-5-yl), 2-aminopyrimidin-5-yl, 2-methoxypyrimidin-5-yl, 2-Hydroxypyrimidin-5-yl, pyrazin-2-yl, 6-hydroxypyrazin-2-yl, and 3-fluoro-4-isopropoxyphenyl;
r, m, n, p are as defined above.
A third group of preferred compounds are compounds of formula (I), wherein
R1And R2Combine to form an oxo group (═ O);
R3selected from H, methyl or ethyl;
R4is H;
R5selected from substituted or unsubstituted (C)3-C6) Heterocycloalkyl, substituted or unsubstituted aryl; and substituted or unsubstituted heteroaryl;
cy is 1H-pyrazolo [3,4-d]Pyrimidin-1-yl optionally and independently substituted with one or more groups selected from: halogen; -NR 12R13；(C1-C6) Alkyl, substituted or unsubstituted aryl; and substituted or unsubstituted heteroaryl;
or a pharmaceutically acceptable salt or solvate thereof.
An even more preferred group of compounds are compounds of formula (I), wherein
R1And R2Combine to form an oxo group (═ O);
R3selected from H, methyl or ethyl;
R4is H;
R5is selected from phenyl; 5-thiazolyl; 2-,3-, 4-or 5-thienyl, 1,2,3, 6-tetrahydropyridin-4-yl and 1,2,5, 6-tetrahydropyridin-3-yl; optionally substituted with one or more groups selected from (C)1-C6) Alkyl, -C (O) NR10R11、COOR14Substituted or unsubstituted (C)1-C6) Aminoalkyl radical, (C)1-C6) Alkanoyl, heteroaryl (C)1-C6) Alkyl, (C)3-C7) Cycloalkyl (C)1-C6) Alkyl and NR10R11；
Cy is 1H-pyrazolo [3,4-d]Pyrimidin-1-yl optionally substituted with one or more groups independently selected from halogen, -NR12R13、(C1-C6) Alkyl, phenyl and heteroaryl; the phenyl and heteroaryl groups are thus further optionally and independently substituted by one OR more groups selected from OR7Halogen, -NR12R13、-C(O)NR12R13、-NR7C(O)R9、(C1-C6) Alkyl, (C)1-C6) -haloalkyl, (C)1-C6) A hydroxyalkyl group;
R7、R9、R10、R11、R12、R13、R14m, n, q and as defined above,
or a pharmaceutically acceptable salt or solvate thereof.
A fourth group of preferred compounds are compounds of formula (I), wherein
R1And R2Combine to form an oxo group (═ O);
R3selected from H, methyl or ethyl;
R4is H;
R5is selected from (C)5-C7) Cycloalkenyl group, (C)2-C6) Aminoalkynyl, substituted or unsubstituted (C)3-C6) A heterocycloalkyl group; substituted or unsubstituted aryl and substituted or unsubstituted heteroaryl;
cy is a 9H-purin-6-yl group, optionally substituted with one or more groups selected from: halogen; -NR12R13；(C1-C6) Alkyl, substituted or unsubstituted aryl; and substituted or unsubstituted heteroaryl;
R12、R13m, n, q and as defined above;
or a pharmaceutically acceptable salt or solvate thereof.
A fifth group of preferred compounds are compounds of formula (I), wherein
R1And R2Combine to form an oxo group (═ O);
R3selected from H, methyl or ethyl;
R4is H;
R5is selected from (C)5-C7) Cycloalkenyl group, (C)2-C6) Aminoalkynyl, substituted or unsubstituted (C)3-C6) A heterocycloalkyl group; substituted or unsubstituted aryl and substituted or unsubstituted heteroaryl;
cy is a pyrimidin-4-yl group, optionally substituted with one or more groups independently selected from: halogen, -NR12R13、-CN、(C1-C6) An alkyl group;
R7、R9、R10、R11、R12、R13、R14m, n, q and as defined above;
or a pharmaceutically acceptable salt or solvate thereof.
According to specific embodiments, the present invention provides the following compounds:
3- ((6-amino-9H-purin-9-yl) methyl) -4-phenyl-1H-isochromen-1-one;
3- ((6-amino-9H-purin-9-yl) methyl) -4- (3-fluorophenyl) -1H-isochromen-1-one;
3- ((6-amino-9H-purin-9-yl) methyl) -4- (2-fluorophenyl) -1H-isochromen-1-one;
3- ((6-amino-9H-purin-9-yl) methyl) -4-m-tolyl-1H-isochromen-1-one;
3- (1- (6-amino-9H-purin-9-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (6-amino-9H-purin-9-yl) ethyl) -4-m-tolyl-1H-isochromen-1-one;
3- (1- (6-amino-9H-purin-9-yl) ethyl) -4- (3-fluorophenyl) -1H-isochromen-1-one;
3- (1- (6-amino-9H-purin-9-yl) ethyl) -4- (3- (dimethylamino) phenyl) -1H-isochromen-1-one;
3- (1- (6-amino-9H-purin-9-yl) ethyl) -4- (3- (morpholinosulfonyl) phenyl) -1H-isochromen-1-one;
3- ((9H-purin-6-ylsulfanyl) methyl) -4-phenyl-1H-isochromen-1-one;
3- ((9H-purin-6-ylsulfanyl) methyl) -4- (2-fluorophenyl) -1H-isochromen-1-one;
3- ((9H-purin-6-ylsulfanyl) methyl) -4-m-tolyl-1H-isochromen-1-one;
3- (1- (9H-purin-6-ylsulfanyl) ethyl) -4-m-tolyl-1H-isochromen-1-one;
3- (1- (9H-purin-6-ylsulfanyl) ethyl) -4- (3-fluorophenyl) -1H-isochromen-1-one;
3- (1- (4-amino-1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (6-methylpyridin-3-yl) -1H-isochromen-1-one;
3- (1- (4-amino-1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (3- (morpholinosulfonyl) phenyl) -1H-isochromen-1-one;
3- (1- (4-amino-1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (thiazol-5-yl) -1H-isochromen-1-one;
3- (1- (4-amino-1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (2-methylpyridin-4-yl) -1H-isochromen-1-one;
3- (1- (4-amino-1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-benzyl-1H-isochromen-1-one;
3- ((9H-purin-6-ylamino) methyl) -4- (3-fluorophenyl) -1H-isochromen-1-one;
3- (1- (9H-purin-6-ylamino) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (9H-purin-6-ylamino) ethyl) -4- (1,2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one;
3- (1- (9H-purin-6-ylamino) ethyl) -4- (3, 6-dihydro-2H-pyran-4-yl) -1H-isochromen-1-one;
3- (1- (9H-purin-6-ylamino) propyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (9H-purin-6-ylamino) ethyl) -4- (4- (2-morpholinoethoxy) phenyl) -1H-isochromen-1-one;
4-amino-8- (1- (1-oxo-4-phenyl-1H-isochromen-3-yl) ethyl) pyrido [2,3-d ] pyrimidin-5 (8H) -one;
3- (1- (9H-purin-6-ylamino) ethyl) -4- (5- (morpholinomethyl) thiophen-2-yl) -1H-isochromen-1-one;
3- ((9H-purin-6-ylamino) methyl) -4-phenyl-1H-isochromen-1-one;
3- ((9H-purin-6-ylamino) methyl) -4- (2-fluorophenyl) -1H-isochromen-1-one;
3- ((9H-purin-6-ylamino) methyl) -4-m-tolyl-1H-isochromen-1-one;
3- (1- (9H-purin-6-ylamino) ethyl) -4- (3-fluorophenyl) -1H-isochromen-1-one;
3- (1- (9H-purin-6-ylamino) ethyl) -4-m-tolyl-1H-isochromen-1-one;
3- (1- (9H-purin-6-ylamino) ethyl) -4- (3- (dimethylamino) phenyl) -1H-isochromen-1-one;
3- ((4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) methyl) -4-phenyl-1H-isochromen-1-one;
3- ((4-amino-3- (1H-indazol-5-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) methyl) -4-phenyl-1H-isochromen-1-one;
3- ((4-amino-3- (3-methyl-1H-indazol-5-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) methyl) -4-phenyl-1H-isochromen-1-one;
3- ((4-amino-3- (1H-indazol-6-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) methyl) -4-phenyl-1H-isochromen-1-one;
3- ((4-amino-3- (3-fluoro-4-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) methyl) -4-phenyl-1H-isochromen-1-one
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-m-tolyl-1H-isochromen-1-one;
3- (1- (4-amino-3- (1H-pyrazol-4-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-m-tolyl-1H-isochromen-1-one;
3- (1- (6-amino-9H-purin-9-yl) ethyl) -4- (6-methoxypyridin-3-yl) -1H-isochromen-1-one;
3- ((4-amino-3-iodo-1H-pyrazolo [3,4-d ] pyrimidin-1-yl) methyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3-iodo-1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3-iodo-1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-m-tolyl-1H-isochromen-1-one;
3- (1- (1H-pyrazolo [3,4-d ] pyrimidin-4-ylamino) ethyl) -4-phenyl-1H-isochromen-1-one;
4-amino-6- (1- (1-oxo-4-phenyl-1H-isochromen-3-yl) ethylamino) pyrimidine-5-carbonitrile;
3- (1- (9H-purin-6-ylamino) ethyl) -4- (1-methyl-1, 2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1-methyl-1, 2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1-methyl-1, 2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one single enantiomer 1;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1-methyl-1, 2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one single enantiomer 2;
3- (1- (9H-purin-6-ylamino) ethyl) -4- (4- (morpholinomethyl) phenyl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (4- (morpholinomethyl) phenyl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (5- (morpholinomethyl) thiophen-2-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (5- (morpholinomethyl) thiophen-2-yl) -1H-isochromen-1-one single enantiomer 1;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (5- (morpholinomethyl) thiophen-2-yl) -1H-isochromen-1-one single enantiomer 2;
3- (1- (9H-purin-6-ylamino) ethyl) -4-cyclohexenyl-1H-isochromen-1-one;
3- (1- (4-amino-3- (2-aminopyrimidin-5-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3- (pyrazin-2-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3- (pyridazin-4-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3- (pyridin-4-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3- (2-methoxypyrimidin-5-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3- (2-hydroxypyrimidin-5-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3- (5-methoxypyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3- (pyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3- (2-aminothiazol-5-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3- (6-methoxypyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3- (6-hydroxypyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
n- (5- (4-amino-1- (1- (1-oxo-4-phenyl-1H-isochromen-3-yl) ethyl) -1H-pyrazolo [3,4-d ] pyrimidin-3-yl) thiazol-2-yl) acetamide;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1-methyl-1, 2,5, 6-tetrahydropyridin-3-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1,2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1- (pyridin-4-ylmethyl) -1,2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1- (cyclopropylmethyl) -1,2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (3- ((dimethylamino) methyl) phenyl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (3- ((dimethylamino) methyl) phenyl) -1H-isochromen-1-one enantiomer 1;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (3- ((dimethylamino) methyl) phenyl) -1H-isochromen-1-one enantiomer 2;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (5- (piperidin-1-ylmethyl) thiophen-2-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (5- (piperidin-1-ylmethyl) thiophen-2-yl) -1H-isochromen-1-one single enantiomer 1;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (5- (piperidin-1-ylmethyl) thiophen-2-yl) -1H-isochromen-1-one single enantiomer 2;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (4- (4-methylpiperazine-1-carbonyl) phenyl) -1H-isochromen-1-one;
3- (3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -1-oxo-1H-isochromen-4-yl) -N- (2- (dimethylamino) ethyl) benzamide;
4- (1-acetyl-1, 2,3, 6-tetrahydropyridin-4-yl) -3- (1- (4-amino-3- (5-hydroxypyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -1H-isochromen-1-one;
4- (1-acetyl-1, 2,3, 6-tetrahydropyridin-4-yl) -3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (5- (pyrrolidin-1-ylmethyl) thiophen-2-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (5- ((bis (2-hydroxyethyl) amino) methyl) thiophen-2-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (5- (hydroxymethyl) thiophen-2-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (5- ((4- (2-hydroxyethyl) piperazin-1-yl) methyl) thiophen-2-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (5- ((4- (2-hydroxyethyl) piperazin-1-yl) methyl) thiophen-2-yl) -1H-isochromen-1-one single enantiomer 1;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (5- ((4- (2-hydroxyethyl) piperazin-1-yl) methyl) thiophen-2-yl) -1H-isochromen-1-one single enantiomer 2;
4- ((5- (3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -1-oxo-1H-isochromen-4-yl) thiophen-2-yl) methyl) piperazin-2-one;
5- (3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -1-oxo-1H-isochromen-4-yl) thiophene-2-carboxylic acid;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-benzyl-1H-isochromen-1-one;
4- (1H-pyrazol-4-yl) -3- (1- (thieno [3,2-d ] pyrimidin-4-ylamino) ethyl) -1H-isochromen-1-one;
4- (5- (morpholinomethyl) thiophen-2-yl) -3- (1- (thieno [3,2-d ] pyrimidin-4-ylamino) ethyl) -1H-isochromen-1-one;
4-amino-6- ((1- (4- (5- (morpholinomethyl) thiophen-2-yl) -1-oxo-1H-isochromen-3-yl) ethyl) amino) pyrimidine-5-carbonitrile;
4-phenyl-3- (1- (pyrrolo [2,1-f ] [1,2,4] triazin-4-ylamino) ethyl) -1H-isochromen-1-one;
4-phenyl-3- (1- (pyrido [3,2-d ] pyrimidin-4-ylamino) ethyl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (5- (hydroxymethyl) pyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3- (6- (hydroxymethyl) pyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
n- (5- (4-amino-1- (1- (1-oxo-4-phenyl-1H-isochromen-3-yl) ethyl) -1H-pyrazolo [3,4-d ] pyrimidin-3-yl) pyridin-3-yl) -4-fluorobenzenesulfonamide;
3- (1- (4-amino-3- (5-aminopyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3- (2-aminopyrimidin-4-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3- (6-hydroxypyrazin-2-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3- (5-hydroxypyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (8-methyl-8-azabicyclo [3.2.1] oct-2-en-3-yl) -1H-isochromen-1-one hydrochloride;
3- (1- (4-amino-3- (3-fluoro-4-isopropoxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3- (5-fluoropyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3- (3-chloro-5-fluorophenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3- (5- (methylsulfonyl) pyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3- (6- (methylsulfonyl) pyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3- (5-fluoro-6-hydroxypyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3- (5-hydroxy-6-methylpyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3- (5- (trifluoromethyl) pyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3- (5-hydroxy-3-pentafluorothio) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
5- (4-amino-1- (1- (1-oxo-4-phenyl-1H-isochromen-3-yl) ethyl) -1H-pyrazolo [3,4-d ] pyrimidin-3-yl) nicotinonitrile;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (4-aminocyclohex-1-en-1-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (trifluoromethyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3-methyl-1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-7H-pyrrolo [2,3-d ] pyrimidin-7-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (2, 6-diamino-9H-purin-9-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
4-phenyl-3- (1- (thieno [2,3-d ] pyrimidin-4-ylamino) ethyl) -1H-isochromen-1-one;
4-phenyl-3- (1- (thieno [3,2-d ] pyrimidin-4-ylamino) ethyl) -1H-isochromen-1-one;
2-amino-N- (1- (1-oxo-4-phenyl-1H-isochromen-3-yl) ethyl) pyrazolo [1,5-a ] pyrimidine-3-sulfonamide;
3- (1- (4-amino-3- (1H-pyrazol-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3- (1H-indazol-4-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3- (3-amino-1H-indazol-5-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3- (3-hydroxy-5- (trifluoromethoxy) phenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (6-amino-9H-purin-9-yl) ethyl) -4- (5- (morpholinomethyl) thiophen-2-yl) -1H-isochromen-1-one;
3- (1- (9H-purin-6-ylamino) ethyl) -4- (6-methoxypyridin-3-yl) -1H-isochromen-1-one;
3- (1- (9H-purin-6-ylamino) ethyl) -4- (thiazol-5-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -2H-pyrazolo [3,4-d ] pyrimidin-2-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (3- (dimethylamino) phenyl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (thiazol-5-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (2-aminothiazol-5-yl) -1H-isochromen-1-one;
3- (1- (9H-purin-6-ylamino) ethyl) -4- (1H-pyrazol-4-yl) -1H-isochromen-1-one;
4-amino-6- ((1- (1-oxo-4- (1H-pyrazol-4-yl) -1H-isochromen-3-yl) ethyl) amino) pyrimidine-5-carbonitrile;
3- (1- (9H-purin-6-ylamino) ethyl) -4- (2-aminopyrimidin-5-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (4- (piperazin-1-ylmethyl) phenyl) -1H-isochromen-1-one;
3- (1- (9H-purin-6-ylamino) ethyl) -4- (4- (piperazin-1-ylmethyl) phenyl) -1H-isochromen-1-one;
3- (1- (4-amino-1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (4- (piperazin-1-ylmethyl) phenyl) -1H-isochromen-1-one;
3- (4-amino-1- ((4-phenyl-1H-isochromen-3-yl) methyl) -1H-pyrazolo [3,4-d ] pyrimidin-3-yl) -5-fluorophenol;
5- (4-amino-1- ((4-phenyl-1H-isochromen-3-yl) methyl) -1H-pyrazolo [3,4-d ] pyrimidin-3-yl) pyridin-3-ol;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (morpholinomethyl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1-benzyl-1, 2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (5-hydroxypyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1-benzyl-1, 2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one;
4- (3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -1-oxo-1H-isochromen-4-yl) -5, 6-dihydropyridine-1 (2H) -carboxylic acid benzyl ester;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (5- (piperazin-1-ylmethyl) thiophen-2-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (5- ((4-methylpiperazin-1-yl) methyl) thiophen-2-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (5- (3- (dimethylamino) propyl) thiophen-2-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (3- (3- (dimethylamino) propyl) phenyl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (3- (3- (dimethylamino) propyl) phenyl) -1H-isochromen-1-one single enantiomer 1;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (3- (3- (dimethylamino) propyl) phenyl) -1H-isochromen-1-one single enantiomer 2;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (4- (3- (dimethylamino) propyl) phenyl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (4- (3- (dimethylamino) propyl) phenyl) -1H-isochromen-1-one single enantiomer 1;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (4- (3- (dimethylamino) propyl) phenyl) -1H-isochromen-1-one single enantiomer 2;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (4- ((4-methylpiperazin-1-yl) methyl) phenyl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (4- (pyrrolidin-1-ylmethyl) phenyl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (4- (piperidin-1-ylmethyl) phenyl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (3- ((4-methylpiperazin-1-yl) methyl) phenyl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (3- (piperidin-1-ylmethyl) phenyl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (2,2,6, 6-tetramethyl-1, 2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (5-hydroxypyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (2,2,6, 6-tetramethyl-1, 2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1- (4- (dimethylamino) butanoyl) -1,2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1- (2- (dimethylamino) acetyl) -1,2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1- (1-methylpiperidine-4-carbonyl) -1,2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1- (1-isopropylpiperidin-4-yl) -1,2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1- (azetidine-3-carbonyl) -1,2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1- (1-methylazetidin-3-carbonyl) -1,2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-chloro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1,2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-hydroxy-5- (trifluoromethyl) phenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1,2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-chloro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1-methyl-1, 2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-hydroxy-5- (trifluoromethyl) phenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1-methyl-1, 2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1- (azetidin-3-yl) -1,2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1- (1- (cyclopropylmethyl) azetidin-3-yl) -1,2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (3- (dimethylamino) prop-1-yn-1-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (5-hydroxy-4-methylpyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3- (5-hydroxy-2-methylpyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3- (5-hydroxy-6- (trifluoromethyl) pyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -7-methyl-4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -7-chloro-4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -6-chloro-4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -6-fluoro-4-phenyl-1H-isochromen-1-one;
4- (1-acetyl-1, 2,3, 6-tetrahydropyridin-4-yl) -3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -1H-isochromen-1-one single enantiomer 1;
4- (1-acetyl-1, 2,3, 6-tetrahydropyridin-4-yl) -3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -1H-isochromen-1-one single enantiomer 2;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (thiazol-5-yl) -1H-isochromen-1-one single enantiomer 1;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (thiazol-5-yl) -1H-isochromen-1-one single enantiomer 2;
3- (1- (4-amino-3- (5-hydroxypyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one single enantiomer 1;
3- (1- (4-amino-3- (5-hydroxypyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one single enantiomer 2;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1-benzyl-1, 2,5, 6-tetrahydropyridin-3-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1-benzyl-1, 2,5, 6-tetrahydropyridin-3-yl) -1H-isochromen-1-one single enantiomer 1;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1-benzyl-1, 2,5, 6-tetrahydropyridin-3-yl) -1H-isochromen-1-one single enantiomer 2;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (pyridin-2-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1,2,5, 6-tetrahydropyridin-3-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1- (1-methylpiperidin-4-yl) -1,2,5, 6-tetrahydropyridin-3-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1- (azetidin-3-yl) -1,2,5, 6-tetrahydropyridin-3-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1-isopropyl-1, 2,5, 6-tetrahydropyridin-3-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (5-hydroxypyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1-methyl-1, 2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one single enantiomer 1;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one single enantiomer 2;
and pharmaceutically acceptable salts thereof.
The compounds of formula (I), including all of the compounds listed above, can generally be prepared according to the procedures outlined in the schemes shown below using generally known methods. Residue M is a 1, 2-phenylene-diyl group, and is optionally substituted by one or more r (p).
Scheme 1
α -aryl-or α -heteroaryl-benzoates of the formula (I') in which R1 and R2 combine to form an oxo group (C ═ O) are reacted with alkynes of the formula (II) under suitable Sonogashira cross-coupling conditions in the presence of palladium catalysts as described in "Transition Metals for organic Synthesis", 2 nd edition, 1, 211-one 229, 2004.
Alkynes (II), such as prop-2-yn-1-ol and but-3-yn-2-ol, are commercially available or derivatives thereof, such as t-butyl but-3-yn-2-ylcarbamate, may be prepared according to well-known methods.
Synthesis 3, 400-2The reaction cyclizes intermediate (III) in the presence of a suitable base to the corresponding halide of formula (IV). By reaction with boric acid or a suitable ester R5B(OR)2Suzuki coupling of (VIII) further converts compound (IV) to (V). Boric acid and esters of formula (VIII) are commercially available.
Compounds of formula (VII) can optionally be prepared from compounds of formula (V) by the synthetic route illustrated in scheme 1, which correspond to compounds of formula (I) wherein Z ═ NH and m ═ 1.
When the group Y represents a hydroxyl moiety, by reaction with a suitable halogenating agent, e.g. PBr3The reaction converts compound (V) to (VI), wherein the group X represents a suitable leaving group, such as a halogen atom.
When group Y represents hydrogen, compound (V) is converted to (VI) by reaction with N-bromosuccinimide, wherein group X represents a suitable leaving group, for example a halogen atom.
Compound (VI) is finally reacted with a nitrogen-or sulfur-based nucleophile (IX) such as 9H-purin-6-amine, 9H-purin-6-thiol hydrate, tert-butyl 9-trityl-9H-purin-6-ylcarbamate, 1H-pyrazolo [3,4-d ] pyrimidin-4-amine to give compound (I).
When the group Y represents an amino group protected by a suitable protecting group, such as BOC, compound (V) is deprotected under acidic conditions and reacted with a suitable halide derivative, Cy-Cl (X), to give a compound of formula (VII).
This scheme provides synthetic routes for the preparation of examples 1-35, 43-45, 47, 50, 80-84, 104-.
Using a similar approach as described in scheme 1, compound (XII), which corresponds to compound (I) wherein m ═ 0(Z is absent), and Cy is optionally substituted 1H-pyrazolo [3,4-d ] pyrimidinyl, can be synthesized as outlined in scheme 2 from compound (VI) and commercially available 3-iodo-1H-pyrazolo [3,4-d ] pyrimidin-4-amine to give compound (XI), which can be further converted to compound (XII) by Suzuki coupling. Boric acid and esters of formula (VIII) are commercially available.
This scheme provides a synthetic route for the preparation of the compounds of examples 35-42, 46, 48-49, 51, 54, 58, 63, 72-73, 79, 111, 118, 120, 125, 133, 161, 165 and 175.
Scheme 2
In another embodiment of the invention, it is possible to use the compounds (XI) according to scheme 2a by Stille, Suzuki cross-coupling reactions (scheme 2a, step 3a), using suitable organotin or organoboron reagents (VIII) and palladium catalysts or by compounds (VI), by nucleophilic substitution, using nitrogen-based catalystsNucleophiles (IXa) of (e.g. optionally substituted 3-iodo-1H-pyrazolo [3, 4-d)]Pyrimidin-4-amine (scheme 2a, step 4a)) to produce compound (XII). Can be prepared by a Suzuki, Stille or Sonogashira cross-coupling reaction (scheme 2a, step 1a), followed by appropriate halogenating reagents such as PBr 3Substituted hydroxy (scheme 2a, step 1b) compound (VI) is prepared from compound (IV), wherein X is a suitable leaving group (Lg), e.g. a halogen atom. Some compounds (XII) may contain protected hydroxyl or amino groups, which are then removed under well-known procedures (scheme 2a, steps 3b and 4 b).
This scheme provides a synthetic route for the preparation of the compounds of examples 52-53, 56-57, 59-62, 68-71, 85-103, 112-.
Scheme 2a
According to scheme 3, the compounds (XI) wherein R5 is (C) containing a suitable secondary amine-NH3-C6) Heterocycloalkyl group) in a similar manner, compounds of formula (XIa) are prepared wherein R1 and R2 combine to form an oxo group (C ═ O). Compound (XIa) may be converted to a compound of formula (XIIa) by two reaction sequences consisting of the following reactions, wherein Rx is a suitable substituent as described above: amine derivatization (step 1) using a suitable carbonyl compound under reductive amination conditions, or by amide coupling with a suitable carboxylic acid, followed by Suzuki cross-coupling with a suitable boronic acid (step 2). To prepare some of the compounds (XIIa), the process can be performed in front-to-back inversion.
This scheme provides a synthetic route for the preparation of the compounds of examples 65-66, 150-.
Scheme 3
In one embodiment of the present invention, compound (IV) can be prepared in A manner analogous to scheme 1 by reaction with the appropriate boronic acid or ester CHO-A-B (ORz)2Or by deprotection of compound (Vb) under acidic conditions (scheme 4, step 1) to prepare compound (Va), wherein R1 and R2 combine to form an oxo group (C ═ O), and Y represents a hydroxy group and R5 is an aryl or heteroaryl group a; which is partially substituted with an aldehyde. By reaction with a suitable boronic acid or ester containing a protected aldehyde (RwO)2CH-A-B(ORz)2The compound of formula (Vb) is prepared from compound (IV). Compound (Va) can be converted to (XIIb) by reductive amination with a suitable primary or secondary amine (scheme 4, step 2a), followed by introduction of a suitable leaving group (Lg), e.g. a bromine atom (scheme 4, step 2b) and finally reaction with a suitable nucleophile (IX) (scheme 4, step 2 c). Compound (XIIb) may contain protected hydroxyl groups, which are subsequently removed under well-known procedures (scheme 4, step 2 d).
This scheme provides a synthetic route for the preparation of the compounds of examples 134-135, 139-142.
Scheme 4
Alternatively, according to scheme 5, compound (Vb) (wherein R1 and R2 combine to form an oxo group (C ═ O) and Y represents a hydroxy group) can be converted to compound (XIIc) by conversion of Y to a suitable leaving group (Lg), e.g. a bromine atom (step a), followed by nucleophilic substitution with a suitable nitrogen-based nucleophile (IX) (step b), deprotection of the carbonyl moiety under acidic conditions (step C), and finally reductive amination using a suitable amine in the presence of a reducing agent, e.g. sodium triacetoxyborohydride.
This scheme provides a synthetic route for the preparation of the compounds of examples 67, 73-77, 143.
Scheme 5
In another embodiment of the invention, compound (Va) may be converted to compound (Vc) by a Wittig reaction with a suitable triphenylphosphonium bromide (scheme 6, step 1), wherein R1 and R2 combine to form an oxo group (C ═ O) and Y represents a hydroxy group. Compound (Vc) can then be converted to compound (XIId) by converting Y to a suitable leaving group such as a bromine atom (scheme 6, step 2a), followed by nucleophilic substitution with a suitable nitrogen-based nucleophile (IX) (scheme 6, step 2b) and finally reducing the double bond with a suitable reducing agent such as triethylsilane in the presence of Pd/C.
This scheme provides a synthetic route for the preparation of the compounds of examples 137 and 138.
Scheme 6
According to scheme 7, compound (Vd) can be converted to compound (XIIe) by Pinnick oxidation (structural Applications of named reactions in Organic Synthesis, Laszlo Kurti, barbarbarara Czako, Elsevier, academic press, 2005) wherein R1 and R2 combine to form an oxo group (C ═ O), wherein R5 is a is heteroaryl spacer a. Compound (Vd) can be prepared from compound (Vb) according to steps a, b and c of scheme 5. This scheme provides a synthetic route for the preparation of the compound of example 78.
Scheme 7
In one embodiment of the present invention, compound (XIIf) may be prepared from compound (Ve) according to scheme 8, wherein R1 and R2 combine to form oxo (C ═ O)). Compound (Ve) can be converted to an aldehyde by ozonization (step a) and then to an amine by reductive amination using a suitable secondary amine such as morpholine in the presence of a reducing agent such as sodium triacetoxyborohydride (step b). The Y group represents a hydroxyl group and is then converted to a suitable leaving group (Lg) and then displaced with a nitrogen-based nucleophile (IX) (step d) followed by partial deprotection of the hydroxyl group (step e). Compound Ve can be prepared from IV by Stille coupling with vinyl tributyl-tin and Pd catalyst according to scheme 2, step 1 a.
This scheme provides a synthetic route for the preparation of the compound of example 130.
Scheme 8
In one embodiment of the present invention, compound (Vf) may be prepared according to scheme 9 from compound (XIII), e.g. a commercially available isochroman-4-one, wherein R1 ═ R2 ═ R3 ═ R4 ═ H. Can be prepared by reaction with a halogenating agent such as POCl3The reaction converts compound (XIII) to chloride (XIV) (scheme 9, step 1). Compound (XIV) can be converted to (XV) by Suzuki coupling with a suitable boronic acid (scheme 9, step 2) and finally to compound (Vf) by reduction with a hydride reagent such as sodium borohydride.
Scheme 9
Compounds of formula (Vf) can be converted to compounds (XIIg) by synthetic methods analogous to those reported in scheme 2a, steps 2 and 3, by converting Y to the appropriate Lg (scheme 9, step 4a), followed by nucleophilic substitution with a suitable nucleophile such as 3-iodo-1H-pyrazolo [3,4-d ] pyrimidin-4-amine and final Suzuki coupling (scheme 9, steps 4b, c), wherein R1 ═ R2 ═ R3 ═ R4 ═ H.
This scheme provides a synthetic route for preparing the compounds of examples 128 and 129.
Enantiomerically pure compounds 46a/b, 49a/b, 67a/b, 68a/b, 76a/b, 137a/b, 138a/b, 166a/b, 167a/b, 168a/b, 169a/b, 176a/b can be prepared from the corresponding racemates by chiral separation of the parent racemate or the final protected analogue followed by a suitable protection step.
The compounds of the invention are inhibitors of kinase activity, particularly PI 3-kinase activity. In general, compounds that are inhibitors of PI3K may be useful in the treatment of a number of disorders related to the PI3K enzymatic mechanism.
In one embodiment, disorders that may be treated with the compounds of the present invention include: a respiratory disease selected from idiopathic chronic cough, cough-variant asthma, cough associated with breast tumor or lung cancer, viral or post-viral cough, Upper Airway Cough Syndrome (UACS); or cough after nasal drip; or cough associated with gastroesophageal reflux disease (acid and non-acid reflux), asthma, chronic bronchitis, Chronic Obstructive Pulmonary Disease (COPD), interstitial lung disease (e.g., Idiopathic Pulmonary Fibrosis (IPF)), congestive heart disease, sarcoidosis, infection (e.g., pertussis), asthma, Chronic Obstructive Pulmonary Disease (COPD), and Idiopathic Pulmonary Fibrosis (IPF)); viral infections (including viral respiratory infections and viral respiratory diseases exacerbated; non-viral respiratory infections including aspergillosis and leishmaniasis; allergic diseases including allergic rhinitis and atopic dermatitis; autoimmune diseases, including rheumatoid arthritis and multiple sclerosis; inflammatory diseases, including inflammatory bowel disease; cardiovascular diseases including thrombosis and atherosclerosis; hematological malignancies; neurodegenerative diseases; pancreatitis; multiple organ failure; renal disease; platelet aggregation; cancer; sperm motility; graft rejection; transplant rejection; lung injury; and pain, including pain associated with rheumatoid arthritis or osteoarthritis, back pain, systemic inflammatory pain, post-herpetic neuralgia, diabetic neuropathy, inflammatory neuropathic pain (trauma), trigeminal neuralgia, and central pain.
In another embodiment, the disorder that can be treated with the compounds of the present invention is selected from idiopathic chronic cough, cough-variant asthma, cough associated with breast tumors or lung cancer, viral or post-viral cough, Upper Airway Cough Syndrome (UACS), post nasal drip cough, cough associated with gastroesophageal reflux disease (both acid and non-acid reflux), asthma, chronic bronchitis, Chronic Obstructive Pulmonary Disease (COPD), and interstitial lung disease (e.g., Idiopathic Pulmonary Fibrosis (IPF).
In another embodiment, the disorder is selected from asthma, Chronic Obstructive Pulmonary Disease (COPD), Idiopathic Pulmonary Fibrosis (IPF), cough, and chronic cough.
The treatment methods of the present invention comprise administering to a patient in need thereof a safe and effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof. As used herein, a "safe and effective amount" when referring to a compound of formula (I) or a pharmaceutically acceptable salt or other pharmaceutically active agent thereof, refers to an amount of the compound that is sufficient to treat a patient's condition, but low enough to avoid serious side effects, as determined by one of ordinary skill in the art by routine means. The compound of formula (I) or a pharmaceutically acceptable salt thereof may be administered once or according to a dosing regimen wherein multiple doses are administered at different time intervals for a specified period of time. Typical daily dosages may vary depending upon the particular route of administration selected.
The invention also provides pharmaceutical compositions of compounds of formula (I) in admixture with one or more pharmaceutically acceptable carriers or excipients, such as those described in Remington's pharmaceutical sciences Handbook, XVII ed., Mack pub., n.y., u.s.a.
The compounds of the invention and their pharmaceutical compositions may be administered to the patient as required, for example, by oral, intranasal, parenteral (subcutaneous, intravenous, intramuscular, intrasternal and by infusion), inhalation, rectal, vaginal, topical, transdermal and by ocular administration.
Various solid oral dosage forms may be used to administer the compounds of the present invention, including solid forms such as tablets, caplets, capsules, caplets, granules, lozenges, and bulk powders. The compounds of the present invention may be administered alone or in combination with various pharmaceutically acceptable carriers, diluents (e.g., sucrose, mannitol, lactose, starch) and excipients known in the art, including, but not limited to, suspending agents, solubilizing agents, buffers, binders, disintegrating agents, preservatives, coloring agents, flavoring agents, lubricants and the like. Time release capsules, tablets and gels are also advantageous for administration of the compounds of the present invention.
Various liquid oral dosage forms may also be used to administer the compounds of the present invention, including aqueous and non-aqueous solutions, emulsions, suspensions, syrups and elixirs. Such dosage forms may also contain suitable inert diluents known in the art (e.g., water), as well as suitable excipients known in the art (e.g., preservatives, wetting agents, sweeteners, flavoring agents) and agents for emulsifying and/or suspending the compounds of the present invention. For example, the compounds of the present invention may be injected intravenously in the form of an isotonic sterile solution. Other formulations are also possible.
Suppositories for rectal administration of the compounds of the invention can be prepared by mixing the compounds with suitable excipients such as cocoa butter, salicylates and polyethylene glycols.
Formulations for vaginal administration may be in the form of creams, gels, pastes, foams or sprays containing in addition to the active ingredient such suitable carriers as are known in the art.
For topical administration, the pharmaceutical compositions may be in the form of creams, ointments, liniments, lotions, emulsions, suspensions, gels, solutions, pastes, powders, sprays and drops suitable for administration to the skin, eye, ear or nose. Topical administration may also include transdermal administration, for example by means of a transdermal patch.
For the treatment of respiratory diseases, the compounds according to the invention are preferably administered by inhalation.
Inhalable formulations include inhalable powders, propellant-containing metered dose aerosols or propellant-free inhalable formulations.
For administration as a dry powder, single or multi-dose inhalers known from the prior art may be used. In this case, the powder may be filled in gelatin, plastic or other capsules, cartridges or blister packs or in a reservoir.
A diluent or carrier that is generally non-toxic and chemically inert to the compounds of the present invention (e.g., lactose) or any other additive suitable for improving the absorbable fraction may be added to the powdered compounds of the present invention.
Inhalation aerosols comprising a propellant gas, such as a hydrofluoroalkane, may comprise the compounds of the invention in solution or in dispersed form. The propellant-driven formulation may also comprise other ingredients, such as co-solvents, stabilizers or optionally other excipients.
Propellant-free inhalable formulations comprising the compounds according to the invention may be in the form of solutions or suspensions in aqueous, alcoholic or hydroalcoholic media, and they may be prepared by means of spray or ultrasonic nebulizers known from the prior art or by means of soft mist nebulizers, for example To be delivered.
The compounds of the invention may be administered as the sole active agent or in combination with other pharmaceutical active ingredients (combination) including those currently used in the treatment of respiratory disorders, e.g. β 2-agonists, antimuscarinics, corticosteroids, inhibitors of the mitogen-activated protein kinase class (P38MAP kinase), inhibitors of the nuclear factor κ -B kinase subunit β (IKK2), inhibitors of Human Neutrophil Elastase (HNE), phosphodiesterase 4(PDE4) inhibitors, leukotriene modulators, non-steroidal anti-inflammatory drugs (NSAIDs) and mucus regulators.
The dosage of the compounds of the invention depends on a variety of factors including the particular disease to be treated, the severity of the symptoms, the route of administration, the frequency of dosage intervals, the particular compound used, the potency, toxicological profile and pharmacokinetic profile of the compound.
Advantageously, the compounds of formula (I) can be administered at a dose of, for example, 0.001 to 1000 mg/day, preferably 0.1 to 500 mg/day.
When they are administered by the inhalation route, the dose of the compound of formula (I) is advantageously comprised between 0.001 and 500 mg/day, preferably between 0.1 and 200 mg/day.
The following examples illustrate the invention without limiting its scope.
Preparation of intermediates and examples
Chemical names of compounds were generated using Structure To Name Enterprise 10.0 Cambridge software.
Abbreviations
Et2O ═ diethyl ether; et (Et)3N ═ triethylamine; DCE ═ 1, 2-dichloroethane; TEA ═ triethylamine; DCC ═ N, N' -dicyclohexylcarbodiimide; HOBt ═ hydroxybenzotriazole; HATU ═ N, N-dimethyl (3H- [1,2, 3)]Triazolo [4,5-b]Pyridin-3-yloxy) methylammonium hexafluorophosphate; HBTU ═ N, N '-tetramethyl-O- (1H-benzotriazol-1-yl) uronium hexafluorophosphate, O- (benzotriazol-1-yl) -N, N' -tetramethyluronium hexafluorophosphate; EDC ═ 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride; DMAP ═ 4-dimethylaminopyridine; DMF ═ dimethylformamide; EtOAc ═ ethyl acetate; RT ═ room temperature; THF ═ tetrahydrofuran; DCM ═ dichloromethane; MeOH ═ methanol; EtOH ═ ethanol; LHMDS ═ lithium bis (trimethylsilyl) amide; m-CPBA ═ m-chloroperbenzoic acid(ii) a TFA ═ trifluoroacetic acid; LC-MS ═ liquid chromatography/mass spectrometry; HPLC ═ high pressure liquid chromatography; MPLC-medium pressure liquid chromatography; SFC ═ supercritical chromatography; dppf ═ 1,1' -bis (diphenylphosphino) ferrocene; X-Phos-Pd-G2 ═ chloro (2-dicyclohexylphosphino-2 ',4',6 '-triisopropyl-1, 1' -biphenyl) [2- (2 '-amino-1, 1' -biphenyl ] ]Palladium (II); S-Phos-Pd-G2 ═ chloro (2-dicyclohexylphosphino-2 ',6' -dimethoxy-1, 1' -biphenyl) [2- (2' -amino-1, 1' -biphenyl)]Palladium (II); DIEA or DIPEA ═ N, N-diisopropylethylamine; MeCN ═ acetonitrile; MTBE ═ tert-butyl methyl ether; ac2O ═ acetic anhydride; AcCl ═ acetyl chloride; HBTU ═ N, N '-tetramethyl-O- (1H-benzotriazol-1-yl) uronium hexafluorophosphate, O- (benzotriazol-1-yl) -N, N' -tetramethyluronium hexafluorophosphate; TBDMSCl ═ tert-butyl (chloro) dimethylsilane; DMSO ═ dimethyl sulfoxide; boc2O-di-tert-butyl dicarbonate; UPLC ═ ultra performance liquid chromatography.
General experimental details
And (3) NMR characterization:
this was done using a Varian MR-400 spectrometer operating at 400MHz (proton frequency)1H-NMR spectroscopy, the spectrometer was equipped with: the self-isolation type digital optical fiber sensor comprises a self-isolation z-gradient coil 5mm 1H/nX broadband probe for reverse detection, a deuterium digital lock channel unit and an orthogonal digital detection unit with a sensor offset frequency shift; or using Agilent VNMRS-500 or Bruker Avance400 spectrometer1H-NMR spectrum. Chemical shifts are reported as values in ppm relative to Trimethylsilane (TMS) as an internal standard. Coupling constants (J values) are given in hertz (Hz) and the diversity is reported using the following abbreviations (s ═ singlet; d ═ doublet; t ═ triplet; q ═ quartet; m ═ multiplet, br ═ broad, nd ═ undetermined).
LC/UV/MS analysis method
LC/MS retention times affected by. + -. 0.5min experimental error were evaluated.
LC/UV/MS-method 1
LC instrument: HPLC Alliance Waters (or equivalent)
Column: kinetex 2.6 mu m C18100A 100 x 4.6mm (Phenomenex)
Column temperature (. degree. C.): 50.0
Mobile phase: HCOONH40.025M pH3 (A); acetonitrile (B)
Flow rate (ml/min): 2.0 (split inflow MS 1: 10)
End time (min): 17.0
Gradient:
| time (min) | ％A | ％B |
| 0.00 | 80.0 | 20.0 |
| 10.00 | 20.0 | 80.0 |
| 12.00 | 20.0 | 80.0 |
| 14.00 | 80.0 | 20.0 |
| 17.00 | 80.0 | 20.0 |
And (4) UV detection: a passage 1245 nm; channel 2254 nm
Injection volume (ul): 5.00
Sample solvent: acetonitrile
MS instrument: waters Quattro Micro API (or equivalent) polarity ES +
Capillary (kV)3.20
Cone voltage (V)20.00
Extractor (V)2.00
RF lens (V)0.3
Polarity ES-
Capillary (kV)3.20
Cone voltage (V)20.00
Extractor (V)3.00
RF lens (V)0.3
Source temperature (. degree. C.) 110
Desolvation temperature (. degree.C.) 210
Cone voltage gas flow rate (L/Hr)150
Desolvated gas stream (L/Hr)650
Scan duration (seconds): 1.00
Mid scan delay (seconds): 0.10
The mass range is as follows: 125-1000
LC/UV/MS-method 2
LC instrument: acquisty Waters UPLC (or equivalent)
Column: kinetex 1.7 μm XB-C18100A 100 x 2.1mm (Phenomenex)
Column temperature (. degree. C.) 50.0
Mobile phase: HCOONH40.025M pH3 (A); acetonitrile + 0.1% formic acid (B)
Flow rate (ml/min)0.65 (split inflow MS 1: 3)
End time (min)10.0
Gradient:
| time (min) | ％A | ％B |
| 0.00 | 80.0 | 20.0 |
| 5.50 | 20.0 | 80.0 |
| 7.50 | 20.0 | 80.0 |
| 8.00 | 80.0 | 20.0 |
| 10.00 | 80.0 | 20.0 |
And (4) UV detection: DAD
UV acquisition range (nm): 210-400
Injection volume (ul) -2.00
Sample solvent: acetonitrile
MS instrument: waters ZQ (or equivalent)
Polarity ES +
Capillary (kV)3.00
Cone voltage (V)20.00
Extractor (V)3.00
RF lens (V)1.0
Polarity ES-
Capillary (kV)3.00
Cone voltage (V)20.00
Extractor (V)3.00
RF lens (V)1.0
Source temperature (. degree. C.) 110
Desolvation temperature (. degree.C.) 210
Conical Voltage Gas flow Rate (L/Hr)150
Desolvated gas stream (L/Hr)650
The mass range is as follows: 100-950
Scan time (sec): 0.32
LC/UV/MS-method 3
LC instrument: acquisty Waters UPLC (or equivalent)
Column: kinetex 1.7 μm PFP 100A 100X 2.1mm (Phenomenex)
Column temperature (. degree. C.) 55.0
Mobile phase: HCOONH40.025M pH3 (A); acetonitrile (B)
Flow rate (ml/min)0.45 (split inflow MS 1: 3)
End time (min)10.0
Gradient:
| time (min) | ％A | ％B |
| 0.00 | 85.0 | 15.0 |
| 5.00 | 55.0 | 45.0 |
| 5.50 | 20.0 | 80.0 |
| 6.50 | 20.0 | 80.0 |
| 7.00 | 85.0 | 15.0 |
| 10.0 | 85.0 | 15.0 |
And (4) UV detection: DAD
UV acquisition range (nm): 210-400
Injection volume (ul) -2.00
Sample solvent: acetonitrile
MS instrument: waters ZQ (or equivalent)
Polarity ES +
Capillary (kV)3.00
Cone voltage (V)20.00
Extractor (V)3.00
RF lens (V)1.0
Polarity ES-
Capillary (kV)3.00
Cone voltage (V)20.00
Extractor (V)3.00
RF lens (V)1.0
Source temperature (. degree. C.) 110
Desolvation temperature (. degree.C.) 210
Conical Voltage gas flow (L/Hr)150
Desolvated gas stream (L/Hr)650
The mass range is as follows: 100-950
Scan time (sec): 0.32
LC/UV/MS-method 4
LC instrument: acquisty Waters UPLC (or equivalent) interface 2996 PDA detector
Column: acquity UPLC BEH c181.7um 50x2.1mm
Column temperature (. degree. C.) 30.0
Mobile phase: 95: 5H 2O: ACN + (0.1% HCOOH) (a); 5: 95H 2O: ACN + (0.1% HCOOH) (B)
Flow rate (ml/min)0.6 (split inflow MS 1: 6)
End time (min)3.5
Gradient:
| time (min) | ％A | ％B |
| 0.00 | 100 | 0 |
| 0.50 | 100 | 0 |
| 2.20 | 0.0 | 100.0 |
| 2.70 | 0.0 | 100.0 |
| 2.90 | 100 | 0 |
And (4) UV detection: BPI detection (start wavelength nm 210, end wavelength nm 400, sample rate spectrum/sec 20)
Injection volume (ul) -1.00
Sample solvent: DMSO, DMSO: MeOH: ratio of CAN 1: 3: 3
MS instrument: waters ZQ (or equivalent)
Polarity ES
Capillary (kV)3.20
Cone voltage (V)25.00
Extractor (V)3.00
RF lens (V)0.1
Polarity ES-
Capillary (kV)3.40
Cone voltage (V)24.00
Extractor (V)2.00
RF lens (V)0.2
Source temperature (. degree. C.) 130
Desolvation temperature (. degree.C.) 400
Conical Voltage gas flow (L/Hr)80
Desolvated gas stream (L/Hr)800
The mass range is as follows: 60-1200
Scan time (sec): 0.4
LC/UV/MS-method 5
LC instrument: acquisty Waters UPLC (or equivalent) interface 2996 PDA detector
Column: acquity UPLC BEH c181.7um 50x2.1mm
Column temperature (. degree. C.) 30.0
Mobile phase: 95: 5H 2O: ACN + (0.1% HCOOH) (a); 5: 95H 2O: ACN + (0.1% HCOOH) (B)
Flow rate (ml/min)0.6 (split inflow MS 1: 6)
End time (min)3.5
Gradient:
| time (min) | ％A | ％B |
| 0.00 | 100 | 0 |
| 0.50 | 100 | 0 |
| 10.0 | 0.0 | 100.0 |
| 11.0 | 0.0 | 100.0 |
| 12.0 | 100 | 0 |
And (4) UV detection: BPI detection (start wavelength nm 210, end wavelength nm 400, sample rate spectrum/sec 20)
Injection volume (ul) -1.00
Sample solvent: DMSO, DMSO: MeOH: ratio of ACN 1: 3: 3
MS instrument: waters ZQ (or equivalent)
Polarity ES
Capillary (kV)3.20
Cone voltage (V)25.00
Extractor (V)3.00
RF lens (V)0.1
Polarity ES-
Capillary (kV)3.40
Cone voltage (V)24.00
Extractor (V)2.00
RF lens (V)0.2
Source temperature (. degree. C.) 130
Desolvation temperature (. degree.C.) 400
Conical Voltage gas flow (L/Hr)80
Desolvated gas stream (L/Hr)800
The mass range is as follows: 60-1200
Scan time (sec): 0.4
LC/UV/MS-method 6
LC instrument: acquisty Waters UPLC (or equivalent) interface 2996 PDA detector. Column: AcquisyteUPLC CSH C181.7um 130A 50 X2.1mm. Column temperature (. degree. C.) 50.0. Mobile phase: HCOONH40.025M pH 3 (A); ACN + 0.1% HCOOH (B). Flow rate (ml/min)0.35 (split flow into MS 1: 3). End time (min)10
Gradient:
| time (min) | ％A | ％B |
| 0.00 | 80 | 20 |
| 5.50 | 20 | 80 |
| 7.5 | 20 | 80 |
| 8 | 80 | 20 |
| 10 | 80 | 20 |
And (4) UV detection: BPI detection (start wavelength nm 210, end wavelength nm 400, sample rate spectrum/sec 20). Injection volume (ul) -2.00. Sample solvent: h 2O/ACN 80/20
LC/UV/MS-method 7
LC instrument: acquisty Waters UPLC (or equivalent) interface 2996 PDA detector. Column: kinetex1.7u PFP 100A 100X 2.1mm (Phenomenex). Column temperature (. degree.C.) 55.0. Mobile phase: HCOONH40.025M pH 3 (A); ACN/MeOH 50/50 (B). Flow rate (ml/min)0.45 (split flow into MS 1: 3). End time (min)10
Gradient:
| time (min) | ％A | ％B |
| 0.0 | 85 | 15 |
| 5.0 | 55 | 45 |
| 5.5 | 20 | 80 |
| 6.5 | 20 | 80 |
| 7.0 | 85 | 15 |
And (4) UV detection: BPI detection (start wavelength nm 210, end wavelength nm 400, sample rate spectrum/sec 20). Injection volume (ul) -2.00. Sample solvent: ACN
LC/UV/MS-method 8
LC instrument: acquisty Waters UPLC (or equivalent) interface 2996 PDA detector. Column: kinetex1.7u PFP 100A 100X 2.1mm (Phenomenex). Column temperature (. degree. C.) 55 ℃. Mobile phase: HCOONH40.025M pH 3 (A); ACN/MeOH 85/15 (B). Flow rate (ml/min)0.45 (split flow into MS 1: 3). End time (min)10
Gradient:
| time (min) | ％A | ％B |
| 0.0 | 85 | 15 |
| 5.0 | 55 | 45 |
| 5.5 | 20 | 80 |
| 6.5 | 20 | 80 |
| 7.0 | 85 | 15 |
And (4) UV detection: BPI detection (start wavelength nm 210, end wavelength nm 400, sample rate spectrum/sec 20). Injection volume (ul) -2.00. Sample solvent: ACN
LC/UV/MS-method 9
LC instrument: acquisty Waters UPLC (or equivalent) interface 2996 PDA detector. Column: AcquisyteUPLC BEH C181.7um 50X2.1mm. column temperature (. degree. C.) 40.0. Mobile phase: 95: 5H 2O: ACN + (0.1% HCOOH) (a); 5: 95H 2O: ACN + (0.1% HCOOH) (B). Flow rate (mL/min)1 mL/min. End time (min) 2.
Gradient:
| time (min) | ％A | ％B |
| 0.00 | 99 | 1 |
| 1.50 | 0.1 | 99.9 |
| 1.90 | 0.1 | 99.9 |
| 2.00 | 99 | 1 |
And (4) UV detection: BPI detection (start wavelength nm 210, end wavelength nm 400, sample rate spectrum/sec 20). Injection volume (ul) -1.00
LC/UV/MS-method 10
LC instrument: acquisty Waters UPLC (or equivalent) interface 2996 PDA detector. Column: AcquisyteUPLC BEH C181.7um 50X2.1mm. column temperature (. degree. C.) 40.0. Mobile phase: 95: 5H 2O: ACN + (0.1% HCOOH) (a); 5: 95H 2O: ACN + (0.1% HCOOH) (B). Flow rate (mL/min)1 mL/min. End time (min)4
Gradient:
| time (min) | ％A | ％B |
| 0.00 | 99 | 1 |
| 3.50 | 0.1 | 99.9 |
| 3.90 | 0.1 | 99.9 |
| 4.00 | 99 | 1 |
And (4) UV detection: BPI detection (start wavelength nm 210, end wavelength nm 400, sample rate spectrum/sec 20). Injection volume (ul) -1.00
LC/UV/MS-method 11
LC instrument: acquisty Waters UPLC (or equivalent) interface 2996 PDA detector. Column: phenomenex kinetex 1.7um C8100 x2.1mm. column temperature (C) 55.0. Mobile phase: ammonium formate salt 25mM pH 3 (a): ACN + 0.1% (B). The flow rate (mL/min) was 0.5 mL/min. End time (min)10
Gradient:
| time (min) | ％A | ％B |
| 0.0 | 99 | 1 |
| 0.5 | 99 | 1 |
| 3.0 | 70 | 30 |
| 6.5 | 50 | 50 |
| 7.5 | 20 | 80 |
| 8.0 | 20 | 80 |
| 8.10 | 99 | 1 |
| 10.0 | 99 | 1 |
And (4) UV detection: BPI detection (start wavelength nm 210, stop UV detection: BPI detection (start wavelength nm 210, stop wavelength nm 400, sample rate spectrum/sec 20) injection volume (ul) -1.00
LC/UV/MS-method 12
LC instrument: UPLC/MS (ES +/ES-) AcquisytTM system coupled with ZQ mass spectrometer. Column: acquity UPLCCSH C18 column (50 mm. times.2.1 mm i.d.1.7 μm particle size). Column temperature (. degree. C.) was 40.0. Mobile phase: 0.1% v/v aqueous HCOOH solution (A); 0.1% v/v HCOOH in acetonitrile (B). Flow rate (ml/min) 1. End time (min)2.0
Gradient:
| time (min) | ％A | ％B |
| 0.00 | 97 | 3 |
| 1.50 | 0.1 | 99.9 |
| 1.90 | 0.1 | 99.9 |
| 2.00 | 97 | 3 |
UV detection range: 210nm-350 nm. The acquisition rate Hz is 20. Injection mode: the needle overfills part of the loop. DAD-MS Rt offset: 0.01min
LC/UV/MS-method 13
LC instrument: UPLC/MS (ES +/ES-) AcquisytTM system coupled with ZQ mass spectrometer. Column: acquity UPLCCSH C18 column (50 mm. times.2.1 mm i.d.1.7 μm particle size). Column temperature (. degree. C.) was 40.0. Mobile phase: 0.1% v/v aqueous HCOOH solution (A); 0.1% v/v HCOOH in acetonitrile (B). Flow rate (ml/min) 1. End time (min)2.0
Gradient:
| time (min) | ％A | ％B |
| 0.00 | 97 | 3 |
| 1.50 | 0.1 | 99.9 |
| 1.90 | 0.1 | 99.9 |
| 2.00 | 97 | 3 |
UV detection range: 210nm-350 nm. The acquisition rate Hz is 40. Injection mode: the needle overfills part of the loop. DAD-MS Rt offset: 0.01min
LC/UV/MS-method 14
LC instrument: a UPLC/MS (ES +/ES-) AcquisytTM system coupled to a Waters SQD2 mass spectrometer. Column: acquity UPLC CSH C18 column (50mm x 2.1mm i.d.1.7 μm particle size). Column temperature (. degree. C.) was 40.0. Mobile phase: 0.1% v/v aqueous HCOOH solution (A); 0.1% v/v HCOOH in acetonitrile (B). Flow rate (ml/min) 1. End time (min)2.0
Gradient:
| time (min) | ％A | ％B |
| 0.00 | 97 | 3 |
| 1.50 | 0.1 | 99.9 |
| 1.90 | 0.1 | 99.9 |
| 2.00 | 97 | 3 |
UV detection range: 210nm-350 nm. The acquisition rate Hz is 40. Injection mode: the needle overfills part of the loop. DAD-MS Rt offset: 0.01min
LC/UV/MS-method 15
LC instrument: a UPLC/MS (ES +/ES-) AcquisytTM system coupled to a Waters SQD2 mass spectrometer. Column: acquity UPLC CSH C18 column (50mm x 2.1mm i.d.1.7 μm particle size). Column temperature (. degree. C.) was 40.0. Mobile phase: H2O + 0.1% ammonia solution pH10 (a); acetonitrile (B). Flow rate (ml/min) 1. End time (min)2.0
Gradient:
| time (min) | ％A | ％B |
| 0.00 | 97 | 3 |
| 1.50 | 0.1 | 99.9 |
| 1.90 | 0.1 | 99.9 |
| 2.00 | 97 | 3 |
UV detection range: 210nm-350 nm. The acquisition rate Hz is 40. Injection mode: the needle overfills part of the loop. The scanning time limit: 0.10 second
LC/UV/MS-method 16
LC instrument: UPLC/PDA/MS AcquisytTM system coupled to Waters SQD mass spectrometer. Column: AcquisytUPLC BEH C18 column (50 mm. times.2.1 mm i.d.1.7 μm particle size). Column temperature (. degree. C.) was 40.0. Mobile phase: 10mM aqueous ammonium bicarbonate solution (A) adjusted to pH10 with ammonia; acetonitrile (B). Flow rate (ml/min) 1. End time (min)2.0
Gradient:
| time (min) | ％A | ％B |
| 0.00 | 97 | 3 |
| 1.50 | 0.1 | 99.9 |
| 1.90 | 0.1 | 99.9 |
| 2.00 | 97 | 3 |
UV detection range: 210nm-350 nm. The acquisition rate Hz is 40. Injection mode: the needle overfills part of the loop. The scanning time limit: 0.10 second
Analytical chirality for chiral compounds
The enantiomeric excess of the chiral compound was determined by chiral HPLC analysis using HPLC Agilent1100 equipped with a 6-position switching valve, DAD and CD detector. The following method was used:
Method A1: column: chiralpak AD-H (25 x 0.46cm), 5 um; mobile phase: n-hexane/(2-propanol + 0.1% isopropylamine) 75/25% v/v; flow rate: 1.0 mL/min; and D, DAD: 220 nm.
Method A2: column: whelk O-1(R, R) (25X 0.46cm), 5 um; mobile phase: n-hexane/(ethanol/methanol 1/1) 40/60% v/v; flow rate: 1.0 mL/min; and D, DAD: 220 nm.
Method A3: column: chiralpak IA (25 × 0.46cm), 5 um; mobile phase: n-hexane/(2-propanol/methanol 1/1) 60/40% v/v; flow rate: 1.0 mL/min; and D, DAD: 220 nm.
Method A4: column: chiralpak IC (25 × 0.46cm), 5 um; mobile phase: n-hexane/(2-propanol/methanol 1/1+ 0.1% isopropylamine) 90/10% v/v; flow rate: 1.0 mL/min; and D, DAD: 220 nm.
Method A5: column: chiralpak AD-H (25 x 0.46cm), 5 um; mobile phase: n-hexane/(2-propanol + 0.1% isopropylamine) 80/20% v/v; flow rate: 1.0 mL/min; and D, DAD: 220 nm; CD: 240 nm.
Method A6: column: whelk O-1(R, R) (25X 0.46cm), 10 um; mobile phase: n-hexane/(2-propanol + 0.1% isopropylamine) 40/60% v/v; flow rate: 1.0 mL/min; and D, DAD: 220 nm.
Method A7: column: whelk O-1(R, R) (25X 0.46cm), 10 um; mobile phase: n-hexane/(ethanol/dichloromethane 9/1+ 0.1% isopropylamine) 40/60% v/v; flow rate: 1.0 mL/min; and D, DAD: 280 nm.
Method A8: column: whelk 0-1(R, R) (25X 0.46cm), 5 um; mobile phase: n-hexane/(ethanol/dichloromethane 9/1+ 0.1% isopropylamine) 70/30% v/v; flow rate: 1.0 mL/min; and D, DAD: 280 nm.
Method A9: column: chiralpak IC (25 × 0.46cm), 5 um; mobile phase: n-hexane/(ethanol + 0.1% isopropylamine) 70/30% v/v; flow rate: 0.8 mL/min; and D, DAD: 220 nm.
Method A10: column: chiralpak AS-H (25X 0.46cm), 5 um; mobile phase: n-hexane/(2-propanol/methanol + 0.1% isopropylamine) 85/15% v/v; flow rate: 1.0 mL/min; and D, DAD: 220 nm.
Method A11: column: chiralpak AD-H (25 x 0.46cm), 5 um; mobile phase: n-hexane/(ethanol/methanol 1/1+ 0.1% isopropylamine) 80/20% v/v; flow rate: 0.8 mL/min; and D, DAD: 220 nm.
Method A12: column: chiralpak AD-H (25 x 0.46cm), 5 um; mobile phase: n-hexane/(ethanol + 0.1% isopropylamine) 75/25% v/v; flow rate: 0.8 mL/min; and D, DAD: 220 nm.
Method A13: column: whelk O-1(R, R) (25X 0.46cm), 10 um; mobile phase: n-hexane/(ethanol/methanol 1/1+ 0.1% isopropylamine) 75/25% v/v; flow rate: 1.0 mL/min; and D, DAD: 220 nm.
Method A14: column: whelk O-1(R, R) (25X 0.46cm), 10 um; mobile phase: n-hexane/(ethanol + 0.1% isopropylamine) 50/50% v/v; flow rate: 0.8 mL/min; and D, DAD: 220 nm.
Method A15: column: chiralpak IC (25 × 0.46cm), 5 um; mobile phase: n-hexane/(2-propanol/methanol 1/1+ 0.1% isopropylamine) 60/40% v/v; flow rate: 1.0 mL/min; and D, DAD: 220 nm.
Method A16: column: chiralpak AD-H (25 x 0.46cm), 5 um; mobile phase: n-hexane/(ethanol + 0.1% isopropylamine) 80/20% v/v; flow rate: 1.0 mL/min; and D, DAD: 220 nm.
Preparative reverse phase HPLC conditions
Preparative HPLC-method 1
Waters Micromass ZQ/sample manager 2767
A photodiode array detector 2996;
column: XTerra Prep MS C18 column (5 μm, 19X 150mm, Waters)
Flow rate: 20ml/min with MS detection
UV wavelength: 254 nm.
Mobile phase: solvent (water: MeCN: HCOOH 95: 5: 0.05); solvent B (Water: MeCN: HCOOH 5: 95: 0.05)
Gradient:
| time (min) | ％A | ％B |
| 0.00 | 100.0 | 0.00 |
| 1.00 | 100 | 0.00 |
| 10.00 | 0.00 | 100.0 |
| 11.00 | 0.00 | 100.0 |
| 12.00 | 100.0 | 0.00 |
Preparative HPLC-method 2
Column: waters Symmetry Prep C1817 um 19x300
Flow rate: 20ml/min
Mobile phase: 90% H2O, 10% acetonitrile, 0.05% tf (a); 10% H2O, 90% acetonitrile, 0.05% TF (B)
Gradient:
| time (min) | ％A | ％B |
| 0.00 | 95 | 5 |
| 2.5 | 95 | 5 |
| 22 | 0 | 100 |
| 30 | 0 | 100 |
Preparative HPLC-method 3
Waters Micromass ZQ/sample manager 2767
Photodiode array detector: 2996
Column: XTERRA Prep MS C1810 um 19x300
Flow rate: 20ml/min
Mobile phase: h 2O, 0.1% tfa (a); acetonitrile, 0.1% TF (B)
Gradient:
| time (min) | ％A | ％B |
| 0.00 | 90 | 10 |
| 2 | 90 | 10 |
| 23 | 0 | 100 |
| 30 | 0 | 100 |
Conditions are as follows:
| time (min) | ％A | ％B |
| 30.5 | 90 | 10 |
| 32 | 90 | 10 |
Preparative HPLC-method 4
Waters Fractionlynx with ZQ MS detector. Column: XSelect CSH Prep.C185um OBD30 x 100 mm. Flow rate: 43 ml/min. UV wavelength: 210nm-350 nm. Ionization mode: positive electrospray (ES +). Mobile phase: solvent a (H2O + 0.1% HCOOH); solvent B (acetonitrile)
Gradient:
| time (min) | ％A | ％B |
| 0.00 | 97.0 | 3.0 |
| 10.00 | 50.0 | 50.0 |
| 10.5 | 0.0 | 100.0 |
| 14.5 | 0.0 | 100.0 |
| 15.0 | 97.0 | 3.0 |
Chiral preparative HPLC for chiral compounds
Chiral resolution was performed using either a semi-preparative Waters 600 system or a semi-preparative Agilent 1100 system, the conditions being reported in the examples.
If the starting materials are prepared as unimodal, they are either commercially available, known in the literature or readily available to the skilled person using standard procedures.
Flash chromatography was performed using the Isolera MPLC system (manufactured by Biotage), using pre-packed silica gel or reverse phase column (provided by Biotage).
Many of the compounds described in the following examples have been prepared from stereochemically pure starting materials, e.g., 95% ee.
If indicated, the stereochemistry of the compounds in the indicated examples was presumed to be based on: the absolute configuration at the resolved stereocenter of the starting material is maintained throughout any subsequent reaction conditions.
In the following methods, after each starting material, the number of the compound referred to is sometimes provided. It is provided merely to assist one of ordinary skill in the chemical arts. The starting materials may not necessarily be prepared from the batch concerned.
When referring to the use of "analogous" or "analogous" methods, as will be appreciated by those skilled in the art, such methods may involve minor changes, such as reaction temperature, reagent/solvent amounts, reaction time, work-up conditions, or chromatographic purification conditions.
Preparation of an intermediate:
intermediate A1
4-bromo-3- (hydroxymethyl) -1H-isochromen-1-one
Step 1.2- (3-hydroxyprop-1-ynyl) benzoic acid methyl ester (intermediate 1.1)
Methyl 2-iodobenzoate (25g, 95mmol), prop-2-yn-1-ol (8.02g, 143mmol) and TEA (26.6ml, 191mmol) were added to Pd (PPh)3)4(0.220g, 0.191mmol), copper (I) iodide (0.073g, 0.382mmol) in DMF (60 ml). The resulting mixture was stirred at 90 ℃ for 5h and then at 40 ℃ overnight. The reaction mixture was poured into EtOAc (600mL) and washed with brine (600 mL). The organic phase is then concentrated with Et2O (600ml) rinse the dark oil. The mixture was filtered and then concentrated to give the title compound as a dark brown oil (13.0 g). The compound was used in the next step without any further purification and characterization.
UPLC-MS: 1.53min, [ M + H-18] +. (method 4)
Step 2.4-bromo-3- (hydroxymethyl) -1H-isochromen-1-one
Methyl 2- (3-hydroxyprop-1-ynyl) benzoate (intermediate 1.1, 6g, 31.5mmol), copper (II) bromide (14.09g, 63.1mmol) and pyridine (5.10ml, 63.1mmol) were reacted in refluxing acetonitrile (100ml) for 1 h. The solvent was then removed in vacuo and the crude product was washed with DCM (200ml) and filtered. Using 100g of SNAP silica gel column with DCM and Et2The resulting crude product was purified by O gradient to give the title compound (2.5g, 31%).
1H NMR(400MHz，DMSO-d6)ppm 8.21(dd，1H)，7.94-8.05(m，1H)，7.77-7.90(m，1H)，7.64-7.76(m，1H)，5.71(t，J＝6.39Hz，1H)，4.54(d，J＝6.17Hz，2H)。UPLC-MS：3.16min，[M+H]+. (method 5).
Intermediate A2
4-bromo-3- (1-hydroxyethyl) -1H-isochromen-1-one
Step 1.2- (3-hydroxybut-1-ynyl) benzoic acid methyl ester (intermediate 2.1)
Pd (PPh)3)4(1.9g, 0.006eq) and CuI (2.6g, 0.05eq) were suspended in DMF (350 ml). Methyl 2-iodobenzoate (70.8g, 1eq), but-3-yn-2-ol (32ml, 1.5eq), and TEA (75ml, 2eq) were added under nitrogen and the mixture was stirred at 60 ℃ for 6h and at RT overnight. The precipitated solid was filtered off, the mother liquor diluted with saturated aqueous NaCl and extracted with EtOAc (2 times). With Na2SO4The collected organic phase was dried, filtered and concentrated under reduced pressure to give the title compound as a light brown thick oil (47.7 g).
Step 2.4-bromo-3- (1-hydroxyethyl) -1H-isochromen-1-one
Methyl 2- (3-hydroxybut-1-ynyl) benzoate (intermediate 2.1, 20.6g, 100.98mmol) was dissolved in DCE (200ml) and CuBr was added at RT2(45g, 201.96mmol) and dicyclohexylamine hydrochloride (2.2g, 10.1 mmol). The mixture was heated to 65 ℃ for 2h under a nitrogen atmosphere. Thereby usingThe mixture was filtered through a pad and washed with DCM. The mother liquor was concentrated in vacuo and the residue was purified by silica gel column chromatography (Hex/EtOAc 8: 2). The solid residue was reacted with Hex/Et2O (1:1) was triturated together to give the title compound as a pale yellow powder (8.7g, 32%).
1H NMR(400MHz，DMSO-d6)ppm 8.20(d，J＝7.50Hz，1H)，7.99(m，1H)，7.85(d，J＝8.38Hz，1H)，7.71(m，1H)，5.66(d，J＝5.29Hz，1H)，5.12(m，1H)，1.38(d，J＝6.62Hz，3H)。UPLC-MS：1.83min，267[M+H]+ (method 1).
Intermediate A3
1- (4-bromo-1-oxo-1H-isochromen-3-yl) ethylcarbamic acid tert-butyl ester
Step 1. methanesulfonic acid but-3-yn-2-yl ester (intermediate 3.1)
But-3-yn-2-ol (25ml, 317mmol) was dissolved in DCM (6vol) under a nitrogen atmosphere. TEA (66ml, 475.5mmol) was added and the mixture was cooled to 5 ℃. Methanesulfonyl chloride (29ml, 380.4mmol) was added dropwise. The reaction was then warmed to RT and stirring continued overnight. The reaction mixture was then poured into water and extracted with DCM (2 times). With NaHCO3Washing the collected organic phase with saturated solution, and adding Na2SO4Drying, filtration and concentration in vacuo gave the title compound as a thick oil (34 g).
Step 2. tert-butyl but-3-yn-2-ylcarbamate (intermediate 3.2)
Butane-3-yn-2-yl methanesulfonate (intermediate 3.1, 34g) in 28-30% NH4Aqueous OH (31ml) was stirred vigorously at RT overnight. DCM (200ml) was added and the phases separated. With Na2SO4DryingAnd (4) organic phase filtering. The residual organic phase was heated to 40 ℃ until the vapors no longer gave a positive base test result. To this solution were added TEA (47.7ml) and di-tert-butyl dicarbonate (50g, 1eq) and stirring was maintained for 20 h. The reaction was stopped with water and the product was extracted with DCM. With Na2SO4The collected organic phase was dried, filtered and concentrated under reduced pressure to give the title compound (26.3g, 68%).
Step 3.2- (3- (tert-Butoxycarbonylamino) but-1-ynyl) benzoic acid methyl ester (intermediate 3.3)
Will be in DMF (30ml) and Et3Pd (Ph) in N (27ml)3P)4(683mg, 0.591mmol), CuI (938mg, 49.2mmol), methyl 2-iodobenzoate (25.8g, 98.49mmol), tert-butyl but-3-yn-2-ylcarbamate (intermediate 3.2, 25g, 147.75mmol) were combined in DMF (125ml) and the mixture was heated to 60 ℃ under a nitrogen atmosphere for 12 h. The mixture was quenched with water and the product was extracted with EtOAc. With Na2SO4The collected organic phase was dried, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (Hex/EtOAc 9:1) to give the title compound (15.3g, 51%).
Step 4.1- (4-bromo-1-oxo-1H-isochromen-3-yl) ethylcarbamic acid tert-butyl ester
In N2Methyl 2- (3- (tert-butoxycarbonylamino) but-1-ynyl) benzoate (intermediate 3.3, 5.6g, 18.48mmol) was dissolved in DCE (110ml) under an atmosphere; dicyclomethylamine hydrochloride (401mg, 1.85mmol) and CuBr were added2(8.3g, 37 mmol). The mixture was stirred at 70 ℃ for 3h and at RT overnight. Then useThe mixture was filtered through a pad, washed with DCM and the mother liquor concentrated in vacuo. The crude product was purified by silica gel column chromatography (DCM/EtOAc 8: 2) to give a solid residue, which was purified with hexane (4vol) and Et2Trituration of O (a few drops) together afforded the title compound (2.3g, 34%).
1H NMR(400MHz，DMSO-d6) d ppm 8.19(d, J ═ 7.72Hz, 1H), 7.92-8.06(m, 1H), 7.83(d, J ═ 7.94Hz, 1H), 7.65-7.75(m, 1H), 7.58(d, J ═ 5.73Hz, 1H), 4.76-5.13(m, 1H), 1.12-1.49(m, 12H). UPLC-MS (method 1).
Intermediate A4
4-bromo-3- (1-hydroxyethyl) -7-methyl-1H-isochromen-1-one
Following the procedure used for the synthesis of intermediate A2, from 2-iodo-5-methyl-benzoic acid methyl ester (2.0g, 7.2mmol) and but-3-yn-2-ol (0.850ml, 10.87mmol) the title compound was obtained (1.04g, 3.67mmol, 53%).
UPLC-MS: 0.96min, 283.1-285.1[ M + H ] +, method 13.
Intermediate A5
4-bromo-7-chloro-3- (1-hydroxyethyl) -1H-isochromen-1-one
Step 1.5-chloro-2-iodobenzoic acid methyl ester (intermediate A5.1)
5-chloro-2-iodobenzoic acid (3.0g, 10.62mmol), SO were sprayed with a hot gunCl2A solution of (12mL) and DMF (0.6mL) was warmed moderately until the mixture became homogeneous (15 min). The solution was maintained at 23 ℃ for a further 30min and then concentrated. MeOH (24mL) was added to the crude residue and the solution was maintained at 23 ℃ for 30 min. The solution was concentrated and the residue was purified by Biotage silica gel column flash chromatography (cyclohexane-cyclohexane: EtOAc ═ 85:15) to give methyl 5-chloro-2-iodobenzoate (3.02g, 10.20mmol, 96%).
UPLC-MS: 1.17min, 296.6[ M + H ] +, method 12.
And 2. step 2.
Following the procedure used for the synthesis of intermediate A2, from methyl 5-chloro-2-iodobenzoate (intermediate A5.1, 3.02g, 10.18mmol) and but-3-yn-2-ol (1.2ml, 15.28mmol) the title compound was obtained as a yellow solid (1.4g, 4.61 mmol).
UPLC-MS: 0.97min, 302.7-304.7[ M + H ] +, method 12.
Intermediate A6
4-bromo-6-chloro-3- (1-hydroxyethyl) -1H-isochromen-1-one
Step 1.4-chloro-2-iodobenzoic acid methyl ester (intermediate A6.1)
Following the procedure used for the synthesis of intermediate a5.1, from 4-chloro-2-iodobenzoic acid (2.00g, 7.08mmol) the title compound was obtained as a white solid (2.07g, 6.98mmol, 99%).
UPLC-MS: 1.20min, 297.0[ M + H ] +, method 13.
And 2. step 2.
Following the procedure used for the synthesis of intermediate A2, from methyl 4-chloro-2-iodobenzoate (intermediate A6.1, 2.07g, 6.98mmol) and but-3-yn-2-ol (0.821ml, 10.47mmol) the title compound was obtained as a tan solid (1.388g, 4.58mmol)
UPLC-MS: 1.00min, 303.0-305.0[ M + H ] +, method 13.
Intermediate A7
4-bromo-6-fluoro-3- (1-hydroxyethyl) -1H-isochromen-1-one
Step 1.4-fluoro-2-iodobenzoic acid methyl ester (intermediate A7.1)
Methyl 4-fluoro-2-iodobenzoate was obtained from 4-fluoro-2-iodobenzoic acid (intermediate a7.1, 1.153g, 4.11mmol, 99%) according to the procedure used for the synthesis of intermediate a 5.1.
UPLC-MS: 1.09min, 281.0[ M + H ] +, method 13.
And 2. step 2.
From methyl 4-fluoro-2-iodobenzoate (intermediate A7.1, 1.153g, 4.11mmol) and but-3-yn-2-ol (0.484ml, 6.176mmol) according to the method for the synthesis of intermediate A2, the title compound (1g, 3.48mmol) was obtained
UPLC-MS: 0.89min, 287.0-289.0[ M + H ] +, method 13.
Intermediate A8
4-bromo-3- (1- ((tert-butyldimethylsilyl) oxy) ethyl) -1H-isochromen-1-one
4-bromo-3- (1-hydroxyethyl) -1H-isochromen-1-one (intermediate A2, 5g, 18.658mmol) was dissolved in DCM (50 ml); imidazole (2.54g, 37.3mmol) and tert-butyl (chloro) dimethylsilane (5.624g, 37.3mmol) were added and the mixture was stirred at RT for 1 h. The mixture was washed with brine, the organic phase was dried over sodium sulfate and the solvent was removed under reduced pressure to give the crude product which was purified by flash chromatography on a silica gel Biotage column (cyclohexane: EtOAc ═ 95:5-60:40) to give the title compound as a white solid (6.5g, 16.97mmol, 91%).
UPLC-MS: 1.58min, 383.3-385.3[ M + H ] +, method 13.
Intermediate B1
3- (1-hydroxyethyl) -4-phenyl-1H-isochromen-1-one
4-bromo-3- (1-hydroxyethyl) -1H-isochromen-1-one (intermediate A2, 0.77g, 2.88mmol), phenylboronic acid (0.63g, 5.18mmol), Pd (PPh)3)4(0.199g, 0.173mmol) and Cs2CO3(1.49g, 4.6mmol) was dissolved in DMF (10.7ml) and heated at 120 ℃ for 20min under microwave irradiation. The reaction mixture was then diluted with AcOEt (100mL) and filtered. The organic phase was washed 2 times with 0.5M aqueous HCl, with water and with saturated NaHCO3Washed 2 times and 1 time with saturated aqueous NaCl. With Na2SO4The organic phase obtained is dried, filtered and concentrated. The crude product was finally purified using a Biotage Si 50g Ultra gradient with hexanes and EtOAc. The title compound was recovered as a dark pink solid (0.53g, 1.99mmol, 69.3%).
UPLC-MS: 1.83min, 267[ M + H ] +, method 1.
Intermediate B2
3- (hydroxymethyl) -4-phenyl-1H-isochromen-1-one
Prepared in a similar manner to intermediate B1 from 4-bromo-3- (hydroxymethyl) -1H-isochromen-1-one (intermediate A1, 0.8g, 3.14mmol), phenylboronic acid (0.467g, 3.83mmol), Pd (PPh)3)4(0.181g, 0.379mmol) and Cs2CO3(3.29g, 10.11mmol) was used as the starting material to prepare the title compound, yielding the title compound (210mg, 36%).
UPLC-MS: 1.80min, 271[ M + H ] +, method 4.
Intermediate B3
4- (3-fluorophenyl) -3- (hydroxymethyl) -1H-isochromen-1-one
4-bromo-3- (hydroxymethyl) -1H-isochromen-1-one (intermediate A1, 0,5g, 1.960mmol), 3-fluorophenylboronic acid (0.4g, 2.86mmol), Pd (PPh) were used in a similar manner to intermediate B23)4(0.136g, 0.118mmol) and Cs2CO3(0.96g, 2.94mmol) to give the title compound (0.21g, 40%).
UPLC-MS: 1.80min, 271[ M + H ] +, method 4.
Intermediate B4
4- (2-fluorophenyl) -3- (hydroxymethyl) -1H-isochromen-1-one
4-bromo-3- (hydroxymethyl) -1H-isochromen-1-one (intermediate A1, 0.6g, 2.352mmol), 2-fluorophenylboronic acid (0.494g, 3.53mmol), Pd (PPh) were used in a similar manner to intermediate B23)4(0.136g, 0.118mmol) and Cs2CO3(0.77g, 2.35mmol) to give the title compound (0.21g, 33%).
UPLC-MS: 1.80min, 271[ M + H ] +, method 4.
Intermediate B5
3- (hydroxymethyl) -4-m-tolyl-1H-isochromen-1-one
4-bromo-3- (hydroxymethyl) -1H-isochromen-1-one (intermediate A1, 0.5g, 1.96mmol), m-tolylboronic acid (0.400g, 2.94mmol), Pd (PPh) were used in a similar manner to intermediate B2 3)4(0.113g, 0.098mmol) and Cs2CO3(0.64g, 1.96mmol) to give the title compound (0.21g, 40%).
UPLC-MS: 1.89min, 267[ M + H ] +, method 4.
Intermediate B6
3- (1-hydroxyethyl) -4-m-tolyl-1H-isochromen-1-one
4-bromo-3- (1-hydroxyethyl) -1H-isochromen-1-one (intermediate A2, 0.52g, 1.93mmol), m-tolylboronic acid (0.45g, 3.28mmol), Pd (PPh) were used in a similar manner to intermediate B13)4(0.11g, 0.096mmol) and Cs2CO3(0.81g, 2.5mmol) to give the title compound (0.3g, 55%).
UPLC-MS: 5.18min, 281[ M + H ] +, method 5
Intermediate B7
4- (3-fluorophenyl) -3- (1-hydroxyethyl) -1H-isochromen-1-one
4-bromo-3- (1-hydroxyethyl) -1H-isochromen-1-one (intermediate A2, 0.8g, 2.97mmol), 3-fluorophenylboronic acid (0.62g, 4.46mmol), Pd (PPh) were used in a similar manner to intermediate B13)4(0.17g, 0.149mmol) and Cs2CO3(0.97g, 2.9mmol) to give the title compound (0.39g, 46%).
UPLC-MS: 1.85min, 285[ M + H ] +, method 5.
Intermediate B8
4- (3- (dimethylamino) phenyl) -3- (1-hydroxyethyl) -1H-isochromen-1-one
4-bromo-3- (1-hydroxyethyl) -1H-isochromen-1-one (intermediate A2, 0.5g, 1.86mmol), 3- (dimethylamino) phenylboronic acid (0.46g, 2.79mmol), Pd (PPh) were used in a similar manner to intermediate B1 3)4(0.11g, 0.093mmol) and Cs2CO3(0.78g, 2.41mmol) to give the title compound (0.2g, 35%).
UPLC-MS: 1.87min, 350[ M + H + ACN ] +, method 4.
Intermediate B9
3- (1-hydroxyethyl) -4- (3- (morpholinosulfonyl) phenyl) -1H-isochromen-1-one
4-bromo-3- (1-hydroxyethyl) -1H-isochromen-1-one (intermediate A2, 0.3g, 1.11mmol), 3- (4-morpholinosulfonyl) phenylboronic acid (0.45g, 1.67mmol), Pd (PPh) were used in a similar manner to intermediate B13)4(0.064g, 0.056mmol) and Cs2CO3(0.47g, 1.45mmol) to give the title compound (0.163g, 35%).
UPLC-MS: 1.70min, 416[ M + H ] +, method 4.
Intermediate B10
3- (1-hydroxyethyl) -4- (6-methylpyridin-3-yl) -1H-isochromen-1-one
In a similar manner to intermediate B1, 4-bromo-3- (1-hydroxyethyl) -1H-isochromen-1-one (intermediate A2, 0.55g, 2.04mmol), 6-methylpyridin-3-ylboronic acid (0.550g, 2.0mmol), Pd (PPh)3)4(0.118g, 0.102mmol) and Cs2CO3(1.06g, 3.27mmol) to give the title compound (0.163g, 28%)
UPLC-MS: 1.25min, 282[ M + H ] +, method 4.
Intermediate B11
3- (1-hydroxyethyl) -4- (thiazol-5-yl) -1H-isochromen-1-one
4-bromo-3- (1-hydroxyethyl) -1H-isochromen-1-one (intermediate A2, 0.1g, 0.372mmol), 6-methyl-2- (thiazol-5-yl) -1,3,6, 2-dioxazaboroxin-4, 8-dione (0.134g, 0.56mmol), Pd (PPh)3)4(0.021g, 0.019mmol) and Cs2CO3(0.182g, 0.56mmol) in DMF (1mL) was heated under microwave irradiation at 120 deg.C1h and 15 min. Then 6-methyl-2- (thiazol-5-yl) -1,3,6, 2-dioxazaboroxin-4, 8-dione (0.134g, 0.56mmol), Pd (PPh) were added3)4(0.021g, 0.019mmol) and Cs2CO3(0.182g, 0.56mmol) and the resulting mixture was then allowed to react at 100 ℃ for 5 h. The crude product was purified by reverse phase chromatography with a Biotage C1830 g SNAP column (phase a, water 95%, ACN 4.9%, formic acid 0.1%; phase B, ACN 99.9%, formic acid 0.1%) to give the title compound (102 mg).
UPLC-MS: 1.48min, 274[ M + H ] +, method 4
Intermediate B12
4- (3- (1-hydroxyethyl) -1-oxo-1H-isochromen-4-yl) -5, 6-dihydro-pyridine-1 (2H) -carboxylic acid tert-butyl ester
In a similar manner to intermediate B1 using 4-bromo-3- (1-hydroxyethyl) -1H-isochromen-1-one (intermediate A2, 0.55g, 2.04mmol), 4- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -5, 6-dihydropyridine-1 (2H) -carboxylic acid tert-butyl ester (0.79g, 2.55mmol), Pd (PPh) 3)4(118mg, 0.102mmol) and Cs2CO3(1.0g, 3.27mmol) the title compound was prepared as a pale yellow oil (0.135g, 18%).
UPLC-MS: 1.91min, 371[ M + H ] +, method 4
Intermediate B13
3- (1-hydroxyethyl) -4- (2-methylpyridin-4-yl) -1H-isochromen-1-one
In a similar manner to intermediate B1, 4-bromo-3- (1-hydroxyethyl) -1H-isochromen-1-one (center)Intermediate A2, 0.5g, 1.86mmol), 2-methylpyridin-4-ylboronic acid (0.38g, 2.79mmol), Pd (PPh)3)4(0.107g, 0.093mmol) and Cs2CO3(0.78g, 2.41mmol) the title compound was prepared. The crude product was purified by reverse phase chromatography with a Biotage C18SNAP30g column (phase a, water 95%, ACN 4.9%, formic acid 0.1%; phase B, ACN 99.9%, formic acid 0.1%) to give the title compound (0.25g, 48%).
UPLC-MS: 1.20min, 282[ M + H ] +, method 4.
Intermediate B14
4-benzyl-3- (1-hydroxyethyl) -1H-isochromen-1-one
3- (1-hydroxyethyl) -1H-isochromen-1-one (intermediate A2, 0.6g, 2.23mmol), 2-benzyl-4, 4,5, 5-tetramethyl-1, 3, 2-dioxaborolan (0.730g, 3.3mmol), Pd (PPh) were used in a similar manner to intermediate B13)4(0.129g, 0.111mmol) and Cs2CO3(1.07g, 3.12mmol) the title compound was prepared to give the title crude compound (0.62g) which was used without further purification.
UPLC-MS: 1.82min, 281[ M + H ] +, method 4.
Intermediate B15
3- (1-hydroxyethyl) -4- (4- (2-morpholinoethoxy) phenyl) -1H-isochromen-1-one
In a similar manner to intermediate B1 using 4-bromo-3- (1-hydroxyethyl) -1H-isochromen-1-one (intermediate A2, 0.36g, 1.35mmol), 4- (2- (4- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) phenoxy) ethyl) -morpholine (0.45g, 1.35mmol)、Pd(PPh3)4(0.078g, 0.068mmol) and Cs2CO3(0.53g, 1.6mmol) the title compound was prepared. The crude product was filtered, diluted with 1M aqueous HCl (3ml) and purified by reverse phase chromatography on a Biotage C18 SNAP 120g column (phase a, water 95%, ACN 4.9%, formic acid 0.1%); phase B, ACN 99.9%, formic acid 0.1%) to give the title compound (0.179g, 33%).
UPLC-MS: 1.36min, 396[ M + H ] +, method 4
Intermediate B16
3- (1-hydroxyethyl) -4- (5- (morpholinomethyl) thiophen-2-yl) -1H-isochromen-1-one
4-bromo-3- (1-hydroxyethyl) -1H-isochromen-1-one (intermediate A2, 0.4g, 1.49mmol), 4- ((5- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) thiophen-2-yl) methyl) morpholine (0.598g, 1.93mmol), Pd (PPh) were used in a similar manner to intermediate B13)4(0.086g, 0.074mmol) and Cs 2CO3(0.678g, 2.1mmol) the title compound was prepared. The crude product was purified by reverse phase chromatography with a Biotage C18SNAP 60g column (phase a, water 95%, ACN 4.9%, formic acid 0.1%); phase B, ACN 99.9%, formic acid 0.1%) to give the title compound (0.26g, 48%).
UPLC-MS: 1.21min, 372[ M + H ] +, method 4.
Intermediate B17
3- (1-hydroxyethyl) -4- (6-methoxypyridin-3-yl) -1H-isochromen-1-one
In a similar manner to intermediate B1, 4-bromo-3- (1-hydroxyethyl) -1H-isochromen-1-Ketone (intermediate A2, 0.6g, 2.23mmol), 6-methoxypyridin-3-ylboronic acid (0.443g, 2.90mmol), Pd (PPh)3)4(0.206g, 0.178mmol) and Cs2CO3(1.02g, 3.1mmol) the title compound was prepared. The crude product was purified by reverse phase chromatography using a Biotage 30g C18SNAP column (phase a, water 95%, acetonitrile 4.9%, formic acid 0.1%; phase B, acetonitrile 99.9%, formic acid 0.1%) to give the title compound (0.25g, 38%).
UPLC-MS: 1.69min, 298[ M + H ] +, method 4.
Intermediate B18
4- (3, 6-dihydro-2H-pyran-4-yl) -3- (1-hydroxyethyl) -1H-isochromen-1-one
4-bromo-3- (1-hydroxyethyl) -1H-isochromen-1-one (intermediate A2, 0.50g, 1.86mmol), 2- (3, 6-dihydro-2H-pyran-4-yl) -4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan (0.51g, 2.42mmol), Pd (PPh) were used in a similar manner to intermediate B1 3)4(0.11g, 0.093mmol) and Cs2CO3(0.30mg, 0.93mmol) the title compound was prepared to give the title compound (0.13g, 26%) as an orange oil.
UPLC-MS: 1.52min, 273[ M + H ] +, method 4
Intermediate B24
3- (1-hydroxyethyl) -4- (3- (4,4,5, 5-tetramethyl-1, 3-dioxolan-2-yl) phenyl) -1H-isochromen-1-one
4-bromo-3- (1-hydroxyethyl) -1H-isochromen-1-one (intermediate A2, 4.05G, 15.05mmol), 4,5, 5-tetramethyl-2- (3- (4,4,5, 5-tetramethyl-1, 3-dioxolan-2-yl) phenyl) -1,3, 2-dioxaborolane (intermediate G2, 5G, 15.05mmol), X-Phos-Pd-G2(1.184G, 1.505mmol) and K3PO4·H2O (9.81g, 30.1mmol) was dispersed in THF (42mL), deoxygenated under an argon atmosphere, then water (42mL) was added and the mixture stirred at rt overnight. The reaction was diluted with AcOEt (250ml), washed with 0.2M aqueous HCl (250ml), 1 time with saturated aqueous NaCl and Na2SO4Drying and removing the solvent under reduced pressure. The crude product was purified by flash chromatography on silica gel using a Biotage 100G +50G SNAP gradient with hexanes and AcOEt to give the title compound (5.4G, 91%) as a dark oil.
UPLC-MS：1.19min，377.2[(M-H2O)+H]Method 9
Intermediate B31
(5- (3- (1-bromoethyl) -1-oxo-1H-isochromen-4-yl) thiazol-2-yl) carbamic acid tert-butyl ester
To a solution of 4-bromo-3- (1-hydroxyethyl) -1H-isochromen-1-one (intermediate a2, 500mg, 1.858mmol), Pd-bis (diphenylphosphine) chloride (130mg, 0,186mmol), cesium fluoride (847mg, 5,57mmol) in 1, 4-dioxane (5ml) was added commercial tert-butyl (5- (tributylstannyl) thiazol-2-yl) carbamate (1.182g, 2,42mmol), and the mixture was stirred at RT for 4H, at 80 ℃ for 1H. The reaction system is partitioned between NH4Between Cl (100ml) and AcOEt (30ml), the organic phase was washed with brine, dried and evaporated in vacuo. Reverse phase chromatography with a Biotage C18 SNAP 30g column (phase a, water 95%, ACN 4.9%, formic acid 0.1%); phase B, ACN 95%, water 5%, formic acid 0.1%) to give the title compound (255mg, 35.3%)
1H NMR(400MHz，DMSO-d6)ppm 8.24-8.35(m，1H)，7.82-7.93(m，1H)，7.53-7.76(m，8H)，7.38-7.47(m，1H)，7.10-7.23(m，1H)，6.82-6.99(m，1H)，5.46-5.70(m，1H)，4.12-4.33(m，1H)，1.48(s，4H)，1.26-1.39(m，11H)
Intermediate B34
(4-phenyl-1H-isochromen-3-yl) methanol
4-phenyl-1H-isochromene-3-carbaldehyde (intermediate G21, 410mg, 1,735mmol) was suspended in MeOH (1ml) and added dropwise at RT to a solution of sodium tetrahydroborate (65,7mg, 1,735mmol) in MeOH (17,3 ml). The mixture was partitioned between AcOEt/NH4Cl 5% (2ml/2ml), the organic phase was separated and Na was used2SO4Drying and evaporation in vacuo gave the crude product which was taken on to the next step without any further purification.
1H NMR(400MHz，DMSO-d6)d ppm 7.34-7.48(m，3H)，7.21-7.29(m，2H)，7.05-7.19(m，3H)，6.37-6.56(m，1H)，5.09(s，2H)，4.85-4.96(t，1H)，3.74-3.85(d，2H)
Intermediate B37
5- (3- (1-hydroxyethyl) -1-oxo-1H-isochromen-4-yl) thiophene-2-carbaldehyde
3- (1-hydroxyethyl) -4- (5- (4,4,5, 5-tetramethyl-1, 3-dioxolan-2-yl) thiophen-2-yl) -1H-isochromen-1-one (intermediate B29, 940mg, 2.347mmol) was dissolved in 20ml of MeCN and 20ml of HCl 1M was added. The clear yellow solution was stirred at rt overnight. The mixture was diluted with 50ml of water, 200ml of EtOAc were added and then stirred for 20 min. The phases are separated and the reaction is carried out with 100ml NaHCO3The saturated solution washed the organic phase 1 time. After separation of the phases, the organic phase was dried over sodium sulfate, filtered and concentrated to dryness to give 5- (3- (1-hydroxyethyl) -1-oxo-1H-isochromen-4-yl) thiophene-2-carbaldehyde (700mg, 2.331mmol, 99% yield) as a yellow loose solid.
UPLC-MS: 0.83min, 300.98[ M + H ] +, method 9.
Intermediate B38
Benzyl 4- ((5- (3- (1-hydroxyethyl) -1-oxo-1H-isochromen-4-yl) thiophen-2-yl) methyl) piperazine-1-carboxylate
In a 250ml round bottom flask, 5- (3- (1-hydroxyethyl) -1-oxo-1H-isochromen-4-yl) thiophene-2-carbaldehyde (intermediate B37) (780mg, 2.60mmol) was dissolved in 30ml DCM, followed by the addition of acetic acid (0.446ml, 7.79mmol) and benzyl piperazine-1-carboxylate (1.503ml, 7.79 mmol). After a few minutes, sodium triacetoxyborohydride (2.7) was added 5g, 12.99mmol), the mixture was stirred at r.t. The mixture was poured into 100ml DCM and 100NaHCO3Saturated solution, then phases were separated and the organic phase was concentrated to dryness to give a brown oil, which was immediately purified by chromatography eluting with a hexane/EtOAc mixture to give the title compound (903mg, 1.790mmol, 68.9% yield) as a yellow oil.
UPLC-MS: 0.79min, 505.12[ M + H ] +, method 9
Intermediate B39
3- (1-hydroxyethyl) -4- (5- ((4-methylpiperazin-1-yl) methyl) thiophen-2-yl) -1H-isochromen-1-one
The title compound was prepared from 5- (3- (1-hydroxyethyl) -1-oxo-1H-isochromen-4-yl) thiophene-2-carbaldehyde (intermediate B37) (800mg, 2.66mmol)), 1-methylpiperazine (913 μ l, 7.99mmol) in a similar manner to intermediate B38 to give the title compound (412mg, 1.072mmol, 40.2% yield) as a yellow oil.
UPLC-MS: 0.56min, 385.13[ M + H ] +, method 9.
Intermediate B40
4- (5- (3- (dimethylamino) prop-1-en-1-yl) thiophen-2-yl) -3- (1-hydroxyethyl) -1H-isochromen-1-one
In a 100ml 3-neck round bottom flask, (2- (dimethylamino) ethyl) triphosphium bromide (1569mg, 3,79mmol) was charged and suspended in 15ml dioxane under an argon atmosphere. A solution of potassium bis (trimethylsilyl) amide in 0.5M toluene (6,0ml, 3,03mmol) was added dropwise and the yellow/orange color appeared. The mixture was stirred for a further 20min and then dissolved dropwise in 5ml of dioxane 5- (3- (1-hydroxyethyl) -1-oxo-1H-isochromen-4-yl) thiophene-2-carbaldehyde (intermediate B37, 455mg, 1,515mmol) was added to a 0.5M toluene solution (6,0ml, 3,03mmol), and the yellow/orange color appeared. The mixture was stirred for a further 20min, then 5- (3- (1-hydroxyethyl) -1-oxo-1H-isochromen-4-yl) thiophene-2-carbaldehyde (intermediate B37, 455mg, 1,515mmol) dissolved in 5ml dioxane was added. The mixture is stirred for a further 2h at rt and then poured into 100ml NH4The solution was saturated with Cl and dried 1 time over sodium sulfate. The solvent was removed and the crude product was purified by chromatography eluting with a mixture of DCM \ 20% MeOH in DCM to give 4- (5- (3- (dimethylamino) prop-1-en-1-yl) thiophen-2-yl) -3- (1-hydroxyethyl) -1H-isochromen-1-one (220mg, 0,619mmol, 40, 9% yield) as a yellow oil.
UPLC-MS: 0.62min, 356.15[ M + H ] +, method 9
Intermediate B41
53- (3- (1-hydroxyethyl) -1-oxo-1H-isochromen-4-yl) benzaldehyde
The title compound was prepared from 3- (1-hydroxyethyl) -4- (3- (4,4,5, 5-tetramethyl-1, 3-dioxolan-2-yl) phenyl) -1H-isochromen-1-one (intermediate B24, 1,268mmol, 500mg) in analogy to intermediate B37 to yield the title compound (400mg, 1,359mmol, 107%) as a yellow oil.
UPLC-MS: 0.86min, [ M + H ] +, method 10.
Intermediate B42
4- (3- (3- (dimethylamino) prop-1-en-1-yl) phenyl) -3- (1-hydroxyethyl) -1H-isochromen-1-one
The title compound was prepared from 3- (3- (1-hydroxyethyl) -1-oxo-1H-isochromen-4-yl) benzaldehyde (intermediate B41, 373mg, mmol) and (2- (dimethylamino) ethyl) triphenylphosphonium bromide (3.17mmol, 1.313mg) in analogy to intermediate B40 to give the title compound (280mg, 0.801mmol, 63.2% yield) as a yellow oil.
UPLC-MS: 1.09min, 350.20[ M + H ] +, method 10.
Intermediate 44
4- (3- (1-hydroxyethyl) -1-oxo-1H-isochromen-4-yl) benzaldehyde
The title compound was prepared from 3- (1-hydroxyethyl) -4- (4- (4,4,5, 5-tetramethyl-1, 3-dioxolan-2-yl) phenyl) -1H-isochromen-1-one (intermediate B43, 1,83g, 4,64mmol) in analogy to intermediate B37 to yield the title compound (1,12g, 3,81mmol, 82% yield) as a yellow oil.
UPLC-MS：1.42min，277.17[M+H-H2O]+, method 10.
Intermediate B45
4- (4- (3- (dimethylamino) prop-1-en-1-yl) phenyl) -3- (1-hydroxyethyl) -1H-isochromen-1-one
The title compound was prepared from 4- (3- (1-hydroxyethyl) -1-oxo-1H-isochromen-4-yl) benzaldehyde (intermediate B44, 480mg, 1.63mmol) and (2- (dimethylamino) ethyl) triphenylphosphonium bromide (4.08mmol, 1.69mg) in analogy to intermediate B40 to give the title compound (210mg, 0,601mmol, 36, 8% yield) as a yellow oil.
UPLC-MS: 0.49min, 350.21[ M + H ] +, method 9.
Intermediate B46
3- (1-hydroxyethyl) -4- (4- ((4-methylpiperazin-1-yl) methyl) phenyl) -1H-isochromen-1-one
The title compound was prepared from 4- (3- (1-hydroxyethyl) -1-oxo-1H-isochromen-4-yl) benzaldehyde (intermediate B44, 320mg, 1.087mmol) and 1-methylpiperazine (373 μ l, 3.26mmol) in analogy to intermediate B38 to give the desired product as 3- (1-hydroxyethyl) -4- (4- ((4-methylpiperazin-1-yl) methyl) phenyl) -1H-isochromen-1-one (428mg, 1.131mmol, 104% yield) as a yellow oil.
UPLC-MS: 0.52min, 379.33[ M + H ] +, method 9.
Intermediate B47
3- (1-hydroxyethyl) -4- (4- (pyrrolidin-1-ylmethyl) phenyl) -1H-isochromen-1-one
The title compound was prepared from 4- (3- (1-hydroxyethyl) -1-oxo-1H-isochromen-4-yl) benzaldehyde (intermediate B44, 320mg, 1.087mmol) and pyrrolidine (232mg, 3.26mmol) in analogy to example B37 to give the title compound (353mg, 1.010mmol, 93% yield) as a yellow oil.
UPLC-MS: 0.55min, 350.25[ M + H ] +, method 9
Intermediate B48
3- (1-hydroxyethyl) -4- (4- (piperidin-1-ylmethyl) phenyl) -1H-isochromen-1-one
The title compound was prepared from 4- (3- (1-hydroxyethyl) -1-oxo-1H-isochromen-4-yl) benzaldehyde (intermediate B44, 650mg, 2.209mmol) and piperidine (654 μ l, 6.63mmol) in analogy to intermediate B37 to give the title compound (720mg, 1.981mmol, 90% yield) as a yellow oil.
UPLC-MS: 0.58min, 364.25[ M + H ] +, method 9
Intermediate B49
3- (1-hydroxyethyl) -4- (3- ((4-methylpiperazin-1-yl) methyl) phenyl) -1H-isochromen-1-one
The title compound was prepared from 3- (3- (1-hydroxyethyl) -1-oxo-1H-isochromen-4-yl) benzaldehyde (intermediate B42, 530mg, 1.801mmol) and 1-methylpiperazine (617 μ l, 5.40mmol) in analogy to intermediate B37 to give the title compound (341mg, 0.900mmol, 50% yield) as a yellow oil.
UPLC-MS: 0.57min, 379.28[ M + H ] +, method 9
Intermediate B52
(3- (3- (1-hydroxyethyl) -1-oxo-1H-isochromen-4-yl) prop-2-yn-1-yl) (methyl) carbamic acid benzyl ester
To a solution of 4-bromo-3- (1-hydroxyethyl) -1H-isochromen-1-one (intermediate A2, 2.65G, 9.84mmol) in dry DMF (30ml) was added benzyl methyl (prop-2-yn-1-yl) carbamate (intermediate G22, 4G, 19.68mmol), copper (I) iodide (0.562G, 2.95mmol), tetrakis (triphenylphosphine) palladium (0) (1.946ml, 1.476mmol), benzyl methyl (prop-2-yn-1-yl) carbamate (4G, 19.68mmol) and triethylamine (4.04ml, 29.5 mmol). The resulting suspension was heated to 95 ℃ overnight. The solvent was then removed under reduced pressure. The product was purified by Biotage Si 10g eluting with a heptane and EtOAc gradient to give the title compound (3.524g, 9.00mmol, 91% yield) as a light yellow oil.
UPLC-MS: 1.08min, 392[ M + H ] +, method 9
Intermediate B53
3- (1-hydroxyethyl) -7-methyl-4-phenyl-1H-isochromen-1-one
To 4-bromo-3- (1-hydroxyethyl) -7-methyl-1H-isochromen-1-one (intermediate a4, 1.04g, 3.67mmol) in dioxane/H2To a solution in O (5:1) (60ml) were added phenylboronic acid (0.67g, 5.51mmol) and Na2CO3(0.779g, 7.34mmol) followed by the addition of Pd (dppf) Cl2(0.269g, 0.367mmol), the resulting mixture was heated at 90 ℃ for 2h, 1N HCl (pH. apprxeq.2) was added, and the mixture was partitioned between EtOAc and water. The organic phase was washed 2 times with brine and dried over sodium sulfate. The solvent was removed in vacuo and the crude product was purified by flash chromatography using a Biotage silica gel column (cyclohexane: EtOAc ═ 95:5-40:60) to give the title compound as a yellow oil (0.835g, 2.98mmol, 81%).
UPLC-MS: 1.12min, 281.2[ M + H ] +, method 13
Intermediate B54
7-chloro-3- (1-hydroxyethyl) -4-phenyl-1H-isochromen-1-one
The title compound was prepared in analogy to intermediate B53 from 4-bromo-7-chloro-3- (1-hydroxyethyl) -1H-isochromen-1-one (intermediate a5, 1.4g, 4.61mmol) and phenylboronic acid (0.844g, 6.92mmol) to yield the title compound as a yellow foam (0.540g) and was used without any additional purification.
UPLC-MS: 1.11min, 300.8[ M + H ] +, method 12
Intermediate B55
6-chloro-3- (1-hydroxyethyl) -4-phenyl-1H-isochromen-1-one
The title compound was prepared in analogy to intermediate B53 from 4-bromo-6-chloro-3- (1-hydroxyethyl) -1H-isochromen-1-one (intermediate a6, 0.973g, 3.205mmol) and phenylboronic acid (0.586g, 4.808mmol) to yield the title compound as a white foam (0.430g, 1.43mmol, 45%).
UPLC-MS: 1.12min, 301.1[ M + H ] +, method 13
Intermediate B56
6-fluoro-3- (1-hydroxyethyl) -4-phenyl-1H-isochromen-1-one
The title compound was prepared in analogy to intermediate B53 from 4-bromo-6-fluoro-3- (1-hydroxyethyl) -1H-isochromen-1-one (intermediate a7, 0.95g, 3.31mmol) and phenylboronic acid (0.605g, 4.96mmol) to give the title compound (0.736g) which was used without any additional purification.
UPLC-MS: 1.04min, 285.2[ M + H ] +, method 13
Intermediate B58
3- (1-hydroxyethyl) -4- (pyridin-2-yl) -1H-isochromen-1-one
Step 1.3- (1- ((tert-butyldimethylsilyl) oxy) ethyl) -4- (pyridin-2-yl) -1H-isochromen-1-one (intermediate B58.1)
To a solution of degassed 4-bromo-3- (1- ((tert-butyldimethylsilyl) oxy) ethyl) -1H-isochromen-1-one (intermediate A8, 2g, 5.22mmol) in toluene (30ml) was added PdCl 2(PPh3)2(0.183g, 0.261mmol) followed by 2- (tributylstannyl) pyridine (4.55ml, 10.44mmol) and the resulting mixture was heated to reflux overnight. The mixture was cooled to RT and then filtered through celite. The solvent was removed in vacuo, the crude product was dissolved in AcOEt, saturated aqueous KF was added and the mixture was stirred for 2 h. The organic phase was separated, dried over sodium sulfate and evaporated to dryness. The crude product was purified by flash chromatography on a silica gel Biotage column (cyclohexane-cyclohexane: EtOAc ═ 20:80) to give 3- (1- ((tert-butyldimethylsilyl) oxy) ethyl) -4- (pyridin-2-yl) -1H-isochromen-1-one as a light yellow oil (intermediate B58.1, 0.508g, 1.33 mmol).
UPLC-MS: 1.42min, 382.4[ M + H ] +, method 13
And 2. step 2.
A1.0M THF solution of TBAF (1.43mL, 1.43mmol) was added dropwise to a solution of 3- (1- ((tert-butyldimethylsilyl) oxy) ethyl) -4- (pyridin-2-yl) -1H-isochromen-1-one (intermediate B58.1, 0.394g, 1.3mmol) in THF (15 mL). The resulting mixture was stirred at RT for 2 h. The mixture was diluted with DCM and water was added. The mixture was extracted 2 times with DCM and the combined organic layers were dried over sodium sulphate and evaporated to dryness.
The crude product was purified by flash chromatography on a silica gel Biotage column (cyclohexane-cyclohexane: EtOAc ═ 10:90) to give the title compound (0.284g, 1.06mmol)
UPLC-MS: 0.67min, 268.2[ M + H ] +, method 13
Intermediate B59
3- (1-hydroxyethyl) -4- (morpholinomethyl) -1H-isochromen-1-one
Step 1.3- (1- ((tert-butyldimethylsilyl) oxy) ethyl) -4-vinyl-1H-isochromen-1-one (intermediate B59.1)
To a solution of degassed 4-bromo-3- (1- ((tert-butyldimethylsilyl) oxy) ethyl) -1H-isochromen-1-one (intermediate A8, 6g, 15.7mmol) in toluene (120ml) was added PdCl2(PPh3)2(0.6g, 079mmol) followed by tributyl (1-ethoxyvinyl) tin (5ml, 17.2mmol) and the resulting mixture was heated to reflux overnight. At room temperature PdCl is added2(PPh3)2(0.6g), the mixture was refluxed for 30 minutes. The mixture was cooled to room temperature and then filtered through celite. The filtrate was evaporated to dryness and the crude product was purified by Biotage silica gel column flash chromatography (cyclohexane-cyclohexane: EtOAc ═ 80:20) to give 3- (1- ((tert-butyldimethylsilyl) oxy) ethyl) -4-vinyl-1H-isochromen-1-one as a light yellow oil (intermediate B59.1, 5.8 g).
UPLC-MS: 1.51min, 331.1[ M + H ] +, method 12.
Step 2.3- (1- ((tert-butyldimethylsilyl) oxy) ethyl) -1-oxo-1H-isochromene-4-carbaldehyde (intermediate B59.2)
A slow stream of O3 in O2 was passed through a cooled solution of 3- (1- ((tert-butyldimethylsilyl) oxy) ethyl) -4-vinyl-1H-isochromen-1-one (intermediate B59.1, 2.967g crude) in DCM (100ml) at-78 deg.C for 1.5H. By N2Excess O3 was purged by bubbling followed by PPh addition3(2.316g, 8.83mmol) in DCM (20 ml). The solution was allowed to reach 25 ℃ and stirred overnight. The solvent was removed in vacuo and the crude material was purified by Biotage silica gel column flash chromatography (cyclohexane-cyclohexane: EtOAc ═ 90:10) to give 3- (1- ((tert-butyldimethylsilyl) oxy) ethyl) -1-oxo-1H-isochromene-4-carbaldehyde (intermediate B59.2, 1.453g, 4.38 mmol).
UPLC-MS: 1.43min, 333.1[ M + H ] +, method 12.
Step 3.3- (1- ((tert-butyldimethylsilyl) oxy) ethyl) -4- (morpholinomethyl) -1H-isochromen-1-one (intermediate B59.3)
To a solution of 3- (1- ((tert-butyldimethylsilyl) oxy) ethyl) -1-oxo-1H-isochromene-4-carbaldehyde (intermediate B59.2, 0.5g, 1.5mmol) and morpholine (0.12ml, 1.35mmol) in DCM (20ml) was added dry Na2SO4The mixture was stirred at RT for 10 min. Sodium triacetoxyborohydride (0.286g, 2.25mmol) and acetic acid (0.09ml, 1.5mmol) were added and the reaction mixture was stirred at RT 24. The reaction was stopped by addition of 2M HCl (3ml), the heterogeneous mixture was filtered and the filtrate was purified by flash chromatography on silica NH column (cyclohexane: EtOAc: 90:10-80:20) to give 3- (1- ((tert-butyldimethylsilyl) oxy) ethyl) -4- (morpholinomethyl) -1H-isochromen-1-one (intermediate B59.3, 0.183g, 0.45mmol, 30%).
UPLC-MS: 1.27min, 404.5[ M + H ] +, method 14.
Step 4
3- (1- ((tert-butyldimethylsilyl) oxy) ethyl) -4- (morpholinomethyl) -1H-isochromen-1-one (intermediate B59.3, 0.203g, 0.503mmol) was dissolved in THF (7 mL); TBAF (1.0M in THF, 0.33ml, 0.33mmol) was added dropwise and the resulting mixture was stirred at room temperature for 2 h. Water (10ml) was added and the mixture was extracted 2 times with DCM; the combined organic layers were dried over sodium sulfate and evaporated to dryness. The crude product was purified by reverse phase chromatography using a C18 Biotage column ((H)2O:CH3CN 95:5-80:20, containing 0.1% HCOOH) to give the title compound (0.145 g). The product was used in the next step without further purification.
UPLC-MS: 0.41min, 290.4[ M + H ] +, method 14.
Intermediate C1
3- (bromomethyl) -4-phenyl-1H-isochromen-1-one
Triphenylphosphine (250mg, 0.953mmol) was added to a solution of 3- (hydroxymethyl) -4-phenyl-1H-isochromen-1-one (intermediate B2, 185mg, 0.733mmol) and perbromomethane (316mg, 0.953mmol) in DCM (3.7ml, 57.5mmol), and the resulting mixture was stirred at RT for 3 days with several additions of reagents to complete (4 aliquots in total). The reaction was dissolved with MeOH, then concentrated under reduced pressure. The crude product was purified using a biotage silica 50g SNAP column with a hexane and EtOAc gradient to give the title compound (0.11g, 47.6%).
UPLC-MS: 2.21min, 358[ M + H + ACN ] +, method 4.
Intermediate C2
3- (bromomethyl) -4- (2-fluorophenyl) -1H-isochromen-1-one
Triphenylphosphine (0.127g, 0.488mmol)) was added to a solution of 4- (2-fluorophenyl) -3- (hydroxymethyl) -1H-isochromen-1-one (intermediate B4, 40mg, 0.148mmol) and perbromomethane (0.161g, 0.488mmol) in DCM (0.7ml, 57.5mmol) and the resulting mixture was stirred at RT for 24H. The reaction was dissolved with MeOH, then concentrated under reduced pressure. The crude product was purified on a Biotage Silica 50g SNAP column with a hexane and EtOAc gradient to give the title compound (37mg, 75%).
UPLC-MS: 2.21min, 358[ M + H + ACN ] +, method 4.
Intermediate C3
3- (bromomethyl) -4-m-tolyl-1H-isochromen-1-one
Triphenylphosphine (270mg, 1.030mmol) and perbromomethane (342mg, 1.030mmol) were added to a solution of 3- (hydroxymethyl) -4-m-tolyl-1H-isochromen-1-one (intermediate B5, 211mg, 0.792mmol) in DMF (2ml) and the resulting mixture was stirred at RT. The reaction was then purified by reverse phase chromatography using a Biotage C1830 g SNAP column (phase a, water 95%, ACN 4.5%, formic acid 0.5%; phase B, ACN 99.5%, formic acid 0.5%) to give the title compound (192mg, 73.6%).
UPLC-MS: 2.32min, 372[ M + H + ACN ] +, method 4
Intermediate C4
3- (bromomethyl) -4- (3-fluorophenyl) -1H-isochromen-1-one
Triphenylphosphine (265mg, 1.01mmol) and perbromomethane (335mg, 1.010mmol) were added to a solution of 4- (3-fluorophenyl) -3- (hydroxymethyl) -1H-isochromen-1-one (intermediate B3, 210mg, 0.777mmol) in DCM (6.5ml) and the mixture was stirred at RT overnight. Triphenylphosphine (265mg, 1.010mmol) and perbromomethane (335mg, 1.01mmol) were then added, and stirring was maintained at RT for 6 h. The reaction was then diluted with MeOH (1ml) and purified directly on a 50g biotage silica gel column eluting with a gradient of hexane and EtOAc to give the title compound (154mg, 59.5%) as a yellow solid.
UPLC-MS: 2.20min, 334.6[ M + H ] +, method 4
Intermediate C5
3- (1-bromoethyl) -4-m-tolyl-1H-isochromen-1-one
Triphenylphosphine (365mg, 1.39mmol) and perbromomethane (461mg, 1.39mmol) were added to a solution of 3- (1-hydroxyethyl) -4-m-tolyl-1H-isochromen-1-one (intermediate B6, 300mg, 1.07mmol) in DCM (2.2ml) and the resulting mixture was stirred at RT. The reaction product was then purified using silica gel Biotage SNAP 50g, eluting with a gradient of hexanes and EtOAc to give the title compound (67mg, 18.3%).
UPLC-MS: 2.39min, 386[ M + H + ACN ] +, method 4.
Intermediate C6
3- (1-bromoethyl) -4- (3-fluorophenyl) -1H-isochromen-1-one
1M PBr3To a solution of 4- (3-fluorophenyl) -3- (1-hydroxyethyl) -1H-isochromen-1-one (intermediate B7, 390mg, 1.372mmol) in DCM (7.8ml) was added 2.332mmolThe mixture was stirred at RT for 2 h. The reaction mixture was then evaporated under reduced pressure and purified using silica gel Biotage SNAP 100g eluting with a gradient of hexane and EtOAc to give the title compound (227mg, 47.7%).
UPLC-MS: 2.27min, 348[ M + H ] +, method 4
Intermediate C7
3- (1-bromoethyl) -4-phenyl-1H-isochromen-1-one
In a similar manner to intermediate C6 at RT was used 3- (1-hydroxyethyl) -4-phenyl-1H-isochromen-1-one (intermediate B1, 2.6g, 9.76mmol), 1M PBr3Was dissolved in DCM (17.5ml, 17.5mmol) (30ml) to prepare the title compound. The crude product was purified using a Biotage Silica SNAP 100g gradient with hexanes and AcOEt to give the title compound (1.64g, 51%).
UPLC-MS: 2.28min, 331[ M + H + ACN ] +, method 4
Intermediate C8
3- (1-bromoethyl) -4- (3- (dimethylamino) phenyl) -1H-isochromen-1-one
In a similar manner to intermediate C6 at RT was used 4- (3- (dimethylamino) phenyl) -3- (1-hydroxyethyl) -1H-isochromen-1-one (intermediate B8, 200mg, 0.64mmol), 1M PBr 3The title compound was prepared as a solution of (1.1ml, 1.1mmol) in DCM (2 ml). The crude product was purified using a gradient with Biotage Silica SNAP 50g with DCM and MeOH to give the title compound (300 mg).
UPLC-MS: 292[ M-Br ] +, method 2, for 6.9 min.
Intermediate C9
3- (1-bromoethyl) -4- (3- (morpholinosulfonyl) phenyl) -1H-isochromen-1-one
In a similar manner to intermediate C6 at RT was used 3- (1-hydroxyethyl) -4- (3- (morpholinosulfonyl) phenyl) -1H-isochromen-1-one (intermediate B9, 163mg, 0.392mmol), 1M PBr3The title compound was prepared (0.68ml, 0.667mmol) in DCM (3.2 ml). The crude product was triturated with DMF to give the title compound (93mg, 49%). The solution was purified by reverse phase chromatography using a Biotage C18 SNAP 30g column (phase a, water 95%, ACN 4.9%, formic acid 0.1%); phase B, ACN 99.9%, formic acid 0.1%), yielding a further compound (68mg, 36.2%).
UPLC-MS: 5.75min, 479[ M + H ] +, method 5.
Intermediate C10
3- (1-bromoethyl) -4- (6-methylpyridin-3-yl) -1H-isochromen-1-one hydrobromide
Intermediate B10(163mg, 0.579mmol), 1M PBr in a mixture of DCM (2ml) and DMF (3ml) was used at RT in a similar manner to intermediate C6 using 3- (1-hydroxyethyl) -4- (6-methylpyridin-3-yl) -1H-isochromen-1-one intermediate B10(163mg, 0.579mmol) 3(0.87mL, 0.87mmol) the title compound was prepared. The crude product was purified by reverse phase chromatography using a Biotage C18 SNAP 60g column (phase a, water 95%, ACN 4.9%, formic acid 0.1%); phase B, ACN 99.9%, formic acid 0.1%). To the aqueous fraction was added 50% HBr water, and concentrated under reduced pressure to give the title compound (320 mg).
UPLC-MS: 1.73min, 344[ M + H ] +, method 4.
Intermediate C11
4- (3- (1-bromoethyl) -1-oxo-1H-isochromen-4-yl) -5, 6-dihydropyridine-1 (2H) -carboxylic acid tert-butyl ester
1M PBr3(0.36ml, 0.363mmol) of DCM solution was added dropwise to a solution of 4- (3- (1-hydroxyethyl) -1-oxo-1H-isochromen-4-yl) -5, 6-dihydropyridine-1 (2H) -carboxylic acid tert-butyl ester (intermediate B12, 135mg, 0.363mmol) in DCM (5 ml). The reaction mixture was then dried under reduced pressure and dissolved in DCM (5 ml). Di-tert-butyl dicarbonate (0.34ml, 1.454mmol) and DMAP (89mg, 0.727mmol) were added simultaneously and the mixture was stirred at RT overnight. The reaction mixture was purified directly using a Biotage SNAP 50g silica gel column with a hexane and EtOAc gradient to give the title compound (100mg, 63.3%) as a pale yellow solid.
UPLC-MS: 2.30min, 435[ M + H ] +, method 4.
Intermediate C12
3- (1-bromoethyl) -4- (thiazol-5-yl) -1H-isochromen-1-one
1M PBr at RT3(0.5ml, 0.50mmol) of DCM solution was added dropwise to 3- (1-hydroxyethyl) -4- (thiazol-5-yl) -1H-isochromen-1-one (intermediate B11, 102mg, 0.372mmol) in DCM (4ml) and ACN (2 ml). The reaction mixture was then concentrated under reduced pressure and purified directly using a Biotage SNAP 12g column with a hexane and EtOAc gradient to give the title compound (56mg, 45%).
UPLC-MS: 1.90min, 337[ M + H ] +, method 4.
Intermediate C13
4-benzyl-3- (1-bromoethyl) -1H-isochromen-1-one
1M PBr at RT3To 4-benzyl-3- (1-hydroxyethyl) -1H-isochromen-1-one (intermediate B14, 625mg, 2.230mmol) in DCM (2mL) was added a solution of DCM (0.105mL, 1.105 mmol). The reaction mixture was purified directly using a biotage snap 25g column with a hexane and EtOAc gradient to give the title compound (22mg, 2.9%).
UPLC-MS: 2.19min, 345[ M + H ] +, method 4.
Intermediate C14
3- (1-bromoethyl) -4- (2-methylpyridin-4-yl) -1H-isochromen-1-one
1M PBr at RT3To a solution of 3- (1-hydroxyethyl) -4- (2-methylpyridin-4-yl) -1H-isochromen-1-one (intermediate B13, 140mg, 0.498mmol) in DCM (846 μ L, 0.846 mmol). The reaction mixture was concentrated under reduced pressure and directly purified by reverse phase chromatography using a Biotage C18 SNAP 60g column (phase a, water 95%, ACN 4.5%, formic acid 0.5%); phase B, ACN 99.5%, formic acid 0.5%) to give the title compound (80mg, 45%).
UPLC-MS: 1.60min, 345[ M + H + ACN ] +, method 4.
Intermediate C15
3- (1-bromoethyl) -4- (4- (2-morpholinoethoxy) phenyl) -1H-isochromen-1-one hydrobromide
1M PBr at RT3To 3- (1-hydroxyethyl) -4- (4- (2-morpholinoethoxy) phenyl) -1H-isochromen-1-one (intermediate B15, 180mg, 0.455mmol) in DCM (2mL) was added a solution of DCM (1.82mL, 1.82 mmol). Then using Et2The reaction mixture was diluted with O and filtered. Reuse of Et2The residue was washed with O and finally dried to give the title compound (150mg, 61%) as a pink solid.
UPLC-MS: 1.58min, 457[ M + H + ACN ] +, method 4.
Intermediate C16
3- (1-bromoethyl) -4- (5- (morpholinomethyl) thiophen-2-yl) -1H-isochromen-1-one hydrobromide
1M PBr at RT3To a solution of 3- (1-hydroxyethyl) -4- (5- (morpholinomethyl) thiophen-2-yl) -1H-isochromen-1-one (intermediate B16, 258mg, 0.695mmol) in DCM (2.6 mL). The reaction mixture was concentrated under reduced pressure and the crude product crystallized from boiling DCM/MeOH 4/1(5ml) to give the title compound as a colorless solid (138mg, 38.6%)
UPLC-MS: 3.60min, 354[ M-Br ] +, method 5.
Intermediate C17
3- (1-bromoethyl) -4- (6-methoxypyridin-3-yl) -1H-isochromen-1-one
1M PBr at RT3To a solution of 3- (1-hydroxyethyl) -4- (6-methoxypyridin-3-yl) -1H-isochromen-1-one (intermediate B17, 225mg, 0.76mmol) in DCM (9mL) was added a solution of DCM (1.80mL, 1.80 mmol). The reaction mixture was directly purified using a Biotage SNAP 25g silica gel column with a hexane and EtOAc gradient to giveTo the title compound (105mg, 38.5%).
UPLC-MS: 2.12min, 361[ M + H ] +, method 4.
Intermediate C18
3- (1-chloroethyl) -4-phenyl-1H-isochromen-1-one
3- (1-hydroxyethyl) -4-phenyl-1H-isochromen-1-one (intermediate B1, 300mg, 1.27mmol), methanesulfonyl chloride (516mg, 4.51mmol), TEA (0.62mmol, 4.5ml) in DCM (10ml) was reacted at RT. The solvent was then removed under reduced pressure to give the title compound (180 mg). It was used in the next step without any further purification.
UPLC-MS: 2.25min, 285[ M + H ] +, method 4.
Intermediate C19
3- (1-bromoethyl) -4- (3, 6-dihydro-2H-pyran-4-yl) -1H-isochromen-1-one
1M PBr3To 4- (3, 6-dihydro-2H-pyran-4-yl) -3- (1-hydroxyethyl) -1H-isochromen-1-one (intermediate B18, 130mg, 0.477mmol) in DCM (2mL), stirred at RT for 2H, then the reaction mixture was adsorbed on a silica gel pad, purified directly on a Biotage silica gel column, eluting with a gradient of hexane and AcOEt to give the title compound (94mg, 58.7%) as an off-white solid.
UPLC-MS: 1.98min, 377[ M + H ] +, method 4.
Intermediate C20
3- (1-bromopropyl) -4-phenyl-1H-isochromen-1-one
Step 1.2- (pent-1-ynyl) benzoic acid methyl ester (intermediate C20.1)
Pent-1-yne (5.6mL, 57.2mmol) was added at RT to a solution in DMF (5mL) and Et3Methyl 2-iodobenzoate (5.6mL, 38.2mmol), copper (I) iodide (1.0g, 5.72mmol) and Pd (PPh) in N (1mL)3)4(1.3g, 1.14mmol) for 3 h.
The reaction was partitioned between Et2Between O and 0.1M aqueous HCl. With 0.1M aqueous HCl, NaHCO3The resulting organic phase was washed with aqueous solution and brine. Then using Na2SO4The organic phase was dried and dried under reduced pressure to give methyl 2- (pent-1-ynyl) benzoate (7.8g) as a dark oil. The crude product was used in the next step without further purification.
UPLC-MS: 2.14min, 202[ M + H ] +, method 4.
Step 2.4-iodo-3-propyl-1H-isochromen-1-one (intermediate C20.2)
Methyl 2- (pent-1-ynyl) benzoate (intermediate C20.1, 3g, 14.83mmol), iodine (11.29g, 44.5mmol) and sodium bicarbonate (3.74g, 44.5mmol) were reacted in acetonitrile (50mL) at RT for 20 min. With Et2The reaction mixture was diluted with O and 20% Na2S2O3Aqueous solution and brine. Then using Na2SO4The organic phase was dried and concentrated under reduced pressure. The crude product was dissolved in Et 2O, purification on a silica gel pad afforded the title compound (3.54g, 76%) as a brown oil.
UPLC-MS: 2.27min, 315[ M + H ] +, method 4.
Step 3.4-phenyl-3-propyl-1H-isochromen-1-one (intermediate C20.3)
4-iodo-3-propyl-1H-isochromen-1-one (intermediate C20.2, 0.80g, 2.55mmol), phenylboronic acid (0.3 mmol)9g，3.18mmol)、Pd(PPh3)4(0.15g, 0.127mmol) and Cs2CO3(1.16g, 3.57mmol) was reacted in DMF (10ml) at 80 ℃ for 3 h. The reaction was partitioned between AcOEt and 1M aqueous HCl, washed with 1M aqueous HCl, brine, and Na2SO4Drying, and concentrating under reduced pressure. The resulting crude product was purified on a silica gel 50G Biotage column, eluting with a gradient of hexane and AcOEt to give the title compound (0.11G, 15.9%) as a pale yellow oil.
UPLC-MS: 2.35min, 265[ M + H ] +, method 4.
Step 4.3- (1-Bromopropyl) -4-phenyl-1H-isochromen-1-one
4-phenyl-3-propyl-1H-isochromen-1-one (intermediate C20.3, 107mg, 0.405mmol), N-bromosuccinimide (86mg, 0.486mmol), benzoyl peroxide (25mg, 0.1mmol) were added to CCl4(2mL) at 100 ℃ for 2 h. The reaction mixture was then purified on a Biotage50g silica gel column with a hexane and EtOAc gradient to give the title compound (101mg, 72.7%) as a light yellow oil.
UPLC-MS: 2.37min, 344[ M + H ] +, method 4.
Intermediates and compounds D1
3- ((4-amino-3-iodo-1H-pyrazolo [3,4-d ] pyrimidin-1-yl) methyl) -4-phenyl-1H-isochromen-1-one
Commercially available 3-iodo-1H-pyrazolo [3,4-d]Pyrimidin-4-amine (124mg, 0.476mmol) was added to a solution of 3- (bromomethyl) -4-phenyl-1H-isochromen-1-one (intermediate C1, 100mg, 0.317mmol) in DMF (1 mL). Adding K2CO3(65.8mg, 0.476mmol) and the resulting mixture was stirred at 50 ℃ for 2 h. The resulting crude product was purified directly by reverse phase chromatography using a Biotage C18SNAP30g column (phase a, water 95%, ACN 5%, formic acid 0.01%); phase B, ACN 95%, water 5%, formic acid 0.01%) to give the title compound (138mg, 88%).
UPLC-MS: 1.92min, 496[ M + H ] +, method 4.
Intermediates D2a and D2b
3- (1- (4-amino-3-iodo-1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one and 3- (1- (4-amino-3-iodo-2H-pyrazolo [3,4-d ] pyrimidin-2-yl) ethyl) -4-phenyl-1H-isochromen-1-one
Commercially available 3-iodo-1H-pyrazolo [3,4-d]Pyrimidin-4-amine (270mg, 1.033mmol) and 3- (1-bromoethyl) -4-phenyl-1H-isochromen-1-one (intermediate C7, 500mg, 1.519mmol) were dissolved in DMF (4 mL). Adding K2CO3(315mg, 2.278mmol) and the resulting mixture was stirred at 80 ℃ for 1 h. The mixture was poured into water (50ml) and extracted with AcOEt (10ml X3). The organic phase was dried and evaporated under reduced pressure. The crude product was purified by reverse phase chromatography using a Biotage C18SNAP 60g column (phase a, water 95%, ACN 4.9%, formic acid 0.1%); phase B, ACN 95%, water 5%, formic acid 0.1%) to give 3- (1- (4-amino-3-iodo-1H-pyrazolo [3, 4-d) ]Pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one (intermediate D2a, 78mg, 10%) and 3- (1- (4-amino-3-iodo-2H-pyrazolo [3, 4-D)]Pyrimidin-2-yl) ethyl) -4-phenyl-1H-isochromen-1-one (intermediate D2b, 20mg, 2, 6%).
Intermediate d2a. uplc-MS: 1.85min, 510[ M + H ] +, method 4.
Intermediate d2b. uplc-MS: 1.95min, 510[ M + H ] +, method 4.
Intermediates and compounds D3
3- (1- (4-amino-3-iodo-1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-m-tolyl-1H-isochromen-1-one
In a similar manner to intermediate D2 from commercially available 3-iodo-1H-pyrazolo [3,4-D]Pyrimidin-4-amine (125mg, 0.481mmol) and K2CO3(66.4mg, 0.481mmol) and 3- (1-bromoethyl) -4-m-tolyl-1H-isochromen-1-one (intermediate C5, 110mg, 0)320mmol) in DMF (1.1mL) to give the title compound (120mg, 71.5%).
UPLC-MS: 5.87min, 523[ M + H ] +, method 5.
The intermediates D4-D22, D29-34 found in the table below can be prepared analogously to the procedure described for compound D2 starting from the suitable intermediates (Int.) reported below and 3-iodo-1H-pyrazolo [3,4-D ] pyrimidin-4-amine.
(continuation)
(continuation)
(continuation)
(continuation)
Intermediate D23
3- (1- (4-amino-3-iodo-1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1,2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one hydrochloride
Tert-butyl 4- (3- (1- (4-amino-3-iodo-1H-pyrazolo [3,4-D ] pyrimidin-1-yl) ethyl) -1-oxo-1H-isochromen-4-yl) -5, 6-dihydropyridine-1 (2H) -carboxylate (intermediate D14, 1.5g, 2.441mmol) was suspended in 1, 4-dioxane (5ml) and 4M HCl in dioxane (15.26ml, 61.0mmol) was added. The mixture was stirred at RT for 4h, then volatiles were removed under reduced pressure to give the title compound (1.56g) as a pale pink powder.
UPLC-MS: 0.47min, 515[ M + H ] +, method 9
Intermediate D24
3- (1- (4-amino-3-iodo-1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1- (1-isopropylpiperidin-4-yl) -1,2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one formate salt
A mixture of 3- (1- (4-amino-3-iodo-1H-pyrazolo [3,4-D ] pyrimidin-1-yl) ethyl) -4- (1,2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one hydrochloride (intermediate D23, 0.1g, 0.182mmol), 1-isopropylpiperidin-4-one (0.064ml, 0.438mmol), DIPEA (0.032ml, 0.182mmol) and spatula in DCM (3ml) sodium sulfate was stirred at RT for 10 min. Acetic acid (0.061mL, 1.09mmol) was then added, followed by sodium triacetoxyborohydride (154mg, 0.726 mmol). The resulting suspension was stirred at RT for 3h, then the reaction was stopped by addition of 2N HCl. The heterogeneous mixture was concentrated under reduced pressure. Purification by RP-flash chromatography (Biotage 30g C18 column, gradient elution 100:0-60:40A/B, 15 CV; A: water/MeCN 95/5+ 0.01% HCOOH, B: water/MeCN 5/95+ 0.01% HCOOH) gave the title compound (99.8mg, 0.146mmol, 80% yield) as a light yellow powder.
UPLC-MS: 0.71min, 640[ M + H ] +, method 9
Intermediate D25
3- (1- (4-amino-3-iodo-1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1- (4- (dimethylamino) butanoyl) -1,2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one formate salt
DIPEA (0.253mL, 1.452mmol) was added to a stirred suspension of 3- (1- (4-amino-3-iodo-1H-pyrazolo [3,4-D ] pyrimidin-1-yl) ethyl) -4- (1,2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one hydrochloride (intermediate D23, 0.200g, 0.363mmol), HATU (0.166g, 0.436mmol) and 4-dimethylaminobutyric acid hydrochloride (0.073g, 0.436mmol) in THF/DMF 5:1(6mL) at 0 ℃. The mixture was stirred at 0 ℃ for 15min and then at RT for an additional 45 min. The reaction mixture was poured into saturated aqueous sodium bicarbonate solution and extracted with DCM. The collected organic phases were concentrated under reduced pressure and the crude product was purified by RP-flash chromatography (Biotage 30g C18 column, gradient elution 100: 0-70: 30A/B, 15 CV; A: water/MeCN 95/5+ 0.01% HCOOH, B: water/MeCN 5/95+ 0.01% HCOOH) to give the title compound (0.153g, 0.227mmol, 62.6% yield) as a white powder.
UPLC-MS: 0.67min, 628[ M + H ] +, method 9
Intermediate D26
3- (1- (4-amino-3-iodo-1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1- (2- (dimethylamino) acetyl) -1,2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one formate salt
According to the procedure used for the synthesis of intermediate D25, from 3- (1- (4-amino-3-iodo-1H-pyrazolo [3,4-D ] pyrimidin-1-yl) ethyl) -4- (1,2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one hydrochloride (intermediate D23, 0.200g, 0.363mmol) and 2- (dimethylamino) acetic acid (0.045g, 0.436mmol), the title compound was obtained (0.127g, 0.197mmol, 54.2% yield) as a white powder.
UPLC-MS: 0.64min, 600[ M + H ] +, method 9
Intermediate D27
3- (1- (4-amino-3-iodo-1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1- (1-methylpiperidine-4-carbonyl) -1,2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one formate salt
Following the procedure used for the synthesis of intermediate D25, from 3- (1- (4-amino-3-iodo-1H-pyrazolo [3,4-D ] pyrimidin-1-yl) ethyl) -4- (1,2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one hydrochloride (intermediate D23, 0.200g, 0.363mmol) and 1-methylpiperidine-4-carboxylic acid (0.062g, 0.436mmol) the title compound was obtained (99.4mg, 0.145mmol, 39.9% yield) as a white powder.
UPLC-MS: 0.65min, 640[ M + H ] +, method 9
Intermediate D28
3- (4- (3- (1- (4-amino-3-iodo-1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -1-oxo-1H-isochromen-4-yl) -1,2,3, 6-tetrahydropyridine-1-carbonyl) azetidine-1-carboxylic acid tert-butyl ester
Following the procedure used for the synthesis of intermediate D25, from 3- (1- (4-amino-3-iodo-1H-pyrazolo [3,4-D ] pyrimidin-1-yl) ethyl) -4- (1,2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one hydrochloride (intermediate D23, 0.200g, 0.363mmol) and Boc-azetidine-3-carboxylic acid (0.088g, 0.436mmol) the title compound was obtained (0.252g, 0.361mmol, 99% yield).
UPLC-MS: 1.04min, 698[ M + H ] +, method 9
Intermediate E1
1- (1-oxo-4-phenyl-1H-isochromen-3-yl) ethylcarbamic acid tert-butyl ester
The title compound was prepared from tert-butyl 1- (4-bromo-1-oxo-1H-isochromen-3-yl) ethylcarbamate (intermediate A3, 500mg, 1.358mmol), phenylboronic acid (215mg, 1.765mmol) in analogy to intermediate B1 to give the title compound (112mg, 22.6%) as a light yellow oil.
UPLC-MS: 2.24min, 731.0[2M + H ] +, method 4.
Intermediate E2
3- (1-aminoethyl) -4-phenyl-1H-isochromen-1-one hydrochloride
1- (1-oxo-4-phenyl-1H-isochromen-3-yl) ethylcarbamic acid tert-butyl ester (intermediate E1, 11)2mg, 0.306mmol) was dissolved in DCM (3ml) and a 4M HCl solution in dioxane (3ml, 12.00mmol) was added and the mixture was stirred at rt for 4 h. By adding Et 2The reaction was stopped with O (50mL) and the mixture was dried under reduced pressure to give the title compound.
UPLC-MS: 1.43min, 265.8[ M + H ] +, method 4.
Intermediate E3
3- (1-aminoethyl) -4- (5- (morpholinomethyl) thiophen-2-yl) -1H-isochromen-1-one dihydrochloride
The title compound was prepared from tert-butyl (1- (4-bromo-1-oxo-1H-isochromen-3-yl) ethyl) carbamate (intermediate A3, 1g, 2.72mmol), 4- ((5- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) thiophen-2-yl) methyl) morpholine (0.840g, 2.72mmol) in analogy to intermediate B1 to give (before drying, small amount of 1M aqueous HCl) the title compound (587mg, 71.2%) as a light yellow solid.
UPLC-MS：0.39min，354.1[(M+H)-NH3)]Method 9
Intermediate E4
3- (1-aminoethyl) -4- (1H-pyrazol-4-yl) -1H-isochromen-1-one dihydrochloride
The title compound was prepared in analogy to intermediate B1 from tert-butyl 1- (4-bromo-1-oxo-1H-isochromen-3-yl) ethylcarbamate (intermediate A3, 500mg, 1.358mmol), 1H-pyrazol-4-ylboronic acid (198mg, 1.765 mmol). After purification, 37% aqueous HCl (1ml) was added to the collected fractions, and concentrated to obtain the title compound (158mg, 35.5% yield).
UPLC-MS: 1.44min, 256, 511[ M + H ] +, method 3.
Intermediate F1
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (3- (4,4,5, 5-tetramethyl-1, 3-dioxolan-2-yl) phenyl) -1H-isochromen-1-one
Reacting 3- (1- (4-amino-3-iodo-1H-pyrazolo [3, 4-d)]Pyrimidin-1-yl) ethyl) -4- (3- (4,4,5, 5-tetramethyl-1, 3-dioxolan-2-yl) phenyl) -1H-isochromen-1-one (intermediate D8, 1.14g, 1.788mmol), (3-fluoro-5-hydroxyphenyl) boronic acid (0.558g, 3.58mmol), K2CO3(0.494g, 3.58mmol) and PdCl2(dppf) (0.196g, 0.268mmol) was reacted in dioxane (30ml) at 120 ℃ overnight. The reaction was diluted with DCM (100ml), filtered to remove the solid and the filtrate was evaporated under reduced pressure. The crude product was purified by flash chromatography on silica gel using Biotage100G SNAP with DCM and MeOH gradient to give the title compound (819mg, 73.7%) as a light brown solid.
UPLC-MS: 1.23min, 622.2[ M + H ] +, method 9
Intermediate F2
3- (3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -1-oxo-1H-isochromen-4-yl) benzaldehyde
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (3- (4,4,5, 5-tetramethyl-1, 3-dioxolan-2-yl) phenyl) -1H-isochromen-1-one (intermediate F1, 819mg, 1.317mmol) was dissolved in MeCN (10mL) and 2M aqueous HCl (30mL) and the mixture was stirred at RT for 3H. The reaction was diluted with DCM (300mL) and washed with water (200 mL). The aqueous layer was extracted 2 times with DCM, the combined organic fractions were dried by phase separator and the solvent was evaporated under reduced pressure to give the title compound (598mg, 87% yield).
UPLC-MS: 0.97min, 522.1[ M + H ] +, method 9
Intermediate F3 and examples
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (5- (4,4,5, 5-tetramethyl-1, 3-dioxolan-2-yl) thiophen-2-yl) -1H-isochromen-1-one
The title compound was prepared from 3- (1- (4-amino-3-iodo-1H-pyrazolo [3,4-D ] pyrimidin-1-yl) ethyl) -4- (5- (4,4,5, 5-tetramethyl-1, 3-dioxolan-2-yl) thiophen-2-yl) -1H-isochromen-1-one (intermediate D13, 385mg, 0.598mmol), (3-fluoro-5-hydroxyphenyl) boronic acid (187mg, 1.197mmol) in a similar manner to intermediate F1 to give the title compound (254mg, 67.6%) as a pale yellow solid.
UPLC-MS: 5.13min, 627.9[ M + H ] +, method 6
Intermediate F4
5- (3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -1-oxo-1H-isochromen-4-yl) thiophene-2-carbaldehyde
The title compound was prepared from 3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (5- (4,4,5, 5-tetramethyl-1, 3-dioxolan-2-yl) thiophen-2-yl) -1H-isochromen-1-one (intermediate F3, 254mg, 0.405mmol) in MeCN (4mL)/2M aqueous HCl (4mL) in a similar manner to intermediate F2 to give the title compound (201mg, 94%) as a pale yellow solid.
UPLC-MS: 1.73min, 528.1[ M + H ] +, method 9.
Intermediate F5
3- (1- (4-amino-3- (3- (benzyloxy) -5-fluorophenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (3- (4,4,5, 5-tetramethyl-1, 3-dioxolan-2-yl) phenyl) -1H-isochromen-1-one
In a 100ml round-bottom flask, 3- (3- (benzyloxy) -5-fluorophenyl) -1H-pyrazolo [3,4-d]Pyrimidin-4-amine (intermediate G18, 400mg, 1,194mmol) was dissolved in 7ml dry DMF and K was added2CO3(254mg, 1,837 mmol). After stirring for 5min, a solution of 3- (1-bromoethyl) -4- (3- (4,4,5, 5-tetramethyl-1, 3-dioxolan-2-yl) phenyl) -1H-isochromen-1-one (intermediate C26, 420mg, 0,918mmol) in 7ml dry DMF was added and the clear brown mixture was heated at 60 ℃ for 30 min. The mixture was cooled to r.t., then 30ml HCl 0.5M and 50ml EtOAc were added and stirred for 15 min. The phases were separated and the crude material was purified by chromatography eluting with DCM/MeOH in DCM (80/20) to give the title compound (820mg) as a brown oil. The material was used in the next step without any further purification.
UPLC-MS: 1.48min, 712.28[ M + H ] +, method 9
Intermediate F6
3- (3- (1- (4-amino-3- (3- (benzyloxy) -5-fluorophenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -1-oxo-1H-isochromen-4-yl) benzaldehyde
3- (1- (4-amino-3- (3- (benzyloxy) -5-fluorophenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (3- (4,4,5, 5-tetramethyl-1, 3-dioxolan-2-yl) phenyl) -1H-isochromen-1-one (intermediate F5, 654mg, 0,919mmol) was dissolved in MeCN, then 2M HCl was added and the mixture was stirred overnight. DCM (25ml) and water (25ml) were added and the phases separated and the aqueous phase washed with 10ml of DCM. The collected organic phases were concentrated to give the title compound (490mg, 0.801mmol, 87% yield) as a brown oil.
UPLC-MS: 1.28min, 612.15[ M + H ] +, method 9
Intermediate G1
3- (3-fluoro-5-methoxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-4-amine
Reacting 3-iodo-1H-pyrazolo [3,4-d]Pyrimidin-4-amine (1.52g, 5.82mmol), 3-fluoro-5-methoxyphenylboronic acid (1.4g, 8.24mmol), Pd (dppf) Cl2(0.18g, 0.246mmol) and 8.7mL of 1M aqueous NaOH (8.73mmol) were reacted in DMF (13mL) under Ar for 48h at 120 ℃. The reaction was stopped by addition of 1M aqueous HCl (10mL), dried under reduced pressure, and the dark crude oil was purified by flash chromatography on silica gel using a Biotage 100G SNAP gradient with DCM and iPrOH to give the title compound (275mg, 18.2%) as a light yellow solid.
UPLC-MS: 0.54min, 260.0[ M + H ] +, method 9
Intermediate G2
4,4,5, 5-tetramethyl-2- (3- (4,4,5, 5-tetramethyl-1, 3-dioxolan-2-yl) phenyl) -1,3, 2-dioxaborolane
(3-formylphenyl) boronic acid (5g, 33.3mmol), 2, 3-dimethylbutane-2, 3-diol (19.70g, 167mmol) and p-toluenesulphonic acid monohydrate (0.317g, 1.667mmol) were dissolved in toluene (278mL) and refluxed using a dean-Stark apparatus for 3h until the reaction was complete. The mixture was dried under reduced pressure, the residue diluted with AcOEt (250mL), washed 3 times with copious amounts of water, 1 time with saturated aqueous NaCl (250mL), Na2SO4Drying and removing the solvent under reduced pressure. The crude product was purified by flash chromatography on silica gel using a Biotage100G +50G SNAP gradient with hexanes and AcOEt to give the title compound (8.5G, 77%) as a white solid.
1H NMR(400MHz，DMSO-d6)d ppm 7.72(s，1H)，7.63(d，J＝7.06Hz，1H)，7.55(d，J＝7.50Hz，1H)，7.31-7.44(m，1H)，5.91(s，1H)，1.24-1.37(s，18H)，1.18(s，6H)。
Intermediate G3
4- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1,2,3, 6-tetrahydropyridine hydrochloride
Tert-butyl 4- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -5, 6-dihydropyridine-1 (2H) -carboxylate (2g, 6.47mmol) was suspended in MTBE (8.1mL) and 2M HCl in Et2O solution (24 mL). The reaction was stirred at rt overnight, the white precipitate formed collected by filtration and washed with Et2O wash to afford the title compound (1.434g, 90%).
UPLC-MS: 0.51min, 210.3[ M + H ] +, method 9.
Intermediate G4
1- (4- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -5, 6-dihydropyridin-1 (2H) -yl) ethanone
4- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1,2,3, 6-tetrahydropyridine hydrochloride (intermediate G3, 1.4G, 5.70mmol) was suspended in DCM (15mL) at 0 deg.C, followed by the addition of TEA (2.384mL, 17.10mmol) and AcCl (0.405mL, 5.70 mmol). The reaction was warmed to rt and stirred for a further 30min, then the reaction volume was reduced to 1/3 of the original volume and the residue was diluted with AcOEt (150 ml). The organic phase is washed 2 times with water, 1 time with 0,2M HCl in water, 1 time with saturated NaCl in water, Na2SO4Drying and removal of the solvent under reduced pressure gave the title compound (1.24g, 87% yield) as a pale yellow solid.
UPLC-MS: 0.83min, 252.3[ M + H ] +, method 9.
Intermediate G5
4,4,5, 5-tetramethyl-2- (5- (4,4,5, 5-tetramethyl-1, 3-dioxolan-2-yl) thiophen-2-yl) -1,3, 2-dioxaborolan
The title compound was prepared from (5-formylthiophen-2-yl) boronic acid (2.5G, 16.03mmol) in analogy to intermediate G2 to give the desired product (3.93G, 72.5%) as a yellow-white solid.
1H NMR(400MHz，DMSO-d6)d ppm 7.38(d，J＝3.53Hz，1H)，7.22(s，1H)，6.11(s，1H)，0.98-1.52(m，24H)。
Intermediate G6
N- (5-bromopyridin-3-yl) -4-fluorobenzenesulfonamide
To a solution of 5-bromopyridin-3-amine (3g, 17.34mmol) in anhydrous EtOH (15ml) was added 4-fluorobenzene-1-sulfonyl chloride (0.989g, 5.08mmol) and the reaction was stirred overnight. Ethanol was removed under reduced pressure and the residue diluted with DCM (40ml) and saturated NaHCO3Washing is carried out for 1 time. With Na2SO4The organic phase was dried, filtered and concentrated. The product was purified directly by reverse phase chromatography using a Biotage C1830g column (phase a, water 95%, ACN 5%, formic acid 0.1%); phase B, ACN 95%, water 5%, formic acid 0.1%) to give the title compound (158mg, 24.4% yield) as a pale yellow solid.
UPLC-MS: 0.96min, 331[ M + H ] +, method 9.
Intermediate G7
5-Bromopyridin-3-ylcarbamic acid tert-butyl ester
To a solution of 5-bromopyridin-3-amine (3g, 17.34mmol) in dry DCM (25ml) was added di-tert-butyl dicarbonate (3.78g, 17.34mmol) with stirring. The resulting solution was cooled to 0 deg.C and a solution of sodium bis (trimethylsilyl) amide in 1M THF (17.34ml, 17.34mmol) was added dropwise over a period of 20 min. The solution was stirred overnight. The solution was then diluted with DCM (40mL) and saturated NaHCO3Washing is carried out for 1 time. With Na2SO4The organic phase was dried, filtered and concentrated. The crude product was finally purified using a BiotageSi 50g gradient with hexane and EtOAc. The title compound (2.60g, 54.9% yield) was recovered as a white solid.
UPLC-MS: 1.07min, 273[ M + H ] +, method 9.
Intermediate G8
4-chloro-2-amino (biscarbamate) -pyrimidines
To a solution of 4-chloropyrimidin-2-amine (1.0g, 7.72mmol) in dry DCM (30ml) were added N, N-dimethylpyridin-4-amine (0.094g, 0.772mmol) and N-ethyl-N, N-isopropylpropan-2-amine (2.494g, 19.30 mmol). After 10min di-tert-butyl dicarbonate (1.685g, 7.72mmol) was added and the resulting solution was stirred overnight. The reaction was diluted with DCM (40mL) and saturated NaHCO3Washing is carried out for 1 time. With Na2SO4The organic phase obtained is dried, filtered and concentrated. The crude product was finally purified with a Biotage Si50g gradient with hexanes and EtOAc. The title compound (0.53g, 69.3%) was recovered as a white solid.
UPLC-MS: 1.24min, 330[ M + H ] +, method 9.
Intermediate G9
4-fluoro-N- (5- (trimethylstannyl) pyridin-3-yl) benzenesulfonamide
To a solution of N- (5-bromopyridin-3-yl) -4-fluorobenzenesulfonamide (intermediate G6, 158mg, 0.477mmol) in dry dioxane (6ml) was added 1,1,1,2,2, 2-hexamethyldisiloxane (0.340ml, 1.637mmol) and Pd (PPh)3)4(105mg, 0.091 mmol). The solution was heated to 100 ℃ and stirred overnight. The solvent was removed in vacuo and the crude product finally purified with a biotage si10g gradient with hexanes and EtOAc. The title compound (165mg, 83% yield) was recovered as a white solid.
UPLC-MS: 0.99min, 417[ M + H ] +, method 9.
Intermediate G10
(5- (Trimethylstannyl) pyridin-3-yl) carbamic acid tert-butyl ester
The title compound was prepared in analogy to intermediate G9 using tert-butyl (5-bromopyridin-3-yl) carbamate (intermediate G7, 600mg, 2.197mmol), 1,1,1,2,2, 2-hexamethyl-dimethylstannane (0.820ml, 3.95mmol) to give the title compound (165mg, 0.398mmol, 83% yield) as a white solid.
UPLC-MS: 0.82min, 359[ M + H ] +, method 9.
Intermediate G11
4-trimethylstannanyl-2-amino- (biscarbamate) -pyrimidine
The title compound was prepared in analogy to intermediate G9 using 4-chloro-2-amino (biscarbamate) -pyrimidine (intermediate G8, 400mg, 1.21mmol), 1,1,1,2,2, 2-hexamethyldisiloxane (0.503ml, 2.43mmol) to yield the title compound (398mg, 69.8%) as a white solid.
UPLC-MS: 1.42min, 460[ M + H-Boc ] +, method 9.
Intermediate G12
Trifluoromethanesulfonic acid 4- (((benzyloxy) carbonyl) amino) cyclohex-1-en-1-yl ester
To a solution of sodium bis (trimethylsilyl) amide in 1M THF (22.24mL, 22.24mmol) pre-cooled to-78 deg.C in dry THF (25mL) was slowly added a solution of benzyl (4-oxocyclohexyl) carbamate (2.5g, 10.11mmol) in dry THF (25 mL). The solution was stirred at-78 ℃ for 30min, then a solution of 1,1, 1-trifluoro-N-phenyl-N- (trifluoromethylsulfonyl) methanesulfonamide (7.58g, 21.23mmol) in dry THF (25mL) was added. The reaction system is Stirring at-78 deg.C for 10min, and then bringing to room temperature. The reaction was diluted with ether (200mL), and the organic layer was washed with 1M aqueous NaOH (100 mL). With Na2SO4The organic phase was dried, filtered and the solvent removed under reduced pressure. The product was purified by Biotage Si 50g with a heptane and EtOAc gradient to give the title compound (1.43g, 3.77mmol, 37.3% yield) as a white solid.
UPLC-MS: 1.25min, 380[ M + H ] +, method 9
Intermediate G13
(4- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) cyclohex-3-en-1-yl) carbamic acid benzyl ester
To a solution of 4- (((benzyloxy) carbonyl) amino) cyclohex-1-en-1-yl trifluoromethanesulfonate (1.4G, 3.69mmol) in dry DMF was added 4,4,4',4',5,5,5',5' -octamethyl-2, 2' -bis (1,3, 2-dioxaborolane) (intermediate G12, 0.984G, 3.88mmol), Pd (dppf) Cl2·CH2Cl2(0.904g, 1.107mmol) and potassium acetate (1.07g, 11.07mmol), the reaction was stirred at 80 ℃ overnight. DMF was removed and the residue was diluted with ethyl acetate (200 mL). The organic phase was washed with brine (100mL), Na2SO4Drying, filtering and removing the solvent under reduced pressure. The product was purified by Biotage Si 50g with a heptane and EtOAc gradient to give the title compound (0.984g, 15%) as a yellow oil.
UPLC-MS: 1.28min, 358[ M + H ] +, method 9.
Intermediate G14
3-iodo-N, 1-bis (tetrahydro-2H-pyran-2-yl) -1H-pyrazolo [3,4-d ] pyrimidin-4-amine
To a solution of 3-iodo-1H-pyrazolo [3,4-d ] pyrimidin-4-amine (1g, 3.83mmol) in dry DMF (15mL) was added 3, 4-dihydro-2H-pyran (1.103mL, 11.49mmol) and 4-methylbenzenesulfonic acid monohydrate (0.208mL, 0.958 mmol). The solution was stirred at 90 ℃ for 5 days. The solvent was removed and the product purified by a Biotage Si 25g gradient with heptane and ethyl acetate to give the title compound (320mg, 19.5% yield).
UPLC-MS: 0.68min, 430[ M + H ] +, method 9
Intermediate G15
N, 1-bis (tetrahydro-2H-pyran-2-yl) -3- (trifluoromethyl) -1H-pyrazolo [3,4-d ] pyrimidin-4-amine
To a solution of 3-iodo-N, 1-bis (tetrahydro-2H-pyran-2-yl) -1H-pyrazolo [3,4-d ] pyrimidin-4-amine (intermediate G14, 322mg, 0.75mmol, 31.6% yield) in dry DMF 6(mL) were added copper (I) iodide (429mg, 2.250mmol) and methyl 2, 2-difluoro-2- (fluorosulfonyl) acetate (0.096mL, 0.750mmol), and the solution was stirred at 80 ℃ for 18H. DMF was removed under reduced pressure and the product was purified by a Biotage Si 25g gradient with heptane and EtOAc to give the title compound (88mg, 31.6% yield).
UPLC-MS: (mixture of diastereomers) 1.16 and 1.18min, 372[ M + H ] +, method 9.
Intermediate G16
3- (trifluoromethyl) -1H-pyrazolo [3,4-d ] pyrimidin-4-amines
To a solution of N, 1-bis (tetrahydro-2H-pyran-2-yl) -3- (trifluoromethyl) -1H-pyrazolo [3,4-d ] pyrimidin-4-amine (intermediate G15, 88mg, 0.237mmol) in EtOH (5mL) was added water (0.5 mL). 2,2, 2-trifluoroacetic acid (0,3ml, 0.237mmol) was then added and the mixture was stirred at 80 ℃ overnight. The solvent was removed to give the title compound (45mg, 93%) as a yellow solid.
UPLC-MS: 0.47min, 204[ M + H ] +, method 9.
Intermediate G17
3-methoxyphenyl sulfide pentafluoride (sulfate) -5-boronic acid
Step 1.2- (3-methoxy-5- (pentafluoro-l 6-thio) phenyl) -4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolane (intermediate G17.1)
To a solution of 3-methoxyphenylthio pentafluoride (0.5g, 2.135mmol) in dry THF (10mL) were added 4,4' -di-tert-butyl-2, 2' -bipyridine (0.115g, 0.427mmol), 4,4,4',4',5,5,5',5' -octamethyl-2, 2' -bis (1,3, 2-dioxaborolane) (0.488g, 1.922mmol) and Ir2(COD)2OMe2(0.142g, 0.214mmol) and the solution was stirred at 80 ℃ overnight. The solvent was removed and the product was purified by a Biotage Si 25G gradient with heptane and EtOAc to give 2- (3-methoxy-5- (pentafluoro-l 6-thio) phenyl) -4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolane (intermediate G17.1, 621mg, 81% yield) as a yellow solid.
Step 2
To a solution of 2- (3-methoxy-5- (pentafluoro-l 6-thio) phenyl) -4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolane (intermediate G17.1, 621mg, 1.724mmol) in THF (6ml) was added 6N aqueous HCl and the solution was stirred for 3 h. The solvent was removed and the product was directly purified by reverse phase chromatography using a Biotage C18 SNAP column (phase a, water 95%, ACN 5%, formic acid 0.1%); (phase B, ACN 95%, water 5%, formic acid 0.1%) to give the title compound (245mg, 51.1% yield) as a white solid.
UPLC-MS: 0.71min, 279[ M + H ] +, method 9.
Intermediate G18
3- (3- (benzyloxy) -5-fluorophenyl) -1H-pyrazolo [3,4-d ] pyrimidin-4-amine
3-iodo-1H-pyrazolo [3,4-d ] pyrimidin-4-amine (6,10mmol, 1.591g), (3- (benzyloxy) -5-fluorophenyl) boronic acid (4,06mmol, 1.0g), dppf (0,610mmol, 338mg), potassium phosphate anhydrous (10,16mmol, 2.157g) were suspended in dioxane, and the mixture was heated at 180C in an Mw reactor for 25min and then poured into 100ml water. The mixture was stirred overnight and then filtered on a buchner funnel, washed with 30ml water. The crude material was purified by chromatography, eluting with DCM/MeOH in DCM (80/20) to give the title compound (450mg, 1.342mmol, 33% yield) as a tan solid.
UPLC-MS: 1.42min, 336.15[ M + H ] +, method 10
Intermediate G19
4- (trimethylstannanyl) -1H-indazole
To a solution of 4-iodo-1H-indazole (500mg, 2,049mmol) in 1, 4-dioxane (500ml) was added 1,1,1,2,2, 2-hexamethyl dimethylstannane (1.0g, 3,07mmol) and Pd (Ph)3P)4(237mg, 0,250mmol) and the mixture was stirred at 80 ℃ for 18 h. The crude product was purified by reverse phase chromatography using a Biotage C1860 g SNAP column (phase a, water 95%, ACN 5%, formic acid 0.1%); phase B, ACN 95%, water 5%, AAcid 0.1%) to give the title compound (576mg, 66%).
1H NMR(400MHz，DMSO-d6)ppm 12.82-13.13(bs，1H)，7.99(s，1H)，7.45-7.52(m，1H)，7.24-7.32(m，1H)，7.20(s，1H)，0.11-0.58(m，9H)。
Intermediate G20
4-phenyl-1H-isochromene-3-carbaldehyde
Step 1.4-chloro-1H-isochromene-3-carbaldehyde (intermediate G20.1)
To a commercial isochroman-4-one (900mg, 6.07mmol) in DCM (18ml) was added DMF (0.706ml) and POCl successively at r.t. under a nitrogen atmosphere3(1.699ml, 18.23 mmol). The mixture was refluxed for 6h and kept at r.t overnight.
The reaction mixture was diluted with DCM (15ml), washed with water and saturated NaCl and then Na2SO4The crude product obtained was dried, evaporated in vacuo, purified by reverse phase using a Biotage C18 SNAP 30g column (phase a, water 95%, ACN 5%, formic acid 0.1%); phase B, ACN 95%, water 5%, formic acid 0.1%) to give 4-chloro-1H-isochromene-3-carbaldehyde (intermediate G20.1, 716mg, 60, 6% yield)
1H NMR(400MHz，DMSO-d6)d ppm 10.02(s，1H)，7.61-7.81(m，1H)，7.42-7.58(m，2H)，7.20-7.40(m，1H)，5.21(s，2H)。
UPLC-MS: 0.94min, 195[ M + H ] +, method 9
Step 2
Mixing X-phos Pd G2(116mg, 0,147mmol) and phenylboronAcid (538mg, 4,41mmol) was sealed in a closed vessel equipped with a magnetic stirrer and oxygen was removed by Ar/vacuum cycling. Degassed 4-chloro-1H-isochromene-3-carbaldehyde (intermediate G20.1, 716mg, 3,68mmol) in THF (8ml) was added followed by degassed K3PO4H2A solution of O (1,795ml, 7,36mmol) in water (8ml) was added and the resulting mixture was stirred at RT overnight. The reaction mixture was partitioned between 1MHCl (15ml) and the same amount of AcOEt. With Na2SO4The organic layer was dried, evaporated in vacuo and purified by reverse phase chromatography using a Biotage C18SNAP 120g column (phase a, water 95%, ACN 5%, formic acid 0.1%); phase B, ACN 95%, water 5%, formic acid 0.1%) to give the title compound (821mg, 94% yield).
UPLC-MS: 1.12min, 237[ M + H ] +, method 9.
Intermediate G21
3- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -8-azabicyclo [3.2.1] oct-2-ene-8-carboxylic acid benzyl ester
Step a. (1R,5S) -3- (((trifluoromethyl) sulfonyl) oxy) -8-azabicyclo [3.2.1] oct-2-ene-8-carboxylic acid tert-butyl ester (intermediate G21.1)
To a solution of sodium bis (trimethylsilyl) amide in 1M THF (12.43ml, 12.43mmol) previously cooled to-78 deg.C in dry THF (10ml) was slowly added a solution of tert-butyl (1R,5S) -3-oxo-8-azabicyclo [3.2.1] octane-8-carboxylate (2g, 8.88mmol) in dry THF (10ml) and the resulting solution was stirred at-78℃ for 1 h. A solution of 1,1, 1-trifluoro-N-phenyl-N- (trifluoromethylsulfonyl) methanesulfonamide (3.49g, 9.77mmol) in dry THF (10ml) was then added slowly and the reaction stirred at-78C for 30min at room temperature for 1 h. Ethyl acetate (100mL) was added and the organic phase was washed with 1M aqueous NaOH. The organic phase was collected, dried, filtered and the solvent removed under reduced pressure. The product was purified by Biotage Si 50g with heptane and EtOAc gradient to give tert-butyl (1R,5S) -3- (((trifluoromethyl) sulfonyl) oxy) -8-azabicyclo [3.2.1] oct-2-ene-8-carboxylate (2.93g, 8.20mmol, 92% yield) as a white amorphous solid.
UPLC-MS: 1.30min, 258[ M + H-Boc ] +, method 9
Step b. (1R,5S) -3- (((trifluoromethyl) sulfonyl) oxy) -8-azabicyclo [3.2.1] oct-2-ene-8-carboxylic acid tert-butyl ester (intermediate G21.2)
To (1R,5S) -3- (((trifluoromethyl) sulfonyl) oxy) -8-azabicyclo [3.2.1]To a solution of tert-butyl oct-2-ene-8-carboxylate (intermediate G21.1, 2.93G, 8.20mmol) in dry DMF (29ml) were added 4,4,4',4',5,5,5',5' -octamethyl-2, 2' -bis (1,3, 2-dioxaborolane) (2.290G, 9.02mmol), potassium acetate (2.414G, 24.60mmol) and Pd (dppf) Cl2·CH2Cl2(2.009g, 2.460mmol) and the reaction was heated to 80C overnight. The solvent was removed. Purification of the title compound by flash chromatography on silica gel (Snap 50g heptane: ethyl acetate 100:0-7:3) to give (1R,5S) -3- (((trifluoromethyl) sulfonyl) oxy) -8-azabicyclo [ 3.2.1) oxy]Tert-butyl oct-2-ene-8-carboxylate (intermediate G21.2, 1.93G, 5.76mmol, 70.2% yield) as a white solid.
UPLC-MS: 1.34min, 280[ M + H-tBu ] +, method 9
And c, step (c).
To a solution of tert-butyl 3- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -8-azabicyclo [3.2.1] oct-2-ene-8-carboxylate (intermediate G21.2, 1.93G, 5.76mmol) in dry dioxane (20ml) was added a 4M solution of hydrogen chloride in 1,4 dioxane (5.76ml, 23.03mmol) and the solution was stirred for 1 h. The solvent is then removed. The crude product was dissolved in dry DCM (20.00ml), N- (benzyloxycarbonyloxy) succinimide (1.964ml, 6.91mmol) and triethylamine (3.14ml, 23.03mmol) were added and the suspension was stirred for 1 h. The solvent is then removed. The product was purified by Biotage Si 25g with a heptane and EtOAc gradient to give the title compound (810mg, 2.194mmol, 38.1% yield) as a white solid.
UPLC-MS: 1.30min, 258[ M + H-Boc ] +, method 9
Intermediate G22
Methyl (prop-2-yn-1-yl) carbamic acid benzyl ester
To a solution of N-methylpropan-2-yn-1-amine (3.05ml, 36.2mmol) in THF (50ml) and NaHCO3To a solution of saturated aqueous solution (50.0ml) was added N- (benzyloxycarbonyloxy) succinimide (11.31ml, 39.8mmol), and the reaction was stirred overnight. The organic layer was then extracted with ethyl acetate (2 × 100 mL). The organic phase was collected, dried, filtered and the solvent removed under reduced pressure. The product was purified by Biotage Si 10g with a heptane and EtOAc gradient to give the title compound (5g, 24.60mmol, 68.0% yield) as a colourless oil.
UPLC-MS: 0.95min, 204[ M + H ] +, method 9
Intermediate G23
5-methoxy-4-methylpyridin-3-yl) boronic acid
To a solution of 3-bromo-5-methoxy-4-methylpyridine (0.5g, 2.475mmol) in dry 1, 4-dioxane (5ml) was added 4,4,4',4',5,5,5',5' -octamethyl-2, 2' -bis (1,3, 2-dioxaborolane) (0.691g, 2.72mmol), Pd (dppf) Cl2·CH2Cl2(0.606g, 0.742mmol) and potassium acetate (0.729g, 7.42mmol), the solution was heated to 80 ℃. The reaction was stirred overnight. The solvent was then removed and the residue dissolved in THF (5.00ml), 12N aqueous hydrogen chloride (4.12ml, 49.5mmol) was added and stirred for 3 h. The solvent was then removed and the product was purified by C-18 flash chromatography ((H) eluent) 2O/ACN))95:5+0.1％HCOOH)：{(ACN/H2O)95:5+ HCOOH 0.1% }95:5-0:100 to give the title compound as a white solid (350mg, 2.096mmol, 85%).
UPLC-MS: 0.14min, 168[ M + H ] +, method 9
Intermediate G24
(5-methoxy-2-methylpyridin-3-yl) boronic acid
To a solution of 3-bromo-5-methoxy-2-methylpyridine (500mg, 2.475mmol) in dry 1, 4-dioxane (5ml) was added 4,4,4',4',5,5,5',5' -octamethyl-2, 2' -bis (1,3, 2-dioxaborolane) (691mg, 2.72mmol), pd (dppf) Cl2·CH2Cl2(606mg, 0.742mmol) and potassium acetate (729mg, 7.42mmol), the solution was heated to 80 ℃. The reaction was stirred overnight. The solvent was then removed and the residue dissolved in THF (5.00ml), 12N aqueous hydrogen chloride (4.12ml, 49.5mmol) was added and stirred for 3 h. The solvent was then removed and the residue was purified by C-18 flash chromatography (eluent ((H)2O/ACN))95:5+0.1％HCOOH)：{(ACN/H2O)95:5+ HCOOH 0.1% }95:5-0:100 to give the title compound (350mg, 2.096mmol, 85% yield) as a white solid.
UPLC-MS: 0.13min, 168[ M + H ] +, method 9
Intermediate G25
(5- (benzyloxy) -6- (trifluoromethyl) pyridin-3-yl) boronic acid
Step a.3- (benzyloxy) -5-bromo-2-iodopyridine (intermediate G25.1)
To a solution of 5-bromo-2-iodopyridin-3-ol (0.700g, 2.334mmol, prepared according to bioorg.med.chem.lett.2013, 2, 6784) in dry THF (10ml) previously cooled to 0 ℃ was added benzyl alcohol (0.728ml, 7.00mmol), triphenylphosphine (PPh) 3) (2.095ml, 7.00mmol) and diisopropyl azodicarboxylate (1.287ml, 6.54 mmol). The reaction was stirred at room temperature for 4 h. The solvent was removed. The product was purified by Biotage Si 50G with a heptane and EtOAc gradient to give 3- (benzyloxy) -5-bromo-2-iodopyridine (intermediate G25.1, 835mg, 2.141mmol, 92% yield) as a white foam.
UPLC-MS: min, [ M + H ] +, method 9
Step b.3- (benzyloxy) -5-bromo-2- (trifluoromethyl) pyridine (intermediate G25.2)
To a solution of 3- (benzyloxy) -5-bromo-2-iodopyridine (intermediate G25.1, 800mg, 2.051mmol) in dry DMF (10ml) were added copper (I) iodide (2344mg, 12.31mmol) and methyl 2, 2-difluoro-2- (fluorosulfonyl) acetate (0.522ml, 4.10mmol), and the suspension was stirred at 80 ℃ overnight. The solid was then filtered through a pad of celite and the solvent removed. The product was purified by Biotage Si 25G with a heptane and EtOAc gradient to give 3- (benzyloxy) -5-bromo-2- (trifluoromethyl) pyridine (intermediate G25.2, 320mg, 0.964mmol, 47.0% yield) as a white solid.
UPLC-MS: 1.30min, 330[ M + H ] +, method 9
And c, step (c).
To a solution of 3- (benzyloxy) -5-bromo-2- (trifluoromethyl) pyridine (intermediate G25.2, 521mg, 1.569mmol) in dry 1, 4-dioxane (10ml) was added 4,4,4',4',5,5,5',5' -octamethyl-2, 2 '-bis (1,3, 2-dioxaborolane) (438mg, 1.726mmol), [1,1' -bis (diphenylphosphino) ferrocene ]Palladium (II) dichloride (344mg, 0.471mmol) and potassium acetate (462mg, 4.71mmol), and the suspension was stirred at 90 ℃ overnight. HCl 4N (10mL) was then added and the solution was stirred overnight. The solvent was removed under reduced pressure and the eluate was purified by C18 flash chromatography, Snap 30g ((H)2O/ACN))95:5+0.1％HCOOH}：{(ACN/H2O)95:5+ HCOOH 0.1% }95: 5-0: the crude product was purified 100 to give the title compound (400mg, 1.347mmol, 86% yield) as a white solid.
UPLC-MS: 0.97min, 298[ M + H ] +, method 9.
Preparation of the Compounds
Example 1
3- ((6-amino-9H-purin-9-yl) methyl) -4-phenyl-1H-isochromen-1-one
3- (bromomethyl) -4-phenyl-1H-isochromen-1-one (intermediate C1, 35mg, 0.111mmol), commercial 9H-purin-6-amine (30.0mg, 0.222mmol) and K2CO3(30.7mg, 0.222mmol) was dissolved in DMF (1ml) and stirred at RT for 5min and then at 50 ℃ for 10 min. The resulting mixture was then purified directly by reverse phase chromatography using a Biotage C18 SNAP column, adding 2M aqueous HCl (0.5ml) (phase a, water 95%, ACN 5%, formic acid 0.1%); phase B, ACN 95%, water 5%, formic acid 0.1%) to give the title compound (19mg, 46.3%) as a colorless solid.
1H NMR(400MHz，DMSO-d6)ppm 8.16-8.27(m，1H)，8.09(s，2H)，7.74-7.86(m，1H)，7.47-7.70(m，6H)，7.14-7.30(m，2H)，6.94-7.06(m，1H)，5.09(s，2H)。UPLC-MS：2.01min，370.3[M+H]+, method 2.
Example 2
3- ((6-amino-9H-purin-9-yl) methyl) -4- (3-fluorophenyl) -1H-isochromen-1-one
In a similar manner to example 1, 3- (bromomethyl) -4- (3-fluorophenyl) -1H-isochromen-1-one (intermediate C4, 50mg, 0.150mmol), commercial 9H-purin-6-amine (40.6mg, 0.300mmol) and K were used2CO3(41.5mg, 0.300mmol) to give the title compound (36.4mg, 62.6%) as a colorless solid.
1H NMR(400MHz，DMSO-d6)ppm 8.14-8.22(m，1H)8.07(d，J＝9.70Hz，2H)7.74-7.84(m，1H)7.51-7.70(m，2H)7.37-7.46(m，1H)7.27-7.36(m，2H)7.11(bs，2H)6.89-7.04(m，1H)5.07(s，2H)。UPLC-MS：2.09min，388.3[M+H]+, method 2.
Example 3
3- ((6-amino-9H-purin-9-yl) methyl) -4- (2-fluorophenyl) -1H-isochromen-1-one
In a similar manner to example 1, 3- (bromomethyl) -4- (2-fluorophenyl) -1H-isochromen-1-one (intermediate C2, 60mg, 0.180mmol), 9H-purin-6-amine (48.7mg, 0.360mmol) and K were used2CO3(49.8mg, 0.360mmol) the title compound was prepared to give the title compound (45mg,64.5％)。
1H NMR(400MHz，DMSO-d6)8.15-8.24(m，1H)，7.97-8.08(m，2H)，7.75-7.84(m，1H)，7.51-7.72(m，3H)，7.34-7.47(m，2H)，6.86-7.28(m，3H)，4.34(s，2H)。UPLC-MS：1.92min，384.4[M+H]+, method 2.
Example 4
3- ((6-amino-9H-purin-9-yl) methyl) -4-m-tolyl-1H-isochromen-1-one
In a similar manner to example 1, 3- (bromomethyl) -4-m-tolyl-1H-isochromen-1-one (intermediate C3, 64mg, 0.190mmol), 9H-purin-6-amine (52.5mg, 0.39mmol) and K were used2CO3(53.7mg, 0.39mmol) to give the title compound (35mg, 47%).
1H NMR(400MHz，DMSO-d6)ppm 8.17-8.27(m，1H)，8.00-8.14(m，2H)，7.75-7.86(m，1H)，7.57-7.71(m，1H)，7.40-7.50(m，1H)，7.27-7.38(m，3H)，7.11-7.25(s，2H)，6.96-7.09(m，1H)，4.95(s，2H)，2.37(s，3H)。UPLC-MS：2.37min，384.3[M+H]+, method 2.
Example 5
3- (1- (6-amino-9H-purin-9-yl) ethyl) -4-phenyl-1H-isochromen-1-one
In a similar manner to example 1, 3- (1-bromoethyl) -4-phenyl-1H-isochromen-1-one (intermediate C7, 70mg, 0.213mmol), 9H-Purin-6-amine (57.5mg, 0.425mmol) and K2CO3(58.8mg, 0.425mmol) to give the title compound (31mg, 38%).
1H NMR(400MHz，DMSO-d6)ppm 8.40(s，1H)，8.14-8.27(m，1H)，8.10(s，1H)，7.37-7.85(m，7H)，7.17-7.24(bs，2H)，6.88-6.98(m，1H)，5.33-5.49(m，1H)，4.03-4.20(m，1H)，1.75-1.91(d，3H)。UPLC-MS：2.35min，384.4[M+H]+, method 2.
Example 6
3- (1- (6-amino-9H-purin-9-yl) ethyl) -4-m-tolyl-1H-isochromen-1-one
In a similar manner to example 1, 3- (1-bromoethyl) -4-m-tolyl-1H-isochromen-1-one (intermediate C5, 80mg, 0.23mmol), 9H-purin-6-amine (63mg, 0.46mmol) and K were used2CO3(64.4mg, 0.46mmol) to give the title compound (50mg, 54%).
1H NMR(400MHz，DMSO-d6)ppm 8.27-8.38(m，1H)，8.16-8.25(m，1H)，8.00-8.09(m，1H)，7.72-7.83(m，1H)，7.57-7.67(m，1H)，7.36-7.51(m，1H)，7.28-7.36(m，1H)，7.11-7.26(m，4H)，6.88-7.01(m，1H)，5.35-5.52(m，1H)，2.32(s，3H)，1.71-1.92(m，3H)。UPLC-MS：3.32min，398[M+H]+, method 2.
Example 7
3- (1- (6-amino-9H-purin-9-yl) ethyl) -4- (3-fluorophenyl) -1H-isochromen-1-one
In a similar manner to example 1, 3- (1-bromoethyl) -4- (3-fluorophenyl) -1H-isochromen-1-one (intermediate C6, 70mg, 0.202mmol), 9H-purin-6-amine (54.5mg, 0.403mmol) and K were used2CO3(55.7mg, 0.403mmol) to give the title compound (30mg, 37.1%).
1H NMR(400MHz，DMSO-d6)ppm 8.20-8.10(m，3H)，7.95(s，1H)，7.80-7.74(m，2H)，7.65-7.60(m，2H)，7.37-7.20(m，3H)，6.95-6.85(m，1H)，5.50-5.40(m，1H)，1.85-.1.82(m，3H)。UPLC-MS：2.89min，402[M+H]+, method 2.
Example 8
3- (1- (6-amino-9H-purin-9-yl) ethyl) -4- (3- (dimethylamino) phenyl) -1H-isochromen-1-one
In a similar manner to example 1, 3- (1-bromoethyl) -4- (3- (dimethylamino) phenyl) -1H-isochromen-1-one (intermediate C8, 86mg, 0.231mmol), 9H-purin-6-amine (46.8mg, 0.347mmol) and K were used2CO3(47.9mg, 0.347mmol) the title compound was prepared to yield the title compound (36mg, 37%). The resulting mixture was purified directly by reverse phase chromatography using a Biotage C18SNAP column (phase a, water 95%, ACN 5%, formic acid 0.1%); phase B, ACN 95%, water 5%, formic acid 0.1%). The resulting product was further purified by preparative HPLC (method 1) to give the title compound (12mg, 12%).
1H NMR(400MHz，DMSO-d6)ppm 8.38(s，1H)，8.12-8.24(m，1H)，8.03(s，1H)，7.73-7.84(m，1H)，7.54-7.67(m，1H)，7.27-7.42(m，1H)，7.14-7.25(bs，2H)，6.97-7.07(m，1H)，6.87-6.91(m，1H)，6.79-6.86(m, 1H), 6.62-6.71(m, 1H), 5.42-5.62(m, 1H), 2.86 and 2.98(2s, 6H, 3H eacH), 1.75-1.90(m, 3H). UPLC-MS: 4.68min, 427[ M + H ]]+, method 1.
Example 9
3- (1- (6-amino-9H-purin-9-yl) ethyl) -4- (3- (morpholinosulfonyl) phenyl) -1H-isochromen-1-one
In a similar manner to example 1, 3- (1-bromoethyl) -4- (3- (morpholinosulfonyl) phenyl) -1H-isochromen-1-one (intermediate C9, 68mg, 0.142mmol), 9H-purin-6-amine (57.6mg, 0.426mmol) and K were used2CO3(29.5mg, 0.213mmol) to give the title compound (30mg, 37.1%).
The resulting mixture was purified directly by reverse phase chromatography using a Biotage C18 SNAP column (phase a, water 95%, ACN 5%, formic acid 0.1%); phase B, ACN 95%, water 5%, formic acid 0.1%). The resulting product was then purified by preparative HPLC (method 2) to give the compound as chemically pure (10mg, 13%) and in lower purity batches (9mg, 12%).
1H NMR(400MHz，DMSO-d6)ppm 8.45-8.35(m，1H)，8.30-8.20(m，1H)，8.05-7.65(m，6H)，7.45(br s，2H)，6.90(t，1H)，5.30-5.45(m，1H)，3.60-2.6(m，8H)，1.80-1.55(m，1H)。UPLC-MS：3.59min，533[M+H]Method 1
Example 10
3- ((9H-purin-6-ylthio) methyl) -4-phenyl-1H-isochromen-1-one
3- (bromomethyl) -4-phenyl-1H-isochromen-1-one (intermediate C1, 35mg, 0.111mmol), 9H-purine-6-thiol hydrate (18.9mg, 0.111mmol) and K2CO3(15.35mg, 0.111mmol) was dissolved in DMF (1ml) and stirred at RT for 1h 30 min. The reaction mixture was then diluted with water (10ml) and 0.1N aqueous HCl (1 ml). The mixture was extracted with EtOAc and Na2SO4The collected organic phase was dried, filtered and concentrated under reduced pressure. The crude product was purified by preparative HPLC (method 2) to give the title compound (24mg, 56%).
1H NMR(400MHz，DMSO-d6)ppm 8.51(s，1H)，8.33-8.44(m，1H)，8.19-8.30(m，1H)，7.73-7.85(m，1H)，7.59-7.68(m，1H)，7.48-7.60(m，3H)，7.35-7.43(m，2H)，6.91-7.05(m，1H)，4.46(s，2H)。UPLC-MS：2.80min，387.3[M+H]+, method 2.
Example 11
3- ((9H-purin-6-ylthio) methyl) -4- (2-fluorophenyl) -1H-isochromen-1-one
In a similar manner to example 10, 3- (bromomethyl) -4- (2-fluorophenyl) -1H-isochromen-1-one (intermediate C2, 37mg, 0.111mmol), 9H-purine-6-thiol hydrate (18.90mg, 0.111mmol) and K 2CO3(15.35mg, 0.111mmol) to give the title compound (33mg, 73.5%).
1H NMR(400MHz，DMSO-d6)ppm 8.41(m，2H)8.19-8.24(m，1H)7.71-7.83(m，1H)7.51-7.66(m，2H)7.25-7.51(m，3H)6.81-7.02(m，1H)4.27-4.66(s，2H)。UPLC-MS：2.68min，405.3[M+H]+, method 2.
Example 12
3- ((9H-purin-6-ylthio) methyl) -4-m-tolyl-1H-isochromen-1-one
In a similar manner to example 10, 3- (bromomethyl) -4-m-tolyl-1H-isochromen-1-one (intermediate C3, 64mg, 0.194mmol), 9H-purine-6-thiol hydrate (33mg, 0.194mmol) and K2CO3(27mg, 0.111mmol) to give the title compound (64mg, 82%).
1H NMR(400MHz，DMSO-d6)13.01-13.77(bs，1H)，8.46(s，1H)，8.37(s，1H)，8.15-8.22(m，1H)，7.71-7.79(m，1H)，7.55-7.62(m，1H)，7.35-7.42(m，1H)，7.24-7.29(m，1H)，7.10-7.17(m，2H)，6.93-6.99(m，1H)，4.11-4.56(m，2H)，2.28(s，3H)。UPLC-MS：3.15min，401.3[M+H]+, method 2.
Example 13
3- (1- (9H-purin-6-ylthio) ethyl) -4-m-tolyl-1H-isochromen-1-one
In a similar manner to example 10, 3- (1-bromoethyl) -4-m-tolyl-1H-isochromen-1-one (intermediate C5, 67mg, 0.195mmol), 9H-purine-6-thiol hydrate (33.2mg, 0.195mmol)) and K were used2CO3(27mg, 0.111mmol) to give the title compound (70mg, 87%).
1H NMR(400MHz，DMSO-d6)13.5(bs，1H)，8.45-8.39(m，2H)，8.26-8.21 9m，1H)，7.80-7.75(m，1H)，7.65-7.60(m，1H)，7.50-7.20(m，3H)，7.10-6.95(m，2H)，5.40-5.30(m，1H)，2.45(s，1.5H)，2.20(2，1.5H)，1.72-1.67(m，3H)。UPLC-MS：3.32min，415.4[M+H]+, method 2.
Example 14
3- (1- (9H-purin-6-ylsulfanyl) ethyl) -4- (3-fluorophenyl) -1H-isochromen-1-one
In a similar manner to example 10 using 3- (1-bromoethyl) -4- (3-fluorophenyl) -1H-isochromen-1-one (intermediate C6, 70mg, 0.202mmol), 9H-purine-6-thiol hydrate (34.3mg, 0.202mmol) and K 2CO3(27.9mg, 0.202mmol) the title compound was prepared.
The resulting mixture was purified directly by reverse phase chromatography using a Biotage C18 SNAP column (phase a, water 95%, ACN 5%, formic acid 0.1%); phase B, ACN 95%, water 5%, formic acid 0.1%) to give the title compound (53mg, 63%).
1H NMR(400MHz，DMSO-d6)ppm 13.24-13.73(bs，1H)，8.34-8.46(m，2H)，8.25(d，1H)，7.72-7.86(m，1H)，7.65(d，2H)，7.33(d，3H)，7.06(m，1H)，6.98(m，1H)，5.33(m，1H)，1.72(t，，3H)。UPLC-MS：3.04min，419.4[M+H]+, method 2.
Example 15
3- (1- (4-amino-1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (6-methylpyridin-3-yl) -1H-isochromen-1-one
The reaction mixture was washed with 3- (1-bromoethyl) -4- (6-methylpyridin-3-yl) -1H-isochromen-1-one hydrobromide (intermediate C10, 120mg, 0.282mmol), 1H-pyrazolo [3,4-d]Pyrimidin-4-amine (76mg, 0.565mmol) and K2CO3(117mg, 0.847mmol) was stirred in DMF (1.5ml) at 50 ℃ for 2.5 h. The reaction mixture was diluted with 1M aqueous HCl (2ml) and purified by reverse phase chromatography using a Biotage C18 SNAP column (phase a, water 95%, ACN 5%, formic acid 0.1%); phase B, ACN 95%, water 5%, formic acid 0.1%) to give the title compound as a colorless solid (16.7mg, 15%).
1H NMR(400MHz，DMSO-d6)ppm 8.49(br s，0.5H)，8.23-8.20(m，1H)，8.12(br s，0.5H)，8.09(br s，1H)，8.01(br s，1H)，7.80-7.62(m，4H)，7.42-7.38(m，1.5H)，7.25-7.23(m，0.5H)6.90-6.88(d，1H)，5.65-5.55(m，1H)，2.54(m，3H)，，1.81(t，3H)。UPLC-MS：1.04min，399.4[M+H]+, method 2.
Example 16
3- (1- (4-amino-1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (3- (morpholinosulfonyl) phenyl) -1H-isochromen-1-one
In a similar manner to example 15, 3- (1-bromoethyl) -4- (3- (morpholinosulfonyl) phenyl) -1H-isochromen-1-one (intermediate C9, 90mg, 0.188mmol), 1H-pyrazolo [3,4-d ] was used at 80 deg.C ]Pyrimidin-4-amine (50.8mg, 0.376mmol) and K2CO3(52mg, 0.376mmol) was prepared in DMF (1ml) for 2h to give the title compound (54mg, 54%).
1H NMR(400MHz，DMSO-d6)ppm 8.20-8.16(m，1H)，8.03(s，1H)，7.98(s，0.5H)，7.89(s，0.5H)，7.82-7.69(m，4H)，7.61-7.57(m，1H)，7.50-7.46(m，1H)，6.81-6.7(m，1H)，5.70-5.60(q，0.5H)，5.50-5.40(q，0.5H)，3.64-3.46(m，4H)，3.04-2.74(m，4H)，1.80(d，0.5H)，1.71(d，0.5H)。UPLC-MS：1.98min，533.4[M+H]+, method 2.
Example 17
3- (1- (4-amino-1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one
In a similar manner to example 15, 3- (1-bromoethyl) -4-phenyl-1H-isochromen-1-one (intermediate C7, 70mg, 0.213mmol), 1H-pyrazolo [3,4-d ] was used at 80 ℃]Pyrimidin-4-amine (40.2mg, 0.298mmol) and K2CO3(41.1mg, 0.298mmol) in DMF (1ml) was prepared for 4h to give the title compound (28mg, 34%).
1H NMR(400MHz，DMSO-d6)ppm 8.22(d，1H)，8.14(s，1H)，8.10(s，1H)，8.02(s，1H)，7.72-7.81(m，1H)，7.50-7.66(m，3H)，7.41-7.49(m，2H)，7.34-7.40(m，1H)，7.11(d，1H)，6.90(d，1H)，5.60(d，1H)，1.79(d，3H)。UPLC-MS：4.77min，384.2[M+H]+, method 3.
Example 18
3- (1- (4-amino-1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (thiazol-5-yl) -1H-isochromen-1-one
In a similar manner to example 15, 3- (1-bromoethyl) -4- (thiazol-5-yl) -1H-isochromen-1-one (intermediate C12, 56mg, 0.17mmol), 1H-pyrazolo [3,4-d ] was used at 80 deg.C]Pyrimidin-4-amines(45mg, 0.33mmol) and K2CO3(46mg, 0.33mmol) the title compound was prepared in DMF (1ml) for 3h to give the title compound (35mg, 54%).
1H NMR(400MHz，DMSO-d6)ppm 9.30(s，1H)，8.22(d，1H)，8.14(s，1H)，8.10(s，1H)，8.04(s，1H)，7.83(s，2H)，7.63-7.69(m，2H)，7.05(d，1H)，5.75(d，1H)，1.81(d，3H)。UPLC-MS：3.09min，391.2[M+H]+, method 3.
Example 19
3- (1- (4-amino-1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (2-methylpyridin-4-yl) -1H-isochromen-1-one
In a similar manner to example 15, 3- (1-bromoethyl) -4- (2-methylpyridin-4-yl) -1H-isochromen-1-one (intermediate C14, 98mg, 0.285mmol), 1H-pyrazolo [3,4-d ] was used at 50 ℃]Pyrimidin-4-amine (57.7mg, 0.43mmol) and K2CO3(59mg, 0.42mmol) the title compound was prepared in DMF (2ml) for 6h to give the title compound (3.4mg, 3%).
1H NMR(400MHz，DMSO-d6)ppm 8.57(d，0.5H)，8.38(d，0.5H)，8.25-8.22(m，1H)，8.11(s，0.5H)，8.05(s，0.5H)，8.01(d，1H)，7.80-7.62(m，4H)，7.27-7.25(m，1H)，6.90-6.87(m，1.5H)，6.68(br s，0.5H)，5.70-5.60(m，1H)，3.15-3.05(m，1H)1.81-1.75(m，3H)，1.18(t，3H)。UPLC-MS：3.02min，399.3[M+H]+, method 3.
Example 20
3- (1- (4-amino-1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-benzyl-1H-isochromen-1-one
4-benzyl-3- (1-bromoethyl) -1H-isochromen-1-one (intermediate C13, 22mg, 0.064mmol), 1H-pyrazolo [3,4-d ] was used at 55 ℃ in a similar manner to example 15]Pyrimidin-4-amine (12.99mg, 0.096mmol) and K2CO3(13.29mg, 0.096mmol) the title compound was prepared in DMF (1ml) for 2h to give the title compound (9mg, 35%).
1H NMR(400MHz，DMSO-d6)ppm 8.15-8.23(m，1H)，8.08-8.13(m，2H)，7.70-8.22(m，4H)，6.97-7.24(m，6H)，6.20-6.31(m，1H)，4.18-4.44(m，2H)，1.78-1.89(m，3H)。UPLC-MS：4.55min，398.1[M+H]+, method 3.
Example 21
3- ((9H-purin-6-ylamino) methyl) -4- (3-fluorophenyl) -1H-isochromen-1-one
Tert-butyl 9-trityl-9H-purin-6-ylcarbamate (93mg, 0.195mmol) and 50% NaH mineral oil dispersion (9.4mg, 0.195mmol) were dissolved in DMF (0.2ml) at 0 ℃. A solution of 3- (bromomethyl) -4- (3-fluorophenyl) -1H-isochromen-1-one (intermediate C4, 50mg, 0.150mmol) in DMF (0.6mL) was then added. The reaction mixture was stirred at 0 ℃ for 5min and at RT for 15 min. The reaction mixture was then diluted with EtOAc (20mL), washed with 0.2M aqueous HCl, saturated aqueous NaCl, and Na 2SO4Dried and concentrated under reduced pressure to give a crude yellow oil.
The crude oil was reacted with TFA (1.5ml) in DCM (2ml) for 1 h. Then diluted with DCM and dried under reduced pressure to give a yellow oil. It was purified by reverse phase chromatography using a Biotage C1830 g SNAP column (phase a, water 95%, ACN 5%, formic acid 0.1%); phase B, ACN 95%, water 5%, formic acid 0.1%) to give the title compound (20mg, 34.4%) as a colorless solid.
1H NMR(400MHz，DMSO-d6)8.08-8.23(m，4H)，7.72-7.81(m，1H)，7.51-7.63(m，2H)，7.25-7.38(m，3H)，6.97(d，1H)，3.35-3.43(m，2H)。UPLC-MS：2.25min，388.3[M+H]+, method 2.
Example 22
3- (1- (9H-purin-6-ylamino) ethyl) -4-phenyl-1H-isochromen-1-one
The title compound was prepared in analogy to example 21 from tert-butyl 9-trityl-9H-purin-6-ylcarbamate (139mg, 0.292mmol), 3- (1-bromoethyl) -4-phenyl-1H-isochromen-1-one (intermediate C7, 80mg, 0.243mmol) and a solution of 50% NaH in mineral oil dispersion (16.3mg, 0.340mmol) in DMF to give the title compound (16mg, 17.2%) as light yellow solid.
1H NMR(400MHz，DMSO-d6)ppm 13.08(s，1H)，8.10(m，3H)，7.85-7.95(m，1H)，7.72-7.81(m，1H)，7.46-7.64(m，5H)，7.30-7.43(m，1H)，6.85-6.99(m，1H)，4.78-5.22(m，1H)，1.51(d，3H)。UPLC-MS：2.52min，384.5[M+H]+, method 2.
Example 22a (enantiomer 1) and example 22b (enantiomer 2): 3- (1- (9H-purin-6-ylamino) ethyl) -4-phenyl-1H-isochromen-1-one single enantiomer
The racemate 3- (1- (9H-purin-6-ylamino) ethyl) -4-phenyl-1H-isochromen-1-one (example 22, 0.145g, 0.378mmol) was dissolved in ethanol/methanol 1/1(34mL) and subjected to chiral resolution by chiral preparative chromatography. Conditions are as follows: column: chiralpak AD-H (25X2.0cm), 5. mu.l; mobile phase: n-hexane/(2-propanol + 0.1% isopropylamine) 75/25% v/v; flow rate: 19 ml/min; and D, DAD detection: 220 nm; and (3) injection: 19mg (per injection).
Evaporation of the fraction containing the first eluting enantiomer gave compound 22a (first eluting enantiomer, 0.052g, 0.135 mmol). Chiral HPLC (method a 1): rt ═ 6.0min, ee > 99%;
1H NMR(400MHz，DMSO-d6)ppm 12.92(br.s.，1H)，8.06-8.27(m，3H)，7.86-7.94(m，1H)，7.73-7.81(m，1H)，7.48-7.64(m，5H)，7.37-7.43(m，1H)，6.95(d，1H)，5.06(br.s.，1H)，1.53(d，3H)。UPLC-MS：0.85min，384.2[M+H]+, method 13.
Evaporation of the fraction containing the second eluting enantiomer gave compound 22b (second eluting enantiomer, 0.032g, 0.083mmol), chiral HPLC (method a 1): rt ═ 13.0min, ee > 99%;
1H NMR(400MHz，DMSO-d6)ppm 12.78(br.s.，1H)，8.09-8.24(m，3H)，7.82-7.90(m，1H)，7.74-7.81(m，1H)，7.49-7.64(m，5H)，7.38-7.44(m，1H)，6.95(d，1H)，5.06(br.s.，1H)，1.53(d，3H)。UPLC-MS：0.83min，384.2[M+H]+, method 13.
Example 23
3- (1- (9H-purin-6-ylamino) ethyl) -4- (1,2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one hydrochloride
The title compound was prepared in analogy to example 21 from 9-trityl-9H-purin-6-ylcarbamic acid tert-butyl ester (71mg, 0.149mmol) and a solution of 4- (3- (1-bromoethyl) -1-oxo-1H-isochromen-4-yl) -5, 6-dihydropyridine-1 (2H) -carboxylic acid tert-butyl ester (intermediate C11, 54mg, 0.124mmol) and 50% NaH mineral oil dispersion (18mg, 0.34mmol) in DMF.
It was purified by reverse phase chromatography using a Biotage C18 SNAP column (phase a, water 95%, ACN 5%, formic acid 0.1%); phase B, ACN 95%, water 5%, formic acid 0.1%). Fractions saturated with the desired product were collected, added to 1M aqueous HCl (5ml), and concentrated under reduced pressure to give the title compound (24mg, 45%) as a colorless solid.
1H NMR(400MHz，DMSO-d6)ppm 9.3-9.0(m，2H)，8.5-8.3(m，1H)，8.20-8.14(m，1H)，7.89-7.85(m，1H)，7.7-760(m，2H)，6.05-5.92(m，1H)，5.60-5.40(m，1H)，3.79-3.25(m，6H)，1.61-1.59(m，3H)。UPLC-MS：2.29min，389.5[M+H]+, method 3.
Example 24
3- (1- (9H-purin-6-ylamino) ethyl) -4- (3, 6-dihydro-2H-pyran-4-yl) -1H-isochromen-1-one
The title compound was prepared in analogy to example 21 from tert-butyl 9-trityl-9H-purin-6-ylcarbamate (214mg, 0.449mmol) and a solution of 3- (1-bromoethyl) -4- (3, 6-dihydro-2H-pyran-4-yl) -1H-isochromen-1-one (intermediate C19, 94mg, 0.280mmol) and 50% NaH mineral oil dispersion (18mg, 0.34mmol) in DMF. It was purified by reverse phase chromatography using a Biotage C18 SNAP column (phase a, water 95%, ACN 5%, formic acid 0.1%); phase B, ACN 95%, water 5%, formic acid 0.1%). Fractions saturated with the desired product were collected, added to 1M aqueous HCl (5ml), and concentrated under reduced pressure to give the title compound (6.3mg, 5.8%) as a colorless solid.
1H NMR(400MHz，DMSO-d6)ppm 12.94(s，0.8H)，12.08(s，0.2H)，8.20-8.12(m，3H)，7.89-7.84(m，2H)，7.63-7.45(m，3H)，5.98-5.94(m，1H)，5.75-5.50(m，1H)。4.30-4.21(m，2H)，3.96-3.90(m，2H)，2.23-2.26(m，2H)，1.59-1.55(m，3H)。UPLC-MS：3.29min，390[M+H]+, method 3.
Example 25
3- (1- (9H-purin-6-ylamino) propyl) -4-phenyl-1H-isochromen-1-one
The title compound was prepared in analogy to example 21 from a solution of tert-butyl 9-trityl-9H-purin-6-ylcarbamate (167mg, 0.350mmol), 3- (1-bromopropyl) -4-phenyl-1H-isochromen-1-one (intermediate C20, 100mg, 0.291mmol) and 50% NaH mineral oil dispersion (21mg, 0.870mmol) in DMF at 55 ℃.
The crude product was purified by reverse phase chromatography using a column consisting of two Biotage C1830 g SNAP columns (phase a, water 95%, ACN 5%, formic acid 0.1%); phase B, ACN 95%, water 5%, formic acid 0.1%) was purified by preparative HPLC (method 1) to give the title compound (2.8mg, 2.4%) as a colorless solid.
1H NMR(400MHz，DMSO-d6)ppm 12.64-13.09(bs，1H)，8.05-8.26(m，3H)，7.70-7.88(m，2H)，7.46-7.63(m，5H)，7.36(d，1H)，6.92(d，1H)，4.66-5.08(m，1H)，1.83-2.03(m，2H)，0.84(t，3H)。UPLC-MS：5.04min，398.5[M+H]+, method 3.
Example 26
3- (1- (9H-purin-6-ylamino) ethyl) -4- (4- (2-morpholinoethoxy) phenyl) -1H-isochromen-1-one formate salt
The title compound was prepared in analogy to example 21 from tert-butyl 9-trityl-9H-purin-6-ylcarbamate (140mg, 0.29mmol), 3- (1-bromoethyl) -4- (4- (2-morpholinoethoxy) phenyl) -1H-isochromen-1-one hydrobromide (intermediate C15, 100mg, 0.185mmol) and a solution of 50% NaH mineral oil dispersion (27mg, 0.550mmol) in DMF at 65 ℃ to yield the title compound (16.9mg, 16.3%) as a colorless solid.
1H NMR(400MHz，DMSO-d6)ppm 12.77-13.04(bs，1H)，8.15(m，4H)，7.82-7.92(m，1H)，7.71-7.81(m，1H)，7.52-7.62(m，1H)，7.34-7.52(m，1H)，7.22-7.35(m，1H)，7.11(m，2H)，6.80-7.02(m，1H)，4.80-5.28(m，1H)，4.17(m，2H)，3.51-3.76(m，4H)，2.75(m，2H)，2.51-2.60(m，4H)，1.50(d，J＝7.06Hz，3H)。UPLC-MS：3.94min，513.1[M+H]+, method 3.
Example 27
4-amino-8- (1- (1-oxo-4-phenyl-1H-isochromen-3-yl) ethyl) pyrido [2,3-d ] pyrimidin-5 (8H) -one
3- (1-bromoethyl) -4-phenyl-1H-isochromen-1-one (intermediate C7, 50mg, 0.15mmol), 4-aminopyrido [2,3-d ] pyrimidin-5 (8H) -one (37mg, 0.23mmol), potassium carbonate (31.5mg, 0.23mmol) in DMF (0.5ml) was reacted at 80 ℃. The crude product was purified by reverse phase chromatography using a Biotage C1830 g SNAP column (phase a, water 95%, ACN 5%, formic acid 0.1%); phase B, ACN 95%, water 5%, formic acid 0.1%) to give the title compound (32mg, 51%).
1H NMR(400MHz，DMSO-d6)ppm 9.48-9.56(m，1H)，8.16-8.27(m，1H)，8.06-8.13(m，2H)，7.98-8.04(m，1H)，7.72-7.81(m，1H)，7.58-7.66(m，1H)，7.46-7.57(m，1H)，7.33-7.44(m，3H)，7.07-7.16(m，1H)，6.85-6.94(m，1H)，6.13-6.20(m，2H)，1.51-1.81(m，3H)。UPLC-MS：4.7min，411.1[M+H]+, method 5.
Example 28
3- (1- (9H-purin-6-ylamino) ethyl) -4- (5- (morpholinomethyl) thiophen-2-yl) -1H-isochromen-1-one
9-Tribenzyl-9H-purin-6-ylcarbamic acid tert-butyl ester (102mg, 0.213mmol) was added to a solution of 50% NaH mineral oil dispersion (5.12mg, 0.213mmol) in N, N-dimethylformamide (0.5ml), and the mixture was stirred at RT for 30 min.
3- (1-bromoethyl) -4- (5- (morpholinomethyl) thiophen-2-yl) -1H-isochromen-1-one hydrobromide (intermediate C16, 100mg, 0.194mmol) was suspended in DMF (0.5ml, 0.194mmol) and reacted with 50% NaH mineral oil dispersion (5.1mg, 0.213mmol) at RT for 15 min. The resulting solution was added to the previously prepared mixture, kept stirring at 60 ℃ for 1h, at RT for 3h and then at 80 ℃ for 3 h. The reaction mixture was poured into brine and extracted with EtOAc. The collected organic phase was dried and concentrated. The resulting crude material was purified by reverse phase chromatography using Biotage C1860g SNAP column, 37% aqueous HCl (1ml) was added to the collected fractions, and concentrated. The resulting material was further purified by reverse phase chromatography using a Biotage C1830 g SNAP column. The resulting material was finally purified on silica gel with a gradient of DCM and 2-propanol, modified with 0.5% TEA to give the title compound (9mg, 9.5%).
1H NMR(400MHz，DMSO-d6)ppm 12.64-13.05(bs，1H)，8.36-8.44(m，1H)，8.11-8.19(m，1H)，8.01-8.10(m，2H)，7.86-7.99(m，1H)，7.74-7.83(m，1H)，7.47-7.63(m，1H)，7.12-7.22(m，1H)，6.98-7.11(m，2H)，5.11-5.34(m，1H)，3.35-3.79(m，6H)，2.33-2.42(m，4H)，1.41-1.57(m，3H)。UPLC-MS：3.43min，489.1[M+H]+, method 3.
Example 29
3- ((9H-purin-6-ylamino) methyl) -4-phenyl-1H-isochromen-1-one
Step 1. (1-oxo-4-phenyl-1H-isochromen-3-yl) methyl (9-trityl-9H-purin-6-yl) carbamic acid tert-butyl ester
Tert-butyl 9-trityl-9H-purin-6-ylcarbamate (63.6mg, 0.133mmol) in DMF (0.1ml) was added to a solution of 50% NaH in a suspension in mineral oil (5.33mg, 0.133mmol) in DMF (0.1ml) and 3- (bromomethyl) -4-phenyl-1H-isochromen-1-one (intermediate C1, 35mg, 0.111mmol) in DMF (0.2ml) was added and the resulting mixture was warmed to RT at 0 ℃. The reaction mixture was then poured into water and extracted with EtOAc. The collected organic phase was dried and concentrated under reduced pressure. The resulting crude product (80mg) was used in the next step without any further purification and characterization.
Step 2
Tert-butyl (1-oxo-4-phenyl-1H-isochromen-3-yl) methyl (9-trityl-9H-purin-6-yl) carbamate (80mg, 0.112mmol) was dissolved in DCM (0.7mL) and TFA (1mL) for 45min at RT. The solvent was then removed and the crude product was purified directly by preparative HPLC (method 1) to give the title compound (17mg, 40.9%).
1H NMR(400MHz，DMSO-d6)12.51-13.14(bs，1H)，8.15-8.25(m，1H)，8.04-8.13(m，2H)，7.92-8.04(bs，1H)，7.68-7.77(m，1H)，7.54-7.61(m，1H)，7.38-7.54(m，5H)，6.91-7.00(m，1H)，4.54(s，2H)。UPLC-MS：2.17min，370.3[M+H]+, method 2.
Example 30
3- ((9H-purin-6-ylamino) methyl) -4- (2-fluorophenyl) -1H-isochromen-1-one
The title compound was prepared from tert-butyl 9-trityl-9H-purin-6-ylcarbamate (60mg, 0.180mmol), 3- (bromomethyl) -4- (2-fluorophenyl) -1H-isochromen-1-one (intermediate C2, 99mg, 0.207mmol) in analogy to example 29 to yield 35mg, 50%.
1H NMR(400MHz，DMSO-d6)12.78-12.99(bs，1H)，7.93-8.23(m，4H)，7.68-7.84(m，1H)，7.27-7.63(m，5H)，6.88-7.00(m，1H)，4.44(s，2H)。UPLC-MS：2.08min，388.3[M+H]+, method 2.
Example 31
3- ((9H-purin-6-ylamino) methyl) -4-m-tolyl-1H-isochromen-1-one
The title compound was prepared from tert-butyl 9-trityl-9H-purin-6-ylcarbamate (102mg, 0.214mmol), 3- (bromomethyl) -4-m-tolyl-1H-isochromen-1-one (intermediate C3, 64mg, 0.194mmol) in analogy to example 29 to yield 33mg, 39%.
1H NMR(400MHz，DMSO-d6)ppm 12.91(bs，1H)7.91-8.27(m，4H)7.74(t，1H)7.57(t，1H)7.33-7.42(m，1H)7.13-7.30(m，3H)6.97(d，1H)4.39(s，2H)2.36(s，3H)。UPLC-MS：2.51min，384.3[M+H]+, method 2.
Example 32
3- (1- (9H-purin-6-ylamino) ethyl) -4- (3-fluorophenyl) -1H-isochromen-1-one
The title compound was prepared from tert-butyl 9-trityl-9H-purin-6-ylcarbamate (132mg, 0.276mmol), 3- (1-bromoethyl) -4- (3-fluorophenyl) -1H-isochromen-1-one (intermediate C6, 87mg, 0.251mmol) in analogy to example 29 to yield 23mg, 28%.
1H NMR(400MHz，DMSO-d6)ppm 12.82-13.02(bs，1H)，8.02-8.27(m，3H)，7.86-8.01(m，1H)，7.73-7.83(m，1H)，7.52-7.66(m，2H)，7.15-7.45(m，3H)，6.88-7.00(m，1H)，4.89-5.18(m，1H)，1.56(m，3H)。UPLC-MS：2.59min，402.4[M+H]+, method 2.
Example 33
3- (1- (9H-purin-6-ylamino) ethyl) -4-m-tolyl-1H-isochromen-1-one
The title compound was prepared from tert-butyl 9-trityl-9H-purin-6-ylcarbamate (138mg, 0.28mmol), 3- (1-bromoethyl) -4-m-tolyl-1H-isochromen-1-one (intermediate C5, 90mg, 0.26mmol) in analogy to example 29 to yield 36mg, 35%.
1H NMR(400MHz，DMSO-d6)ppm 12.79-13.01(m，1H)，8.05-8.31(m，3H)，7.81-7.98(m，1H)，7.68-7.80(m，1H)，7.50-7.66(m，1H)，7.25-7.48(m，3H)，7.11-7.21(m，1H)，6.88-7.00(m，1H)，4.95-5.21(m，1H)，2.35(m，3H)，1.44-1.58(m，3H)。UPLC-MS：3.49min，398.1[M+H]+, method 2.
Example 34
3- (1- (9H-purin-6-ylamino) ethyl) -4- (3- (dimethylamino) phenyl) -1H-isochromen-1-one
The title compound was prepared from tert-butyl 9-trityl-9H-purin-6-ylcarbamate (127mg, 0.26mmol), 3- (1-bromoethyl) -4- (3- (dimethylamino) phenyl) -1H-isochromen-1-one (intermediate C8, 90mg, 0.242mmol) in analogy to example 28 to yield 6mg, 6%.
1H NMR(400MHz，DMSO-d6)ppm 12.61-13.18(bs，1H)，8.53(s，1H)，8.13(m，3H)，7.75(m，1H)，7.60(m，1H)，7.28-7.39(m，1H)，6.97-7.10(m，1H)，6.72-6.88(m，2H)，6.57-6.71(m，1H)，4.91-5.28(m，1H)，2.78(s，3H)，2.97(s，3H)，1.43-1.58(m，3H)。UPLC-MS：4.75min，427[M+H]+, method 1.
Example 35
3- ((4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) methyl) -4-phenyl-1H-isochromen-1-one
Reacting 3- ((4-amino-3-iodo-1H-pyrazolo [3, 4-d)]Pyrimidin-1-yl) methyl) -4-phenyl-1H-isochromen-1-one (intermediate D1, 50mg, 0.101mmol), 3-fluoro-5-hydroxyphenylboronic acid (32mg, 0.201mmol), Cs2CO3(69mg，0.202mmol)、Pd(PPh3)4(9.3mg, 8.0umol) was reacted in DMF (0.5mL) under mw irradiation at 110 ℃. The resulting crude product was purified directly by reverse phase using a Biotage C1830 g SNAP gradient with water and acetonitrile to give the title compound (6mg, 12%).
1H NMR(400MHz，DMSO-d6)ppm 10.5(br s，1H)，8.54(s，1H)，8.24(s，1H)，8.22-8.20(m，1H)，7.83-7.79(m，1H)，7.66-7.62(m，1H)，7.55-7.47(m，6H)，7.04-7.02(m，1H)，6.89(br s，1H)，6.84-6.82(m，1H)，6.66-6.64(m，1H)，5.26(s，2H)。UPLC-MS：5.56min，479.9[M+H]+, method 1.
Example 43
3- (1- (1H-pyrazolo [3,4-d ] pyrimidin-4-ylamino) ethyl) -4-phenyl-1H-isochromen-1-one
3- (1-aminoethyl) -4-phenyl-1H-isochromen-1-one hydrochloride (intermediate E1, 141mg, 0.136mmol), 4-chloro-1H-pyrazolo [3,4-d ] pyrimidine (42.0mg, 0.272mmol) and DIEA (95. mu.L, 0.543mmol) were stirred in tert-butanol (800. mu.L) at 80 ℃ for 3H. The reaction was stopped by addition of 1mL of 1M aqueous HCl and the resulting crude product was purified directly by reverse phase chromatography using Biotage C1830 g SNAP with a gradient of water and acetonitrile to give the title compound (29mg, 56%).
1H NMR(400MHz，DMSO-d6)ppm 13.10-13.60(bs，1H)，8.46-8.70(m，1H)，8.15-8.30(m，2H)，8.05-8.13(m，1H)，7.68-7.85(m，1H)，7.47-7.65(m，5H)，7.32-7.43(m，1H)，6.85-7.06(m，1H)，4.92-5.13(m，1H)，1.42-1.63(m，3H)。UPLC-MS：4.69min，384.1[M+H]+, method 3.
Example 44
4-amino-6- (1- (1-oxo-4-phenyl-1H-isochromen-3-yl) ethylamino) pyrimidine-5-carbonitrile
The title compound was prepared in a similar manner to example 43 from (intermediate E1, 52mg, 0.172mmol) and 4-amino-6-chloropyrimidine-5-carbonitrile (53.3mg, 0.345mmol) to give the title compound (30mg, 19.8%).
1H NMR(400MHz，DMSO-d6)ppm 8.17-8.25(m，1H)，7.89(s，1H)，7.72-7.79(m，1H)，7.57-7.63(m，1H)，7.40-7.56(m，5H)，7.31-7.38(m，1H)，7.15-7.27(m，2H)，6.86-6.94(m，1H)，4.79-5.00(m，1H)，1.43(d，J＝7.06Hz，3H)。UPLC-MS：4.88min，384.1[M+H]+, method 3.
Example 45
3- (1- (9H-purin-6-ylamino) ethyl) -4- (1-methyl-1, 2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one hydrochloride
3- (1-bromoethyl) -4- (1-methyl-1, 2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one hydrobromide (C21, 100mg, 0.233mmol), tert-butyl 9-trityl-9H-purin-6-ylcarbamate (120mg, 0.251mmol) and NaH (33.6mg, 0.699mmol) were reacted at 65 ℃ for 2H under a nitrogen atmosphere. The reaction was diluted with 15mL EtOAc, washed 3 times with 10mL 0.1M aqueous HCl, 1 time with saturated aqueous NaCl and the solvent evaporated to give an oil. The crude product was dissolved in TFA/DCM (3mL +3mL), stirred at RT for 3h, and quenched by addition of 1M aqueous HCl (1 mL). The resulting mixture was purified directly by reverse phase chromatography on a Biotage C1830 g SNAP gradient with water and acetonitrile. To the combined fractions from flash chromatography was added 1M aqueous HCl (5mL) and the mixture was dried under pressure to give the title compound as a yellow solid (13.6mg, 13.3% yield).
1H NMR(400MHz，DMSO-d6)ppm 10.77(br.s.，1H)，8.31-8.55(m，2H)，8.08-8.26(m，1H)，7.72-8.02(m，2H)，7.42-7.72(m，2H)，5.86-6.06(m，1H)，5.53(br.s.，1H)，4.00-4.40(m，4)、2.48-2.98(m，5H)，1.61(d，J＝7.06Hz，3H)。UPLC-MS：3.89min，403.1[M+H]+, method 7.
Example 46a (enantiomer 1) and example 46b (enantiomer 2): 3-1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1-methyl-1, 2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one hydrochloride
Single enantiomers
The racemate 3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1-methyl-1, 2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one hydrochloride (example 46, 0.250g, 0.456mmol) was dissolved in ethanol/methanol 1/1(18ml) and subjected to chiral resolution by chiral preparative chromatography: two methods were identified for obtaining each enantiomer separately, and both methods were applied. First process conditions: column: chiralpak IA (25 X2.0cm), 5 μm; mobile phase: n-hexane/(2-propanol/methanol 1/1) 60/40% v/v; flow rate: 18 ml/min; and D, DAD detection: 220 nm; loop circuit: 750 mu l of the solution; and (3) injection: 10 mg/injection. second method conditions: column: whelk 0-1 (25X 2.0cm), 10 μm; mobile phase: n-hexane/(2-propanol/methanol 1/1) 65/35% v/v; flow rate: 18 ml/min; and D, DAD detection: 220 nm; loop circuit: 750 mu l of the solution; and (3) injection: 10 mg/injection.
The fraction containing the first eluted enantiomer obtained using the first method was evaporated (second elution using the second method), 1.25M HCl in MeOH was added and the volatiles were removed under reduced pressure to give compound 46a (49mg, 0.089 mmol). Chiral HPLC (method a 3): rt ═ 6.1min, ee ═ 93%;
1H NMR(400MHz，DMSO-d6)ppm 10.10-11.00(m，2H)，8.23-8.45(m，1H)，8.10-8.23(m，1H)，7.79-8.01(m，2H)，7.45-7.72(m，1H)，7.02-8.78(m，2H)，6.82-7.01(m，2H)，6.65-6.76(m，1H)，5.43-6.35(m，2H)，2.91-3.04(m，3H)，2.54-4.22(m，6H)，1.82-2.04(m，3H)。UPLC-MS：0.58min，513.3[M+H]Method 13
The fraction saturated with the second eluted enantiomer from the first procedure (first elution by the second procedure) was evaporated, 1.25M HCl in MeOH was added and the volatiles were removed under reduced pressure to give compound 46b (45mg, 0.082 mmol). Chiral HPLC (method a 3): rt ═ 7.7min, ee > 99%;
1H NMR(400MHz，DMSO-d6)ppm 10.08-11.44(m，2H)，8.32-8.56(m，1H)，8.13-8.23(m，1H)，7.80-8.03(m，2H)，7.48-7.74(m，1H)，7.02-8.78(m，2H)，6.84-7.01(m，2H)，6.69-6.79(m，1H)，5.47-6.42(m，2H)，2.90-3.03(m，3H)，2.55-4.29(m，6H)，1.85-2.04(m，3H)。UPLC-MS：0.58min，513.3[M+H]+, method 13.
Example 47
3- (1- (9H-purin-6-ylamino) ethyl) -4- (4- (morpholinomethyl) phenyl) -1H-isochromen-1-one hydrochloride
The title compound was prepared from 3- (1-bromoethyl) -4- (4- (morpholinomethyl) phenyl) -1H-isochromen-1-one hydrobromide (C22, 100mg, 0.196mmol) in analogy to example 21 to yield the title compound (5.8mg, 5.7%).
1H NMR(400MHz，DMSO-d6)ppm 10.43-11.09(m，1H)，8.22(d，J＝7.94Hz，3H)，7.27-7.93(m，7H)，6.95(d，J＝8.38Hz，1H)，4.80-5.18(m，1H)，4.44(br.s.，2H)，3.67-4.15(m，6H)，3.18(m，2H)，1.56(d，J＝7.06Hz，3H)。UPLC-MS：5.69min，483.1[M+H]+, method 7.
Example 49a (enantiomer 1) and example 49b (enantiomer 2)3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (5- (morpholinomethyl) thiophen-2-yl) -1H-isochromen-1-one hydrochloride the single enantiomer
The racemate 3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (5- (morpholinomethyl) thiophen-2-yl) -1H-isochromen-1-one hydrochloride (example 49, 0.137g, 0.21mmol) was dissolved in ethanol (7ml) and subjected to chiral resolution by chiral preparative chromatography. Conditions are as follows: column: whelk O-1(R, R) (25X2.0cm), 10 μm; mobile phase: n-hexane/(ethanol/methanol 1/1) 40/60% v/v; flow rate: 18 ml/min; and D, DAD detection: 220 nm; loop circuit: 750 mu l of the solution; and (3) injection: 15mg (per injection).
The first eluted enantiomer was further purified by preparative HPLC-method 4; the residue was treated with 1.25M HCl in MeOH and the volatiles removed in vacuo to give compound 49 (first eluting enantiomer, 27.8mg, 0.044 mmol). Chiral HPLC (method a 2): rt 8.2min, ee 96.8%;
1H NMR(400MHz，DMSO-d6)ppm 11.10(br.s.，1H)，10.29(br.s.，1H)，8.17-8.27(m，2H)，7.79-7.90(m，1H)，7.65-7.71(m，1H)，7.41-7.49(m，1H)，6.96-7.22(m，2H)，6.90-6.94(m，1H)，6.83-6.88(m，1H)，6.74-8.17(m，2H)，6.71(dt，1H)，5.91-5.98(m，1H)，4.62(br.s.，2H)，3.60-4.14(m，4H)，3.25-3.43(m，2H)，3.02-3.20(m，2H)，1.87(d，3H)。UPLC-MS：0.63min，599.2[M+H]method 13
Fractions containing the second eluting enantiomer were evaporated, 1.25M HCl in MeOH was added and volatiles were removed in vacuo to give compound 49b (second eluting enantiomer, 33mg, 0.052 mmol). Chiral HPLC (method a 2): rt is 9.5min, ee is 98.2%;
1H NMR(400MHz，DMSO-d6)ppm 11.28(br.s.，1H)，10.30(br.s.，1H)，8.20-8.26(m，2H)，7.79-7.90(m，1H)，7.65-7.71(m，1H)，7.42-7.50(m，1H)，6.96-7.22(m，2H)，6.90-6.94(m，1H)，6.83-6.88(m，1H)，6.74-8.17(m，2H)，6.71(dt，1H)，5.90-5.98(m，1H)，4.61(br.s.，2H)，3.41-4.10(m，4H)，3.24-3.42(m，2H)，3.01-3.20(m，2H)，1.87(d，3H)。UPLC-MS：0.65min，599.1[M+H]+, method 12.
Example 50
3- (1- (9H-purin-6-ylamino) ethyl) -4-cyclohexenyl-1H-isochromen-1-one
The title compound was prepared from 3- (1-bromoethyl) -4-cyclohexenyl-1H-isochromen-1-one (intermediate C24, 88mg, 0.264mmol) in analogy to example 21 to yield the title compound (4.7mg, 4.6%) as a white solid.
1H NMR(400MHz，DMSO-d6)ppm 12.71-13.16(bs，1H)，8.02-8.29(m，3H)，7.75-7.94(m，2H)，7.31-7.66(m，2H)，5.37-5.93(m，2H)，1.88-2.29(m，4H)，1.64-1.87(m，4H)，1.43-1.61(m，3H)。UPLC-MS：6.34min，388.2[M+H]+, method 7.
Example 51
3- (1- (4-amino-3- (2-aminopyrimidin-5-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one
Reacting 3- (1- (4-amino-3-iodo-1H-pyrazolo [3, 4-d)]Pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one (intermediate D2a, 60mg, 0.118mmol), K 2CO3(32.6mg, 0.236mmol), 2-aminopyrimidin-5-ylboronic acid (32.7mg, 0.236mmol) and Pd (dppf) Cl2(4.31mg, 5.89. mu. mol) was reacted in 2ml dioxane, purged with argon and heated at 120 ℃ overnight. The reaction was stopped by addition of 1M aqueous HCl (2ml) and purified directly by reverse phase chromatography using Biotage C1830g SNAP with a gradient of water and acetonitrileThe resulting mixture (2mL of 2M aqueous HCl was added before drying) to give the title compound (16.2mg, 28.9%) as a yellow solid.
1H NMR(400MHz，DMSO-d6)ppm 8.47(s，2H)，8.22(s，2H)，7.86-8.05(m，1H)，7.77(m，1H)，7.63(m，1H)，7.53(m，1H)，7.42(m，3H)，7.20-7.33(m，1H)，7.06-7.18(m，1H)，6.89(d，J＝7.94Hz，1H)，5.74(d，J＝7.06Hz，1H)，1.85(d，J＝7.06Hz，3H)。UPLC-MS：4.56min，477.1[M+H]+, method 7.
Example 52
3- (1- (4-amino-3- (pyrazin-2-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one
Reacting 3- (1- (4-amino-3-iodo-1H-pyrazolo [3, 4-d)]Pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one (intermediate D2a, 60mg, 0.118mmol), 2- (tributylstannyl) pyrazine (0.072mL, 0.236mmol), Pd (PPh)3)4(13.61mg, 0.012mmol) and LiCl (4.99mg, 0.118mmol) were reacted in dioxane (1.2mL), purged under argon atmosphere, and heated at 120 ℃ overnight. The reaction was stopped by addition of 1M aqueous HCl (2ml) and the solid was collected by filtration. The crude solid was purified directly by reverse phase chromatography using a Biotage C1830g SNAP gradient with water and acetonitrile (5 mL of 2M aqueous HCl was added before drying). The obtained white solid Et 2Trituration in O afforded (13.5mg, 24.8%) the title compound.
1H NMR(400MHz，DMSO-d6)ppm 9.27-9.50(m，1H)，9.00-9.13(m，1H)，8.75-8.83(m，1H)，8.66-8.75(m，1H)，8.40-8.61(m，1H)，8.18-8.30(m，1H)，7.97-8.15(m，2H)，7.73-7.85(m，1H)，7.35-7.69(m，5H)，7.17-7.31(m，1H)，6.78-6.97(m，1H)，5.66-5.84(m，1H)，1.73-2.00(m，3H)。UPLC-MS：6.04min，462.1[M+H]+, method 7.
Example 53
3- (1- (4-amino-3- (pyridazin-4-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one
The title compound was prepared from 3- (1- (4-amino-3-iodo-1H-pyrazolo [3,4-D ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one (intermediate D2a, 60mg, 0.118mmol) and 4- (tributylstannyl) pyridazine (87mg, 0.236mmol) in analogy to example 52 to give the title compound (34.5mg, 63.5%).
1H NMR(400MHz，DMSO-d6)ppm 9.46(dd，J＝2.21，1.32Hz，1H)，9.38(dd，J＝5.51，1.10Hz，1H)，8.15-8.29(m，2H)，7.91(dd，J＝5.29，2.65Hz，1H)，7.77(m，2H)，7.63(m，1H)，7.48-7.57(m，1H)，7.35-7.47(m，3H)，7.17(d，J＝7.50Hz，1H)，6.89(d，J＝7.94Hz，1H)，5.80(d，J＝7.06Hz，1H)，1.88(d，J＝7.06Hz，3H)。UPLC-MS：4.80min，462.1[M+H]+, method 7.
Example 54
3- (1- (4-amino-3- (pyridin-4-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one
The title compound was prepared from 3- (1- (4-amino-3-iodo-1H-pyrazolo [3,4-D ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one (intermediate D2a, 60mg, 0.118mmol), pyridin-4-ylboronic acid (29.0mg, 0.236mmol) in analogy to example 51 to give the title compound (10mg, 18.4%).
1H NMR(400MHz，DMSO-d6)ppm 8.89(d，J＝6.17Hz，2H)，8.21(s，2H)，7.98(d，J＝5.29Hz，2H)，7.70-7.82(m，1H)，7.58-7.66(m，1H)，7.48-7.58(m，1H)，7.34-7.47(m，4H)，7.16(d，J＝7.06Hz，1H)，6.89(d，J＝8.38Hz，1H)，5.70-5.91(m，1H)，1.87(d，J＝7.06Hz，3H)。UPLC-MS：5.34min，461.1[M+H]+, method 7.
Example 55
3- (1- (4-amino-3- (2-methoxypyrimidin-5-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one
The title compound was prepared from 3- (1- (4-amino-3-iodo-1H-pyrazolo [3,4-D ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one (intermediate D2a, 60mg, 0.118mmol), 2-methoxypyrimidin-5-ylboronic acid (36.3mg, 0.236mmol) in analogy to example 51 to give the title compound (36mg, 62.2%) as a light yellow solid.
1H NMR(400MHz，DMSO-d6)ppm 8.74(s，2H)，8.17-8.27(m，1H)，8.12(s，1H)，7.71-7.83(m，1H)，7.58-7.66(m，1H)，7.49-7.57(m，1H)，7.32-7.48(m，3H)，7.11-7.28(m，2H)，6.74-6.96(m，1H)，5.63-5.82(m，1H)，4.00(s，3H)，1.85(d，J＝7.06Hz，3H)。UPLC-MS：6.20min，492.2[M+H]+, method 7.
Example 56
3- (1- (4-amino-3- (2-hydroxypyrimidin-5-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one
Reacting 3- (1- (4-amino-3- (2-methoxy pyrimidine-5-yl) -1H-pyrazolo [3, 4-d)]Pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one (example 55, 23.3mg, 0.047mmol) is dissolved in HBr 33% CH3The COOH solution (1ml, 0.047mmol) was stirred at rt for 1 h. The reaction was then diluted with water (3mL) and the resulting mixture was purified by reverse phase chromatography using Biotage C1830g SNAP with a gradient of water and acetonitrile to give the title compound (20mg, 88%) as a white solid.
1H NMR(400MHz，DMSO-d6) ppm of 8.29-8.54(m, 2H), 8.17-8.26(m, 1H), 7.98-8.11(m, 1H), 7.72-7.81(m, 1H), 7.58-7.66(m, 1H), 7.49-7.57(m, 1H), 7.33-7.47(m, 3H), 7.11-7.23(m, 3H), 6.79-6.94(m, 1H), 5.27-5.79(m, 1H), 1.71-1.87(m, 3H). UPLC-MS: 4.67min (40%) and 4.71min (60%), 478.2[ M + H ]]+, method 7.
Example 57
3- (1- (4-amino-3- (5-methoxypyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one
The title compound was prepared in a similar manner to example 52 from 3- (1- (4-amino-3-iodo-1H-pyrazolo [3,4-D ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one (intermediate D2a, 60mg, 0.118mmol) and 3-methoxy-5- (tributylstannyl) pyridine (94mg, 0.236mmol) to give the title compound (31.3mg, 54.2%).
1H NMR(400MHz，DMSO-d6)ppm 8.32-8.47(m，2H)，8.18-8.25(m，1H)，8.05-8.16(m，1H)，7.72-7.82(m，1H)，7.32-7.67(m，6H)，7.15-7.23(m，1H)，6.96-7.11(m，1H)，6.81-6.93(m，1H)，5.59-5.83(m，1H)，3.95(s，3H)，1.76-1.92(m，3H)。UPLC-MS：6.29min，491.3[M+H]+, method 7.
Example 58
3- (1- (4-amino-3- (pyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one
The title compound was prepared in a similar manner to example 51 from 3- (1- (4-amino-3-iodo-1H-pyrazolo [3,4-D ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one (intermediate D2a, 60mg, 0.118mmol), pyridin-3-ylboronic acid (29.0mg, 0.236mmol) to give the title compound (42.7mg, 79%) as a pale yellow solid.
1H NMR(400MHz，DMSO-d6)ppm 8.94(s，1H)，8.83(d，J＝3.97Hz，1H)，8.17-8.36(m，3H)，7.78(m，3H)，7.63(t，1H)，7.50-7.57(m，1H)，7.35-7.48(m，3H)，7.29(s，2H)，7.04(m，1H)，6.90(d，J＝7.94Hz，1H)，5.80(d，J＝7.06Hz，1H)，1.78-1.95(m，3H)。UPLC-MS：6.06min，461.3[M+H]+, method 7.
Example 59
3- (1- (4-amino-3- (2-aminothiazol-5-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one hydrochloride
The title compound was prepared from 3- (1- (4-amino-3-iodo-1H-pyrazolo [3,4-D ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one (intermediate D2a, 47.7mg, 0.094mmol) and 4- (tributylstannyl) thiazol-2-ylcarbamate (92mg, 0.187mmol) in analogy to example 52 to give the title compound (24mg, 49.5%).
1H NMR(400MHz，DMSO-d6)ppm 8.78-9.18(m，1H)，8.13-8.27(m，2H)，7.71-8.04(m，2H)，7.59-7.69(m，1H)，7.50-7.58(m，2H)，7.36-7.49(m，3H)，7.15(d，J＝7.50Hz，1H)，6.90(d，J＝7.94Hz，1H)，5.70(d，J＝7.06Hz，1H)，1.82(d，J＝7.06Hz，3H)。UPLC-MS：5.84min，482.1[M+H]+, method 7.
Example 60
3- (1- (4-amino-3- (6-methoxypyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one
The title compound was prepared in a similar manner to example 52 from 3- (1- (4-amino-3-iodo-1H-pyrazolo [3,4-D ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one (intermediate D2a, 60mg, 0.118mmol) and 2-methoxy-5- (tributylstannyl) pyridine (94mg, 0.236mmol) to give the title compound (35.1mg, 60.7%).
1H NMR(400MHz，DMSO-d6)ppm 8.30-8.44(m，1H)，8.07-8.24(m，2H)，7.85-7.99(m，1H)，7.71-7.82(m，1H)，7.51-7.66(m，3H)，7.28-7.48(m，4H)，7.09-7.23(m，1H)，6.80-7.05(m，2H)，5.47-5.80(m，1H)，3.93(s，3H)，1.85(d，J＝7.50Hz，3H)。UPLC-MS：6.44min，491.2[M+H]+, method 7.
Example 61
3- (1- (4-amino-3- (6-hydroxypyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one
3- (1- (4-amino-3- (6-methoxypyridin-3-yl) -1H-pyrazolo [3, 4-d)]Pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one (example 60, 28mg, 0.057mmol) is dissolved in HBr 33% CH3The COOH solution (2ml, 36.8mmol) was stirred at 60 ℃ for 4 h. The reaction was then diluted with 3ml of water and the resulting mixture was purified by reverse phase chromatography using a Biotage C1830 g SNAP gradient with water and acetonitrile to give the title compound (6.9mg, 25.4%) as a pale yellow solid.
1H NMR(400MHz，DMSO-d6)ppm 8.11-8.30(m，2H)，7.77(t，J＝7.06Hz，2H)，7.32-7.68(m，8H)，7.14(d，J＝7.50Hz，1H)，6.89(d，J＝7.94Hz，1H)，6.48(d，J＝9.70Hz，1H)，5.72(d，J＝7.06Hz，1H)，1.84(d，J＝7.06Hz，3H)。UPLC-MS：5.18min，477.2[M+H]+, method 7.
Example 62
N- (5- (4-amino-1- (1- (1-oxo-4-phenyl-1H-isochromen-3-yl) ethyl) -1H-pyrazolo [3,4-d ] pyrimidin-3-yl) thiazol-2-yl) acetamide
Reacting 3- (1- (4-amino-3- (2-aminothiazole-5-yl) -1H-pyrazolo [3, 4-d)]Pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one hydrochloride (example 59, 23mg, 0.044mmol) was dissolved in TEA (124. mu.L, 0.888mmol)/Ac2O (419. mu.L, 4.44mmol), stir at rt for 1 h. The reaction mixture was quenched by addition of 1M HCl (1ml) by using a Biotage C1830 g SNAP gradient with water and acetonitrile Purification by reverse phase chromatography gave the title compound (2.3mg, 9.9%) as a whitish solid.
1H NMR(400MHz，DMSO-d6)ppm 8.16-8.20(d，1H，J＝8.0Hz)、8.04(s，1H)，7.73(d，2H，J＝8.0Hz)、7.73(d，1H，J＝8.0Hz)、7.54-7.58(m，2H)，7.33-7.44(m，5H)，6.83-6.89(m，1H)，5.65(q，1H，J＝8.0Hz)、2.14(s，3H)，1.79(d，3H，J＝8.0Hz)。UPLC-MS：3.81min，524.1[M+H]+, method 6.
Example 64
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1,2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one hydrochloride
Step 1.4- (3- (1- (4-amino-3-iodo-1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -1-oxo-1H-isochromen-4-yl) -5, 6-dihydropyridine-1 (2H) -carboxylate
Reacting 4- (3- (1- (4-amino-3-iodo-1H-pyrazolo [3, 4-d)]Pyrimidin-1-yl) ethyl) -1-oxo-1H-isochromen-4-yl) -5, 6-dihydropyridine-1 (2H) -carboxylic acid tert-butyl ester (intermediate D14, 120mg, 0.195mmol), 3-fluoro-5-hydroxyphenylboronic acid (67.0mg, 0.430mmol), Pd (dppf) Cl2(21.44mg, 0.029mmol) and K2CO3(59.4mg, 0.430mmol) was reacted in 3.2mL dioxane at 120 ℃ overnight under an argon atmosphere. The reaction mixture was diluted with AcOEt (100mL), washed with 0.5M aqueous HCl (100mL), washed with saturated aqueous NaCl, and Na2SO4Drying and evaporating to dryness. The crude product was purified by flash chromatography on silica gel using a Biotage 25G SNAP gradient with heptane and AcOEtThe resulting product was 4- (3- (1- (4-amino-3-iodo-1H-pyrazolo [3, 4-d)]Pyrimidin-1-yl) ethyl) -1-oxo-1H-isochromen-4-yl) -5, 6-dihydropyridine-1 (2H) -carboxylate (83mg, 71%) as a solid.
UPLC-MS: 1.19min, 599.0[ M + H ] +, method 9
And 2. step 2.
Reacting 4- (3- (1- (4-amino-3-iodo-1H-pyrazolo [3, 4-d)]Pyrimidin-1-yl) ethyl) -1-oxo-1H-isochromen-4-yl) -5, 6-dihydropyridine-1 (2H) -carboxylate (150mg, 0.244mmol) in dioxane (2.4ml) and 4M HCl in dioxane (2.4ml) was reacted at rt for 3H, then Et was added2O, a pale yellow precipitate was collected by filtration to give the title compound (135mg, 99%).
1H NMR(400MHz，DMSO-d6)ppm 10.13-10.36(bs，1H)，8.96-9.35(bs，2H)，8.38(s，1H)，8.29(s，1H)，8.08-8.21(m，1H)，7.83-7.96(m，1H)，7.66(m，2H)，6.95(m，2H)，6.62-6.75(m，1H)，6.18(m，1H)，6.01-6.09(m，1H)，3.66-3.83(m，2H)，2.96-3.38(m，2H)，2.56-2.74(m，2H)，1.79-2.00(d，3H)。UPLC-MS：2.17min，499.0[M+H]Method 6
Example 65
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1- (pyridin-4-ylmethyl) -1,2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one hydrochloride
Reacting 3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3, 4-d)]Pyrimidin-1-yl) ethyl) -4- (1,2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one hydrochloride (example 64, 37mg, 0.064mmol), nicotinaldehyde (8.89mg, 0.083mmol) and DIE (11.15. mu.l, 0.064mmol) were dissolved in DCM (1.23ml) and a spatula tip of anhydrous Na was added2SO4The mixture was stirred at rt for 10min, then AcOH (10.97. mu.l, 0.192mmol) and NaBH (OAc) were added3(27.1mg, 0.128 mmol). The resulting mixture was stirred at rt for 1h then quenched with 2M aqueous HCl (1ml), filtered to remove insoluble material and the purified filtrate was taken up by reverse phase chromatography using Biotage C1860 g SNAP with a gradient of water and acetonitrile to give (before drying, small amount of 1M aqueous HCl added) the title compound (10.6mg, 26.5%) as a white solid.
1H NMR(400MHz，DMSO-d6)ppm 11.56-11.67(m，br 1H)，10.20-10.34(m，br 1H)，8.78-8.82(m，2H)，8.10-8.30(m，2H)，7.74-8.00(m，3H)，7.48-7.68(m，2H)，6.82-6.97(m，2H)，6.67-7.73(m，1H)，6.15-7.73(m，1H)，6.15-6.35(m，1H)，5.97-6.10(m，1H)，4.50-4.68(m，2H)，3.90-3.97(m，3H)，3.35-3.50(m，2H)，3.04-3.27(m，1H)，2.52-2.68(m，1H)，1.81-1.99(m，3H)。UPLC-MS：2.34min，590.0[M+H]+, method 6.
Example 66
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1- (cyclopropylmethyl) -1,2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one hydrochloride
The title compound was prepared in analogy to example 65 from 3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1,2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one hydrochloride (example 64, 50mg, 0.086mmol) and cyclopropanecarboxaldehyde (7.26mg, 0.104mmol) to yield the title compound (28mg, 55.1% yield) as a white solid.
1H NMR(400MHz，DMSO-d6)ppm 10.09-10.51(m，2H br)、8.27-8.43(m，1H)，8.12-8.26(m，1H)7.80-7.95(m，1H)，7.61-7.70(m，1H)，6.82-6.98(m，2H)，6.64-6.73(m，1H)，6.04-6.30(m，2H)，3.85-4.17(m，3H)，3.71-3.82(m，1H)，3.23-3.33(m，1H)，3.08-3.22(m，2H)，2.71-2.96(m，2H)，2.52-2.65(m，1H)，1.82-2.04(m，3H)，1.17-1.27(m，1H)，0.67-0.76(m，2H)，0.44-0.53(m，2H)。UPLC-MS：2.56min，533.0[M+H]+, method 6.
Example 67
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (3- ((dimethylamino) methyl) phenyl) -1H-isochromen-1-one hydrochloride
3- (3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3, 4-d)]Pyrimidin-1-yl) ethyl) -1-oxo-1H-isochromen-4-yl) benzaldehyde (intermediate F2, 598mg, 1.147mmol), AcOH (131. mu.l, 2.293mmol) and 2M dimethylamine (1.15ml) in THF was dissolved in DCM (23ml) and a spoonful of anhydrous Na was added2SO4The mixture was stirred at rt for 15min, then NaB (OAc) was added3H (1215mg, 5.73 mmol). The resulting mixture was stirred at r t to complete conversion and then quenched with 1M aqueous HCl (5ml), filtered to remove insoluble material and the filtrate was purified by reverse phase chromatography using biotage c 1860 g SNAP with a gradient of water and acetonitrile to give (before drying, small amount of 1M aqueous HCl added) the title compound (460mg, 68.3% yield) as a white solid.
1H NMR(400MHz，DMSO-d6)d ppm 10.74(br.s.，1H)，10.59(br.s.，1H)，10.31(br.s.，2H)，8.33(s，1H)，8.19-8.29(m，3H)，7.48-7.85(m，11H)，7.41(t，J＝7.50Hz，1H)，7.04(d，J＝7.94Hz，1H)，6.78-6.97(m，6H)，6.66-6.77(m，2H)，5.85(q，J＝7.06Hz，1H)，5.60-5.75(m，1H)，4.07-4.44(m，4H)，2.78(t，J＝4.41Hz，6H)，2.72(d，J＝4.85Hz，3H)，2.65(d，J＝4.85Hz，3H)，1.77-1.94(m，6H)。UPLC-MS：2.52min，551.1[M+H]+, method 6.
Example 67a (enantiomer 1) and example 67b (enantiomer 2)3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (3- ((dimethylamino) methyl) phenyl) -1H-isochromen-1-one
The racemate 3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (3- ((dimethylamino) methyl) phenyl) -1H-isochromen-1-one hydrochloride (example 67, 0.558g, 0.95mmol) was dissolved in 10ml of ethanol and subjected to chiral resolution by chiral preparative chromatography. Conditions are as follows: column: chiralpak AD-H (25x3cm), 5 um; mobile phase: n-hexane/(ethanol + 0.1% isopropylamine) 75/25% v/v; flow rate: 32 ml/min; and D, DAD detection: 220 nm; loop circuit: 540 μ l; and (3) injection: 30mg (per injection).
Fractions containing the enantiomer of the first eluate were evaporated to dryness to give compound 67a (0.214g, 0.39 mmol). Chiral HPLC (method a 12): rt 7.1min, ee > 99%.
1H NMR(500MHz，DMSO-d6)ppm 10.19(s，1H)，8.18-8.29(m，1H)，8.03-8.12(m，1H)，7.72-7.83(m，1H)，7.57-7.68(m，1H)，7.22-7.52(m，4H)，6.75-7.01(m，5H)，6.66(d，1H)，5.64-5.80(m，1H)，3.06-3.57(m，2H)，1.93-2.28(m，6H)，1.81(d，3H)。UPLC-MS：0.63-.065min，551.4[M+H]+, method 13.
Fractions containing the enantiomer of the second eluent were evaporated to dryness to give compound 67b (second eluted enantiomer, 0.200g, 0.37 mmol). Chiral HPLC (method a 12): rt 11.0min, ee 98.4%.
1H NMR(500MHz，DMSO-d6)ppm 10.19(s，1H)，8.18-8.29(m，1H)，8.03-8.12(m，1H)，7.72-7.83(m，1H)，7.57-7.68(m，1H)，7.22-7.52(m，4H)，6.75-7.01(m，5H)，6.66(d，1H)，5.64-5.80(m，1H)，3.06-3.57(m，2H)，1.93-2.28(m，6H)，1.81(d，3H)。UPLC-MS：0.62-0.65min，551.4[M+H]+, method 13.
Example 68
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (5- (piperidin-1-ylmethyl) thiophen-2-yl) -1H-isochromen-1-one hydrochloride
Reacting 3- (1- (4-amino-3-iodo-1H-pyrazolo [3, 4-d)]Pyrimidin-1-yl) ethyl) -4- (5- (piperidin-1-ylmethyl) thiophen-2-yl) -1H-isochromen-1-one hydrochloride (intermediate D9, 100mg, 0.154mmol), 3-fluoro-5-hydroxyphenylboronic acid (48.1mg, 0.308mmol), S-Phos-Pd-G2(11.10mg, 0.015mmol) and K3PO4(151mg, 0.462mmol) in THF (1.2ml) and water (0.3ml) was reacted under an argon atmosphere at 80 ℃ under mw irradiation for 30 min. The reaction was stopped by addition of 1M aqueous HCl (2ml) and the mixture was purified by reverse phase chromatography using a Biotage C1860 g SNAP gradient with water and acetonitrile to give (before drying, a small amount of 1M aqueous HCl was added) the title compound (70mg, 71.7% yield) as a whitish solid.
1H NMR(400MHz，DMSO-d6)ppm 10.11-10.46(bs，2H)，8.15-8.31(m，2H)，7.77-8.03(m，1H)，7.67(t，J＝7.50Hz，1H)，7.43(br.s.，1H)，7.18(d，J＝7.94Hz，2H)，6.92(s，1H)，6.85(d，J＝8.82Hz，1H)，6.70(d，J＝11.03Hz，1H)，5.94(d，J＝7.06Hz，1H)，4.53(d，J＝3.53Hz，2H)，3.38(m，H)，2.87(d，J＝11.47Hz，2H)，1.61-1.99(m，8H)，1.37(d，J＝11.91Hz，1H)。UPLC-MS：2.82min，597.0[M+H]+, method 6.
Example 68a (enantiomer 1) and example 68b (enantiomer 2): 3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (5- (piperidin-1-ylmethyl) thiophen-2-yl) -1H-isochromen-1-one hydrochloride as a single enantiomer
The racemate 3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (5- (piperidin-1-ylmethyl) thiophen-2-yl) -1H-isochromen-1-one hydrochloride (example 68, 0.053g, 0.0837mmol) was dissolved in 3.5ml of ethanol and subjected to chiral resolution by chiral preparative chromatography. Conditions are as follows: column: whelk O-1(R, R) (25 x 2cm), 10 um; mobile phase: n-hexane/(ethanol + 0.1% isopropylamine) 55/45% v/v; flow rate: 14 ml/min; and D, DAD detection: 220 nm; loop circuit: 500 mul; and (3) injection: 7.5mg (per injection).
The fractions containing the enantiomer of the first eluent were evaporated to dryness, 1M HCl was added and the volatiles were removed under reduced pressure. The residue was purified by reverse phase chromatography using a C18 column (H)2O:CH3CN 95:5-50:50, containing 0.1% HCOOH); before drying, 2ml of 1N HCl was added and the volatiles removed under reduced pressure to give compound 68a as a white solid (first eluting enantiomer, 0.0149g, 0.0235 mmol). Chiral HPLC (method a 14): rt 8.4min, ee 99%.
1H NMR(400MHz，DMSO-d6)ppm 10.21-10.48(m，2H)，8.20-8.29(m，2H)，7.82-7.90(m，1H)，7.65-7.72(m，1H)，7.41-7.48(m，1H)，7.00-7.33(m，2H)，6.90-6.95(m，1H)，6.83-6.89(m，1H)，6.71(dt，1H)，6.65-8.50(m，2H)，5.91-5.99(m，1H)，4.49-4.61(m，2H)，3.26-3.88(m，2H)，2.80-2.96(m，2H)，1.66-1.93(m，8H)，1.30-1.47(m，1H)。UPLC-MS：0.68min，597.5[M+H]+, method 13.
The fractions containing the enantiomer of the second eluent were evaporated to dryness, 1M HCl was added and the volatiles were removed under reduced pressure. The residue was purified by reverse phase chromatography using a C18 column (H) 2O:CH3CN 95:5-50:50, containing 0.1% HCOOH); before drying, 2ml of 1N HCl was added and the volatiles were removed under reduced pressure to give compound 68b asWhite solid (second eluting enantiomer, 0.013g, 0.02 mmol). Chiral HPLC (method a 14): rt 10.1min, ee 97.8%.
1H NMR(400MHz，DMSO-d6)ppm 10.21-10.62(m，2H)，8.20-8.30(m，2H)，7.82-7.90(m，1H)，7.65-7.72(m，1H)，7.40-7.50(m，1H)，7.00-7.33(m，2H)，6.90-6.95(m，1H)，6.83-6.89(m，1H)，6.71(dt，1H)，6.65-8.50(m，2H)，5.91-5.99(m，1H)，4.48-4.61(m，2H)，3.24-4.00(m，2H)，2.80-2.96(m，2H)，1.66-1.94(m，8H)，1.30-1.47(m，1H)。UPLC-MS：0.67min，597.5[M+H]+, method 13.
Examples 69-71, 85-86, 93-102, 113-114, 121, 128-129, 131-132, 146-149, 152-153, 159-160 found in the following table can be prepared in a similar manner to compound 68 starting with the appropriate reagents reported below.
The process used for examples 99, 101, 159 and 160 required a further deprotection step consisting of reaction in dry dichloromethane (5ml) with a molar excess of 1M boron tribromide in DCM at room temperature, followed by quenching with EtOH at 0 ℃ and finally a suitable chromatographic purification step.
Example 72
4- (1-acetyl-1, 2,3, 6-tetrahydropyridin-4-yl) -3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -1H-isochromen-1-one
The title compound was prepared in analogy to example 51 from 4- (1-acetyl-1, 2,3, 6-tetrahydropyridin-4-yl) -3- (1- (4-amino-3-iodo-1H-pyrazolo [3,4-D ] pyrimidin-1-yl) ethyl) -1H-isochromen-1-one (intermediate D12, 1.08g, 1.941mmol), (3-fluoro-5-hydroxyphenyl) boronic acid (0.605g, 3.88mmol) to yield the title compound (456mg, 43.5%) as a light gray solid.
1H NMR(400MHz，DMSO-d6)ppm 10.02-10.36(s，1H)，8.12-8.33(m，2H)，7.81-7.99(m，1H)，7.57-7.70(m，1H)，7.37-7.50(m，1H)，6.80-6.96(m，2H)，6.54-6.71(m，1H)，6.18-6.31(m，1H)，5.97-6.12(m，1H)，5.16-5.33(m，1H)，4.17-4.30(m，1H)，4.04-4.15(m，1H)，3.86-4.04(m，1H)，3.48-3.82(m，2H)，3.20(s，3H)，2.01-2.05(d，3H)，1.75-1.95(m，2H)。UPLC-MS：3.13min，540.9[M+H]+, method 6.
Example 73
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (5- (pyrrolidin-1-ylmethyl) thiophen-2-yl) -1H-isochromen-1-one hydrochloride
The title compound was prepared in analogy to example 67 from 5- (3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -1-oxo-1H-isochromen-4-yl) thiophene-2-carbaldehyde (intermediate F4, 57mg, 0.108mmol) and pyrrolidine (18.04 μ l, 0.216mmol) to yield (before drying, small amounts of 1m hci in water) the title compound (37.8mg, 56.5%) as a pale solid.
1H NMR(400MHz，DMSO-d6)ppm 10.49(br.s.，1H)，10.25(br.s.，1H)，8.10-8.43(m，2H)，7.79-7.90(m，1H)，7.61-7.75(m，1H)，7.41(d，J＝3.09Hz，1H)，7.15(d，J＝7.94Hz，2H)，6.91(s，1H)，6.80-6.88(m，1H)，6.69(dt，J＝11.03，2.21Hz，1H)，5.92(d，J＝7.06Hz，1H)，4.61(br.s.，2H)，3.44(m，2H)，3.10(br.s.，2H)，2.04(m，2H)，1.70-1.95(m，7H)。UPLC-MS：2.62min，582.9[M+H]+, method 6.
Example 74
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (5- ((bis (2-hydroxyethyl) amino) methyl) thiophen-2-yl) -1H-isochromen-1-one, formate salt
The title compound was prepared in a similar manner to example 67 from 5- (3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -1-oxo-1H-isochromen-4-yl) thiophene-2-carbaldehyde (intermediate F4, 60mg, 0.114mmol) and 2,2' -azanediyl diethanol (23.92mg, 0.227mmol) to give the title compound (3mg, 3.98%).
1H NMR(400MHz，DMSO-d6)ppm 10.22(s，1H)，9.68-10.01(m，1H)，8.22(d，J＝7.94Hz，1H)，8.14(s，1H)，7.77-7.92(m，1H)，7.66(t，J＝7.50Hz，1H)，7.46(br.s.，1H)，7.16(d，J＝7.94Hz，2H)，6.91(s，1H)，6.83(d，J＝8.82Hz，1H)，6.67(d，J＝11.03Hz，1H)，5.93(d，J＝7.06Hz，1H)，5.38(br.s.，2H)，4.70(br.s.，2H)，3.82(br.s.，4H)，3.32(s，2H)，1.85(d，J＝7.06Hz，3H)。UPLC-MS：2.40min，616.9[M+H]+, method 6.
Example 75
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (5- (hydroxymethyl) thiophen-2-yl) -1H-isochromen-1-one
The title compound was isolated during the experiment used to produce example 74, and was recovered, particularly during the final rapid purification step, as the second eluting compound (8.5mg, 14.1%).
1H NMR(400MHz，DMSO-d6)ppm 10.19(s，1H)，8.14(m，2H)，7.76-7.94(m，1H)，7.47-7.71(m，1H)，7.12-7.26(m，1H)，7.00(m，2H)，6.89-6.93(m，1H)，6.79-6.87(m，1H)，6.56-6.76(m，1H)，5.76-6.09(m，1H)，5.34-5.65(m，1H)，4.52-4.78(m，2H)，1.88(d，J＝7.06Hz，3H)。UPLC-MS：3.43min，529.9[M+H]+, method 6.
Example 76
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (5- ((4- (2-hydroxyethyl) piperazin-1-yl) methyl) thiophen-2-yl) -1H-isochromen-1-one hydrochloride
The title compound was prepared in analogy to example 67 from 5- (3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -1-oxo-1H-isochromen-4-yl) thiophene-2-carbaldehyde (intermediate F4, 60mg, 0.114mmol) and 2- (piperazin-1-yl) ethanol (29.6mg, 0.227mmol) to give the desired product (48.6mg, 63.0%) as a whitish solid.
1H NMR(400MHz，DMSO-d6)ppm 9.85-10.43(m，1H)，8.09-8.32(m，2H)，7.78-7.91(m，1H)，7.66(t，J＝7.50Hz，1H)，7.20(d，J＝7.94Hz，2H)，6.96-7.11(m，1H)，6.81-6.96(m，2H)，6.61-6.76(m，1H)，5.85-6.04(m，1H)，3.45-3.83(m，10H)，3.21(br.s.，4H)，1.87(d，J＝7.06Hz，3H)。UPLC-MS：2.66min，642.0[M+H]+, method 6.
Example 76a (enantiomer 1) and example 76b (enantiomer 2)3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (5- ((4- (2-hydroxyethyl) piperazin-1-yl) methyl) thiophen-2-yl) -1H-isochromen-1-one hydrochloride
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (5- ((4- (2-hydroxyethyl) piperazin-1-yl) methyl) thiophen-2-yl) -1H-isochromen-1-one hydrochloride (example 76, 0.040g) was dissolved in ethanol/methanol 1/1(4ml) and chiral resolution was performed by chiral preparative chromatography. Conditions are as follows: column: chiralpak AD-H (25 x 2.0cm), 5 um; mobile phase: n-hexane/(ethanol + 0.1% isopropylamine) 80/20% v/v; flow rate: 17 ml/min; and D, DAD detection: 220 nm; loop circuit: 300 mu l; and (3) injection: 3mg (per injection).
The fraction containing the enantiomer of the first eluate was evaporated to dryness and purified by reverse phase flash chromatography using a C18 column (H)2O+0.1％HCOOH:CH3CN + 0.1% HCOOH ═ 95:5 to 50: 50); the residue was treated with 1.25M HCl in MeOH and evaporated to dryness to give example 76a (first eluting enantiomer, 12mg, 0.0168 mmol). Chiral HPLC (method a 16): rt 12.1min, ee>99％。
1H NMR(500MHz，DMSO-d6)ppm 10.27(br.s.，1H)，8.25(s，1H)，8.21(d，1H)，7.81-7.86(m，1H)，7.66(t，1H)，6.94-7.52(m，5H)，6.91(s，1H)，6.85(d，1H)，6.67-6.74(m，1H)，5.96(q，1H)，4.75-5.50(m，1H)，3.56-4.22(m，10H)，3.06-3.37(m，4H)，1.87(d，3H)。UPLC-MS：0.65min，642.2[M+H]+, method 13.
The fraction containing the enantiomer of the second eluent was evaporated to dryness and purified by reverse phase flash chromatography using a C18 column (H)2O+0.1％HCOOH:CH3CN + 0.1% HCOOH ═ 95:5 to 50: 50); the residue was treated with 1.25M HCl in MeOH and evaporated to dryness to give example 76b (second eluting enantiomer, 0.014g, 0.0196 mmol). Chiral HPLC (method a 16): rt 14.6min, ee >99％。
1H NMR(400MHz，DMSO-d6)ppm 10.26(br.s.，1H)，8.19-8.26(m，2H)，7.81-7.86(m，1H)，7.63-7.69(m，1H)，6.94-7.52(m，5H)，6.89-6.93(m，1H)，6.82-6.88(m，1H)，6.67-6.73(m，1H)，5.96(q，1H)，4.75-5.55(m，1H)，3.48-4.30(m，10H)，3.07-3.36(m，4H)，1.87(d，3H)。UPLC-MS：0.65min，642.2[M+H]+, method 13.
Example 77
4- ((5- (3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -1-oxo-1H-isochromen-4-yl) thiophen-2-yl) methyl) piperazin-2-one hydrochloride
The title compound was prepared in analogy to example 67 from 5- (3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -1-oxo-1H-isochromen-4-yl) thiophene-2-carbaldehyde (intermediate F4, 60mg, 0.114mmol) and piperazin-2-one (22.77mg, 0.227mmol) to give the desired product (29.8mg, 40.4%) as a light yellow solid.
1H NMR(400MHz，DMSO-d6)ppm 10.27(br.s.，1H)，8.15-8.59(m，3H)，7.57-7.98(m，2H)，7.41(br.s.，1H)，6.79-7.30(m，4H)，6.54-6.75(m，1H)，5.96(q，J＝7.06Hz，1H)，4.59(br.s.，2H)，3.46(br.s.，6H)，1.87(d，J＝7.06Hz，3H)。UPLC-MS：3.11min，612.0[M+H]+, method 6.
Example 78
5- (3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -1-oxo-1H-isochromen-4-yl) thiophene-2-carboxylic acid
Reacting 5- (3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3, 4-d)]Pyrimidin-1-yl) ethyl) -1-oxo-1H-isochromen-4-yl) thiophene-2-carbaldehyde (intermediate F4, 19mg, 0.036mmol) was dissolved in 2-methyl-2-butene (300. mu.l, 2.83mmol) and t-butanol (0.5ml) and then NaClO was added2(32.6mg, 0.360mmol) and K2H2PO4(49.0mg, 0.360mmol) in 0.5mL of water and the mixture stirred at rt for 2 h. The reaction was stopped by addition of 2M aqueous HCl (1ml) and the crude product was purified by reverse phase chromatography using Biotage C1830 g SNAP with a gradient of water and acetonitrile to give the title compound (8mg, 40.9%) as a white solid.
1H NMR(400MHz，DMSO-d6)ppm 12.87-13.46(bs，1H)，9.96-10.54(bs，1H)，8.17-8.27(m，1H)，8.12(s，1H)，7.77-7.87(m，1H)，7.59-7.72(m，2H)，7.13(m，4H)，6.91(s，1H)，6.79-6.86(m，1H)，6.59-6.71(m，1H)，5.75-6.06(m，1H)，1.87(d，J＝7.06Hz，3H)。UPLC-MS：3.52min，543.9[M+H]+, method 6.
Example 79
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-benzyl-1H-isochromen-1-one
The title compound was prepared from 3- (1- (4-amino-3-iodo-1H-pyrazolo [3,4-D ] pyrimidin-1-yl) ethyl) -4-benzyl-1H-isochromen-1-one (intermediate D15, 45mg, 0.086mmol), (3-fluoro-5-hydroxyphenyl) boronic acid (26.8mg, 0.172mmol) in analogy to example 51 to yield the title compound (13mg, 19.8%) as a light brown solid.
1H NMR(400MHz，DMSO-d6)ppm 10.06(s，1H)，8.14-8.27(m，2H)，7.70-7.81(m，1H)，7.38-7.63(m，2H)，7.02-7.12(m，3H)，6.89-6.98(m，2H)，6.84-6.89(m，1H)，6.75-6.82(m，1H)，6.59-6.69(m，1H)，6.21-6.45(m，1H)，4.12-4.43(m，2H)，1.68-1.96(d，3H)。UPLC-MS：4.22min，507.9[M+H]+, method 6.
Example 80
4- (1H-pyrazol-4-yl) -3- (1- (thieno [3,2-d ] pyrimidin-4-ylamino) ethyl) -1H-isochromen-1-one hydrochloride
3- (1-aminoethyl) -4- (1H-pyrazol-4-yl) -1H-isochromen-1-one dihydrochloride (intermediate E4, 62mg, 0.189mmol), 4-chlorothieno [3,2-d ] pyrimidine (45.1mg, 0.264mmol), TEA (0.092ml, 0.661mmol) in t-BuOH (1.1ml) was reacted at 85 ℃ for 2H and at 90 ℃ for 4H. The reaction was stopped by addition of 1M aqueous HCl (2ml) and purified by reverse phase chromatography using a Biotage C1860 g SNAP gradient with water and acetonitrile to give the title compound (24mg, 29.8%) as a pale yellow solid.
1H NMR(400MHz，DMSO-d6)ppm 10.08(br.s.，1H)，8.81(s，1H)，8.46(d，J＝5.29Hz，1H)，8.19(d，J＝7.50Hz，1H)，7.82(m，3H)，7.62(t，J＝7.50Hz，1H)，7.48(m，1H)，7.22(d，J＝7.94Hz，1H)，5.32(t，J＝6.84Hz，1H)，1.60(d，3H)。UPLC-MS：0.63min，390.0[M+H]Method 9
Example 81
4- (5- (morpholinomethyl) thiophen-2-yl) -3- (1- (thieno [3,2-d ] pyrimidin-4-ylamino) ethyl) -1H-isochromen-1-one hydrochloride
The title compound was prepared from 3- (1-aminoethyl) -4- (5- (morpholinomethyl) thiophen-2-yl) -1H-isochromen-1-one dihydrochloride (intermediate E3, 80mg, 0.180mmol) and 4-chlorothieno [3,2-d ] pyrimidine (43.1mg, 0.253mmol) in analogy to example 80 to yield the title compound (52mg, 53.3%) as a light yellow solid.
1H NMR(400MHz，DMSO-d6)ppm 11.12-11.64(bs，1H)，9.50-9.92(bs，1H)，8.79(br.s.，1H)，8.38(d，J＝4.85Hz，1H)，8.20(d，J＝7.94Hz，1H)，7.84(m，1H)，7.57-7.72(m，1H)，7.39-7.56(m，2H)，7.33(m 1H)，7.22(d，J＝7.94Hz，1H)，5.27(t，1H)，4.65(br.s.，2H)，3.68-4.13(m，4H)，3.15(m，4H)，1.62(d，J＝7.06Hz，3H)。UPLC-MS：6.38min，504.9[M+H]+, method 8.
Example 82
4-amino-6- ((1- (4- (5- (morpholinomethyl) thiophen-2-yl) -1-oxo-1H-isochromen-3-yl) ethyl) amino) pyrimidine-5-carbonitrile hydrochloride
The title compound was prepared in analogy to example 80 from 3- (1-aminoethyl) -4- (5- (morpholinomethyl) thiophen-2-yl) -1H-isochromen-1-one dihydrochloride (intermediate E3, 60mg, 0.135mmol) and 4-amino-6-chloropyrimidine-5-carbonitrile (29,3mg, 0,189mmol) to give the title compound (4mg, 5.6%) as a whitish solid.
1H NMR(400MHz，DMSO-d6)ppm 11.24(br.s.，1H)，8.22(d，J＝7.94Hz，1H)，8.14(br.s.，1H)，8.08(s，1H)，7.83(t，J＝7.72Hz，1H)，7.64(t，J＝7.50Hz，3H)，7.49(d，J＝3.09Hz，1H)，7.24(d，J＝3.53Hz，1H)，7.17(d，J＝8.38Hz，1H)，5.11(t，J＝6.84Hz，1H)，4.63(br.s.，2H)，4.00(d，J＝11.91Hz，2H)，3.78(d，J＝7.50Hz，2H)，3.32(d，J＝11.91Hz，2H)，3.12(br.s.，2H)，1.49(d，J＝7.06Hz，3H)。UPLC-MS：1.87min，488.9[M+H]+，
Method 6
Example 83
4-phenyl-3- (1- (pyrrolo [2,1-f ] [1,2,4] triazin-4-ylamino) ethyl) -1H-isochromen-1-one
The title compound was prepared from 3- (1-aminoethyl) -4-phenyl-1H-isochromen-1-one hydrochloride (intermediate E2, 50mg, 0.166mmol) and 4-chloropyrrolo [2,1-f ] [1,2,4] triazine (35.6mg, 0.232mmol) in analogy to example 80 to give the title compound (50.3mg, 79%) as a whitish solid.
1H NMR(400MHz，DMSO-d6)ppm 8.53(d，J＝6.62Hz，1H)，8.20(d，J＝7.94Hz，1H)，7.72-7.87(m，2H)，7.45-7.66(m，6H)，7.39(d，J＝7.06Hz，1H)，7.04(d，J＝3.97Hz，1H)，6.93(d，J＝8.38Hz，1H)，6.47-6.69(m，1H)，5.01(t，J＝7.06Hz，1H)，1.52(d，J＝7.06Hz，3H)。UPLC-MS：5.15min，383.0[M+H]+, method 6.
Example 84
4-phenyl-3- (1- (pyrido [3,2-d ] pyrimidin-4-ylamino) ethyl) -1H-isochromen-1-one
The title compound was prepared from 3- (1-aminoethyl) -4-phenyl-1H-isochromen-1-one hydrochloride (intermediate E2, 50mg, 0.166mmol) and 4-chloropyrido [3,2-d ] pyrimidine (38.4mg, 0.232mmol) in analogy to example 80 to give the desired product (2.2mg, 3.4%).
1H NMR(400MHz，DMSO-d6)ppm 8.81-9.08(m，2H)，8.54(s，1H)，8.10-8.24(m，2H)，7.91(dd，J＝8.38，3.97Hz，1H)，7.71-7.80(m，1H)，7.45-7.64(m，5H)，7.41(d，J＝7.50Hz，1H)，6.93(d，J＝7.94Hz，1H)，5.11(t，J＝7.06Hz，1H)，1.60(d，J＝7.06Hz，3H)。UPLC-MS：4.13min，395.0[M+H]+, method 6.
Example 87
N- (5- (4-amino-1- (1- (1-oxo-4-phenyl-1H-isochromen-3-yl) ethyl) -1H-pyrazolo [3,4-d ] pyrimidin-3-yl) pyridin-3-yl) -4-fluorobenzenesulfonamide
The title compound was prepared in analogy to example 52 from 4-fluoro-N- (5- (trimethylstannyl) pyridin-3-yl) benzenesulfonamide (intermediate G9, 98mg, 0.236mmol) and 3- (1- (4-amino-3-iodo-1H-pyrazolo [3,4-D ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one (intermediate D2a, 60mg, 0.118mmol) to yield the title compound (13mg, 0.021mmol, 17.4% yield) as a white solid.
1H NMR(400MHz，DMSO-d6)ppm 10.94(br.s.，1H)，8.50(d，J＝1.76Hz，1H)，8.43(d，J＝2.21Hz，1H)，8.17-8.27(m，2H)，7.81-7.96(m，3H)，7.73-7.80(m，3H)，7.63(m，1H)，7.48-7.58(m，1H)，7.33-7.47(m，5H)，7.12(d，J＝7.06Hz，1H)，6.89(d，J＝8.38Hz，1H)，5.77(q，J＝7.06Hz，1H)，1.85(d，J＝7.06Hz，4H)。UPLC-MS：1.13min，634[M+H]+, method 9.
Example 88
3- (1- (4-amino-3- (5-aminopyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one hydrochloride
The title compound was prepared in analogy to example 52 from 3- (1- (4-amino-3-iodo-1H-pyrazolo [3,4-D ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one (intermediate D2a, 80mg, 0.157mmol) and tert-butyl 5- (trimethylstannyl) pyridin-3-ylcarbamate (intermediate G10, 112mg, 0.314 mmol) to yield the title compound (14mg, 0.029mmol, 18.74% yield) as a white powder.
1H NMR(400MHz，DMSO-d6)ppm 8.01-8.28(m，4H)，7.76(d，J＝14.11Hz，2H)，7.63(m，1H)，7.53(m，1H)，7.32-7.47(m，3H)，7.14(m，1H)，6.90(d，J＝7.94Hz，1H)，6.47-6.80(m，1H)，5.78(d，J＝7.06Hz，1H)，1.85(d，J＝7.06Hz，3H)。UPLC-MS：0.80min，476[M+H]+, method 9.
Example 89
3- (1- (4-amino-3- (2-aminopyrimidin-4-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one hydrochloride
To a solution of triphenylphosphine (16.48mg, 0.063mmol) in dry dioxane (6mL) was added 3- (1- (4-amino-3-iodo-1H-pyrazolo [3,4-D ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one (intermediate D2a, 80mg, 0.157mmol), copper (I) iodide (5.98mg, 0.031mmol), 4-trimethylstannyl-2-amino-bis (tert-butylcarbamate) -pyrimidine (intermediate G11, 144mg, 0.314mmol), bis (dibenzylideneacetone) palladium (0) (18.06mg, 0.031mmol), and lithium chloride (19.98mg, 0.471 mmol). Nitrogen was bubbled through the solution for 10min, the solution was heated to 80 ℃ and stirred overnight. The solvent was removed under reduced pressure and the product was purified by Biotage Si 10g eluting with a gradient of DCM and ethanol to give a yellow solid. This compound was dissolved in 4M HCl in 1,4 dioxane. Stir at rt for 8 h. The solvent was removed and the product was purified directly by reverse phase chromatography using a Biotage C1810 g column (phase a, water 95%, ACN 5%, formic acid 0.1%); (phase B, ACN 95%, water 5%, formic acid 0.1%) to give the title compound (6.6mg, 7.5%) as a white powder.
1H NMR(400MHz，DMSO-d6)ppm 8.51-8.71(bs，1H)，8.39-8.49(m，1H)，8.33(s，1H)，8.22(d，J＝7.50Hz，1H)，7.72-7.87(m，1H)，7.59-7.70(m，2H)，7.51-7.58(m，1H)，7.31-7.50(m，4H)，7.19(d，J＝7.06Hz，1H)，6.91(d，J＝7.94Hz，1H)，5.68-5.85(m，1H)，4.27-4.39(m，1H)，1.89(d，J＝7.06Hz，3H)。UPLC-MS：3.91min，477[M+H]+, method 6.
Example 90
3- (1- (4-amino-3- (6-hydroxypyrazin-2-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one
The title compound was prepared in analogy to example 89 from 3- (1- (4-amino-3-iodo-1H-pyrazolo [3,4-D ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one (intermediate D2a, 120mg, 0.236mmol) and 2-methoxy-6- (tributylstannyl) pyrazine (188mg, 0.471mmol) to give the title compound (25mg, 36.8%) as a white powder.
1H NMR(400MHz，DMSO-d6)ppm 8.85(s，1H)，8.11-8.23(m，3H)，7.86-7.96(m，1H)，7.68-7.80(m，1H)，7.56-7.63(m，1H)，7.48-7.55(m，1H)，7.33-7.46(m，3H)，7.12-7.24(m，1H)，6.80-6.91(m，1H)，5.65-5.80(m，1H)，1.78-1.97(m，3H)。UPLC-MS：4.08min，478[M+H]+, method 6.
Example 91
3- (1- (4-amino-3- (5-hydroxypyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one
The title compound was prepared in analogy to example 89 from 3- (1- (4-amino-3-iodo-1H-pyrazolo [3,4-D ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one (intermediate D2a, 804mg, 1.579mmol) and 3-methoxy-5- (tributylstannyl) pyridine (1257mg, 3.16mmol) at 120 ℃ overnight. The solvent was removed and the product was purified by Biotage Si 25g using a gradient of DCM and EtOH to give a pale yellow solid (500 mg).
This material was dissolved in dry DCM (10mL) and 1M BBr was added3Was added to the solution of (1) (24ml), and the solution was stirred at room temperature for 48 h. The solution was then cooled to 0 ℃ and dry ethanol (10ml) was added. The solvent was evaporated and the product was purified directly by reverse phase chromatography using a Biotage C18SNAP column Phase a, water 95%, ACN 5%, formic acid 0.1%; (phase B, ACN 95%, water 5%, formic acid 0.1%) to give the title compound as a white solid. The product was then dissolved in a mixture of DCM (100ml) and EtOH (5ml) and saturated NaHCO was used3The aqueous solution washes the solution. The aqueous phase was extracted with DCM (5 × 50 ml). The organic phase was collected, dried and the solvent evaporated to dryness to give the desired product to give the title compound (331mg, 0.695mmol, 68.4% yield) as a white solid.
1H NMR(400MHz，DMSO-d6)ppm 8.21(d，J＝7.94Hz，1H)，8.07(s，1H)，7.75(t，J＝7.50Hz，2H)，7.61(t，J＝7.50Hz，2H)，7.51(d，J＝7.50Hz，1H)，7.42(m，3H)，7.12(d，J＝7.50Hz，1H)，6.88(d，J＝7.94Hz，1H)，6.67-6.83(bs，1H)，5.69(d，J＝7.06Hz，1H)，1.83(d，J＝7.06Hz，3H)。UPLC-MS：3.40min，477[M+H]Method 6
Example 92
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (8-methyl-8-azabicyclo [3.2.1] oct-2-en-3-yl) -1H-isochromen-1-one hydrochloride
Step a.3- (3- (1- (4-amino-3- (3-fluoro-5-methoxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -1-oxo-1H-isochromen-4-yl) -8-azabicyclo [3.2.1] oct-2-ene-8-carboxylic acid benzyl ester
The title compound was prepared from 3- (3-fluoro-5-methoxyphenyl) -1H-pyrazolo [3,4-D ] pyrimidin-4-amine (intermediate G1, 52.4mg, 0.202mmol) and benzyl 3- (3- (1-bromoethyl) -1-oxo-1H-isochromen-4-yl) -8-azabicyclo [3.2.1] oct-2-ene-8-carboxylate (intermediate C40, 100mg, 0.202mmol) according to the procedure described for the synthesis of intermediate D1 to give 3- (3- (1- (4-amino-3- (3-fluoro-5-methoxyphenyl) -1H-pyrazolo [3,4-D ] pyrimidin-1-yl) ethyl) -1-oxo-1H-isochromen- 4-yl) -8-azabicyclo [3.2.1] oct-2-ene-8-carboxylic acid benzyl ester (125mg, 0.186mmol, 92% yield) as a yellow solid.
UPLC-MS: 1.28min, 673[ M + H ] +, method 9
And b, step b.
To a solution of 3- (1- (4-amino-3- (3-fluoro-5-methoxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (8-azabicyclo [3.2.1] oct-2-en-3-yl) -1H-isochromen-1-one (80mg, 0.149mmol) in IPrOH (10ml) and aqueous N HCl (1ml) was added Pd/C5% wet (10mg, 0.149 mmol). The resulting suspension was refluxed under reduced pressure, and the flask was charged with hydrogen (1atm) for 10 min. The catalyst was then filtered off and the solvent was removed under reduced pressure. The crude compound was dissolved in dry DCM (5ml), N-isopropyl-N-methylpropan-2-amine (0.023ml, 0.149mmol), trioxane (0.031ml, 0.297mmol) were added and the solution was stirred at rt. After 30min, sodium triacetoxyborohydride (94mg, 0.446mmol) and acetic acid (0.026ml, 0.446mmol) were added and the reaction stirred for an additional 1 h. The solvent was then removed and the crude product was purified by reverse phase chromatography using a biotage c18 SNAP column (phase a, water 95%, ACN 5%, formic acid 0.1%); (phase B, ACN 95%, water 5%, formic acid 0.1%) to yield 40mg of a white powder.
This material was dissolved in DCM (3ml) and 1M BBr was added3Was added to the solution of (1.448ml) in DCM and the solution was stirred for 4 h. The reaction was then cooled to 0 ℃ and EtOH (1ml) was added. The solvent was removed and the product was purified by flash chromatography on C18 ((H) 2O/ACN))95:5+0.1％HCOOH}：{(ACN/H2O)95:5+ HCOOH 0.1% } 100: 0-0: 100 to give the title compound (13mg, 0.037mmol, 31.2% yield) as a white solid.
1H NMR(600MHz，DMSO-d6)ppm 10.06-10.65(bs，2H)，6.61-8.42(m，11H)，6.23-6.45(m，1H)，5.85-6.12(m，1H)，4.03-4.53(m，2H)，1.79-3.31(m，9H)。UPLC-MS：2.46min，539.1[M+H]+, method 6.
Example 103
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (4-aminocyclohex-1-en-1-yl) -1H-isochromen-1-one hydrochloride
To a solution of 3- (3-fluoro-5-methoxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-4-amine (intermediate G1, 81mg, 0.311mmol) in dry DMF (4ml) was added potassium carbonate (86mg, 0.622mmol) and the suspension was stirred at rt for 10 min. Benzyl (4- (3- (1-bromoethyl) -1-oxo-1H-isochromen-4-yl) cyclohex-3-en-1-yl) carbamate (intermediate C32, 150mg, 0.311mmol) was then added and the solution stirred at 60 ℃ for 2H. The DMF was then removed and the product was purified by Biotage Si 10g gradient with DCM and EtOH. The recovered material (37mg) was reacted with 1M boron tribromide in DCM solution (0.280ml) for 18h in dry DCM (4 ml). The solvent was removed and the product was directly purified by reverse phase chromatography using a Biotage C18 SNAP column (phase a, water 95%, ACN 5%, formic acid 0.1%); (phase B, ACN 95%, water 5%, formic acid 0.1%) to give the title compound (10mg, 34.8% yield) as a white solid.
1H NMR(400MHz，DMSO-d6)ppm 10.21(m，1H)，6.55-8.44(m，13H)，6.00-6.42(m，1H)，5.94(br s，1H)，1.66-2.84(m，10H)。UPLC-MS：2.29min，513.0[M+H]+, method 6.
Example 104
3- (1- (4-amino-3- (trifluoromethyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one
The title compound was prepared in analogy to example 1 using 3- (trifluoromethyl) -1H-pyrazolo [3,4-d ] pyrimidin-4-amine (intermediate G16, 45mg, 0.222mmol)3- (1-bromoethyl) -4-phenyl-1H-isochromen-1-one (intermediate C7, 72.9mg, 0.222mmol) to yield the title compound (15mg, 0.033mmol, 15.0% yield) as a white solid.
1H NMR(400MHz，DMSO-d6)ppm 8.03-8.29(m，2H)，7.77(s，1H)，7.63(s，1H)，7.48-7.55(m，1H)，7.36-7.48(m，3H)，7.14-7.21(m，1H)，6.89(d，J＝7.94Hz，1H)，5.59-5.88(m，1H)，1.82(d，J＝7.06Hz，3H)。UPLC-MS：4.79min，452.03[M+H]+, method 6.
Example 105
3- (1- (4-amino-3-methyl-1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one
The title compound was prepared in analogy to example 1 using 3- (1-bromoethyl) -4-phenyl-1H-isochromen-1-one (intermediate C7, 100mg, 0.304mmol) and 3-methyl-1H-pyrazolo [4,3-d ] pyrimidin-7-amine (45.3mg, 0.304mmol) to give the title compound (20mg, 16.6% yield) as a white powder.
1H NMR(400MHz，DMSO-d6)ppm 8.22(m，3H)，8.15(s，1H)，7.72-7.82(m，1H)，7.63(m，1H)，7.52-7.58(m，1H)，7.32-7.50(m，3H)，7.06-7.13(m，1H)，6.89(d，J＝8.38Hz，1H)，5.48-5.71(m，1H)，2.56(s，3H)，1.78(d，J＝7.06Hz，3H)。UPLC-MS：3.42min，398[M+H]Method 6
Example 106
3- (1- (4-amino-7H-pyrrolo [2,3-d ] pyrimidin-7-yl) ethyl) -4-phenyl-1H-isochromen-1-one
The title compound was prepared in a similar manner to example 1 using 3- (1-bromoethyl) -4-phenyl-1H-isochromen-1-one (intermediate C7, 50mg, 0.15mmol), 7H-pyrrolo [2,3-d ] pyrimidin-4-amine (30.6mg, 0.23mmol) to give the title compound (16mg, 28%).
1H NMR (400MHz, methanol-d)4)ppm 8.30(dd，J＝7.94，0.88Hz，1H)，8.26(s，1H)，7.93(s，1H)，7.66-7.73(m，1H)，7.57(d，J＝7.94Hz，2H)，7.45(d，J＝3.53Hz，2H)，7.37(m，2H)，7.01(dd，J＝14.11，7.94Hz，2H)，6.64(d，J＝3.97Hz，1H)，5.80(q，J＝7.06Hz，1H)，1.78(d，J＝7.06Hz，3H)。UPLC-MS：1.66min，383[M+H]+, method 4.
Example 107
3- (1- (2, 6-diamino-9H-purin-9-yl) ethyl) -4-phenyl-1H-isochromen-1-one
The title compound was prepared from 3- (1-bromoethyl) -4-phenyl-1H-isochromen-1-one (intermediate C7, 50mg, 0.15mmol) and 9H-purine-2, 6-diamine (34mg, 0.23mmol) in analogy to example 1 to yield the title compound (19mg, 31%).
1H NMR(400MHz，DMSO-d6)ppm 8.21(d，J＝7.50Hz，1H)，7.94(s，1H)，7.73-7.85(m，1H)，7.49-7.70(m，5H)，7.42(d，J＝7.06Hz，1H)，6.94(d，J＝7.94Hz，1H)，6.65(br.s.，2H)，5.57(s，2H)，5.20(q，J＝7.35Hz，1H)，1.74(d，J＝7.06Hz，3H)。UPLC-MS：4.61min，399[M+H]+, method 3.
Example 108
4-phenyl-3- (1- (thieno [2,3-d ] pyrimidin-4-ylamino) ethyl) -1H-isochromen-1-one
3- (1-aminoethyl) -4-phenyl-1H-isochromen-1-one hydrochloride (intermediate E250mg, 0.166mmol), 4-chlorothieno [2,3-d ] pyrimidine (36.8mg, 0.215mmol), triethylamine (0.046ml, 0.331mmol) were reacted in 2-methylpropan-2-ol (1ml) at 80 ℃ for 6H and at 50 ℃ for 60H. 4-Chlorothiopheno [2,3-d ] pyrimidine (5.7mg, 0.033mmol) and triethylamine (0.09ml, 0.066mmol) were added to the reaction mixture, and the mixture was reacted at 90 ℃ for 4 hours. The crude product was purified by reverse phase chromatography using a biotage c 1830 g SNAP column (phase a, water 95%, ACN 5%, formic acid 0.1%); phase B, ACN 95%, water 5%, formic acid 0.1%) to give the title compound (31mg, 47%).
1H NMR(400MHz，DMSO-d6)ppm 8.17-8.31(m，3H)，7.73-7.82(m，2H)，7.47-7.63(m，6H)，7.39(d，J＝6.62Hz，1H)，6.94(d，J＝7.94Hz，1H)，5.02(t，J＝7.06Hz，1H)，1.54(d，J＝7.06Hz，3H)。UPLC-MS：5.10min，400[M+H]+, method 6.
Example 109
4-phenyl-3- (1- (thieno [3,2-d ] pyrimidin-4-ylamino) ethyl) -1H-isochromen-1-one
3- (1-aminoethyl) -4-phenyl-1H-isochromen-1-one hydrochloride (intermediate E2, 50mg, 0.166mmol), 4-chlorothieno [3,2-d ] pyrimidine (36.8mg, 0.215mmol), triethylamine (0.046ml, 0.331mmol) were reacted in 2-methylpropan-2-ol (1ml) at 50 ℃ for 24H and 80 ℃ for 27H. 4-Chlorothiopheno [2,3-d ] pyrimidine (5.7mg, 0.033mmol) and triethylamine (0.09ml, 0.066mmol) were added to the reaction mixture, and the mixture was reacted at 90 ℃ for 4 hours. The crude product was purified by reverse phase chromatography using a Biotage C1830 g SNAP column (phase a, water 95%, ACN 5%, formic acid 0.1%); phase B, ACN 95%, water 5%, formic acid 0.1%) to give the title compound (33mg, 50%).
1H NMR(400MHz，DMSO-d6)ppm 8.27-8.37(m，2H)，8.17-8.24(m，1H)，8.06-8.15(m，1H)，7.68-7.83(m，1H)，7.49-7.65(m，5H)，7.28-7.45(m，2H)，6.85-7.01(m，1H)，4.84-5.09(m，1H)，1.40-1.58(m，3H)。UPLC-MS：3.84min，400[M+H]+, method 6.
Example 110
2-amino-N- (1- (1-oxo-4-phenyl-1H-isochromen-3-yl) ethyl) pyrazolo [1,5-a ] pyrimidine-3-carboxamide
3- (1-aminoethyl) -4-phenyl-1H-isochromen-1-one hydrochloride (intermediate E2, 50mg, 0.166mmol), 2-aminopyrazolo [1,5-a ] pyrimidine-3-carboxylic acid (32.5mg, 0.182mmol), HOBt (30,4mg, 0,199mmol), HBTU (17,25mg, 0,033mmol) and DIPEA (61, 0. mu.l, 0,349mmol) were reacted in DCM (1ml) at RT for 18H. The crude product was purified by reverse phase chromatography using a Biotage C1830 g SNAP column (phase a, water 95%, ACN 5%, formic acid 0.1%); phase B, ACN 95%, water 5%, formic acid 0.1%) to give the title compound (20mg, 28%).
1H NMR(400MHz，DMSO-d6)8.86-8.97(m，1H)，8.48-8.62(m，1H)，8.19-8.31(m，1H)，7.97-8.12(m，1H)，7.72-7.83(m，1H)，7.46-7.66(m，5H)，7.34-7.45(m，1H)，6.98-7.09(m，1H)，6.85-6.96(m，1H)，6.36(s，2H)，4.70-5.02(m，1H)，1.32-1.57(d，3H)。UPLC-MS：4.29min，426[M+H]Method 6
Example 112
3- (1- (4-amino-3- (1H-indazol-4-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one
The title compound was prepared in analogy to example 52 from 3- (1- (4-amino-3-iodo-1H-pyrazolo [3,4-D ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one (intermediate D2a, 50mg, 0.098mmol) and 4- (trimethylstannyl) -1H-indazole (intermediate G19, 41,4mg, 0,147mmol) to yield the title compound (19mg, 39%).
1H NMR(400MHz，DMSO-d6)ppm 13.30(s，1H)，8.13-8.27(m，2H)，8.04-8.12(m，1H)，7.70-7.79(m，1H)，7.35-7.68(m，7H)，7.25-7.34(m，1H)，7.12-7.24(m，1H)，6.82-6.96(m，1H)，5.67-5.82(m，1H)，1.68-1.97(m，3H)。UPLC-MS：3.92min，500[M+H]Method 6
Example 115
3- (1- (6-amino-9H-purin-9-yl) ethyl) -4- (5- (morpholinomethyl) thiophen-2-yl) -1H-isochromen-1-one
The 3- (1-bromoethyl) -4- (5- (morpholinomethyl) thiophen-2-yl) -1H-isochromen-1-one hydrobromide salt (intermediate C16,52mg, 0.113mmol), N6, N6-bis (tert-butoxycarbonyl) -adenine (74.1mg, 0.221mmol), K2CO3(30.5mg, 0.221mmol) was reacted in DMF (0.5ml) at 80 ℃. The crude product was purified by reverse phase chromatography using a Biotage C1830 g SNAP column (phase a, water 95%, ACN 5%, formic acid 0.1%); phase B, ACN 95%, water 5%, formic acid 0.1%), material was recovered and then treated with HCl 37%, then evaporated to give the crude product, which was then purified by reverse phase chromatography using a Biotage C1812g SNAP column (phase a, water 95%, ACN 5%, formic acid 0.1%); phase B, ACN 95%, water 5%, formic acid 0.1%) to give the title compound (25mg, 69%).
1H NMR(400MHz，DMSO-d6)ppm 11.07-11.37(bs，1H)，8.65(s，1H)，8.31(s，2H)，8.21(d，J＝7.50Hz，1H)，7.95(m，1H)，7.80-7.87(m，1H)，7.62-7.72(m，1H)，7.52(d，J＝2.65Hz，1H)，7.29(d，J＝3.09Hz，1H)，7.22(d，J＝7.94Hz，1H)，5.68(d，J＝7.06Hz，1H)，4.65(s，2H)，4.01(m，4H)，3.81(m，4H)，3.28-3.40(m，4H)，3.15(m，4H)，1.92(d，J＝7.06Hz，3H)。UPLC-MS：3.49min，489[M+H]+, method 3.
Example 116
3- (1- (9H-purin-6-ylamino) ethyl) -4- (6-methoxypyridin-3-yl) -1H-isochromen-1-one
The title compound was prepared from tert-butyl 9-trityl-9H-purin-6-ylcarbamate (82mg, 0.172mmol) and 3- (1-bromoethyl) -4- (thiazol-5-yl) -1H-isochromen-1-one (intermediate C17, 100mg, 0.194mmol) in analogy to example 21 to yield the title compound (45mg, 45.9%).
1H NMR(400MHz，DMSO-d6)ppm 9.00-9.40(bs，1H)，8.46(br.s.，2H)，8.14-8.35(m，2H)，7.72-7.96(m，2H)，7.63(m，1H)，7.00(m，2H)，5.08(m，1H)，3.95(s，3H)，1.42-1.73(m，3H)。UPLC-MS：3.757min，415[M+H]Method 3
Example 117
3- (1- (9H-purin-6-ylamino) ethyl) -4- (thiazol-5-yl) -1H-isochromen-1-one
The title compound was prepared from tert-butyl 9-trityl-9H-purin-6-ylcarbamate (169mg, 0.353mmol) and 3- (1-bromoethyl) -4- (6-methoxypyridin-3-yl) -1H-isochromen-1-one (intermediate C12, 108mg, 0.321mmol) in analogy to example 21 to yield the title compound (12mg, 17.7%).
1H NMR(400MHz，DMSO-d6)ppm 9.55(br.s.，1H)，9.40(s，1H)，8.45-8.64(m，2H)，8.21(d，J＝7.50Hz，1H)，8.11(s，1H)，7.84(t，J＝7.72Hz，1H)，7.65(t，J＝7.50Hz，1H)，7.09(d，J＝7.94Hz，1H)，5.16(m，1H)，1.62(d，J＝7.06Hz，3H)。UPLC-MS：4.14min，391[M+H]+, method 7.
Example 122
3- (1- (9H-purin-6-ylamino) ethyl) -4- (1H-pyrazol-4-yl) -1H-isochromen-1-one
The title compound was prepared from 3- (1-aminoethyl) -4- (1H-pyrazol-4-yl) -1H-isochromen-1-one dihydrochloride (intermediate E4, 50mg, 0.152mmol) 6-chloro-9H-purine (35.3mg, 0.229mmol) in a similar manner to example 43 to give the title compound (19mg, 30.4%).
1H NMR(400MHz，DMSO-d6)ppm 9.13-9.44(bs，1H)，8.52(d，J＝12.79Hz，2H)，8.16(d，J＝7.50Hz，1H)，7.73-7.88(m，3H)，7.58(m，1H)，7.19(d，J＝7.94Hz，1H)，5.07-5.40(m，1H)，1.54(d，J＝7.06Hz，3H)。UPLC-MS：3.60min，374[M+H]+, method 7.
Example 123
4-amino-6- ((1- (1-oxo-4- (1H-pyrazol-4-yl) -1H-isochromen-3-yl) ethyl) amino) pyrimidine-5-carbonitrile hydrochloride
The title compound was prepared from 3- (1-aminoethyl) -4- (1H-pyrazol-4-yl) -1H-isochromen-1-one dihydrochloride (intermediate E4, 50mg, 0.152mmol), 4-amino-6-bromopyrimidine-5-carbonitrile (39.4mg, 0.198mmol) in a similar manner to example 43 to give the title compound (19mg, 30.4%).
1H NMR(400MHz，DMSO-d6)ppm 8.19(d，J＝7.06Hz，1H)，8.01(s，1H)，7.77(m，4H)，7.60(t，J＝7.72Hz，1H)，7.41(br.s.，2H)，7.20(d，J＝7.94Hz，1H)，5.12(t，J＝6.84Hz，1H)，1.44(d，J＝7.06Hz，3H)。UPLC-MS：3.84min，374[M+H]+, method 7.
Example 124
3- (1- (9H-purin-6-ylamino) ethyl) -4- (2-aminopyrimidin-5-yl) -1H-isochromen-1-one
The title compound was prepared from tert-butyl 9-trityl-9H-purin-6-ylcarbamate (25.8mg, 0.054mmol), 4- (2-aminopyrimidin-5-yl) -3- (1-bromoethyl) -1H-isochromen-1-one (intermediate C34, 17mg, 0.094mmol) in analogy to example 21 to yield the title compound (7mg, 35.6%).
1H NMR(400MHz，DMSO-d6)ppm 9.18-9.44(bs，1H)，8.29-8.55(m，4H)，8.19(d，J＝7.94Hz，1H)，7.81(m，1H)，7.62(m，1H)，6.81-7.43(m，3H)，4.95-5.24(m，1H)，1.60(d，J＝7.06Hz，3H)。UPLC-MS：3.28min，401[M+H]+, method 7.
Example 126
3- (1- (9H-purin-6-ylamino) ethyl) -4- (4- (piperazin-1-ylmethyl) phenyl) -1H-isochromen-1-one dihydrochloride
The title compound was prepared in analogy to example 21 from tert-butyl 9-trityl-9H-purin-6-ylcarbamate (130mg, 0.273mmol), tert-butyl 4- (4- (3- (1-bromoethyl) -1-oxo-1H-isochromen-4-yl) benzyl) piperazine-1-carboxylate (intermediate C35, 17mg, 0.094mmol) to yield the title compound (22.8mg, 17.7%).
1H NMR(400MHz，DMSO-d6)ppm 8.98-9.32(m，1H)，8.33(br.s.，1H)，8.22(d，J＝7.50Hz，1H)，7.51-7.85(m，5H)，7.45(d，J＝6.17Hz，1H)，6.95(d，J＝7.94Hz，1H)，4.86-5.25(m，1H)，2.84-3.24(m，10H)，1.56(d，J＝7.06Hz，3H)。UPLC-MS：4.48min，482[M+H]+, method 7.
Example 127
3- (1- (4-amino-1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (4- (piperazin-1-ylmethyl) phenyl) -1H-isochromen-1-one hydrochloride
Tert-butyl 4- (4- (3- (1-bromoethyl) -1-oxo-1H-isochromen-4-yl) benzyl) piperazine-1-carboxylate (intermediate C35, 60mg, 0.114mmol), 1H-pyrazolo [3,4-d]Pyrimidin-4-amine (30.7mg, 0.228mmol) and K2CO3(31.4mg, 0.228mmol) was stirred in DMF (0.7ml) at 80 ℃ for 3 h. The reaction mixture was diluted with 1M aqueous HCl (1ml) and purified by reverse phase chromatography using a Biotage C18 SNAP 60g column (phase a, water 95%, ACN 5%, formic acid 0.1%); phase B, ACN 95%, water 5%, formic acid 0.1%). To the collected fractions was added 37% aqueous HCl (1ml), and concentrated to give the title compound (26mg, 44%).
1H NMR(400MHz，DMSO-d6)ppm 8.49(br s，0.5H)，8.23-8.20(m，1H)，8.12(br s，0.5H)，8.09(br s，1H)，8.01(br s，1H)，7.80-7.62(m，4H)，7.42-7.38(m，1.5H)，7.25-7.23(m，0.5H)6.90-6.88(d，1H)，5.65-5.55(m，1H)，2.54(m，3H)，，1.81(t，3H)。UPLC-MS：3.78min，482[M+H]+, method 3.
Example 130
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (morpholinomethyl) -1H-isochromen-1-one
Step 1.3- (1- (4-amino-3- (3-fluoro-5-methoxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (morpholinomethyl) -1H-isochromen-1-one
Reacting 3- (3-fluoro-5-methoxyphenyl) -1H-pyrazolo [3,4-d]Pyrimidin-4-amine (intermediate G1, 0.135G, 0.521mmol) and potassium carbonate (0.206G, 1.498mmol) were combined in DMF (5ml) and stirred at r.t. for 10 min. A solution of 3- (1-bromoethyl) -4- (morpholinomethyl) -1H-isochromen-1-one hydrobromide (intermediate C48, 0.215g, 0.496mmol) in DMF (5ml) was then added and the mixture heated at 70 ℃ for 3H. The reaction was stopped by the addition of 1M HCl (2ml) and the solvent was removed in vacuo. The crude product was purified by C18 Biotage column reverse phase chromatography (H) 2O:CH3CN＝80:20-100％CH3CN containing 0.1% HCOOH) to give 3- (1- (4-amino-3- (3-fluoro-5-methoxyphenyl) -1H-pyrazolo [3, 4-d)]Pyrimidin-1-yl) ethyl) -4- (morpholinomethyl) -1H-isochromen-1-one (0.033 g).
UPLC-MS: 0.74min, 531.4[ M + H ] +, method 14.
And 2. step 2.
To 3- (1- (4-amino-3- (3-fluoro-5-methoxyphenyl) -1H-pyrazolo [3, 4-d)]Pyrimidin-1-yl) ethyl) -4- (morpholinomethyl) -1H-isochromen-1-one (0.033g) in dry DCM (2ml) was added 1M BBr3Was added to the solution of DCM (0.93ml, 0.93mmol), and the solution was stirred at room temperature for 2.5 h. The solution was cooled to 0 ℃ and ethanol (8ml) was added. The solution was evaporated to dryness and the crude product was purified by flash chromatography on silica gel-NH column (DCM-DCM: MeOH ═ 90:10) to give the title compound as a light orange solid (5 mg).
1H NMR (400MHz, methanol-d)4)ppm 8.28(s，1H)，8.23-8.27(m，1H)，8.03(d，1H)，7.80-7.85(m，1H)，7.55-7.61(m，1H)，6.90-6.93(m，1H)，6.86-6.90(m，1H)，6.63(dt，1H)，6.54(q，1H)，3.66-4.01(m，2H)，3.49-3.56(m，4H)，2.41-2.48(m，4H)，1.98(d，3H)。UPLC-MS：0.66min，517.4[M+H]+, method 14.
Example 133
4- (3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -1-oxo-1H-isochromen-4-yl) -5, 6-dihydropyridine-1 (2H) -carboxylic acid benzyl ester
The title compound was prepared in analogy to example 51 from 4- (3- (1- (4-amino-3-iodo-1H-pyrazolo [3,4-D ] pyrimidin-1-yl) ethyl) -1-oxo-1H-isochromen-4-yl) -5, 6-dihydropyridine-1 (2H) -carboxylic acid benzyl ester (intermediate D22, 334mg, 0.515mmol) and (3-fluoro-5-hydroxyphenyl) boronic acid (161mg, 1.030 mmol) to yield the title compound (190mg, 58.3%) as a light yellow solid.
1H NMR(400MHz，DMSO-d6)ppm 10.23(br.s.，1H)，8.02-8.50(m，3H)，7.56-7.92(m，3H)，7.38(br.s.，1H)，6.75-7.26(m，4H)，6.57-6.73(m，1H)，5.93(q，J＝7.06Hz，1H)，4.56(br.s.，2H)，3.43(br.s.，7H)，1.84(d，J＝7.06Hz，3H)。UPLC-MS：4.87min，633.0[M+H]Method 6
Example 134
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (5- (piperazin-1-ylmethyl) thiophen-2-yl) -1H-isochromen-1-one dihydrochloride
Step a.4- ((benzyl 5- (3- (1- (4-amino-3- (3-fluoro-5-methoxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -1-oxo-1H-isochromen-4-yl) thiophen-2-yl) methyl) piperazine-1-carboxylate
Benzyl 4- ((5- (3- (1-hydroxyethyl) -1-oxo-1H-isochromen-4-yl) thiophen-2-yl) methyl) piperazine-1-carboxylate (intermediate B38, 903mg, 1.790mmol) was dissolved in 20ml dry and then a solution of tribromophosphine 1M in DCM (2684 μ l, 2.68mmol) was added slowly. The mixture was stirred at r.t. By adding 60ml NaHCO3The reaction was stopped by saturated solution and extracted with DCM (100 ml). The phases were separated to give a milky white organic phase. 30ml of CAN was added and the solution became clear. The mixture was concentrated to dryness to give a yellow-white solid. In a separate 30ml vial, 3- (3-fluoro-5-methoxyphenyl) -1H-pyrazolo [3,4-d]Pyrimidin-4-amine (intermediate G1, 510mg, 1.969mmol) and potassium carbonate (742mg, 5.37mmol) were suspended in 10ml of DMF, the mixture was heated at 60C for 15min, then bromide dissolved in 5ml of dry DMF was added. The reaction was stirred at r.t. for a further 1h and then with NaHCO 3The reaction was quenched with aqueous solution (40ml), extracted with DCM (80ml) and then concentrated to give a brown solid which was purified by chromatography with DCM: MeOH elution afforded 4- ((5- (3- (1- (4-amino-3- (3-fluoro-5-methoxyphenyl) -1H-pyrazolo [3, 4-d)]Pyrimidin-1-yl) ethyl) -1-oxo-1H-isochromen-4-yl) thiophen-2-yl) methyl) piperazine-1-carboxylic acid benzyl ester (460mg, 0.617mmol, 34.5% yield) as a brown solid.
UPLC-MS: 1.99min, 746.09[ M + H ] +, method 10
And b, step b.
In a 250ml round bottom flask equipped with a magnetic stirrer, 4- ((5- (3- (1- (4-amino-3- (3-fluoro-5-methoxyphenyl) -1H-pyrazolo [3, 4-d)]Pyrimidin-1-yl) ethyl) -1-oxo-1H-isochromen-4-yl) thiophen-2-yl) methyl) piperazine-1-carboxylic acid benzyl ester (460mg, 0.617mmol) was suspended in 20ml dry dichloromethane, or 1M tribromoborane in DCM (5ml) was added and the mixture was stirred at r.t. overnight. MeOH (15ml) and 1M HCl (5ml) were added, the mixture was stirred for 30min, then the organic solvent was evaporated and the crude product was purified by reverse phase chromatography with H2O \ MeCN \ HCOOH95: 5: 0.1% and MeCN \ H2O \ HCOOH95: 5: 0.1% elution gave the title compound (254mg, 0.379mmol, 61.4% yield) as a whitish solid.
1H NMR(600MHz，DMSO-d6)ppm 8.16-8.29(m，2H)，8.11(s，1H)，7.75-7.91(m，1H)，7.52-7.69(m，1H)，7.15(d，J＝7.89Hz，1H)，6.76-7.05(m，4H)，6.66(d，J＝10.85Hz，1H)，5.97(q，J＝7.13Hz，1H)，3.67(br.s.，2H)，2.80(br.s.，4H)，2.41(br.s.，4H)，1.84(d，J＝7.23Hz，3H)。UPLC-MS：1.32min，597.99[M+H]Method 10
Intermediate 135
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (5- ((4-methylpiperazin-1-yl) methyl) thiophen-2-yl) -1H-isochromen-1-one dihydrochloride
Step a, 3- (1- (4-amino-3- (3-fluoro-5-methoxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (5- ((4-methylpiperazin-1-yl) methyl) thiophen-2-yl) -1H-isochromen-1-one
The title compound was prepared in analogy to example 134 step a from 3- (1-hydroxyethyl) -4- (5- ((4-methylpiperazin-1-yl) methyl) thiophen-2-yl) -1H-isochromen-1-one (intermediate B39, 411mg, 1.069mmol) and 3- (3-fluoro-5-methoxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-4-amine (intermediate G1, 305mg, 1.176mmol) to give 3- (1- (4-amino-3- (3-fluoro-5-methoxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (5- ((4-methylpiperazin-1-yl) ethyl) ) Methyl) thiophen-2-yl) -1H-isochromen-1-one (420mg, 0.671mmol, 62.8% yield) as a brown solid.
UPLC-MS: 1.52min, 626.17[ M + H ] +, method 10
And b, step b.
The title compound was prepared in analogy to example 134 step b and 3- (1- (4-amino-3- (3-fluoro-5-methoxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (5- ((4-methylpiperazin-1-yl) methyl) thiophen-2-yl) -1H-isochromen-1-one (410mg, 0.655mmol) to yield the title compound (100mg, 0.146mmol, 22.29% yield) as a white solid.
1H NMR(600MHz，DMSO-d6)ppm 10.17(br.s.，1H)，8.21(dd，J＝7.89，0.66Hz，1H)，8.13(s，1H)，8.11(s，1H)，7.78-7.87(m，1H)，7.58-7.70(m，1H)，7.15(d，J＝7.89Hz，1H)，6.85-7.02(m，3H)，6.83(dd，J＝8.88，1.64Hz，1H)，6.66(dt，J＝10.85，2.30Hz，1H)，5.96(d，J＝6.91Hz，1H)，3.71(br.s.，2H)，3.29(s，3H)，2.54-2.77(m，4H)，2.39(br.s.，4H)，1.84(d，J＝6.91Hz，3H)。UPLC-MS：1.29min，611.71[M+H]Method 10
Example 136
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (5- (3- (dimethylamino) propyl) thiophen-2-yl) -1H-isochromen-1-one, formate salt
Step a.3- (1- (4-amino-3- (3- (benzyloxy) -5-fluorophenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (5- (3- (dimethylamino) prop-1-en-1-yl) thiophen-2-yl) -1H-isochromen-1-one.
The title compound was prepared in analogy to example 134 step a from 4- (5- (3- (dimethylamino) prop-1-en-1-yl) thiophen-2-yl) -3- (1-hydroxyethyl) -1H-isochromen-1-one (intermediate B40, 220mg, 0,619mmol) and 3- (3- (benzyloxy) -5-fluorophenyl) -1H-pyrazolo [3,4-d ] pyrimidin-4-amine (intermediate G18, 125mg, 0.37mmol) to give 3- (1- (4-amino-3- (3- (benzyloxy) -5-fluorophenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (5- (3- (dimethylamino) prop-1-en-1-yl) thiophen-2-yl) -1H-isochromen-1-one (180mg, 0,268mmol, 43, 2% yield) as a brown oil.
UPLC-MS: 1.86min, 673.3[ M + H ] +, method 10.
And b, step b.
Reacting 3- (1- (4-amino-3- (3- (benzyloxy) -5-fluorophenyl) -1H-pyrazolo [3, 4-d)]Pyrimidin-1-yl) ethyl) -4- (5- (3- (dimethylamino) prop-1-en-1-yl) thiophen-2-yl) -1H-isochromen-1-one (180mg, 0,268mmol) was dissolved in 15ml EtOH followed by palladium on carbon 5% (28,5mg, 0,268mmol) and then triethylsilane (2ml, 12,52mmol) in an Ar atmosphere. The mixture was filtered, and the crude product was concentrated and purified by C1830 g +12g reverse phase chromatography over H 2O \ MeCN \ HCOOH95: 5: 0.1% and MeCN \ H2O \ HCOOH95: 5: 0.1% the title compound (20mg, 0,032mmol, 11, 85% yield) was obtained as a white solid.
1H NMR(400MHz，DMSO-d6)ppm 8.17-8.29(m，3H)，8.12(s，1H)，7.71-7.87(m，2H)，7.56-7.69(m，1H)，7.37-7.56(m，1H)，7.15(d，J＝8.38Hz，1H)，6.76-6.94(m，4H)，6.66(d，J＝10.58Hz，1H)，5.96(d，J＝7.06Hz，1H)，2.78(t，J＝7.28Hz，2H)，2.28(t，J＝6.84Hz，2H)，2.16(s，6H)，1.84(d，J＝7.06Hz，3H)，1.70(t，2H)。UPLC-MS：0.81min，585.25[M+H]+, method 9.
Example 137
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (3- (3- (dimethylamino) propyl) phenyl) -1H-isochromen-1-one carboxylic acid
Step a.3- (1- (4-amino-3- (3-fluoro-5-methoxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (5- ((4-methylpiperazin-1-yl) methyl) thiophen-2-yl) -1H-isochromen-1-one.
The title compound was prepared in analogy to example 134 step a from 4- (3- (3- (dimethylamino) prop-1-en-1-yl) phenyl) -3- (1-hydroxyethyl) -1H-isochromen-1-one (intermediate B42, 280mg, 0.801mmol) and 3- (3- (benzyloxy) -5-fluorophenyl) -1H-pyrazolo [3,4-d ] pyrimidin-4-amine (intermediate G18, 0.417mmol, 140mg) to yield 3- (1- (4-amino-3- (3-fluoro-5-methoxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (5- ((4-methylpiperazin-1-yl) ethyl -yl) methyl) thiophen-2-yl) -1H-isochromen-1-one (120mg, 0.18mmol, 43% yield) as a yellow oil.
UPLC-MS: 1.91min, 667.27[ M + H ] +, method 10
Step b
The title compound was prepared in analogy to example 136 step b from 3- (1- (4-amino-3- (3- (benzyloxy) -5-fluorophenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (3- (3- (dimethylamino) prop-1-en-1-yl) phenyl) -1H-isochromen-1-one to give the title compound (120mg, 0.18mmol, 43.1% yield) as a white solid.
1H NMR(400MHz，DMSO-d6)ppm 1.46-1.63(m，1H)，1.70-1.88(m，4H)，2.08(br.s.，3H)，2.14-2.25(m，4H)，2.30-2.37(m，1H)，2.40-2.48(m，1H)，2.60-2.72(m，1H)，5.74(sxt，J＝6.98Hz，1H)，6.55-6.99(m，6H)，7.13-7.86(m，6H)，8.02-8.30(m，3H)。UPLC-MS：1.37min，579.3[M+H]+, method 10.
Example 137a (enantiomer 1) and 137b (enantiomer 2): 3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (3- (3- (dimethylamino) propyl) phenyl) -1H-isochromen-1-one single enantiomer
The racemate 3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (3- (3- (dimethylamino) propyl) phenyl) -1H-isochromen-1-one formate (example 137, 0.020g, 0.860mmol) was dissolved in 2ml of ethanol and subjected to chiral resolution by chiral preparative chromatography. Conditions are as follows: column: chiralpak IC (25x 2cm), 5 um; mobile phase: n-hexane/(2-propanol/methanol 1/1+ 0.1% isopropylamine) 65/35 v/v; flow rate: 16 ml/min; and D, DAD detection: 220 nm; loop circuit: 500 mul; and (3) injection: 5mg (per injection).
Fractions containing the enantiomer of the first eluate were evaporated to dryness to give compound 137a (first eluted enantiomer, 6.8mg, 0.0117 mmol). Chiral HPLC (method a 15): rt 7.4min, ee > 99%.
1H NMR (400MHz, methanol-d)4)ppm 8.29-8.33(m，1H)，8.05-8.08(m，1H)，7.64-7.71(m，1H)，7.55-7.61(m，1H)，7.10-7.43(m，3H)，6.87-6.97(m，2H)，6.81-6.87(m，1H)，6.60-6.70(m，2H)，5.85-5.97(m，1H)，2.24-2.73(m，10H)，1.86-1.96(m，4H)，1.61-1.71(m，1H)。UPLC-MS：0.69min，579.4[M+H]+, method 14.
The fraction containing the enantiomer of the second eluate was evaporated to dryness to give compound 137b (second eluted enantiomer, 6.3mg, 0.0109 mmol). Chiral HPLC (method a 15): rt 8.4min, ee 96.4%.
1H NMR (400MHz, methanol-d)4)ppm 8.28-8.33(m，1H)，8.04-8.08(m，1H)，7.63-7.71(m，1H)，7.55-7.61(m，1H)，7.10-7.42(m，3H)，6.86-6.96(m，2H)，6.79-6.86(m，1H)，6.59-6.70(m，2H)，5.85-5.97(m，1H)，2.24-2.72(m，7H)，2.17(s，3H)，1.82-1.94(m，4H)，1.57-1.67(m，1H)。UPLC-MS：0.69min，579.4[M+H]+, method 14.
Example 138
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (4- (3- (dimethylamino) propyl) phenyl) -1H-isochromen-1-one formate salt
Step a.3- (1- (4-amino-3- (3- (benzyloxy) -5-fluorophenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (4- (3- (dimethylamino) prop-1-en-1-yl) phenyl) -1H-isochromen-1-one
The title compound was prepared in analogy to example 134 step a from 4- (4- (3- (dimethylamino) prop-1-en-1-yl) phenyl) -3- (1-hydroxyethyl) -1H-isochromen-1-one (intermediate B45, 220mg, 0.630mmol) and 3- (3- (benzyloxy) -5-fluorophenyl) -1H-pyrazolo [3,4-d ] pyrimidin-4-amine (intermediate G18, 0.63mmol, 211mg) to yield 3- (1- (4-amino-3- (3- (benzyloxy) -5-fluorophenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (4- (3- (dimethylamino) Prop-1-en-1-yl) phenyl) -1H-isochromen-1-one (120mg, 0.18mmol, 43.1% yield) as a yellow oil.
UPLC-MS: 2.01min, 667.21[ M + H ] +, method 10
And b, step b.
The title compound was prepared in analogy to example 136 step b from 3- (1- (4-amino-3- (3- (benzyloxy) -5-fluorophenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (4- (3- (dimethylamino) prop-1-en-1-yl) phenyl) -1H-isochromen-1-one (162mg, 0,243mmol) to give the title compound (17mg, 0.027mmol, 11.20% yield) as a white solid.
1H NMR(600MHz，DMSO-d6)ppm 10.03-10.35(bs，1H)，8.22(dd，J＝7.89，0.92Hz，1H)，8.17(s，1H)，8.07(s，1H)，7.76(t，1H)，7.62(t，1H)，7.32(dd，J＝19.81，1.83Hz，2H)，7.12-7.20(m，1H)，6.97(m，1H)，6.86-6.92(m，2H)，6.78-6.85(m，1H)，6.61-6.68(m，1H)，5.75(d，J＝6.97Hz，1H)，2.61(d，J＝7.70Hz，2H)，2.32(m，2H)，2.20(s，6H，3H eacH)，1.82(d，J＝6.97Hz，3H)，1.68-1.78(m，2H)。UPLC-MS：1.35min，579.2[M+H]Method 10
Example 138a (enantiomer 1) and 138b (enantiomer 2): 3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (4- (3- (dimethylamino) propyl) phenyl) -1H-isochromen-1-one single enantiomer
The racemate 3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (4- (3- (dimethylamino) propyl) phenyl) -1H-isochromen-1-one formate (example 138, 0.010g, 0.016mmol) was dissolved in ethanol/methanol 1/1(3ml) and subjected to chiral resolution by chiral preparative chromatography. Conditions are as follows: column: chiralpak IC (25X 2.0cm), 5 μ; mobile phase: n-hexane/(2-propanol/methanol 1/1+ 0.1% isopropylamine) 60/40% v/v; flow rate: 16 ml/min; and D, DAD detection: 220 nm; loop circuit: 1000 μ l; and (3) injection: 3.3mg (per injection).
Fractions containing the enantiomer of the first eluate were evaporated to dryness to give compound 138a (first eluted enantiomer, 3.1mg, 0.005 mmol). Chiral HPLC (method a 15): rt 7.3min, ee > 99%.
1H NMR (400MHz, methanol-d)4)ppm 8.27-8.32(m，1H)，8.06(s，1H)，7.63-7.69(m，1H)，7.53-7.60(m，1H)，7.28-7.33(m，1H)，7.20-7.26(m，1H)，7.04-7.09(m，1H)，6.93(d，1H)，6.88-6.90(m，1H)，6.81-6.86(m，1H)，6.74-6.78(m，1H)，6.63(dt，1H)，5.93(q，1H)，2.57-2.68(m，4H)，2.44(s，6H)，1.81-1.94(m，5H)。UPLC-MS：0.69min，579.5[M+H]+, method 13.
The fraction containing the enantiomer of the second eluent was evaporated to dryness to give compound 138b (second eluted enantiomer, 3.0mg, 0.005 mmol). Chiral HPLC (method a 15): rt 8.0min, ee 93.2%.
1H NMR (400MHz, methanol-d)4)ppm 8.26-8.32(m，1H)，8.06(s，1H)，7.63-7.69(m，1H)，7.53-7.60(m，1H)，7.27-7.34(m，1H)，7.20-7.26(m，1H)，7.03-7.09(m，1H)，6.93(d，1H)，6.88-6.90(m，1H)，6.81-6.86(m，1H)，6.73-6.78(m，1H)，6.63(dt，1H)，5.92(q，1H)，2.52-2.69(m，4H)，2.41(s，6H)，1.79-1.94(m，5H)。UPLC-MS：0.69min，579.5[M+H]+, method 13.
Example 139
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (4- ((4-methylpiperazin-1-yl) methyl) phenyl) -1H-isochromen-1-one dihydrochloride
Step a.3- (1- (4-amino-3- (3-fluoro-5-methoxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (4- ((4-methylpiperazin-1-yl) methyl) phenyl) -1H-isochromen-1-one
The title compound was prepared in analogy to example 134 step a from 3- (1-hydroxyethyl) -4- (4- ((4-methylpiperazin-1-yl) methyl) phenyl) -1H-isochromen-1-one (intermediate B46, 750mg, 1.982mmol) and 3- (3-fluoro-5-methoxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-4-amine (intermediate G1, 565mg, 2.18mmol) to give 3- (1- (4-amino-3- (3-fluoro-5-methoxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (4- ((4-methylpiperazin-1-yl) methyl) phenyl) -1H-isochromen-1-one (98mg, 0.158mmol, 7.98% yield) as a brown solid.
UPLC-MS: 1.53min, 620.20[ M + H ] +, method 10.
Step b
The title compound was prepared from 3- (1- (4-amino-3- (3-fluoro-5-methoxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (4- ((4-methylpiperazin-1-yl) methyl) phenyl) -1H-isochromen-1-one (98mg, 0.158mmol) in analogy to example 134 step b to yield (48mg, 0.071mmol, 44.7% yield) as a white solid
1H NMR(600MHz，DMSO-d6) ppm 10.09-10.24(bs, 1H), 8.22(dd, J ═ 8.06, 0.82Hz, 1H), 8.04(s, 1H), 7.72-7.80(m, 1H), 7.59-7.66(m, 1H), 7.30-7.44(m, 2H), 7.22(dd, J ═ 7.89, 1.32Hz, 1H), 6.89(d, J ═ 1.97Hz, 4H), 6.62-6.70(m, 1H), 5.76(d, J ═ 7.23Hz, 1H), 3.29(m, 4H), 2.38-2.45(m, 4H), 1.81(d, J ═ 7.23Hz, 3H). UPLC-MS: 1.27min, method 10.
Example 140
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (4- (pyrrolidin-1-ylmethyl) phenyl) -1H-isochromen-1-one hydrochloride
Step a.3- (1- (4-amino-3- (3-fluoro-5-methoxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (4- (pyrrolidin-1-ylmethyl) phenyl) -1H-isochromen-1-one
The title compound was prepared in analogy to example 134 step a from 3- (1-hydroxyethyl) -4- (4- (pyrrolidin-1-ylmethyl) phenyl) -1H-isochromen-1-one (intermediate B47, 380mg, 1.088mmol) and 3- (3-fluoro-5-methoxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-4-amine (intermediate G1, 310mg, 1.197mmol) to give 3- (1- (4-amino-3- (3-fluoro-5-methoxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (4- (pyrrolidin-1-ylmethyl) phenyl) -1H-isochromen-1-one -ketone (170mg, 0.288mmol, 26.5% yield) as a brown solid.
UPLC-MS: 0.80min, 591.25[ M + H ] +, method 9
And b, step b.
The title compound was prepared in analogy to example 134 step b from 3- (1- (4-amino-3- (3-fluoro-5-methoxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (4- (pyrrolidin-1-ylmethyl) phenyl) -1H-isochromen-1-one (170mg, 0.288mmol) to give the title compound (126mg, 0.206mmol, 71.4% yield) as a white solid.
1H NMR(600MHz，DMSO-d6)ppm 10.21(br.s.，2H)，8.24(dd，J＝7.89，0.99Hz，1H)，8.03-8.17(m，2H)，7.44-7.85(m，5H)，7.22(dd，J＝7.73，1.81Hz，1H)，6.80-6.96(m，3H)，6.68(dt，J＝10.85，2.30Hz，1H)，5.73(q，J＝7.13Hz，1H)，4.40(d，J＝5.59Hz，2H)，3.30(m，2H)，3.10(dd，J＝10.69，7.07Hz，2H)，2.07(br.s.，2H)，1.88-2.00(m，2H)，1.83(d，J＝6.91Hz，3H)。UPLC-MS：1.23min，577.27[M+H]+, method 10.
Example 141
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (4- (piperidin-1-ylmethyl) phenyl) -1H-isochromen-1-one hydrochloride
Step a.3- (1- (4-amino-3- (3-fluoro-5-methoxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (4- (piperidin-1-ylmethyl) phenyl) -1H-isochromen-1-one
The title compound was prepared from 3- (1-hydroxyethyl) -4- (4- (piperidin-1-ylmethyl) phenyl) -1H-isochromen-1-one (intermediate B48, 720mg, 1.981mmol) and 3- (3-fluoro-5-methoxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-4-amine (intermediate G1, 565mg, 2.17mmol) in analogy to example 134 step a to yield 3- (1- (4-amino-3- (3-fluoro-5-methoxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (4- (piperidin-1-ylmethyl) phenyl) -1H-isochromen-1-one Ketone (850mg, 1.40mmol, 71.0% yield) as a brown solid.
UPLC-MS: 1.53min, 605.23[ M + H ] +, method 10
And b, step b.
The title compound was prepared from 3- (1- (4-amino-3- (3-fluoro-5-methoxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (4- (piperidin-1-ylmethyl) phenyl) -1H-isochromen-1-one (850mg, 1.406mmol) in analogy to example 134 step b to yield (420mg, 0.67mmol, 47.6% yield) as a white solid.
1H NMR(600MHz，DMSO-d6)ppm 10.10-10.23(m，1H)，8.24(dd，J＝7.89，0.99Hz，1H)，8.05(s，1H)，7.77(t，1H)，7.64(t，1H)，7.31-7.60(m，3H)，7.02-7.16(m，1H)，6.86-6.94(m，2H)，6.77-6.84(m，1H)，6.66(d，J＝10.85Hz，1H)，5.75(d，J＝6.91Hz，1H)，3.72-4.21(m，2H)，2.62-3.16(m，4H)，1.82(m，9H)。UPLC-MS：1.34min，591.17[M+H]+, method 10.
Example 142
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (3- ((4-methylpiperazin-1-yl) methyl) phenyl) -1H-isochromen-1-one
Step a.3- (1- (4-amino-3- (3-fluoro-5-methoxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (3- ((4-methylpiperazin-1-yl) methyl) phenyl) -1H-isochromen-1-one
The title compound was prepared in analogy to example 134 step a from 3- (1-hydroxyethyl) -4- (3- ((4-methylpiperazin-1-yl) methyl) phenyl) -1H-isochromen-1-one (intermediate B49, 350mg, 0.925mmol) and 3- (3-fluoro-5-methoxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-4-amine (intermediate G1, 264mg, 1.017mmol) to give 3- (1- (4-amino-3- (3-fluoro-5-methoxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (3- ((4-methylpiperazin-1-yl) methyl) phenyl) -1H-isochromen-1-one (196mg, 0.316mmol, 34.2% yield) as a brown solid.
UPLC-MS: 0.82min, 620.33[ M + H ] +, method 9.
And b, step b.
The title compound was prepared in analogy to example 134 step b from 3- (1- (4-amino-3- (3-fluoro-5-methoxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (3- ((4-methylpiperazin-1-yl) methyl) phenyl) -1H-isochromen-1-one (196mg, 0.316mmol) to give the title compound (137mg, 0.202mmol, 63.8% yield) as a white solid.
1H NMR(600MHz，DMSO-d6)ppm 10.20(d，J＝10.85Hz，1H)，8.23(ddd，J＝7.89，3.29，0.99Hz，1H)，8.15(s，1H)，8.11(s，1H)，7.73-7.82(m，1H)，7.60-7.69(m，1H)，7.23-7.57(m，4H)，7.03(d，J＝6.91Hz，1H)，6.86-6.94(m，2H)，6.78-6.86(m，1H)，6.68(ddt，J＝10.89，6.54，2.30，2.30Hz，1H)，5.61-5.87(m，1H)，3.34(br.s.，4H)，2.52-3.20(m，9H)，1.76-1.90(m，3H)。UPLC-MS：2.78min，606.1[M+H]+, method 9.
Example 143
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (3- (piperidin-1-ylmethyl) phenyl) -1H-isochromen-1-one formate salt
Step a.3- (1- (4-amino-3- (3- (benzyloxy) -5-fluorophenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (3- (piperidin-1-ylmethyl) phenyl) -1H-isochromen-1-one
The title compound was prepared from 3- (3- (1- (4-amino-3- (3- (benzyloxy) -5-fluorophenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -1-oxo-1H-isochromen-4-yl) benzaldehyde (intermediate F6, 150mg, 0,245mmol) and piperidine (0.486ml, 4.90mmol) in analogy to intermediate B37 to give 3- (1- (4-amino-3- (3- (benzyloxy) -5-fluorophenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (3- (piperidin-1-ylmethyl) phenyl) -1H-isochromen-1-one (103mg, 0,151mmol, 61, 7% yield) as a whitish solid.
UPLC-MS: 2.08min, 667.32[ M + H ] +, method 10
And b, step b.
The title compound was prepared in analogy to example 136 step b from 3- (1- (4-amino-3- (3- (benzyloxy) -5-fluorophenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (3- (piperidin-1-ylmethyl) phenyl) -1H-isochromen-1-one (103mg, 0.151mmol) to yield the title compound (13mg, 13.5%) as a white solid.
1H NMR(400MHz，DMSO-d6)ppm 8.15-8.35(m，3H)，8.06(d，J＝4.41Hz，1H)，7.76(d，J＝7.50Hz，1H)，7.57-7.66(m，1H)，7.39-7.51(m，1H)，7.19-7.37(m，3H)，6.74-7.06(m，5H)，6.66(d，J＝11.03Hz，1H)，5.56-5.83(m，1H)，2.37(m，2H)，2.18(d，3H)，1.71-1.88(m，2H)，1.18-1.65(m，8H)。UPLC-MS：1.38min，591.30[M+H]Method 10
Example 144
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (2,2,6, 6-tetramethyl-1, 2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one hydrochloride
Step a.3- (1- (4-amino-3- (3-fluoro-5-methoxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (2,2,6, 6-tetramethyl-1, 2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one hydrochloride
The title compound was prepared from 3- (1-bromoethyl) -4- (2,2,6, 6-tetramethyl-1, 2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one hydrobromide (intermediate C39, 267mg, 0.567mmol) and intermediate G1(147mg, 0.567mmol) according to the method reported for compound D1 to give 3- (1- (4-amino-3- (3-fluoro-5-methoxyphenyl) -1H-pyrazolo [3,4-D ] pyrimidin-1-yl) ethyl) -4- (2,2,6, 6-tetramethyl-1, 2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one hydrochloride (230mg, 0.380mmol, 67.1% yield) as a pale yellow powder.
UPLC-MS: 0.93min, 568.7[ M + H ] +, method 9
And b, step b.
A solution of 1M boron tribromide in DCM (2.1ml, 2.100mmol) was added to a solution of 3- (1- (4-amino-3- (3-fluoro-5-methoxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (2,2,6, 6-tetramethyl-1, 2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one hydrochloride (226mg, 0.373mmol) in DCM (12ml) and the resulting mixture was stirred at rt for 16H. The suspension was added slowly to EtOH at 0 ℃. The resulting mixture was concentrated under reduced pressure. Purification by RP-flash chromatography (Biotage 30g C18 column, gradient elution 100: 0-60: 40A/B, 15 CV; A: water/MeCN 95/5+ 0.01% HCOOH, B: water/MeCN 5/95+ 0.01% HCOOH) gave the title compound (168mg, 0.284mmol, 76% yield) as a light yellow powder.
1H NMR(600MHz，DMSO-d6)ppm 10.07-10.45(bs，1H)，9.11-9.49(m，1H)，8.43(d，J＝2.30Hz，1H)，8.07-8.25(m，1H)，7.55-8.01(m，3H)，6.95(dt，J＝19.07，1.64Hz，1H)，6.66-6.90(m，2H)，5.94-6.18(m，1H)，2.94(d，J＝18.09Hz，1H)，2.89(m，1H)，1.87-2.06(d，3H)，1.48-1.72(m，12H)。UPLC-MS：2.69min，555.1[M+H]Method 6
Example 145
3- (1- (4-amino-3- (5-hydroxypyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (2,2,6, 6-tetramethyl-1, 2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one dihydrochloride
Step a.3- (1- (4-amino-3- (5-methoxypyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (2,2,6, 6-tetramethyl-1, 2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one hydrochloride
The title compound was prepared from 3- (1-bromoethyl) -4- (2,2,6, 6-tetramethyl-1, 2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one hydrobromide (intermediate C39, 0.100g, 0.212mmol) and 3- (5-methoxypyridin-3-yl) -1H-pyrazolo [3,4-D ] pyrimidin-4-amine (0.051g, 0.212mmol) in analogy to intermediate D1 to give 3- (1- (4-amino-3- (5-methoxypyridin-3-yl) -1H-pyrazolo [3,4-D ] pyrimidin-1-yl) ethyl) -4- (2,2,6, 6-tetramethyl-1, 2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one hydrochloride (56.0mg, 0.095mmol, 44.9% yield) as a white powder.
UPLC-MS: 0.87min, 551.8[ M + H ] +, method 9.
And b, step b.
The title compound was prepared from 3- (1- (4-amino-3- (5-methoxypyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (2,2,6, 6-tetramethyl-1, 2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one hydrochloride (53.7mg, 0.091mmol) according to the method reported for the compound of example 144 to give the title compound (17.6mg, 0.029mmol, 31.6% yield) as a white powder.
1H NMR(600MHz，DMSO-d6)ppm 9.98-10.30(bs，1H)，6.89-8.32(m，10H)，5.88-6.13(m，2H)，3.45(m，1H)，2.96(m，1H)，1.98(d，J＝7.23Hz，3H)，0.98-1.60(m，12H)。UPLC-MS：1.65min，538.1[M+H]+, method 6.
Example 150
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1- (azetidine-3-carbonyl) -1,2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one formate salt
Step a.3- (4- (3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -1-oxo-1H-isochromen-4-yl) -1,2,3, 6-tetrahydropyridine-1-carbonyl) azetidine-1-carboxylic acid tert-butyl ester
The title compound was prepared from tert-butyl 3- (4- (3- (1- (4-amino-3-iodo-1H-pyrazolo [3,4-D ] pyrimidin-1-yl) ethyl) -1-oxo-1H-isochromen-4-yl) -1,2,3, 6-tetrahydropyridine-1-carbonyl) azetidine-1-carboxylate (intermediate D28, 0.251g, 0.360mmol) and 3-fluoro-5-hydroxyphenylboronic acid (0.112g, 0.720mmol) according to the method reported for example 68 to give 3- (4- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -1-oxo-1H-isochromen-4-yl) -1,2,3, 6-tetrahydropyridine-1-carbonyl) azetidine-1-carboxylic acid tert-butyl ester (0.155g, 0.227mmol, 63.2% yield).
UPLC-MS: 1.03min, 682.4[ M + H ] +, method 9
And b, step b.
A 4M HCl solution in 1, 4-dioxane (2ml, 8.00mmol) was added to a stirred solution of tert-butyl 3- (4- (3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -1-oxo-1H-isochromen-4-yl) -1,2,3, 6-tetrahydropyridine-1-carbonyl) azetidine-1-carboxylate (0.153g, 0.224mmol) in 1, 4-dioxane (1 ml). After stirring at rt for 1h, volatiles were removed under reduced pressure. Purification by RP flash chromatography (BiotageIsolera, 30g C18 column, gradient elution 100: 0-80: 20A/B, 15 CV; A: water/MeCN 95/5+ 0.01% HCOOH, B: water/MeCN 5/95+ 0.01% HCOOH) gave the title compound (92.2mg, 0.147mmol, 65.5% yield) as a white powder.
1H NMR(600MHz，DMSO-d6)ppm 6.65-8.34(m，11H)，6.23(m，2H)，3.30-4.29(m，11H)，1.82(d，3H)。UPLC-MS：2.36min，582.0[M+H]+, method 6.
Example 151
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1- (1-methylazetidin-3-carbonyl) -1,2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one formate salt
Reacting 3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3, 4-d)]Pyrimidin-1-yl) ethyl) -4- (1- (azetidine-3-carbonyl) -1,2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one formate (example 150, 71.0mg, 0.113mmol), DIPEA (0.020ml, 0.113mmol), paraformaldehyde (69.3mg, 2.30mmol) and spatula Na2SO4The mixture in DCM (2ml) was stirred at rt for 10 min. Then acetic acid (0.033ml, 0.589mmol) was added followed by sodium triacetoxyborohydride (82.2mg, 0.387 mmol). After stirring at rt 6h, volatiles were removed under reduced pressure. The residue was dissolved in DCM/EtOH95:5(10ml), washed with saturated sodium bicarbonate, then filtered through a phase separator and concentrated under reduced pressure. The residue was dissolved in DCM/MeOH 10: 1(1ml) followed by formaldehyde (0.016ml, 0.170mmol) and AcOH (0.014ml, 0.250 mmol). After 5min stirring, NaBH (OAc) is added3(25mg, 0.118 mmol). The resulting mixture was stirred at rt for 30min, then concentrated under reduced pressure. Purification by RP flash chromatography (Bi) otage Isolera, 12g C18 column, gradient elution 100: 0-60: 40A/B, 15 CV; a: water/MeCN 95/5+ 0.01% HCOOH, B: water/MeCN 5/95+ 0.01% HCOOH) to give the title compound (59.8mg, 0.093mmol, 82% yield) as a white solid.
1H NMR(600MHz，DMSO-d6)ppm 1.90(m，3H)，2.52(t，J＝5.59Hz，2H)，2.75-4.56(m，12H)，5.94-6.35(m，1H)，6.62-6.74(m，1H)，6.76-6.89(m，1H)，6.89-7.01(m，1H)，7.36-7.54(m，1H)，7.57-7.69(m，1H)，7.78-7.91(m，1H)，8.04-8.84(m，2H)，10.04-10.49(m，1H)。UPLC-MS：2.41min，596.1[M+H]+, method 6.
Example 154
3- (1- (4-amino-3- (3-chloro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1-methyl-1, 2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one formate salt
The title compound was prepared from 3- (1- (4-amino-3- (3-chloro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1,2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one formate (example 152, 200mg, 0.357mmol) according to the method reported for example 151 to give the title compound (92.5mg, 0.161mmol, 45.1% yield) as a white powder.
1H NMR(600MHz，DMSO-d6)ppm 6.81-8.40(m，11H)，6.04-6.20(m，1H)，5.96(br.s.，1H)，2.24-3.26(m，9H)，1.79-1.95(d，3H)
UPLC-MS: 2.60min, 529.0[ M + H ] +, method 6
Example 155
3- (1- (4-amino-3- (3-hydroxy-5- (trifluoromethyl) phenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1-methyl-1, 2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one formate salt
The title compound was synthesized according to the method described for preparation example 151 from 3- (1- (4-amino-3- (3-hydroxy-5- (trifluoromethyl) phenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1,2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one formate (example 153, 215mg, 0.362mmol) to give the title compound (115mg, 0.189mmol, 52.3% yield) as a white powder.
1H NMR(600MHz，DMSO-d6)ppm 7.12-8.28(m，11H)，6.58-7.09(m，1H)，5.97(br.s.，1H)，2.30-3.31(m，9H)，1.79-1.98(d，3H)。UPLC-MS：2.85min，563.1[M+H]Method 6
Example 156
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1- (azetidin-3-yl) -1,2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one dihydrochloride
Step a.3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1,2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one formate
Make N2Through 4- (3- (1- (4-amino-3-iodo-1H-pyrazolo [3, 4-d)]Pyrimidin-1-yl) ethyl) -1-oxo-1H-isochromen-4-yl) -5, 6-dihydropyridine-1 (2H) -carboxylic acid tert-butyl ester (intermediate D14, 735mg, 1.197mmol), 3-fluoro-5-hydroxyphenylboronic acid (0.28G, 1.795mmol), S-Phos-Pd-G2(0.130G, 0.179mmol) and potassium phosphate (572mg, 2.695mmol) in THF/water 3: the suspension in 1(8ml) was bubbled for 5 min. The mixture was heated at 85 ℃ for 1h by MW irradiation. A second experiment was performed under the same conditions on practically the same amount of reagent mixture. The two reaction mixtures were combined and concentrated HCl (20ml) was added slowly with stirring. Stirring was continued at rt for 3h, then volatile materials were removed under reduced pressure. Purification by RP flash chromatography (Biotage Isolera, column 60g C18, gradient elution 100: 0-70: 30A/B, 15 CV; A: water/MeCN 95/5+ 0.01% HCOOH, B: water/MeCN 5/95+ 0.01% HCOOH) afforded 3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3, 4-d) ]Pyrimidin-1-yl) ethyl) -4- (1,2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one formate (0.990g, 1.818mmol, 76% yield) as a light yellow powder.
UPLC-MS: 0.67min, 499.2[ M + H ] +, method 2min
Step b.3- (4- (3- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -1-oxo-1H-isochromen-4-yl) -5, 6-dihydropyridin-1 (2H) -yl) azetidine-1-carboxylic acid tert-butyl ester formate salt
The compound was prepared from 3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-D ] pyrimidin-1-yl) ethyl) -4- (1,2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one formate (300mg, 0.551mmol) and tert-butyl 3-oxoazetidine-1-carboxylate (236mg, 1.379mmol) according to the procedure described for the synthesis of intermediate D24 to give 3- (4- (3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-D ] pyrimidin-1-yl) ethyl) -1-oxo-1H-pyrazol-1H -isochromen-4-yl) -5, 6-dihydropyridin-1 (2H) -yl) azetidine-1-carboxylic acid tert-butyl ester formate (74,4mg, 0.106mmol, 19.30% yield).
UPLC-MS: 0.85min, 654.3[ M + H ] +, method 9
And c, step (c).
4.0M HCl in 1, 4-dioxane (0.5ml, 2.000mmol) was added to 3- (4- (3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -1-oxo-1H-isochromen-4-yl) -5, 6-dihydropyridin-1 (2H) -yl) azetidine-1-carboxylic acid tert-butyl ester formate (70mg, 0.100mmol) in 1, 4-dioxane-methanol 3: 1(4 ml). The mixture was stirred at rt for 6h, then the volatiles were removed under reduced pressure. Purification by RP-flash chromatography (Biotage 30g C18 column, gradient elution 100: 0-60: 40A/B, 15 CV; A: water/MeCN 95/5+ 0.01% HCOOH, B: water/MeCN 5/95+ 0.01% HCOOH) gave the title compound (57mg, 0.091mmol, 91% yield) as a white powder.
1H NMR(600MHz，DMSO-d6)ppm 12.70-13.40(br.s.，1H)，10.32(br.s.，1H)，9.73(br.s.，1H)，9.22(br.s.，1H)，6.71-8.58(m，10H)，5.95-6.37(m，2H)，2.51-4.79(m，11H)，1.76-2.04(d，3H)。UPLC-MS：2.32min，554.1[M+H]Method 6
Example 157
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1- (1- (cyclopropylmethyl) azetidin-3-yl) -1,2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one dihydrochloride
The title compound was prepared from 3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-D ] pyrimidin-1-yl) ethyl) -4- (1- (azetidin-3-yl) -1,2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one dihydrochloride (example 156, 35mg, 0.059mmol) and cyclopropanealdehyde (10.19 μ l, 0.136mmol) as described for synthetic intermediate D24 to give the compound (33.8mg, 0.050mmol, 84% yield) as a white powder.
1H NMR(600MHz，DMSO-d6)ppm 0.42(br s，2H)，0.60(br d，J＝4.93Hz，2H)，0.94-1.09(m，1H)，1.95(m，3H)，2.35-4.59(m，11H)，4.62-4.96(m，2H)，6.01-8.79(m，12H)，10.08-10.63(br，1H)，11.18-11.66(br，1H)，12.98-13.61(br，1H)。UPLC-MS：2.68min，608.2[M+H]Method 6
Example 158
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (3- (dimethylamino) prop-1-yn-1-yl) -1H-isochromen-1-one hydrochloride
Benzyl (3- (3- (1- (4-amino-3- (3-fluoro-5-methoxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -1-oxo-1H-isochromen-4-yl) prop-2-yn-1-yl) (methyl) carbamate
The title compound was prepared from 3- (3-fluoro-5-methoxyphenyl) -1H-pyrazolo [3,4-D ] pyrimidin-4-amine (intermediate G1, 165mg, 0.638mmol) and benzyl (3- (3- (1-bromoethyl) -1-oxo-1H-isochromen-4-yl) prop-2-yn-1-yl) (methyl) carbamate (intermediate C41, 290mg, 0.638mmol) according to the procedure described for the synthesis of intermediate D1 to give (3- (3- (1- (4-amino-3- (3-fluoro-5-methoxyphenyl) -1H-pyrazolo [3,4-D ] pyrimidin-1-yl) ethyl) -1-oxo-1H-isochromen-4- Yl) prop-2-yn-1-yl) (methyl) carbamic acid benzyl ester (138mg, 0.218mmol, 34.2% yield) as a yellow solid.
UPLC-MS: 1.27min, 633[ M + H ] +, method 9
And b, step b.
To (3- (3- (1- (4-amino-3- (3-fluoro-5-methoxyphenyl) -1H-pyrazolo [3, 4-d)]Pyrimidin-1-yl) ethyl) -1-oxo-1H-isochromen-4-yl) prop-2-yn-1-yl) (methyl) benzyl carbamate (100mg, 0.158mmol) in DCM (4ml) was added 1M BBr3Was added to the reaction solution (3ml, 10.50mmol) in DCM and the reaction was stirred overnight. The solution was then cooled to 0 ℃ and stirred for 1 h. EtOH (1ml) was then added and the solvent removed under reduced pressure.
The resulting material was reacted with trioxane (0.049ml, 0.472mmol), N-isopropyl-N-methylpropan-2-amine (18.14mg, 0.157mmol) in DCM (4ml) for 30 min. Sodium triacetoxyborohydride (100mg, 0.472mmol) and acetic acid (28.4mg, 0.472mmol) were then added and the reaction stirred for 1 h. The solvent was removed and the crude product was purified by flash chromatography on C18 ((H)2O/ACN))95:5+0.1％HCOOH}：{(ACN/H2O)95:5+ HCOOH 0.1% } 100: 0-0: 100 to give the title compound (5mg, 10.03 μmol, 6.37% yield) as a white solid.
1H NMR(600MHz，DMSO-d6)ppm 10.34-10.50(bs，1H)，10.13-10.27(bs，1H)，8.26-8.31(m，1H)，8.16-8.21(m，1H)，7.89-8.03(m，2H)，7.67-7.78(m，1H)，6.90-6.95(m，1H)，6.83-6.89(m，1H)，6.66-6.76(m，1H)，6.40-6.52(m，1H)，4.35-4.55(m，2H)，2.89-3.02(m，6H)，1.92-1.98(m，3H)。UPLC-MS：2.36min，499.2[M+H]Method 6
Example 161
3- (1- (4-amino-3- (5-hydroxy-6- (trifluoromethyl) pyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one
According to analogy to example 68Using 3- (1- (4-amino-3-iodo-1H-pyrazolo [3, 4-d) ]Pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one (intermediate D2a, 100mg, 0.196mmol) and (5- (benzyloxy) -6- (trifluoromethyl) pyridin-3-yl) boronic acid (intermediate G25, 87mg, 0.295mmol) the title compound was prepared. The resulting crude material was reacted in 2-propanol (10ml) with 1N aqueous HCl (1ml), palladium on carbon 5% wet (0.071mmol) and hydrogen (1 atm). The catalyst was then filtered off through a pad of celite and the solvent was removed. The desired product was purified by C18 flash chromatography (Snap 30g chromatography ((H)2O/ACN))95:5+0.1％HCOOH)：{(ACN/H2O)95:5+ HCOOH 0.1% }100:0-0:100 to give the title compound (20mg, 0.037mmol, 51.8% yield) as a white solid.
1H NMR(400MHz，DMSO-d6)ppm 11.47(br.s.，1H)，6.52-8.35(m，14H)，5.82(d，J＝7.06Hz，1H)，1.83-1.99(d，3H)。UPLC-MS：3.30min，545.1[M+H]+, method 6.
Example 162
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -7-methyl-4-phenyl-1H-isochromen-1-one
3- (1- (4-amino-3-iodo-1H-pyrazolo [3, 4-d)]Pyrimidin-1-yl) ethyl) -7-methyl-4-phenyl-1H-isochromen-1-one (intermediate D29, 0.100g, 0.19mmol), (3-fluoro-5-hydroxyphenyl) boronic acid (0.036g, 0.229mmol) and PPh3(0.030g, 0.114mmol) in a mixture of DMF (10ml), EtOH (4ml) and water (4 ml); adding Na2CO3(0.101g, 0.95mmol), the mixture was degassed under a nitrogen atmosphere. Pd (OAc) is added 2(0.009g, 0.038mmol) and the reaction was heated at 80 ℃ for 15 min. 1M HCl (pH. apprxeq.2) was added and the mixture partitioned between EtOAc and water. The organic phase was extracted with EtOAc, the combined organic layers were washed several times with brine,dried over magnesium sulfate. The solvent was removed and the crude product was purified by flash chromatography using a Biotage silica gel column (DCM-DCM: MeOH ═ 97:3) to give the title compound (0.023g, 0.045mmol, 24%).
1H NMR(400MHz，DMSO-d6)ppm 10.19(s，1H)，8.09(s，1H)，8.03(br.s，1H)，7.47-7.69(m，2H)，7.30-7.47(m，3H)，7.09-7.14(m，1H)，6.89-6.92(m，1H)，6.77-6.86(m，2H)，6.66(dt，1H)，6.00-8.00(m，2H)，5.68-5.76(m，1H)，2.42(s，3H)，1.82(d，3H)。UPLC-MS：1.10min，508.2[M+H]+, method 13.
Example 163
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -7-chloro-4-phenyl-1H-isochromen-1-one
The title compound was prepared in analogy to example 162 from 3- (1- (4-amino-3-iodo-1H-pyrazolo [3,4-D ] pyrimidin-1-yl) ethyl) -7-chloro-4-phenyl-1H-isochromen-1-one (intermediate D30, 0.100g, 0.184mmol) and (3-fluoro-5-hydroxyphenyl) boronic acid (0.034g, 0.22mmol) to yield the title compound (0.05g, 0.094mmol, 51%).
1H NMR(400MHz，DMSO-d6)ppm 10.21(s，1H)，8.18(d，1H)，8.10(s，1H)，7.83(dd，1H)，7.51-7.57(m，1H)，7.37-7.49(m，3H)，7.13-7.17(m，1H)，6.86-6.95(m，2H)，6.81-6.87(m，1H)，6.67(dt，1H)，6.00-8.50(m，2H)，5.70-5.77(m，1H)，1.83(d，3H)。UPLC-MS：1.15min，527.9[M+H]+, method 12.
Example 164
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -6-chloro-4-phenyl-1H-isochromen-1-one
The title compound was prepared in analogy to example 162 from 3- (1- (4-amino-3-iodo-1H-pyrazolo [3,4-D ] pyrimidin-1-yl) ethyl) -6-chloro-4-phenyl-1H-isochromen-1-one (intermediate D31, 0.100g, 0.184mmol) and (3-fluoro-5-hydroxyphenyl) boronic acid (0.034g, 0.221mmol) to yield the title compound as a white solid (0.023g, 0.0463mmol, 24%).
1H NMR(400MHz，DMSO-d6)ppm 10.25(br.s.，1H)，8.22(d，1H)，8.09(s，1H)，7.68(dd，1H)，7.51-7.58(m，1H)，7.36-7.49(m，3H)，7.13-7.18(m，1H)，6.88-6.92(m，1H)，6.80-6.85(m，1H)，6.76(d，1H)，6.63-6.69(m，1H)，6.00-8.00(m，2H)，5.68-5.76(m，1H)，1.82(d，3H)。UPLC-MS：1.17min，528.3[M+H]+, method 14.
Example 165
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -6-fluoro-4-phenyl-1H-isochromen-1-one
3- (1- (4-amino-3-iodo-1H-pyrazolo [3, 4-d) is administered in a sealed vial]Pyrimidin-1-yl) ethyl) -6-fluoro-4-phenyl-1H-isochromen-1-one (intermediate D32, 0.240g, 0.455mmol), (3-fluoro-5-hydroxyphenyl) boronic acid (0.156g, 1mmol) and K2CO3(0.138g, 1mmol) in dioxane/H2O4: 1(10ml) of the mixture was degassed; adding Pd (dppf) Cl2(0.04g, 0.054mmol), the reaction was heated at 120 ℃ for 4 h. 1M HCl (pH 1) was added and the mixture partitioned between EtOAc and water. The aqueous phase was extracted with EtOAc and the combined organic layers were washed with brine and dried over sodium sulfate. The solvent was removed and the crude product was purified by flash chromatography using a Biotage silica gel column (cyclohexane: EtOAc 95:5 to 20:80) and then triturated with CAN to give the title compound as a white solid (0.08g, 0.16 mmol).
1H NMR(400MHz，DMSO-d6)ppm 10.19(s，1H)，8.27-8.33(m，1H)，8.09(s，1H)，7.36-7.57(m，5H)，7.13-7.18(m，1H)，6.86-6.94(m，1H)，6.80-6.86(m，1H)，6.66(dt，1H)，6.49(dd，1H)，6.00-7.80(m，2H)，5.66-5.78(m，1H)，1.83(d，3H)。
UPLC-MS: 1.08min, 512.2[ M + H ] +, method 13.
Examples 166a (enantiomer 1) and 166b (enantiomer 2): 4- (1-acetyl-1, 2,3, 6-tetrahydropyridin-4-yl) -3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -1H-isochromen-1-one single enantiomer
Step 1.4- (1-acetyl-1, 2,3, 6-tetrahydropyridin-4-yl) -3- (1- (4-amino-3- (3- ((tert-butyldimethylsilyl) oxy) -5-fluorophenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -1H-isochromen-1-one (intermediate T.1)
4- (1-acetyl-1, 2,3, 6-tetrahydropyridin-4-yl) -3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -1H-isochromen-1-one
Example 72, 726mg, 1.343mmol was dissolved in N, N-dimethylformamide (7.5ml) and then TBDMSCl (405mg, 2.69mmol) and imidazole (366mg, 5.37mmol) were added. The mixture was stirred at rt for 3h, then 0,3eq TBDMSCl and imidazole were added to complete the reaction. The reaction was diluted with DCM (100ml), washed 2 times with 0.5M aqueous HCl (50ml), filtered through a phase separator and concentrated under reduced pressure. The residue was purified by flash chromatography on silica gel using a Biotage 100G SNAP gradient with DCM and EtOH to give the title compound (635mg, 72.2%) as a pink foam.
UPLC-MS: 1.40min, 655.0[ M + H ] +, method 9
Step 2.4- (1-acetyl-1, 2,3, 6-tetrahydropyridin-4-yl) -3- (1- (4-amino-3- (3- ((tert-butyldimethylsilyl) oxy) -5-fluorophenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -1H-isochromen-1-one single enantiomer (T.2 and T3)
The racemate 4- (1-acetyl-1, 2,3, 6-tetrahydropyridin-4-yl) -3- (1- (4-amino-3- (3- ((tert-butyldimethylsilyl) oxy) -5-fluorophenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -1H-isochromen-1-one (intermediate t.1, 0.759g, 1.16mmol) was dissolved in 14ml of ethanol/methanol 1/1+6ml of n-hexane and subjected to chiral resolution by chiral preparative chromatography. Conditions are as follows: column: chiralpak AS-H (25X 2.0cm), 5. mu.; mobile phase: n-hexane/(2-propanol/methanol 1/1+ 0.1% isopropylamine) 85/15% v/v; flow rate: 17 ml/min; and D, DAD detection: 220 nm; loop circuit: 700 mul; and (3) injection: 26.6 mg/injection.
Fractions containing the enantiomer of the first eluate were evaporated to dryness to give intermediate t.2 (first eluted enantiomer, 0.350g, 0.535 mmol). Chiral HPLC (method a 10): rt 13.6min, ee > 99%. UPLC-MS: 1.32min, 655.5[ M + H ] +, method 13.
The fractions containing the enantiomer of the second eluate were evaporated to dryness to give intermediate T.3 (second eluted enantiomer, 0.365g, 0.557 mmol). Chiral HPLC (method a 10): rt 20.6min, ee > 99%. UPLC-MS: 1.33min, 655.5[ M + H ] +, method 13.
Step 3. example 166a (enantiomer 1): 4- (1-acetyl-1, 2,3, 6-tetrahydropyridin-4-yl) -3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -1H-isochromen-1-one single enantiomer
4- (1-acetyl-1, 2,3, 6-tetrahydropyridin-4-yl) -3- (1- (4-amino-3- (3- ((tert-butyldimethylsilyl) oxy) -5-fluorophenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -1H-isochromen-1-one single enantiomer (intermediate T.2, the enantiomer eluting first under the conditions described above, 0.350g, 0.535mmol) was dissolved in 1MHCl in EtOH (1.7ml) and the mixture was stirred at RT overnight. The volatiles were removed under reduced pressure and the residue was purified by flash chromatography on silica gel (DCM-DCM: MeOH ═ 95:5) to give the title compound as a yellow solid (0.270g, 0.500mmol, 93%). This compound was confirmed to be the second eluting enantiomer under chiral HPLC conditions of method a 11: rt 15.8min, ee 99.4%.
1H NMR(500MHz，DMSO-d6)ppm 10.28(br.s.，1H)，8.27-8.54(m，1H)，8.06-8.25(m，1H)，7.80-7.96(m，1H)，7.56-7.72(m，1H)，7.37-7.52(m，1H)，6.79-6.98(m，2H)，6.65-6.75(m，1H)，6.49-8.83(m，2H)，6.07-6.35(m，1H)，5.27-6.07(m，1H)，3.53-4.33(m，4H)，1.96-2.44(m，5H)，1.62-1.94(m，3H)。UPLC-MS：0.83min，541.3[M+H]+, method 13.
Step 4 example 166b (enantiomer 2)4- (1-acetyl-1, 2,3, 6-tetrahydropyridin-4-yl) -3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -1H-isochromen-1-one single enantiomer
4- (1-acetyl-1, 2,3, 6-tetrahydropyridin-4-yl) -3- (1- (4-amino-3- (3- ((tert-butyldimethylsilyl) oxy) -5-fluorophenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -1H-isochromen-1-one single enantiomer (intermediate T.3, enantiomer of the second eluent under the above conditions, 0.365g, 0.557mmol) was dissolved in 1M EtOH solution of HCl (1.78mL) and the mixture was stirred at RT overnight. The volatiles were removed under reduced pressure and the residue was purified by flash chromatography on silica gel (DCM-DCM: MeOH ═ 95:5) to give the title compound as a yellow solid (0.260g, 0.481mmol, 86%). This compound was confirmed to be the first eluting enantiomer under chiral HPLC conditions of method a 11: rt 13.6min, ee 99.6%.
1H NMR(500MHz，DMSO-d6)ppm 10.28(br.s.，1H)，8.30-8.55(m，1H)，8.12-8.24(m，1H)，7.82-7.94(m，1H)，7.57-7.71(m，1H)，7.40-7.51(m，1H)，6.80-6.97(m，2H)，6.63-6.76(m，1H)，6.49-8.83(m，2H)，6.07-6.32(m，1H)，5.26-6.06(m，1H)，3.59-4.30(m，4H)，1.94-2.48(m，5H)，1.77-1.94(m，3H)。UPLC-MS：0.83min，541.3[M+H]+, method 13.
Examples 167a (enantiomer 1) and 167b (enantiomer 2)
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (thiazol-5-yl) -1H-isochromen-1-one single enantiomer
Step 1.3- (1- (4-amino-3- (3- ((tert-butyldimethylsilyl) oxy) -5-fluorophenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (thiazol-5-yl) -1H-isochromen-1-one (intermediate R.1)
To a solution of 3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (thiazol-5-yl) -1H-isochromen-1-one (260mg, 0.519mmol) in dry DMF (10ml) were added imidazole (0.137ml, 2.078mmol) and TBDMSCl (0.270ml, 1.558mmol), and the solution was stirred overnight. DMF was removed and the residue was dissolved in DCM (50mL) and washed with 0.5N aqueous HCl. The organic phase was dried and the solvent was removed. The crude product was purified by flash chromatography on silica gel (SNAP 25g DCM: EtOH 100:0-95:5) to give 3- (1- (4-amino-3- (3- ((tert-butyldimethylsilyl) oxy) -5-fluorophenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (thiazol-5-yl) -1H-isochromen-1-one (210mg, 0.342mmol, 65.8% yield) as a light yellow solid.
UPLC-MS: 1.44min, 615[ M + H ] +, method 9.
Step 2.3- (1- (4-amino-3- (3- ((tert-butyldimethylsilyl) oxy) -5-fluorophenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (thiazol-5-yl) -1H-isochromen-1-one single enantiomer (intermediates R.2 and R.3)
The racemate 3- (1- (4-amino-3- (3- ((tert-butyldimethylsilyl) oxy) -5-fluorophenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (thiazol-5-yl) -1H-isochromen-1-one (intermediate r.1, 0.210g, 0.34mmol) was dissolved in EtOH (8ml) and subjected to chiral resolution by chiral preparative chromatography. Conditions are as follows: column: whelk O-1(R, R) (25x2cm), 10 um; mobile phase: n-hexane/(2-propanol + 0.1% isopropylamine) 40/60% v/v; flow rate: 18 ml/min; and D, DAD detection: 220 nm; loop circuit: 1000 μ l; and (3) injection: 26mg (per injection).
Fractions containing the enantiomer of the first eluate were evaporated to dryness to give intermediate r.2 (first eluted enantiomer, 0.084g, 0.13 mmol). Chiral HPLC (method a 6): rt ═ 14.2min, ee > 99%.
UPLC-MS: 1.40min, 615.4[ M + H ] +, method 13
Fractions containing the enantiomer of the second eluent were evaporated to dryness to give intermediate r.3 (second eluted enantiomer, 0.089g, 0.14 mmol). Chiral HPLC (method a 6): rt 18.8min, ee 98.6%.
UPLC-MS: 1.41min, 615.4[ M + H ] +, method 13
Step 3 example 167 a: 3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (thiazol-5-yl) -1H-isochromen-1-one single enantiomer
The single enantiomer of 3- (1- (4-amino-3- (3- ((tert-butyldimethylsilyl) oxy) -5-fluorophenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (thiazol-5-yl) -1H-isochromen-1-one (intermediate R2, the enantiomer eluting first under the above conditions, 0.084g, 0.13mmol) was dissolved in 1M EtOH solution of HCl (0.416ml) and the mixture was stirred at RT overnight. The volatiles were removed under reduced pressure and the residue was purified by flash chromatography on silica gel (DCM-DCM: MeOH ═ 95:5) to give the title compound as a pale blue solid (0.043g, 0.086mmol, 66%). This compound was confirmed to be the first eluting enantiomer under chiral HPLC conditions of method a 7: rt 16.0min, ee > 99%.
1H NMR(400MHz，DMSO-d6)ppm 10.20(s，1H)，9.29(s，1H)，8.19-8.25(m，1H)，8.14(s，1H)，7.92(br.s.，1H)，7.80-7.86(m，1H)，7.63-7.70(m，1H)，7.04(d，1H)，6.89-6.92(m，1H)，6.81-6.86(m，1H)，6.64-6.70(m，1H)，5.86(q，1H)，5.75-8.50(m，2H)，1.86(d，3H)。UPLC-MS：0.87min，501.3[M+H]Method 13
Step 4 example 167 b: 3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (thiazol-5-yl) -1H-isochromen-1-one single enantiomer
3- (1- (4-amino-3- (3- ((tert-butyldimethylsilyl) oxy) -5-fluorophenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (thiazol-5-yl) -1H-isochromen-1-one single enantiomer (intermediate R.3, enantiomer from the second eluent under the above conditions, 0.089g, 0.14mmol) was dissolved in 1M EtOH solution of HCl (0.5ml) and the mixture was stirred at RT overnight. The volatiles were removed under reduced pressure and the residue was purified by flash chromatography on silica gel (DCM-DCM: MeOH ═ 95:5) to give the title compound as a pale blue solid (0.048g, 0.096mmol, 74%). This compound was confirmed to be the second eluting enantiomer under chiral HPLC conditions of method a 7: rt 20.1min, ee 98.4%.
1H NMR(400MHz，DMSO-d6)ppm 10.21(s，1H)，9.29(s，1H)，8.20-8.24(m，1H)，8.15(s，1H)，7.92(br.s.，1H)，7.79-7.86(m，1H)，7.63-7.69(m，1H)，7.04(d，1H)，6.89-6.92(m，1H)，6.80-6.86(m，1H)，6.63-6.70(m，1H)，5.87(q，1H)，5.75-8.50(m，2H)，1.86(d，3H)。UPLC-MS：0.90min，501.2[M+H]Method 14
Examples 168a (enantiomer 1) and 178b (enantiomer 2): 3- (1- (4-amino-3- (5-hydroxypyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one hydrochloride as a single enantiomer
Step 1.3- (1- (4-amino-3- (5- ((tert-butyldimethylsilyl) oxy) pyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one (intermediate S.1)
To a stirred mixture of 3- (1- (4-amino-3- (5-hydroxypyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one (example 91, 0.260g, 0.546mmol) and imidazole (0.093g, 1.36mmol) in DMF (5.0ml) was added TBDMSCl (0.206g, 1.36mmol) at RT and the mixture stirred for 2H. The mixture was partitioned between water and DCM, the aqueous phase was extracted with DCM, and the combined organic layers were washed with brine and dried over sodium sulfate. The solvent was removed under reduced pressure. The residue was purified by silica gel column flash chromatography (DCM: MeOH ═ 95:5-90:10) to give 3- (1- (4-amino-3- (5- ((tert-butyldimethylsilyl) oxy) pyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one as a white solid (intermediate S1, 0.178g, 0.301mmol, 55%).
UPLC-MS: 1.41min, 591.4[ M + H ] +, method 13
Step 2.3- (1- (4-amino-3- (5- ((tert-butyldimethylsilyl) oxy) pyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one single enantiomer (intermediates S.2 and S.3)
The racemate 3- (1- (4-amino-3- (5- ((tert-butyldimethylsilyl) oxy) pyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one (intermediate s.1, 0.178g, 0.301mmol) was dissolved in ethanol/DCM 1/1(7ml) and subjected to chiral resolution by chiral preparative chromatography. Conditions are as follows: column: whelk O1(R, R) (25X2.11cm), 10. mu.l; mobile phase: n-hexane/(ethanol + 0.1% isopropylamine + 10% DCM) 70/30% v/v; flow rate: 17 ml/min; and D, DAD detection: 220 nm; loop circuit: 500 mul; and (3) injection: 13.5 mg/injection.
Fractions containing the enantiomer of the first eluate were evaporated to dryness to give intermediate s.2 (first eluted enantiomer, 0.070g, 0.118 mmol). Chiral HPLC (method A8): rt 18.9min, ee > 99%. UPLC-MS: 1.43min, 591.3[ M + H ] +, method 16.
The fraction containing the enantiomer of the second eluent was evaporated to dryness to give intermediate s.3 (second eluted enantiomer, 0.077g, 0.130 mmol). Chiral HPLC (method A8): rt 21.9min, ee > 99%. UPLC-MS: 1.43min, 591.3[ M + H ] +, method 16.
Step 3. example 168a (enantiomer 1): 3- (1- (4-amino-3- (5-hydroxypyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one hydrochloride as a single enantiomer
3- (1- (4-amino-3- (5- ((tert-butyldimethylsilyl) oxy) pyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one single enantiomer (intermediate S.2, the enantiomer eluting first under the above conditions, 0.070g, 0.118mmol) was dissolved in 1M HCl in EtOH (0.38mL) and the mixture was stirred at RT for 5H. The volatiles were removed under reduced pressure and the residue was purified by flash chromatography on silica gel (DCM-DCM: MeOH ═ 95:5) to give the title compound as a white solid (0.059g, 0.115mmol, 97%). This compound was confirmed to be the second eluting enantiomer under chiral HPLC conditions of method a 9: rt 15.4min, ee > 99%.
1H NMR(400MHz，DMSO-d6)11.36(br.s.，1H)，8.39-8.48(m，2H)，8.20-8.29(m，2H)，7.95(br.s.，2H)，7.74-7.86(m，2H)，7.60-7.66(m，1H)，7.50-7.57(m，1H)，7.37-7.49(m，3H)，7.11-7.18(m，1H)，6.90(d，1H)，5.79(q，1H)，1.87(d，3H)。UPLC-MS：0.82min，477.3[M+H]+, method 13.
Step 4. example 168b (enantiomer 2): 3- (1- (4-amino-3- (5-hydroxypyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one hydrochloride as a single enantiomer
3- (1- (4-amino-3- (5- ((tert-butyldimethylsilyl) oxy) pyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one single enantiomer (intermediate S.3, the enantiomer eluting second under the above conditions, 0.077g, 0.130mmol) was dissolved in HCl 1M in EtOH (0.417ml) and the mixture was stirred at RT for 5H. The volatiles were removed under reduced pressure and the residue was purified by flash chromatography on silica gel (DCM-DCM: MeOH ═ 95:5) to give the title compound as a white solid (0.034g, 0.066mmol, 51% yield). This compound was confirmed to be the first eluting enantiomer under chiral HPLC conditions of method a 9: rt 13.2min, ee 95.6%.
1H NMR(400MHz，DMSO-d6)11.47(br.s.，1H)，8.43-8.48(m，2H)，8.28(s，1H)，8.20-8.25(m，1H)，8.04(br.s.，2H)，7.81-7.86(m，1H)，7.74-7.81(m，1H)，7.60-7.66(m，1H)，7.50-7.57(m，1H)，7.37-7.48(m，3H)，7.11-7.18(m，1H)，6.89(d，1H)，5.80(q，1H)，1.87(d，3H)。UPLC-MS：0.82min，477.3[M+H]+, method 13.
Example 169
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1-benzyl-1, 2,5, 6-tetrahydropyridin-3-yl) -1H-isochromen-1-one hydrochloride
The title compound was prepared in analogy to example 162 from 3- (1- (4-amino-3-iodo-1H-pyrazolo [3,4-D ] pyrimidin-1-yl) ethyl) -4- (1-benzyl-1, 2,5, 6-tetrahydropyridin-3-yl) -1H-isochromen-1-one hydrochloride (intermediate D33, 1.828g, 2.86mmol) and (3-fluoro-5-hydroxyphenyl) boronic acid (0.890g, 5.71mmol) to yield the title compound (1.3g, 2.1mmol, 71%).
1H NMR(400MHz，DMSO-d6)ppm 10.11-11.75(m，2H)，8.09-8.41(m，2H)，7.57-8.06(m，5H)，7.37-7.56(m，3H)，7.03-8.55(m，2H)，6.81-7.03(m，2H)，6.63-6.76(m，1H)，5.46-6.34(m，2H)，4.32-4.68(m，2H)，2.07-4.20(m，6H)，1.78-1.98(m，3H)。UPLC-MS：0.69min，589.5[M+H]Method 13
Example 169a (enantiomer 1) and example 169b (enantiomer 2): 3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1-benzyl-1, 2,5, 6-tetrahydropyridin-3-yl) -1H-isochromen-1-one hydrochloride as a single enantiomer
The racemate 3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1-benzyl-1, 2,5, 6-tetrahydropyridin-3-yl) -1H-isochromen-1-one hydrochloride (example 169, 0.25g, 0.4mmol) was dissolved in ethanol (5ml) and subjected to chiral resolution by chiral preparative chromatography. Conditions are as follows: column: whelk 0-1(R, R) (25 x 2cm), 10 um; mobile phase: n-hexane/(ethanol/methanol 1/1+ 0.1% isopropylamine) 75/25% v/v; flow rate: 18 ml/min; and D, DAD detection: 220 nm; loop circuit: 500 mul; and (3) injection: 25mg (per injection).
The fraction containing the first eluted enantiomer was evaporated to dryness, 1.25M HCl in MeOH was added and the volatiles were removed under reduced pressure. The residue was purified by reverse phase chromatography using a C18 column (H)2O:CH3CN 95:5-50:50, containing 0.1% HCOOH); before drying, 21N HCl (2ml) was added and the volatiles were removed under reduced pressure to give compound 169a as a white solid (first eluting enantiomer, 0.077g, 0.0123 mmol). Chiral HPLC (method a 13): rt 16.1min, ee>99％。
1H NMR(400MHz，DMSO-d6)ppm 10.12-12.00(m，2H)，8.06-8.46(m，2H)，7.58-8.03(m，5H)，7.37-7.52(m，3H)，7.00-8.55(m，2H)，6.81-7.00(m，2H)，6.66-6.76(m，1H)，5.49-6.42(m，2H)，4.32-4.68(m，2H)，2.07-4.20(m，6H)，1.80-1.98(m，3H)。UPLC-MS：0.70min，589.5[M+H]+, method 13.
The fraction containing the second eluted enantiomer was evaporated to dryness, 1.25M HCl in MeOH was added and the volatiles were removed under reduced pressure. The residue was purified by reverse phase chromatography using a C18 column (H)2O:CH3CN 95:5-50:50, containing 0.1% HCOOH); before drying, 1N HCl (2ml) was added and the volatiles were removed under reduced pressure to give compound 169b as a white solid (second elution)Enantiomer of (4), 0.072g, 0.115 mmol). Chiral HPLC (method a 13): rt 19.0min, ee 98.2%.
1H NMR(400MHz，DMSO-d6)ppm 10.13-12.15(m，2H)，8.06-8.48(m，2H)，7.58-8.06(m，5H)，7.37-7.51(m，3H)，7.00-8.55(m，2H)，6.81-7.00(m，2H)，6.66-6.77(m，1H)，5.50-6.45(m，2H)，4.32-4.68(m，2H)，2.09-4.23(m，6H)，1.80-1.98(m，3H)。UPLC-MS：0.66min，589.3[M+H]+, method 14.
Example 170
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (pyridin-2-yl) -1H-isochromen-1-one hydrochloride
The title compound was prepared in analogy to example 162 from 3- (1- (4-amino-3-iodo-1H-pyrazolo [3,4-D ] pyrimidin-1-yl) ethyl) -4- (pyridin-2-yl) -1H-isochromen-1-one (intermediate D34) and (3-fluoro-5-hydroxyphenyl) boronic acid to give the title compound.
1H NMR(400MHz，DMSO-d6)ppm 10.29(br.s.，1H)，8.70(br.s.，1H)，8.19-8.29(m，2H)，7.97(br.s.，1H)，7.74-7.84(m，1H)，7.62-7.70(m，1H)，7.43-7.60(m，2H)，6.94(d，1H)，6.89(s，1H)，6.83(d，1H)，6.71(dt，1H)，6.50-8.60(m，2H)，5.81-5.91(m，1H)，1.87(d，3H)。UPLC-MS：0.80min，495.0[M+H]+, method 12.
Example 171
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1,2,5, 6-tetrahydropyridin-3-yl) -1H-isochromen-1-one formate salt
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3, 4-d) is reacted at 0 DEG C]Pyrimidin-1-yl) ethyl) -4- (1-benzyl-1, 2,5, 6-tetrahydropyridin-3-yl) -1H-isochromen-1-one hydrochloride (example 169, 0.5g, 0.799mmol) was dissolved in DCM (3 ml); DIPEA (0.395ml, 2.262mmol) and 1-chloroethyl chloroformate (0.651ml, 6.03mmol) were added with vigorous stirring at 0 ℃ and the reaction was stirred for 5min at 0 ℃ and for 2h at 60 ℃. The reaction was cooled to RT and quenched with 5ml MeOH; the mixture was stirred at 60 ℃ for a further 2.5H and evaporated under reduced pressure to give the crude product which was purified by C-18 column reverse phase flash chromatography (H)2O:CH3CN 95:5-50:50, containing 0.1% HCOOH) to give the title compound (0.068g, 0.125mmol, 16%).
1H NMR(400MHz，DMSO-d6)ppm 10.18-10.30(m，1H)，9.04-9.65(m，2H)，8.25-8.31(m，1H)，8.11-8.22(m，2H)，7.83-7.94(m，1H)，7.70-7.82(m，1H)，7.60-7.70(m，1H)，6.81-6.99(m，2H)，6.62-6.75(m，1H)，6.13-8.50(m，2H)，5.55-6.29(m，2H)，2.35-4.18(m，6H)，1.80-1.95(m，3H)。UPLC-MS：0.60min，499.4[M+H]Method 13
Example 172
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1- (1-methylpiperidin-4-yl) -1,2,5, 6-tetrahydropyridin-3-yl) -1H-isochromen-1-one dihydrochloride
To 3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d]Pyrimidin-1-yl) ethyl) -4- (1,2,5, 6-tetrahydropyridin-3-yl) -1H-isochromen-1-one formate (example 171, 0.03g,0.055mmol), 1-methylpiperidin-4-one (0.08ml, 0.066mmol) and DIPEA (0.009ml, 0.055mmol) in DCM (2ml) were added dry Na2SO4The mixture was stirred at RT for 10 min. They were added in this order AcOH (0.009ml, 0.165mmol) and sodium triacetoxyborohydride (0.023g, 0.11mmol) and the reaction mixture was stirred at RT for 3 h. The reaction was stopped by addition of 2M HCl (2ml), the mixture was filtered and the filtrate was purified by reverse phase chromatography using a Biotage C18 column (water/MeCN 90/10+ 0.1% HCOOH-water/MeCN 5/95+ 0.01% HCOOH). Before drying, 2M HCl (2ml) was added and the mixture was dried under reduced pressure to give the title compound (0.025g, 0.037mmol, 68%).
1H NMR(400MHz，DMSO-d6)ppm 8.58-10.68(m，2H)，8.08-8.40(m，2H)，7.81-7.99(m，1H)，7.59-7.74(m，1H)，7.43-7.55(m，1H)，6.78-6.97(m，2H)，6.56-6.73(m，1H)，6.45-7.75(m，2H)，6.09-6.29(m，1H)，5.26-6.08(m，1H)，2.54-3.29(m，13H)，1.40-2.45(m，8H)。UPLC-MS：0.89min，596.5[M+H]Method 15
Example 173
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1- (azetidin-3-yl) -1,2,5, 6-tetrahydropyridin-3-yl) -1H-isochromen-1-one dihydrochloride
The title compound was prepared from 3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1,2,5, 6-tetrahydropyridin-3-yl) -1H-isochromen-1-one formate (example 171, 0.03g, 0.055mmol) and tert-butyl 3-oxoazetidine-1-carboxylate (0.019g, 0.110mmol) in analogy to example 172 to give the title compound (0.019g, 0.030mmol, 55%).
1H NMR(400MHz, methanol-d)4)ppm 8.43-8.56(m，1H)，8.16-8.30(m，1H)，7.82-7.95(m，1H)，7.75-8.01(m，2H)，7.52-7.72(m，1H)，6.87-7.06(m，2H)，6.64-6.80(m，1H)，5.90-6.58(m，2H)，3.30-4.90(m，9H)，2.41-3.04(m，2H)，1.92-2.22(m，3H)。UPLC-MS：0.78min，554.4[M+H]Method 15
Example 174
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1-isopropyl-1, 2,5, 6-tetrahydropyridin-3-yl) -1H-isochromen-1-one hydrochloride
The title compound was prepared in analogy to example 172 from 3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1,2,5, 6-tetrahydropyridin-3-yl) -1H-isochromen-1-one formate (example 171, 0.061g, 0.122mmol) and acetone (0.010mL, 0.134mmol) to yield the title compound (2mg, 0.003mmol, 3%).
1H NMR (400MHz, methanol-d)4)ppm 8.13-8.53(m，2H)，7.47-8.01(m，3H)，6.85-7.04(m，2H)，6.62-6.78(m，1H)，5.72-6.48(m，2H)，3.38-4.47(m，5H)，2.36-3.09(m，2H)，1.91-2.26(m，3H)，1.38-1.69(m，6H)。UPLC-MS：0.63min，541.5[M+H]Method 13
Example 175
3- (1- (4-amino-3- (5-hydroxypyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1-methyl-1, 2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one hydrochloride
The title compound was prepared from 3- (1- (4-amino-3-iodo-1H-pyrazolo [3,4-D ] pyrimidin-1-yl) ethyl) -4- (1-methyl-1, 2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one formate (intermediate D4, 0.050g) and 5- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) pyridin-3-ol (0.023g, 0.104mmol) in analogy to example 162 to yield the title compound (0.014g, 0.026 mmol).
1H NMR(400MHz，DMSO-d6)ppm 11.03-11.70(m，2H)，8.30-8.59(m，3H)，8.10-8.24(m，1H)，7.76-8.04(m，3H)，7.72-8.29(m，2H)，7.48-7.70(m，1H)，5.44-6.47(m，2H)，2.87-2.99(m，3H)，2.44-4.08(m，6H)，1.81-2.05(m，3H)。UPLC-MS：0.42min，496.4[M+H]Method 13
Examples 176a (enantiomer 1) and 176b (enantiomer 2): 3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one single enantiomer
Step 1.3- (1- (4-amino-3- (3- ((tert-butyldimethylsilyl) oxy) -5-fluorophenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one (intermediate Q.1)
To a stirred mixture of 3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one (example 40, 0.20mmol) and imidazole (40.8mg, 0.60mmol) in DMF (1ml) was added TBDMSCl (45.2mg, 0.30mmol) at RT and the mixture was stirred for 2H. Further imidazole (16mg) and TBDMSCl (36mg) were added to the solution, and the mixture was stirred at the same temperature for additional 2 hours. Imidazole (27mg) and TBDMSCl (121mg) were further added thereto, and stirring was continued at room temperature overnight. The reaction mixture was diluted with DCM and washed with 0.5M HCl. The organic layer was dried over sodium sulfate, filtered, and concentrated. The residue was purified by filtration through silica gel column (DCM-DCM: MeOH ═ 95:5) to give 3- (1- (4-amino-3- (3- ((tert-butyldimethylsilyl) oxy) -5-fluorophenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one (intermediate Q.1, 112mg, 0.184mmol, 92%).
UPLC-MS: 1.57min, 608.4[ M + H ] +, method 13
Step 2.3- (1- (4-amino-3- (3- ((tert-butyldimethylsilyl) oxy) -5-fluorophenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one single enantiomer (intermediates Q.2 and Q.3)
The racemate 3- (1- (4-amino-3- (3- ((tert-butyldimethylsilyl) oxy) -5-fluorophenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one (intermediate Q.1, 0.124g, 0.204mmol) was dissolved in a mixture of DCM (5ml) and ethanol/methanol 1/1(6ml) and subjected to chiral resolution by chiral preparative chromatography. Chiral conditions are as follows: column: chiralpak IC (25X 2.0cm), 5 μm; mobile phase: n-hexane/(2-propanol/methanol 1/1+ 0.1% isopropylamine) 90/10% v/v; flow rate: 17 ml/min; and D, DAD detection: 220 nm; loop circuit: 500 mul; and (3) injection: 5 mg/injection.
Fractions containing the first eluted enantiomer were evaporated to dryness to give intermediate Q.2 (first eluted enantiomer, 40mg, 0.066 mmol). Chiral HPLC (method a 4): rt 15.7min, ee > 99%. UPLC-MS: 1.57min, 608.4[ M + H ] +, method 13
The fraction containing the second eluted enantiomer was evaporated to dryness to give intermediate Q.3 (second eluted enantiomer, 38mg, 0.063 mmol). Chiral HPLC (method a 4): rt 17.8min, ee 99%. UPLC-MS: 1.57min, 608.4[ M + H ] +, method 13
Step 3. example 176a, 3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one single enantiomer
The single enantiomer of 3- (1- (4-amino-3- (3- ((tert-butyldimethylsilyl) oxy) -5-fluorophenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one (intermediate Q.2, the enantiomer eluting first under the above conditions, 40mg, 0.065mmol) was dissolved in 1M HCl in EtOH (0.2ml) and stirred at rt for 5H. The volatiles were removed under reduced pressure and the residue was purified by flash chromatography on silica gel (DCM-DCM: MeOH ═ 97:3) to give the title compound as a white solid (28mg, 0.056mmol, 87%). This compound was confirmed to be the second eluting enantiomer under chiral HPLC conditions of method a 5: rt 12.0min, ee 98.6%.
1H NMR(400MHz，DMSO-d6)ppm 10.28(br.s.，1H)，8.19-8.26(m，2H)，7.75-7.80(m，1H)，7.61-7.66(m，1H)，7.51-7.57(m，1H)，7.37-7.48(m，3H)，7.11-7.16(m，1H)，6.87-6.92(m，2H)，6.82-6.87(m，1H)，6.70(dt，1H)，6.0-8.5(m，2H)，5.76(q，1H)，1.85(d，3H)。UPLC-MS：1.04min，494.3[M+H]+, method 13.
Step 4 example 176b, 3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one single enantiomer
3- (1- (4-amino-3- (3- ((tert-butyldimethylsilyl) oxy) -5-fluorophenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one single enantiomer (intermediate Q.3, the enantiomer eluting second under the above conditions, 0.038g, 0.063mmol) was dissolved in 1M EtOH solution of HCl (0.2mL) and stirred at room temperature for 5H. The volatiles were removed under reduced pressure and the residue was purified by flash chromatography on silica gel (DCM-DCM: MeOH: 97:3) to give the title compound as a white solid (0.025g, 0.05mmol, 80%). This compound was confirmed to be the first eluting enantiomer under chiral HPLC conditions of method a 5: rt 7.3min, ee > 99%.
1H NMR(400MHz，DMSO-d6)ppm 10.24(br.s.，1H)，8.20-8.25(m，1H)，8.16(s，1H)，7.74-7.81(m，1H)，7.60-7.66(m，1H)，7.50-7.57(m，1H)，7.35-7.48(m，3H)，7.12-7.18(m，1H)，6.87-6.93(m，2H)，6.81-6.86(m，1H)，6.68(dt，1H)，6.00-8.50(m，2H)，5.70-5.78(m，1H)，1.84(d，3H)。UPLC-MS：1.04min，494.3[M+H]+, method 13.
Pharmacological Activity of the Compounds of the invention
In vitro assay for PI3K enzyme inhibitory Activity in cell-free assays
Human recombinant proteins PI3K α, PI3K β, PI 3K. gamma. and PI3K were purchased from Millipore Ltd (Billerica, MA.) Compounds were dissolved in DMSO at 0.5mM using ADP-GloTMThe kinase assay (Promega, Madison WI) was tested for activity against PI3Ks at various concentrations according to the manufacturer's instructions.
Briefly, the kinase reaction was performed on 384-well white plates (Greiner Bio-one GmbH, Frickenhausen). Each well was loaded with 0.1. mu.l of test compound and 2.5. mu.l of 2x reaction buffer (40mM Tris pH7.5, 0.5mM EGTA, 0.5mM Na3VO45mM β -glycerophosphate, 0.1mg/ml BSA, 1mM DTT) containing 50. mu.M PI and PS substrates (L- α -phosphatidylinositol sodium salt and L- α -phosphatidyl-L-serine, Sigma-Aldrich, St. Louis MO) and PI3K recombinant protein (PI 3K. gamma. 0.25 ng/. mu.l, PI3K1 ng/. mu.l, PI3K α 0.125 ng/. mu.l, PI3K β 1 ng/. mu.l).
The reaction was initiated by adding 2.5. mu.l of 2 XATP solution to each well (final concentration: PI 3K. gamma. ATP 30. mu.M; PI3KATP 80. mu.M; PI3K α. mu.M ATP 50. mu.M; PI3K β. mu.M ATP 100. mu.M), incubated for 60min at room temperature, then, each kinase reaction system was incubated with 5. mu.l of ADP-GloTMThe reagents were incubated together for 40min to allow depletion of unconsumed ATP. Then adding to each wellKinase detection reagent (10 μ l) to convert ADP to ATP and allow the measurement of newly synthesized ATP using the luciferase/luciferin reaction. After 60min incubation, Wallac was usedThe luminescence signal was measured by a multi-label reader (PerkinElmer, WalthammA).
Curve fitting and IC50 calculations were performed using a 4-parameter logistic model applied in XLfit (IDBS, Guilford, UK) of Microsoft Excel (Microsoft, Redmont, WA).
The results are shown in table 1 below.
Table 1: in vitro assay results for PI3K enzyme inhibitory Activity in cell-free assays
(continuation)
(continuation)
(continuation)
(continuation)
(continuation)
(continuation)
Wherein the compounds are classified according to their potency in terms of their inhibitory activity against PI3K- α, - β, - γ, and-as follows:
+++：IC50<10nM
++: IC50 at 10-1000nM
+：IC50>1000nM
In vitro assay for PI3K enzyme inhibitory Activity in PBMCs assays
Human Peripheral Blood Mononuclear Cells (PBMCs) were purchased from Lonza (Basel, CH), washed, resuspended in RPMI 1640 medium (w/o phenol Red) supplemented with 10% FBS, 2mM glutamine, 100U/mL penicillin and 100. mu.g/mL streptomycin (Life Technologies, Carlsbad CA). PBMCs are treated with 105Cell/well density was plated on 96-well plates coated with 6. mu.g/ml anti-human CD3 antibody (Bioleg stop, San Diego CA).
With different concentrations of PI3K inhibitor supplemented with 10% FBS (10)-12M-10-5M, final DMSO concentration 0.2%) were CO-stimulated with 3. mu.g/ml anti-human CD28 antibody (BD Biosciences, San Jose CA) and incubated at 37 ℃ for 72 hours in an atmosphere of 95% air and 5% CO 2. Quantitative ELISA kits Using paired antibodies (from Life Technologies, Carlsbad CA and R, respectively)&D Systems, Minneapolis MN), the supernatant was assayed for human IL-6 and IL-17 according to the manufacturer's instructions.
IC50 values were determined from concentration-response curves by nonlinear regression analysis using Graph Pad Prism v.6(Graph Pad Software, La JollaCA).
Representative compounds of the invention showed an IC50 of less than 1. mu.M for the PI 3K-subunit.
Claims (15)
1. A compound of formula (I):
wherein:
each R, when present, is independently selected from:
-OR6；
-SR6；
-S(O)q-R8；
-NR10R11；
-a halogen;
-(C1-C6) An alkyl group;
-(C1-C6) A haloalkyl group;
-(C3-C7) A cycloalkyl group;
-(C5-C7) A cycloalkenyl group;
-(C2-C6) An alkenyl group;
-(C2-C6) An alkynyl group;
-substituted or unsubstituted aryl; and
-substituted or unsubstituted heteroaryl;
R1and R2Are both H or combine to form an oxo group (═ O);
R3and R4The same or different, at each occurrence is independently selected from:
-H；
-(C1-C6) An alkyl group; and
-(C1-C6) A haloalkyl group;
R5selected from:
-H；
-OR7；
-SR7；
-S(O)q-R9；
-a halogen;
-NR12R13；
-CN；
-C(O)NR12R13；
-COOR14；
-(C1-C6) An alkyl group;
-(C1-C6) A haloalkyl group;
-(C1-C6) A hydroxyalkyl group;
-(C1-C6) An aminoalkyl group;
-(C3-C7) A cycloalkyl group;
aryl (C)1-C3) An alkyl group;
-(C5-C7) A cycloalkenyl group;
-(C2-C6) An alkenyl group;
-(C2-C6) An alkynyl group;
-(C2-C6) An aminoalkynyl group;
-substituted or unsubstituted (C)3-C6) A heterocycloalkyl group;
-substituted or unsubstituted aryl; and
-substituted or unsubstituted heteroaryl;
R6、R7and R14The same or different, at each occurrence is independently selected from:
-H；
-(C1-C6) An alkyl group;
-(C1-C6) A haloalkyl group;
-(C1-C6) A hydroxyalkyl group;
-(C1-C6) An aminoalkyl group;
aryl (C)1-C6) An alkyl group;
-(C1-C6) An alkanoyl group;
-an arylcarbonyl group; and
aryl (C)2-C4) An alkanoyl group;
R8and R9The same or different, at each occurrence is independently selected from
-(C1-C6) An alkyl group;
-(C1-C6) A haloalkyl group;
-(C1-C6) A hydroxyalkyl group;
-(C1-C6) An aminoalkyl group;
-substituted or unsubstituted aryl;
-substituted or unsubstituted heteroaryl; and
-NR12R13；
R10、R11、R12And R13The same or different, at each occurrence, is independently selected from H, (C)1-C6) Aminoalkyl radical (a)C1-C6) Hydroxyalkyl and (C)1-C6) Alkyl, or R10And R11Or R12And R13May form, together with the nitrogen atom to which they are attached, a 5-to 6-membered heterocyclic group;
z, when present, is an atom or group independently selected each time from O, NH, C (O), NHC (O), C (O) NH, S (O) and S (O)2；
m is 0 or 1;
n is 1 or 2;
p is 0 or an integer from 1 to 3;
q is 1 or 2;
cy is selected from:
-substituted or unsubstituted (C)3-C6) A heterocycloalkyl group;
-substituted or unsubstituted aryl; and
-substituted or unsubstituted heteroaryl;
or a pharmaceutically acceptable salt thereof.
2. The compound of claim 1, having formula (IA)
Wherein n is 1, R3Has the same meaning as above except for H, R4Is H and the absolute configuration of the chiral carbon (×) is (R).
3. The compound of claim 1, having formula (IA)
Wherein n is 1, R3Has the same meaning as above except for H, R4Is H and the absolute configuration of the chiral carbon (×) is (S).
4. The compound of claim 1, present as a mixture of diastereomers.
5. The compound of any one of claims 1-4, wherein:
R1and R2Are both H or combine to form an oxo group (═ O);
R3is selected from H and (C) 1-C6) An alkyl group;
R4is H;
R5is selected from H; halogen; OR (OR)7(ii) a Aryl radical (C)1-C3) An alkyl group; (C)5-C7) Cycloalkenyl group, (C)2-C6) Alkynyl (C)2-C6) Aminoalkynyl, substituted or unsubstituted (C)3-C6) A heterocycloalkyl group; substituted or unsubstituted aryl and substituted or unsubstituted heteroaryl;
R、R7m, n, p, Z and CY are as defined above
And pharmaceutically acceptable salts thereof.
6. The compound of any one of claims 1-4, wherein:
p is 0 or 1;
r is absent or selected from halogen and (C)1-C6) An alkyl group;
R1and R2Are both H or combine to form an oxo group (═ O);
R3selected from H, methyl, ethyl and propyl;
R4is H;
R5is selected from H; fluorine; bromine; a phenyl group; a phenyl methyl group; 2-, 3-or 4-pyridyl; 5-thiazolyl; 2-,3-, 4-or 5-thienyl, 1H-pyrazol-4-yl, 2-,4-, 5-or 6-pyrimidinyl, cyclohexenyl, prop-1-ynyl, 1,2,3, 6-tetrahydropyridin-4-yl, 1,2,5, 6-tetrahydropyridin-3-yl, 8-azabicyclo [3.2.1]Oct-2-en-3-yl and 3, 6-dihydro-2H-pyran-4-yl, optionally substituted with one or more groups selected from halogen, (C)1-C6) Alkyl, OR7、-S(O)q-R9、-C(O)NR10R11、COOR14、(C1-C6) Hydroxyalkyl, substituted or unsubstituted (C)1-C6) Aminoalkyl radical, (C)1-C6) Alkanoyl, substituted or unsubstituted (C)3-C6) Heterocycloalkyl, aryl (C)1-C6) Alkyl, heteroaryl (C)1-C6) Alkyl, (C) 3-C7) Cycloalkyl (C)1-C6) Alkyl and NR10R11；
R7、R9、R10、R11、R14M, n, q, Z and CY are as defined above;
and pharmaceutically acceptable salts thereof.
7. The compound of any one of claims 1-4, wherein:
p is 0;
r is absent;
R1and R2Are both H or combine to form an oxo group (═ O);
R3selected from H, methyl, ethyl and propyl;
R4is H;
R5is selected from phenyl; a phenyl methyl group; 2-, 3-or 4-pyridyl; 5-thiazolyl, 2-,3-, 4-or 5-thienyl, 1,2,3, 6-tetrahydropyridin-4-yl and 3, 6-dihydro-2H-pyran-4-yl, optionally substituted with one or more groups selected from fluoro, bromo, methyl, methoxy, amino, dimethylamino, 4-morpholinosulfonyl, 4- (2-morpholinoethoxy), 4-morpholinomethyl and 4-piperazinylmethyl; piperidin-1-ylmethyl, 4-methylpiperazin-1-carbonyl, (2- (dimethylamino) ethyl) -carbonyl, acetyl, phenylmethyl, phenylmethoxy-carbonyl, 4,5, 5-tetramethyl-1, 3-dioxolan-2-yl, pyrrolidin-1-ylmethyl, bis (2-hydroxyethyl) aminomethyl, hydroxymethyl, dimethylaminomethyl, (dimethylamino) propyl, 4- (2-hydroxyethyl) piperazin-1-yl) methylpiperazin-2-one-1-ylmethyl, cyclopropylmethyl, hydroxycarbonyl, pyridin-4-ylmethyl;
m, n, Z and CY are as defined above;
and pharmaceutically acceptable salts thereof.
8. The compound of any one of claims 1-4, wherein:
R1and R2Are both H or combine to form an oxo group (═ O);
R3selected from H, methyl, ethyl and propyl;
R4is H;
R5is selected from H; halogen; OR (OR)7(ii) a Aryl radical (C)1-C3) An alkyl group; substituted or unsubstituted (C)3-C6) A heterocycloalkyl group; substituted or unsubstituted aryl and substituted or unsubstituted heteroaryl;
z, when present, is an atom or group selected from O, NH, C (O), NH-C (O), C (O) -NH, S, SO and SO2；
CY is heteroaryl selected from 7H-purin-7-yl; 9H-purin-9-yl; 9H-purin-6-yl; 1H-pyrazolo [3,4-d]Pyrimidin-1-yl; 1H-pyrazolo [3,4-d]Pyrimidin-4-yl; 2H-pyrazolo [3,4-d]Pyrimidin-2-yl; 2-,4-, 5-or 6-pyrimidinyl; and 2-pyrazinyl, pyrrolo [2,3-d ]]Pyrimidin-7-yl, pyrazolo [1,5-a ]]Pyrimidin-3-yl, pyrido [3,2-d]Pyrimidin-4-yl, pyrido [2,3-d ]]Pyrimidin-8-yl-5-one, thieno [3,2-d]Pyrimidin-4-yl, thieno [2,3-d ]]Pyrimidin-4-yl all of which are optionally substituted with one or more groups selected from halogen, (C)1-C6) Alkyl, (C)1-C6) Haloalkyl, CN, NR10R11Optionally substituted aryl and optionally substituted heteroaryl selected from the group consisting of phenyl, 1H-indazol-4-yl, 1H-indazol-5-yl, 1H-indazol-6-yl, 2-, 3-, 4-,5-, 6-pyridyl, pyrazol-3-yl, pyrazol-4-yl, pyrazol-5-yl, pyrazin-2-yl, pyrimidin-5-yl, pyridazin-4-yl and 2-,4-, 5-thiazolyl;
R、R7、R10、R11M, n, p are as defined above;
and pharmaceutically acceptable salts thereof.
9. The compound of claim 1, wherein:
R1and R2Combine to form an oxo group (═ O);
R3selected from H, methyl or ethyl;
R4is H;
R5is selected from H; halogen selected from fluorine and bromine; aryl which is phenyl; aryl being phenylmethyl (C)1-C3) An alkyl group; selected from 2-, 3-or 4-pyridyl; 5-thiazolyl, 2-,3-, 4-or 5-thienyl heteroaryl; (C) selected from 1,2,3, 6-tetrahydropyridin-4-yl and 3, 6-dihydro-2H-pyran-4-yl3-C6) Heterocycloalkyl and 4-cyclohexenyl, all of which are optionally substituted with one or more groups selected from fluoro, bromo, methyl, methoxy, dimethylamino, morpholinosulfonyl, morpholinoethoxy, morpholinomethyl, and piperazinylmethyl; 4-methylpiperazin-1-carbonyl, 4- (2-hydroxyethyl) piperazin-1-yl-methyl, piperazin-2-on-1-yl-methyl, pyridin-4-ylmethyl;
z, when present, is an atom or group, each time independently selected from O, NH, C (O), NH-C (O), C (O) -NH, S, SO, and SO2；
CY is heteroaryl selected from 7H-purin-7-yl; 9H-purin-9-yl; 9H-purin-6-yl; 1H-pyrazolo [3,4-d]Pyrimidin-1-yl; 1H-pyrazolo [3,4-d]Pyrimidin-4-yl; 2H-pyrazolo [3,4-d ]Pyrimidin-2-yl; and 2-,4-, 5-or 6-pyrimidinyl; 2-pyrazinyl, all of which are optionally substituted with one or more groups selected from Cl, Br, F, I, methyl, trifluoromethyl, CN; NH (NH)2；NH-CH3；N(CH3)2(ii) a 3-methyl-1H-indazol-5-yl, 1H-indazol-4-yl; 3-fluoro-5-hydroxyphenyl; 1- (3-fluoro-4-hydroxyphenyl); 6-,5-, 4-hydroxypyridin-3-yl, 6-, 5-methoxypyridin-3-yl, 5-aminopyridin-3-yl, 5-fluoropyridin-3-yl, 5-fluoro-6-hydroxypyridin-3-yl 6- (methylsulfonyl) pyridin-3-yl, 5-hydroxy-6-methylpyridin-3-yl, 6-,5- (hydroxymethyl) pyridin-3-yl, 2-aminothiazol-5-yl; 2- (acetylamino) - (thiazol-5-yl), 2-aminopyrimidin-5-yl, 2-methoxypyrimidin-5-yl, 2-hydroxypyrimidine-5-yl, pyrazin-2-yl, 6-hydroxypyrazin-2-yl, and 3-fluoro-4-isopropoxyphenyl;
r, m, n, p are as defined above;
and pharmaceutically acceptable salts thereof.
10. The compound of claim 1 selected from:
3- ((6-amino-9H-purin-9-yl) methyl) -4-phenyl-1H-isochromen-1-one;
3- ((6-amino-9H-purin-9-yl) methyl) -4- (3-fluorophenyl) -1H-isochromen-1-one;
3- ((6-amino-9H-purin-9-yl) methyl) -4- (2-fluorophenyl) -1H-isochromen-1-one;
3- ((6-amino-9H-purin-9-yl) methyl) -4-m-tolyl-1H-isochromen-1-one;
3- (1- (6-amino-9H-purin-9-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (6-amino-9H-purin-9-yl) ethyl) -4-m-tolyl-1H-isochromen-1-one;
3- (1- (6-amino-9H-purin-9-yl) ethyl) -4- (3-fluorophenyl) -1H-isochromen-1-one;
3- (1- (6-amino-9H-purin-9-yl) ethyl) -4- (3- (dimethylamino) phenyl) -1H-isochromen-1-one;
3- (1- (6-amino-9H-purin-9-yl) ethyl) -4- (3- (morpholinosulfonyl) phenyl) -1H-isochromen-1-one;
3- ((9H-purin-6-ylsulfanyl) methyl) -4-phenyl-1H-isochromen-1-one;
3- ((9H-purin-6-ylsulfanyl) methyl) -4- (2-fluorophenyl) -1H-isochromen-1-one;
3- ((9H-purin-6-ylsulfanyl) methyl) -4-m-tolyl-1H-isochromen-1-one;
3- (1- (9H-purin-6-ylsulfanyl) ethyl) -4-m-tolyl-1H-isochromen-1-one;
3- (1- (9H-purin-6-ylsulfanyl) ethyl) -4- (3-fluorophenyl) -1H-isochromen-1-one;
3- (1- (4-amino-1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (6-methylpyridin-3-yl) -1H-isochromen-1-one;
3- (1- (4-amino-1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (3- (morpholinosulfonyl) phenyl) -1H-isochromen-1-one;
3- (1- (4-amino-1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (thiazol-5-yl) -1H-isochromen-1-one;
3- (1- (4-amino-1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (2-methylpyridin-4-yl) -1H-isochromen-1-one;
3- (1- (4-amino-1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-benzyl-1H-isochromen-1-one;
3- ((9H-purin-6-ylamino) methyl) -4- (3-fluorophenyl) -1H-isochromen-1-one;
3- (1- (9H-purin-6-ylamino) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (9H-purin-6-ylamino) ethyl) -4- (1,2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one;
3- (1- (9H-purin-6-ylamino) ethyl) -4- (3, 6-dihydro-2H-pyran-4-yl) -1H-isochromen-1-one;
3- (1- (9H-purin-6-ylamino) propyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (9H-purin-6-ylamino) ethyl) -4- (4- (2-morpholinoethoxy) phenyl) -1H-isochromen-1-one;
4-amino-8- (1- (1-oxo-4-phenyl-1H-isochromen-3-yl) ethyl) pyrido [2,3-d ] pyrimidin-5 (8H) -one;
3- (1- (9H-purin-6-ylamino) ethyl) -4- (5- (morpholinomethyl) thiophen-2-yl) -1H-isochromen-1-one;
3- ((9H-purin-6-ylamino) methyl) -4-phenyl-1H-isochromen-1-one;
3- ((9H-purin-6-ylamino) methyl) -4- (2-fluorophenyl) -1H-isochromen-1-one;
3- ((9H-purin-6-ylamino) methyl) -4-m-tolyl-1H-isochromen-1-one;
3- (1- (9H-purin-6-ylamino) ethyl) -4- (3-fluorophenyl) -1H-isochromen-1-one;
3- (1- (9H-purin-6-ylamino) ethyl) -4-m-tolyl-1H-isochromen-1-one;
3- (1- (9H-purin-6-ylamino) ethyl) -4- (3- (dimethylamino) phenyl) -1H-isochromen-1-one;
3- ((4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) methyl) -4-phenyl-1H-isochromen-1-one;
3- ((4-amino-3- (1H-indazol-5-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) methyl) -4-phenyl-1H-isochromen-1-one;
3- ((4-amino-3- (3-methyl-1H-indazol-5-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) methyl) -4-phenyl-1H-isochromen-1-one;
3- ((4-amino-3- (1H-indazol-6-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) methyl) -4-phenyl-1H-isochromen-1-one;
3- ((4-amino-3- (3-fluoro-4-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) methyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-m-tolyl-1H-isochromen-1-one;
3- (1- (4-amino-3- (1H-pyrazol-4-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-m-tolyl-1H-isochromen-1-one;
3- (1- (6-amino-9H-purin-9-yl) ethyl) -4- (6-methoxypyridin-3-yl) -1H-isochromen-1-one;
3- ((4-amino-3-iodo-1H-pyrazolo [3,4-d ] pyrimidin-1-yl) methyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3-iodo-1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3-iodo-1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-m-tolyl-1H-isochromen-1-one;
3- (1- (1H-pyrazolo [3,4-d ] pyrimidin-4-ylamino) ethyl) -4-phenyl-1H-isochromen-1-one;
4-amino-6- (1- (1-oxo-4-phenyl-1H-isochromen-3-yl) ethylamino) pyrimidine-5-carbonitrile;
3- (1- (9H-purin-6-ylamino) ethyl) -4- (1-methyl-1, 2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1-methyl-1, 2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1-methyl-1, 2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one single enantiomer 1;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1-methyl-1, 2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one single enantiomer 2;
3- (1- (9H-purin-6-ylamino) ethyl) -4- (4- (morpholinomethyl) phenyl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (4- (morpholinomethyl) phenyl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (5- (morpholinomethyl) thiophen-2-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (5- (morpholinomethyl) thiophen-2-yl) -1H-isochromen-1-one single enantiomer 1;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (5- (morpholinomethyl) thiophen-2-yl) -1H-isochromen-1-one single enantiomer 2;
3- (1- (9H-purin-6-ylamino) ethyl) -4-cyclohexenyl-1H-isochromen-1-one;
3- (1- (4-amino-3- (2-aminopyrimidin-5-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3- (pyrazin-2-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3- (pyridazin-4-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3- (pyridin-4-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3- (2-methoxypyrimidin-5-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3- (2-hydroxypyrimidin-5-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3- (5-methoxypyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3- (pyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3- (2-aminothiazol-5-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3- (6-methoxypyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3- (6-hydroxypyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
N- (5- (4-amino-1- (1- (1-oxo-4-phenyl-1H-isochromen-3-yl) ethyl) -1H-pyrazolo [3,4-d ] pyrimidin-3-yl) thiazol-2-yl) acetamide;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1-methyl-1, 2,5, 6-tetrahydropyridin-3-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1,2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1- (pyridin-4-ylmethyl) -1,2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1- (cyclopropylmethyl) -1,2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (3- ((dimethylamino) methyl) phenyl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (3- ((dimethylamino) methyl) phenyl) -1H-isochromen-1-one enantiomer 1;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (3- ((dimethylamino) methyl) phenyl) -1H-isochromen-1-one enantiomer 2;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (5- (piperidin-1-ylmethyl) thiophen-2-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (5- (piperidin-1-ylmethyl) thiophen-2-yl) -1H-isochromen-1-one single enantiomer 1;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (5- (piperidin-1-ylmethyl) thiophen-2-yl) -1H-isochromen-1-one single enantiomer 2;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (4- (4-methylpiperazine-1-carbonyl) phenyl) -1H-isochromen-1-one;
3- (3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -1-oxo-1H-isochromen-4-yl) -N- (2- (dimethylamino) ethyl) benzamide;
4- (1-acetyl-1, 2,3, 6-tetrahydropyridin-4-yl) -3- (1- (4-amino-3- (5-hydroxypyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -1H-isochromen-1-one;
4- (1-acetyl-1, 2,3, 6-tetrahydropyridin-4-yl) -3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (5- (pyrrolidin-1-ylmethyl) thiophen-2-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (5- ((bis (2-hydroxyethyl) amino) methyl) thiophen-2-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (5- (hydroxymethyl) thiophen-2-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (5- ((4- (2-hydroxyethyl) piperazin-1-yl) methyl) thiophen-2-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (5- ((4- (2-hydroxyethyl) piperazin-1-yl) methyl) thiophen-2-yl) -1H-isochromen-1-one single enantiomer 1;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (5- ((4- (2-hydroxyethyl) piperazin-1-yl) methyl) thiophen-2-yl) -1H-isochromen-1-one single enantiomer 2;
4- ((5- (3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -1-oxo-1H-isochromen-4-yl) thiophen-2-yl) methyl) piperazin-2-one;
5- (3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -1-oxo-1H-isochromen-4-yl) thiophene-2-carboxylic acid;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-benzyl-1H-isochromen-1-one;
4- (1H-pyrazol-4-yl) -3- (1- (thieno [3,2-d ] pyrimidin-4-ylamino) ethyl) -1H-isochromen-1-one;
4- (5- (morpholinomethyl) thiophen-2-yl) -3- (1- (thieno [3,2-d ] pyrimidin-4-ylamino) ethyl) -1H-isochromen-1-one;
4-amino-6- ((1- (4- (5- (morpholinomethyl) thiophen-2-yl) -1-oxo-1H-isochromen-3-yl) ethyl) amino) pyrimidine-5-carbonitrile;
4-phenyl-3- (1- (pyrrolo [2,1-f ] [1,2,4] triazin-4-ylamino) ethyl) -1H-isochromen-1-one;
4-phenyl-3- (1- (pyrido [3,2-d ] pyrimidin-4-ylamino) ethyl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (5- (hydroxymethyl) pyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3- (6- (hydroxymethyl) pyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
n- (5- (4-amino-1- (1- (1-oxo-4-phenyl-1H-isochromen-3-yl) ethyl) -1H-pyrazolo [3,4-d ] pyrimidin-3-yl) pyridin-3-yl) -4-fluorobenzenesulfonamide;
3- (1- (4-amino-3- (5-aminopyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3- (2-aminopyrimidin-4-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3- (6-hydroxypyrazin-2-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3- (5-hydroxypyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (8-methyl-8-azabicyclo [3.2.1] oct-2-en-3-yl) -1H-isochromen-1-one hydrochloride;
3- (1- (4-amino-3- (3-fluoro-4-isopropoxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3- (5-fluoropyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3- (3-chloro-5-fluorophenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3- (5- (methylsulfonyl) pyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3- (6- (methylsulfonyl) pyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3- (5-fluoro-6-hydroxypyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3- (5-hydroxy-6-methylpyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3- (5- (trifluoromethyl) pyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3- (5-hydroxy-3-pentafluorothio) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
5- (4-amino-1- (1- (1-oxo-4-phenyl-1H-isochromen-3-yl) ethyl) -1H-pyrazolo [3,4-d ] pyrimidin-3-yl) nicotinonitrile;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (4-aminocyclohex-1-en-1-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (trifluoromethyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3-methyl-1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-7H-pyrrolo [2,3-d ] pyrimidin-7-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (2, 6-diamino-9H-purin-9-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
4-phenyl-3- (1- (thieno [2,3-d ] pyrimidin-4-ylamino) ethyl) -1H-isochromen-1-one;
4-phenyl-3- (1- (thieno [3,2-d ] pyrimidin-4-ylamino) ethyl) -1H-isochromen-1-one;
2-amino-N- (1- (1-oxo-4-phenyl-1H-isochromen-3-yl) ethyl) pyrazolo [1,5-a ] pyrimidine-3-carboxamide;
3- (1- (4-amino-3- (1H-pyrazol-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3- (1H-indazol-4-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3- (3-amino-1H-indazol-5-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3- (3-hydroxy-5- (trifluoromethoxy) phenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (6-amino-9H-purin-9-yl) ethyl) -4- (5- (morpholinomethyl) thiophen-2-yl) -1H-isochromen-1-one;
3- (1- (9H-purin-6-ylamino) ethyl) -4- (6-methoxypyridin-3-yl) -1H-isochromen-1-one;
3- (1- (9H-purin-6-ylamino) ethyl) -4- (thiazol-5-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -2H-pyrazolo [3,4-d ] pyrimidin-2-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (3- (dimethylamino) phenyl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (thiazol-5-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (2-aminothiazol-5-yl) -1H-isochromen-1-one;
3- (1- (9H-purin-6-ylamino) ethyl) -4- (1H-pyrazol-4-yl) -1H-isochromen-1-one;
4-amino-6- ((1- (1-oxo-4- (1H-pyrazol-4-yl) -1H-isochromen-3-yl) ethyl) amino) pyrimidine-5-carbonitrile;
3- (1- (9H-purin-6-ylamino) ethyl) -4- (2-aminopyrimidin-5-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (4- (piperazin-1-ylmethyl) phenyl) -1H-isochromen-1-one;
3- (1- (9H-purin-6-ylamino) ethyl) -4- (4- (piperazin-1-ylmethyl) phenyl) -1H-isochromen-1-one;
3- (1- (4-amino-1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (4- (piperazin-1-ylmethyl) phenyl) -1H-isochromen-1-one;
3- (4-amino-1- ((4-phenyl-1H-isochromen-3-yl) methyl) -1H-pyrazolo [3,4-d ] pyrimidin-3-yl) -5-fluorophenol;
5- (4-amino-1- ((4-phenyl-1H-isochromen-3-yl) methyl) -1H-pyrazolo [3,4-d ] pyrimidin-3-yl) pyridin-3-ol;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (morpholinomethyl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1-benzyl-1, 2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (5-hydroxypyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1-benzyl-1, 2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one;
4- (3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -1-oxo-1H-isochromen-4-yl) -5, 6-dihydropyridine-1 (2H) -carboxylic acid benzyl ester;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (5- (piperazin-1-ylmethyl) thiophen-2-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (5- ((4-methylpiperazin-1-yl) methyl) thiophen-2-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (5- (3- (dimethylamino) propyl) thiophen-2-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (3- (3- (dimethylamino) propyl) phenyl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (3- (3- (dimethylamino) propyl) phenyl) -1H-isochromen-1-one single enantiomer 1;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (3- (3- (dimethylamino) propyl) phenyl) -1H-isochromen-1-one single enantiomer 2;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (4- (3- (dimethylamino) propyl) phenyl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (4- (3- (dimethylamino) propyl) phenyl) -1H-isochromen-1-one single enantiomer 1;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (4- (3- (dimethylamino) propyl) phenyl) -1H-isochromen-1-one single enantiomer 2;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (4- ((4-methylpiperazin-1-yl) methyl) phenyl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (4- (pyrrolidin-1-ylmethyl) phenyl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (4- (piperidin-1-ylmethyl) phenyl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (3- ((4-methylpiperazin-1-yl) methyl) phenyl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (3- (piperidin-1-ylmethyl) phenyl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (2,2,6, 6-tetramethyl-1, 2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (5-hydroxypyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (2,2,6, 6-tetramethyl-1, 2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1- (4- (dimethylamino) butanoyl) -1,2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1- (2- (dimethylamino) acetyl) -1,2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1- (1-methylpiperidine-4-carbonyl) -1,2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1- (1-isopropylpiperidin-4-yl) -1,2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1- (azetidine-3-carbonyl) -1,2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1- (1-methylazetidin-3-carbonyl) -1,2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-chloro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1,2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-hydroxy-5- (trifluoromethyl) phenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1,2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-chloro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1-methyl-1, 2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-hydroxy-5- (trifluoromethyl) phenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1-methyl-1, 2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1- (azetidin-3-yl) -1,2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1- (1- (cyclopropylmethyl) azetidin-3-yl) -1,2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (3- (dimethylamino) prop-1-yn-1-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (5-hydroxy-4-methylpyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3- (5-hydroxy-2-methylpyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3- (5-hydroxy-6- (trifluoromethyl) pyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -7-methyl-4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -7-chloro-4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -6-chloro-4-phenyl-1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -6-fluoro-4-phenyl-1H-isochromen-1-one;
4- (1-acetyl-1, 2,3, 6-tetrahydropyridin-4-yl) -3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -1H-isochromen-1-one single enantiomer 1;
4- (1-acetyl-1, 2,3, 6-tetrahydropyridin-4-yl) -3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -1H-isochromen-1-one single enantiomer 2;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (thiazol-5-yl) -1H-isochromen-1-one single enantiomer 1;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (thiazol-5-yl) -1H-isochromen-1-one single enantiomer 2;
3- (1- (4-amino-3- (5-hydroxypyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one single enantiomer 1;
3- (1- (4-amino-3- (5-hydroxypyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one single enantiomer 2;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1-benzyl-1, 2,5, 6-tetrahydropyridin-3-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1-benzyl-1, 2,5, 6-tetrahydropyridin-3-yl) -1H-isochromen-1-one single enantiomer 1;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1-benzyl-1, 2,5, 6-tetrahydropyridin-3-yl) -1H-isochromen-1-one single enantiomer 2;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (pyridin-2-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1,2,5, 6-tetrahydropyridin-3-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1- (1-methylpiperidin-4-yl) -1,2,5, 6-tetrahydropyridin-3-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1- (azetidin-3-yl) -1,2,5, 6-tetrahydropyridin-3-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1-isopropyl-1, 2,5, 6-tetrahydropyridin-3-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (5-hydroxypyridin-3-yl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4- (1-methyl-1, 2,3, 6-tetrahydropyridin-4-yl) -1H-isochromen-1-one;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one single enantiomer 1;
3- (1- (4-amino-3- (3-fluoro-5-hydroxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) ethyl) -4-phenyl-1H-isochromen-1-one single enantiomer 2;
and pharmaceutically acceptable salts thereof.
11. A pharmaceutical composition comprising a compound as defined in any one of claims 1 to 10, or a pharmaceutically acceptable salt thereof, alone or in combination with one or more active ingredients, in admixture with one or more pharmaceutically acceptable carriers or excipients.
12. A compound according to any one of claims 1 to 10 for use as a medicament.
13. Use of a compound as defined in any one of claims 1 to 10, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for the treatment of a disorder associated with the PI3K enzyme mechanism.
14. The use of claim 13, wherein the disorder associated with the PI3K enzyme mechanism is selected from the group consisting of: respiratory diseases including idiopathic chronic cough, cough variant asthma, cough associated with breast tumors or lung cancer, viral or post-viral cough, Upper Airway Cough Syndrome (UACS) or post nasal drip cough or cough associated with gastroesophageal reflux (acid and non-acid reflux), chronic bronchitis, Chronic Obstructive Pulmonary Disease (COPD), interstitial lung diseases (e.g., Idiopathic Pulmonary Fibrosis (IPF)), congestive heart disease, sarcoidosis or infections (e.g., pertussis), asthma, Chronic Obstructive Pulmonary Disease (COPD) and Idiopathic Pulmonary Fibrosis (IPF)); viral infections (including viral respiratory infections and viral exacerbation of respiratory diseases such as asthma and COPD); non-viral respiratory infections (including aspergillosis and leishmaniasis); allergic diseases (including allergic rhinitis and atopic dermatitis); autoimmune diseases (including rheumatoid arthritis and multiple sclerosis); inflammatory diseases (including inflammatory bowel disease); cardiovascular disease (including thrombosis and atherosclerosis); hematological malignancies; neurodegenerative diseases; pancreatitis; multiple organ failure; renal disease; platelet aggregation; cancer; sperm motility; graft rejection; transplant rejection; lung injury; and pain (including pain associated with rheumatoid arthritis or osteoarthritis, back pain, systemic inflammatory pain, post-herpetic neuralgia, diabetic neuropathy, inflammatory neuropathic pain (trauma), trigeminal neuralgia, and central pain).
15. The method of claim 14, wherein the disorder associated with the PI3K enzyme mechanism is asthma, chronic obstructive pulmonary disease, COPD, Idiopathic Pulmonary Fibrosis (IPF).
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| EP13197986.6 | 2013-12-18 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| HK1226719A1 true HK1226719A1 (en) | 2017-10-06 |
| HK1226719B HK1226719B (en) | 2020-04-24 |
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