WO2024254490A1 - Composés pyrazolo-pyrimidinone destinés à être utilisés dans des méthodes d'inhibition de la kinase wee1 - Google Patents

Composés pyrazolo-pyrimidinone destinés à être utilisés dans des méthodes d'inhibition de la kinase wee1 Download PDF

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WO2024254490A1
WO2024254490A1 PCT/US2024/033065 US2024033065W WO2024254490A1 WO 2024254490 A1 WO2024254490 A1 WO 2024254490A1 US 2024033065 W US2024033065 W US 2024033065W WO 2024254490 A1 WO2024254490 A1 WO 2024254490A1
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compound
methyl
dihydro
cancer
allyl
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Inventor
Hans Mattias JÖNSSON
Nima RAJABI
Filip Alexander PAULSEN
Andrea ÖHLANDER
Björn BORGSTRÖM
Magnus Munck AF ROSENSCHÖLD
Lars Thomas BRIMERT
Caroline Maria WIGERUP
John Henry Van Duzer
David Proia
Lei Shi
Åsa Helén NILSSON
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Acrivon Therapeutics Inc
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Acrivon Therapeutics Inc
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Priority to EP24739831.6A priority Critical patent/EP4724443A1/fr
Priority to IL325136A priority patent/IL325136A/en
Priority to AU2024284667A priority patent/AU2024284667A1/en
Priority to CN202480051896.8A priority patent/CN121752567A/zh
Priority to KR1020267000376A priority patent/KR20260039689A/ko
Publication of WO2024254490A1 publication Critical patent/WO2024254490A1/fr
Priority to MX2025014761A priority patent/MX2025014761A/es
Anticipated expiration legal-status Critical
Priority to CONC2026/0000188A priority patent/CO2026000188A2/es
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D487/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00
    • C07D487/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00 in which the condensed system contains two hetero rings
    • C07D487/04—Ortho-condensed systems
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D519/00—Heterocyclic compounds containing more than one system of two or more relevant hetero rings condensed among themselves or condensed with a common carbocyclic ring system not provided for in groups C07D453/00 or C07D455/00

Definitions

  • Cancer cells frequently have a defective Gl/S checkpoint, often via disrupted p53 activity due to mutations or deletion, or inactivation by viral oncoproteins. Therefore, cancer cells rely heavily on other cell cycle checkpoints, including the G2/M checkpoint, to avoid accumulation of deleterious DNA damage and mitotic catastrophe. As such, cancer cells are hypothesized to be particularly vulnerable to inhibition of proteins that safeguard the entry into mitosis. Matheson, C. J. et al Trends Pharmacol Sci 37, 872-881 (2016).
  • WeelA kinase is a tyrosine kinase belonging to the Weel kinase family, including WeelA kinase, WeelB kinase, and Mytl kinase. Rora, A. G. L. et al J Hematol Oncol 13, 126 (2020).
  • the primary role for this kinase family is to regulate cell cycle progression and entry into mitosis (WeelA kinase and Mytl kinase) or meiosis (WeelB kinase).
  • the key complex regulating mitotic entry is Cdkl/cyclin Bl complex, also known as the mitosispromoting factor.
  • WeelA kinase constrains Cdkl/cyclin Bl complex activity by phosphorylating Cdkl on the inhibitory tyrosine 15 site (Y 15 ). Hence, inhibition of WeelA kinase effectively promotes Cdkl/cyclin Bl complex activity by preventing inhibitory Y 15 phosphorylation. Untimely activation of Cdkl/cyclin B complex promotes premature entry into mitosis with unresolved DNA damages, ultimately leading to mitotic catastrophe and cell death.
  • Cdk2 is the primary Cdk driving DNA replication and inhibition of WeelA kinase leads to excessive DNA replication, leading to exhaustion of nucleotide pools and degradation of the ribonucleotide reductase subunit RRM2 (Pfister, S. X. et al Cancer Cell 28, 557-568 (2015)). Pfister et al. showed that Wee1A kinase inhibition selectively kills H3K36me3-deficient cancer cells through dNTP starvation resulting from RRM2 depletion. The histone methyl transferase SETD2 catalyzes H3K36me3, which promotes RRM2 expression and synthesis of dNTPs.
  • HNSCC head and neck squamous cell carcinoma
  • Wee1A kinase inhibitors are currently being tested in clinical trials (Bukhari, A. B. et al Frontiers Oncol 12, 828684 (2022) and have shown activity in many indications.
  • a phase II study of the Wee1A kinase inhibitor AZD1775 (adavosertib) has shown promising results in women with uterine serous carcinoma.
  • Wee1A kinase inhibitors Given the encouraging signs of clinical activity with Wee1A kinase inhibition, there is an urgent need for novel Wee1A kinase inhibitors with improved potency and selectivity, as well as for compounds that inhibit both Wee1A kinase and Myt1 kinase to maximize the efficacy potential of this target class.
  • the present disclosure provides a compound of formula I: or a pharmaceutically acceptable salt thereof, wherein each of X, Y, Z, R 1 , R 2 , R 3 , R 4 , R 8 , n and m are as defined below and described herein. [0009] In some embodiments, the present disclosure provides a pharmaceutical composition comprising a compound of formula I, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.
  • the present disclosure provides a method of inhibiting Wee1A kinase in a patient or in a biological sample, the method comprising administering to the patient or contacting the biological sample with a compound of formula I, or a pharmaceutically acceptable salt thereof.
  • the present disclosure provides a method of inhibiting both Wee1A kinase and Myt1 kinase in a patient or in a biological sample, the method comprising administering to the patient or contacting the biological sample with a compound of formula I, or a pharmaceutically acceptable salt thereof.
  • the present disclosure provides a method of treating a disease or disorder associated with Wee1A kinase, the method comprising administering to a patient in need thereof a compound of formula I, or a pharmaceutically acceptable salt thereof.
  • the present disclosure provides a method of treating a disease or disorder associated with both Wee1A kinase and Myt1 kinase, the method comprising administering to a patient in need thereof a compound of formula I that has dual activity, or a pharmaceutically acceptable salt thereof.
  • the disease or disorder associated with Wee1A kinase is a cancer.
  • the disease or disorder associated with Wee1A kinase and Myt1 kinase is a cancer.
  • the cancer is selected from a brain cancer, a cervicocerebral cancer, a cardiac cancer, a gastrointestinal cancer, an esophageal cancer, a thyroid cancer, a small cell cancer, a non-small cell cancer, a breast cancer, a lung cancer, a stomach cancer, a gallbladder/bile duct cancer, a liver cancer, a pancreatic cancer, a colon cancer, a rectal cancer, an ovarian cancer, a choriocarcinoma, an uterus body cancer, an uterocervical cancer, a renal pelvis/ureter cancer, a bladder cancer, a prostate cancer, a penis cancer, a testicular cancer, a fetal cancer, Wilms' cancer, a skin cancer, malignant melanoma, a neuroblastoma, an osteos
  • the present disclosure provides inhibitors of Wee1A kinase.
  • such compounds include those of the formulae described herein, or a pharmaceutically acceptable salt thereof, wherein each variable is as defined and described herein.
  • the present disclosure provides a compound having structural formula AA: or a solvate, enantiomer, tautomer, or diastereomer thereof, or a pharmaceutically acceptable salt of any of the foregoing, wherein: each of X, Y and Z is independently CH or N; R 1 is -O-, -NH-, or -N(C 1 -C 3 alkyl)-; each R 2 is independently fluoro, -CN, unsubstituted C1-C5 alkyl, fluoro-substituted C 1 -C 5 alkyl, or cyano-substituted C 1 -C 5 alkyl, wherein two R 2 bound to the same carbon atom are optionally taken together to form a spiro-fused C3-C7 cycloalkyl ring, or two R 2 bound to different carbon atoms are optionally taken together to form a bridged or fused C 3 -C 5 cycloal
  • R represents a point of attachment of R 4 to the compound; each R 5 and each R 6 is independently hydrogen, halo, -CN, -C 1 -C 4 alkyl optionally substituted with halo, -O-(C1-C4 alkyl) optionally substituted with halo, -O-(C1-C4 alkyl) substituted with C 3 -C 6 cycloalkyl, an N-linked saturated 3-7 membered heterocyclyl, a 5-6 membered heteroaryl, or phenyl, wherein no more than two R 6 are other than hydrogen; wherein R 5 and an R 6 on an adjacent ring atom are optionally taken together to form a 4-7 membered saturated heterocyclic or cycloalkyl ring that is fused to R 4 ; R 7 is hydrogen, -C 1 -C 4 alkyl, -C 1 -C 4 alkylene-O-C 1 -C 4 alkyl, -C(O)- C 1 -
  • each of X, Y and Z is independently CH or N; R 1 is -O-, -NH-, or -N(C 1 -C 3 alkyl)-; each R 2 is independently fluoro, -CN, unsubstituted C1-C5 alkyl, fluoro-substituted C 1 -C 5 alkyl, or cyano-substituted C 1 -C 5 alkyl, wherein two R 2 bound to the same carbon atom are optionally taken together to form a spiro-fused C3-C7 cycloalkyl ring, or two R 2 bound to different carbon atoms are optionally taken together to form a bridged C3-C5 cycloalkyl ring, or one R 2 and R 8 are optionally taken together to form a bridged 3-5 member
  • the present disclosure provides a compound having structural formula I: or a solvate, enantiomer, tautomer, or diastereomer thereof, or a pharmaceutically acceptable salt of any of the foregoing, wherein: each of X, Y and Z is independently CH or N; R 1 is -O-, -NH-, or -N(C1-C3 alkyl)-; each R 2 is independently fluoro, -CN, unsubstituted C 1 -C 5 alkyl, fluoro-substituted C1-C5 alkyl, or cyano-substituted C1-C5 alkyl, wherein two R 2 bound to the same carbon atom are optionally taken together to form a spiro-fused C3-C7 cycloalkyl ring, or two R 2 bound to different carbon atoms are optionally taken together to form a bridged C 3 -C 5 cycloalkyl ring, or
  • each R 5 and each R 6 is independently hydrogen, halo, -CN, -C 1 -C 4 alkyl optionally substituted with halo, or -O-(C 1 -C 4 alkyl) optionally substituted with halo, wherein no more than two R 6 are other than hydrogen; wherein R 5 and an R 6 on an adjacent ring atom are optionally taken together to form a 4-7 membered saturated heterocyclic or cycloalkyl ring that is fused to R 4 ; R 7 is hydrogen, -C 1 -C 4 alkyl, or -C 1 -C 4 alkylene-O-C 1 -C 4 alkyl, wherein any C 1 - C4 alkyl or C1-C4 alkylene portion of R 7 is optionally substituted with one or more substituents independently selected from halo and -CN; R 8 is hydrogen or -C1
  • aliphatic groups contain 1-5 aliphatic carbon atoms. In other embodiments, aliphatic groups contain 1-4 aliphatic carbon atoms. In still other embodiments, aliphatic groups contain 1-3 aliphatic carbon atoms, and in yet other embodiments, aliphatic groups contain 1-2 aliphatic carbon atoms.
  • “cycloaliphatic” refers to a monocyclic C 3 -C 6 hydrocarbon that is completely saturated or that contains one or more units of unsaturation, but which is not aromatic, that has a single point of attachment to the rest of the molecule.
  • Suitable aliphatic groups include, but are not limited to, linear or branched, substituted or unsubstituted alkyl, alkenyl, alkynyl groups and hybrids thereof such as (cycloalkyl)alkyl, (cycloalkenyl)alkyl or (cycloalkyl)alkenyl.
  • alkyl as used herein, means a straight or branched hydrocarbon chain that is completely saturated.
  • Exemplary alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, iso-butyl, tert-butyl, etc.
  • heteroatom means one or more of oxygen, sulfur, nitrogen, phosphorus, or silicon (including, any oxidized form of nitrogen, sulfur, phosphorus, or silicon; the quaternized form of any basic nitrogen or; a substitutable nitrogen of a heterocyclic ring, for example N (as in 3,4-dihydro-2H-pyrrolyl), NH (as in pyrrolidinyl) or NR + (as in N-substituted pyrrolidinyl)).
  • unsaturated as used herein, means that a moiety has one or more units of unsaturation.
  • Suitable substituents include those described below for a carbon atom.
  • halogen and “halo” are used interchangeably and mean F, Cl, Br, or I.
  • aryl used alone or as part of a larger moiety as in “aralkyl”, “aralkoxy”, or “aryloxyalkyl”, refers to monocyclic and bicyclic ring systems having a total of five to fourteen ring members, wherein each ring atom is carbon, at least one ring in the system is aromatic and wherein each ring in the system contains three to seven ring members.
  • aryl may be used interchangeably with the term “aryl ring”.
  • aryl refers to an aromatic ring system which includes, but not limited to, phenyl, biphenyl, naphthyl, anthracyl and the like, which may bear one or more substituents. Also included within the scope of the term “aryl”, as it is used herein, is a group in which an aromatic ring is fused to one or more non-aromatic carbocyclic rings. [0030] The term “cycloalkyl ring” refers to a fully saturated carbocyclic ring. [0031] A first ring is “spiro-fused” to a second ring when the two rings share a single ring carbon atom.
  • a first ring is “fused” to a second ring when the two rings share two ring carbon atoms that are directly connected to one another.
  • Example of fused ring systems are: [0033] A first ring is “bridged” to a second ring when the two rings share two ring carbon atoms that are separated by at least one ring carbon atom.
  • a reference to the number of atoms in a bridged ring e.g., one R 2 and R 8 are taken together to form a bridged 3-5 membered ring, refers to the two bridgehead ring atoms plus any additional ring atoms in between those bridgehead atoms that form the ring that is bridged to the existing ring. For example if the ring defined by the structure taken together to form , e number of atoms in the bridged ring is 4 as enumerated in the bridged structure.
  • heteroatom refers to nitrogen, oxygen, or sulfur, and includes any oxidized form of nitrogen or sulfur, and any quaternized form of a basic nitrogen.
  • Heteroaryl groups include, without limitation, thienyl, furanyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, isothiazolyl, thiadiazolyl, pyridyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, indolizinyl, purinyl, naphthyridinyl, and pteridinyl.
  • heteroaryl and “heteroar-”, as used herein, also include groups in which a heteroaromatic ring is fused to one or more aryl or heteroaryl rings such that the resulting bi- or multicyclic ring system as a whole is fully aromatic.
  • heteroaryl may be used interchangeably with the terms “heteroaryl ring”, “heteroaryl group”, or “heteroaromatic”, any of which terms include rings that are optionally substituted.
  • heteroarylkyl refers to an alkyl group substituted by a heteroaryl, wherein the alkyl and heteroaryl portions independently are optionally substituted.
  • heterocycle As used herein, the terms “heterocycle”, “heterocyclyl”, “heterocyclic radical”, and “heterocyclic ring” are used interchangeably and refer to a stable 5- to 7-membered monocyclic or 7- to 10-membered bicyclic heterocyclic moiety that is either saturated or partially unsaturated, and having, in addition to carbon atoms, one or more, preferably one to four, heteroatoms, as defined above.
  • nitrogen includes a substituted nitrogen.
  • the nitrogen may be N (as in 3,4-dihydro-2H-pyrrolyl), NH (as in pyrrolidinyl), or +NR (as in N-substituted pyrrolidinyl).
  • a heterocyclic ring can be attached to its pendant group at any heteroatom or carbon atom that results in a stable structure and any of the ring atoms can be optionally substituted.
  • saturated or partially unsaturated heterocyclic radicals include, without limitation, tetrahydrofuranyl, tetrahydrothiophenyl pyrrolidinyl, piperidinyl, pyrrolidinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, decahydroquinolinyl, oxazolidinyl, piperazinyl, dioxanyl, dioxolanyl, diazepinyl, oxazepinyl, thiazepinyl, morpholinyl, and quinuclidinyl.
  • heterocycle used interchangeably herein, and also include groups in which a heterocyclyl ring is fused to one or more aryl, heteroaryl, or cycloaliphatic rings, such as indolinyl, 3H-indolyl, chromanyl, phenanthridinyl, or tetrahydroquinolinyl.
  • a heterocyclyl group may be mono- or bicyclic.
  • heterocyclylalkyl refers to an alkyl group substituted by a heterocyclyl, wherein the alkyl and heterocyclyl portions independently are optionally substituted.
  • compounds of the disclosure may contain “optionally substituted” moieties.
  • substituted whether preceded by the term “optionally” or not, means that one or more hydrogens of the designated moiety are replaced with a suitable substituent.
  • an “optionally substituted” group may have a suitable substituent at each substitutable position of the group, and when more than one position in any given structure may be substituted with more than one substituent selected from a specified group, the substituent may be either the same or different at every position.
  • Combinations of substituents envisioned by this disclosure are preferably those that result in the formation of stable or chemically feasible compounds.
  • stable refers to compounds that are not substantially altered when subjected to conditions to allow for their production, detection, and, in certain embodiments, their recovery, purification, and use for one or more of the purposes disclosed herein.
  • cyano-substituted Cx-Cy alkyl and fluoro-substituted Cx-Cy alkyl where each of x and y is an integer, refer to the corresponding alkyl having one or more of the indicated substituents in place of hydrogen.
  • Suitable monovalent substituents on R° are independently halogen, -(CH 2 )0-2R ⁇ , -(haloR ⁇ ), -(CH 2 )0-2OH, -(CH 2 )0-2OR ⁇ , -(CH 2 )0-2CH(OR ⁇ )2; O(haloR ⁇ ), -CN, -N 3 , -(CH 2 ) 0-2 C(O)R ⁇ , -(CH 2 ) 0-2 C(O)OH, -(CH 2 ) 0-2 C(O)OR ⁇ , -(CH 2 ) 0-2 SR ⁇ , -(CH 2 ) 0-2 SH, -(CH 2 )0-2NH 2 , -(CH 2 )0-2NHR ⁇ , -(CH 2 )0-2NR ⁇ 2,
  • Suitable divalent substituents that are bound to vicinal substitutable carbons of an “optionally substituted” group include: -O(CR * 2)2-3O-, wherein each independent occurrence of R * is selected from hydrogen, C 1-6 aliphatic which may be substituted as defined below, or an unsubstituted 5-6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.
  • Suitable substituents on the aliphatic group of R * include halogen, -R ⁇ , (haloR ⁇ ), OH, -OR ⁇ , -O(haloR ⁇ ), -CN, -C(O)OH, -C(O)OR ⁇ , -NH 2 , -NHR ⁇ , -NR ⁇ 2 , or -NO2, wherein each R ⁇ is unsubstituted or where preceded by “halo” is substituted only with one or more halogens, and is independently C 1-4 aliphatic, -CH 2 Ph, -O(CH 2 ) 0-1 Ph, or a 5-6- membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.
  • Suitable substituents on the aliphatic group and the substituted 5-6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur of R ⁇ are independently halogen, -R ⁇ , (haloR ⁇ ), -OH, -OR ⁇ , - O(haloR ⁇ ), -CN, -C(O)OH, -C(O)OR ⁇ , -NH 2 , -NHR ⁇ , -NR ⁇ 2 , or NO 2 , wherein each R ⁇ is unsubstituted or where preceded by “halo” is substituted only with one or more halogens, and is independently C 1-4 aliphatic, -CH 2 Ph, -O(CH 2 ) 0-1 Ph, or a 5-6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur
  • the term “pharmaceutically acceptable salt” refers to those salts which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response and the like, and are commensurate with a reasonable benefit/risk ratio.
  • Pharmaceutically acceptable salts are well known in the art. For example, S. M. Berge et al., describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 1977, 66, 1-19, incorporated herein by reference.
  • Pharmaceutically acceptable salts of the compounds of this disclosure include those derived from suitable inorganic and organic acids and bases.
  • Examples of pharmaceutically acceptable, nontoxic acid addition salts are salts of an amino group formed with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid or with organic acids such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid or malonic acid or by using other methods used in the art such as ion exchange.
  • inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid
  • organic acids such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid or malonic acid or by using other methods used in the art such as ion exchange.
  • salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate,
  • Salts derived from appropriate bases include alkali metal, alkaline earth metal, ammonium and N + (C1-4alkyl)4 salts.
  • Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like.
  • Further pharmaceutically acceptable salts include, when appropriate, nontoxic ammonium, quaternary ammonium, and amine cations formed using counterions such as halide, hydroxide, carboxylate, sulfate, phosphate, nitrate, lower alkyl sulfonate and aryl sulfonate.
  • structures depicted herein are also meant to include all isomeric (e.g., enantiomeric, diastereomeric, and geometric (or conformational)) forms of the structure; for example, the R and S configurations for each asymmetric center, Z and E double bond isomers, rotational isomers (atropisomers) and Z and E conformational isomers. Therefore, single stereochemical isomers as well as enantiomeric, diastereomeric, and geometric (or conformational) mixtures of the present compounds are within the scope of the disclosure. Unless otherwise stated, all tautomeric forms of the compounds of the disclosure are within the scope of the disclosure.
  • structures depicted herein are also meant to include compounds that differ only in the presence of one or more isotopically enriched atoms.
  • compounds having the present structures including the replacement of hydrogen by deuterium or tritium, or the replacement of a carbon by a 13 C- or 14 C-enriched carbon are within the scope of this disclosure.
  • Such compounds are useful, for example, as analytical tools, as probes in biological assays, or as therapeutic agents in accordance with the present disclosure.
  • Combinations of substituents and variables envisioned by this disclosure are only those that result in the formation of stable compounds.
  • stable refers to compounds which possess stability sufficient to allow manufacture and which maintains the integrity of the compound for a sufficient period of time to be useful for the purposes detailed herein (e.g., therapeutic or prophylactic administration to a subject).
  • the recitation of a listing of chemical groups in any definition of a variable herein includes definitions of that variable as any single group or combination of listed groups.
  • the recitation of an embodiment for a variable herein includes that embodiment as any single embodiment or in combination with any other embodiments or portions thereof.
  • biological sample includes, without limitation, cell cultures or extracts thereof; biopsied material obtained from a mammal or extracts thereof; and hair, skin, blood, saliva, urine, feces, semen, tears, or other body fluids or extracts thereof.
  • Inhibition of activity of a protein kinase, for example, Wee1A kinase or a mutant thereof, in a biological sample is useful for a variety of purposes that are known to one of skill in the art. Examples of such purposes include, but are not limited to, blood transfusion, organ transplantation, biological specimen storage, and biological assays.
  • a “disease or disorder associated with Wee1A kinase” or, alternatively, “a Wee1A kinase-mediated disease or disorder” means any disease or other deleterious condition in which Wee1A kinase, or a mutant thereof, is known or suspected to play a role.
  • the term “subject”, as used herein, means a mammal and includes human and animal subjects, such as domestic animals (e.g., horses, dogs, cats, etc.).
  • the terms “subject” and “patient” are used interchangeably.
  • the “patient” or “subject” means an animal, preferably a mammal, and most preferably a human.
  • compositions of this disclosure refers to a non- toxic carrier, adjuvant, or vehicle that does not destroy the pharmacological activity of the compound with which it is formulated.
  • Pharmaceutically acceptable carriers, adjuvants or vehicles that may be used in the compositions of this disclosure include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins, such as human serum albumin, buffer substances such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes, such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, cellulose- based substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene-polyoxy
  • compositions are formulated so that a dosage of between 0.01 to about 100 mg/kg, or about 0.1 mg/kg to about 50 mg/kg, and preferably from about 1 mg/kg to about 25 mg/kg, of subject body weight/day of the inhibitor can be administered to a patient receiving these compositions to obtain the desired therapeutic effect.
  • the amount of a compound of the present disclosure in the composition will also depend upon the particular compound in the composition.
  • treatment refers to partially or completely alleviating, inhibiting, delaying onset of, preventing, ameliorating and/or relieving a disorder or condition, or one or more symptoms of the disorder or condition, as described herein.
  • treatment may be administered after one or more symptoms have developed.
  • the term “treating” includes preventing or halting the progression of a disease or disorder.
  • treatment may be administered in the absence of symptoms.
  • treatment may be administered to a susceptible individual prior to the onset of symptoms (e.g., in light of a history of symptoms and/or in light of genetic or other susceptibility factors).
  • the term “treating” includes preventing relapse or recurrence of a disease or disorder.
  • the term “inhibitor” is defined as a compound that binds to and /or inhibits the target protein kinase with measurable affinity.
  • an inhibitor has an IC 50 and/or binding constant of less than about 50 ⁇ M, less than about 1 ⁇ M, less than about 500 nM, less than about 100 nM, less than about 50 nM, less than about 20 nM, or less than about 10 nM.
  • measurable affinity and “measurably inhibit,” as used herein, means a measurable change in Wee1A kinase or Myt1 kinase activity between a sample comprising a compound of the present disclosure, or composition thereof, and an equivalent sample comprising Wee1A kinase or Myt1 kinase, in the absence of said compound, or composition thereof.
  • the terms “dual inhibitor” and “dual Wee1A kinase/Myt1 kinase inhibitor” are used interchangeably and mean a compound disclosed herein that meets one or more of the following criteria: 1) a Myt1 kinase binding activity IC 50 of ⁇ 100 nM (i.e., “A” or “B” rated in Table 3); or 2) a Myt1 kinase target engagement EC 50 of ⁇ 500 nM (i.e., “A” or “B” rated in Table 4). 3.
  • the present disclosure provides a compound having structural formula AA: or a solvate, enantiomer, tautomer, or diastereomer thereof, or a pharmaceutically acceptable salt of any of the foregoing, wherein: each of X, Y and Z is independently CH or N; R 1 is -O-, -NH-, or -N(C1-C3 alkyl)-; each R 2 is independently fluoro, -CN, unsubstituted C 1 -C 5 alkyl, fluoro-substituted C1-C5 alkyl, or cyano-substituted C1-C5 alkyl, wherein two R 2 bound to the same carbon atom are optionally taken together to form a spiro-fused C 3 -C 7 cycloalkyl ring, or two R 2 bound to different carbon atoms are optionally taken together to form a bridged or fused C3-C
  • each R 5 and each R 6 is independently hydrogen, halo, -CN, -C 1 -C 4 alkyl optionally substituted with halo, -O-(C1-C4 alkyl) optionally substituted with halo, -O-(C1-C4 alkyl) substituted with C 3 -C 6 cycloalkyl, an N-linked saturated 3-7 membered heterocyclyl, a 5-6 membered heteroaryl, or phenyl, wherein no more than two R 6 are other than hydrogen; wherein R 5 and an R 6 on an adjacent ring atom are optionally taken together to form a 4-7 membered saturated heterocyclic or cycloalkyl ring that is fused to R 4 ; R 7 is hydrogen, -C1-C4 alkyl, -C1-C4 alkylene-O-C1-C4 alkyl, -C(O)- C1-C4 alkyl,
  • the present disclosure provides a compound having structural formula A: or a solvate, enantiomer, tautomer, or diastereomer thereof, or a pharmaceutically acceptable salt of any of the foregoing, wherein: each of X, Y and Z is independently CH or N; R 1 is -O-, -NH-, or -N(C 1 -C 3 alkyl)-; each R 2 is independently fluoro, -CN, unsubstituted C1-C5 alkyl, fluoro-substituted C 1 -C 5 alkyl, or cyano-substituted C 1 -C 5 alkyl, wherein two R 2 bound to the same carbon atom are optionally taken together to form a spiro-fused C3-C7 cycloalkyl ring, or two R 2 bound to different carbon atoms are optionally taken together to form a bridged C 3 -C 5 cycloalkyl ring
  • each R 5 and each R 6 is independently hydrogen, halo, -CN, -C 1 -C 4 alkyl optionally substituted with halo, -O-(C1-C4 alkyl) optionally substituted with halo, -O-(C1-C4 alkyl) substituted with C3-C6 cycloalkyl, an N-linked saturated 3-7 membered heterocyclyl, a 5-6 membered heteroaryl, or phenyl, wherein no more than two R 6 are other than hydrogen; wherein R 5 and an R 6 on an adjacent ring atom are optionally taken together to form a 4-7 membered saturated heterocyclic or cycloalkyl ring that is fused to R 4 ; R 7 is hydrogen, -C1-C4 alkyl, -C1-C4 alkylene-O-C1-C4 alkyl, -C(O)- C1-C4 alkyl, or a C 3 -C
  • each of X, Y and Z is independently CH or N; R 1 is -O-, -NH-, or -N(C 1 -C 3 alkyl)-; each R 2 is independently fluoro, -CN, unsubstituted C1-C5 alkyl, fluoro-substituted C 1 -C 5 alkyl, or cyano-substituted C 1 -C 5 alkyl, wherein two R 2 bound to the same carbon atom are optionally taken together to form a spiro-fused C 3 -C 7 cycloalkyl ring, or two R 2 bound to different carbon atoms are optionally taken together to form a bridged C3-C5 cycloalkyl ring, or one R 2 and R 8 are optionally taken together to form a bridged 3-5
  • each R 5 and each R 6 is independently hydrogen, halo, -CN, -C1-C4 alkyl optionally substituted with halo, or -O-(C 1 -C 4 alkyl) optionally substituted with halo, wherein no more than two R 6 are other than hydrogen; wherein R 5 and an R 6 on an adjacent ring atom are optionally taken together to form a 4-7 membered saturated heterocyclic or cycloalkyl ring that is fused to R 4 ; R 7 is hydrogen, -C1-C4 alkyl, or -C1-C4 alkylene-O-C1-C4 alkyl, wherein any C1- C4 alkyl or C1-C4 alkylene portion of R 7 is optionally substituted with one or more substituents independently selected from halo and -CN; R 8 is hydrogen or -C 1 -C 4 alkyl; R 9 is hydrogen, halo, -CN, -C1-C4 alkyl optional
  • each of X, Y and Z is independently CH or N.
  • each of X, Y and Z is CH (i.e., the ring comprising X, Y, and Z is phen-2,6-diyl).
  • X is N and each of Y and Z is CH (i.e., the ring is pyridin-2,6-diyl).
  • X and Y is N and Z is CH (i.e., the ring is pyrimidin- 2,6-diyl).
  • each of X and Z is N and Y is CH (i.e., the ring is pyrazin- 2,6-diyl).
  • R 1 is -O-, -NH-, or -N(C1-C3 alkyl)-.
  • R 1 is -O-.
  • R 1 is -NH-.
  • R 1 is -N(CH3)-.
  • R 1 is -N(CH 2 CH3)-.
  • R 1 is -N(CH 2 CH 2 CH3)-.
  • each R 2 is independently fluoro, -CN, unsubstituted C 1 -C 5 alkyl, fluoro-substituted C 1 -C 5 alkyl, or cyano- substituted C1-C5 alkyl, wherein two R 2 bound to the same carbon atom are optionally taken together to form a spiro-fused C 3 -C 7 cycloalkyl ring, or two R 2 bound to different carbon atoms are optionally taken together to form a bridged or fused C3-C5 cycloalkyl ring, or one R 2 and R 8 are optionally taken together to form a bridged or fused 3-5 membered ring.
  • the spiro-fused, fused, or bridged ring in the definition of R 2 refers to the ring formed by taking the two R 2 or the R 2 and R 8 substituents together and how that ring is attached to the ring depicted as in Formula I.
  • Examples of such spiro-fused, fused and bridged ring systems formed by taking two R 2 or one R 2 and one R 8 together include, but are not limited t [0068]
  • R 2 is absent.
  • one R 2 and R 8 are taken together with the ring to which they are bound to form . so e embodiments, one R 2 and R 8 are taken together with the ring to which they are bound to form .
  • R 2 and R 8 are taken together with the ring to which they are bound to form .
  • R 4 is ,
  • R 4 [0071] is .
  • R 5 is hydrogen. In some aspects of either of these embodiments, R 5 is C 1 -C 4 alkyl. In some aspects of this embodiment, R 5 is methyl. In some aspects of this embodiment, R 5 is -CN. In some aspects of either of these embodiments, R 5 is halo. In some aspects of this embodiment, R 5 is chloro. In some aspects of this embodiment, R 5 is fluoro. In some aspects of this embodiment, R 5 is bromo. In some aspects of either of these embodiments, R 5 is -O-(C 1 -C 4 alkyl) optionally substituted with halo.
  • R 5 is -O-(C1-C4 alkyl) optionally substituted with fluoro.
  • R 5 is -OCF 3 .
  • each R 6 is hydrogen.
  • one R 6 is hydrogen and the other R 6 is halo.
  • one R 6 is hydrogen and the other R 6 is chloro.
  • one R 6 is hydrogen and the other R 6 is fluoro.
  • one R 6 is hydrogen and the other R 6 is bromo.
  • one R 6 is hydrogen and the other R 6 is -C 1 -C 4 alkyl optionally substituted with halo.
  • one R 6 is hydrogen and the other R 6 is -C1-C4 alkyl optionally substituted with fluoro. In some aspects of this embodiment, one R 6 is hydrogen and the other R 6 is -CH 3 . In some aspects of this embodiment, one R 6 is hydrogen and the other R 6 is -CF3. In some aspects of this embodiment, one R 6 is hydrogen and the other R 6 is -O(C 1 -C 4 alkyl) optionally substituted with halo. In some aspects of this embodiment, one R 6 is hydrogen and the other R 6 is -O(C1-C4 alkyl) optionally substituted with fluoro. In some aspects of this embodiment, one R 6 is hydrogen and the other R 6 is -OCH3.
  • one R 6 is hydrogen and the other R 6 is -OCH 2 CF 3 . In some aspects of this embodiment, one R 6 is hydrogen and the other R 6 is -CN. In some aspects of this embodiment, R 5 and the R 6 bound to an adjacent ring atom are taken together to form methylenedioxy. In some aspects of this embodiment, one R 6 is hydrogen and the other R 6 is cyclopropylmethoxy. In some aspects of this embodiment, one R 6 is hydrogen and the other R 6 is morpholinyl. In some aspects of this embodiment, one R 6 is methyl and the other R 6 is morpholinyl. In some aspects of this embodiment, one R 6 is hydrogen and the other R 6 is thiomorpholinyl.
  • one R 6 is hydrogen and the other R 6 is 2-methylpropan-1-yloxy. In some aspects of this embodiment, one R 6 is hydrogen and the other R 6 is 1-methyl-1H-pyrazolyl. In some aspects of this embodiment, one R 6 is hydrogen and the other R 6 is phenyl optionally substituted with halo. In some aspects of this embodiment, one R 6 is hydrogen and the other R 6 is ethyloxy optionally substituted with halo.
  • R 4 is any one of phenyl, 3-methylphenyl, 3-chloro-5- bromophenyl, 3,4-dichlorophenyl, 4-bromophenyl, 4-chlorophenyl, 4-fluorophenyl, 4-(1- methylpiperidin-4-yl)-5-methylphenyl, 4-trifluoromethoxyphenyl, 3-methyl-4- trifluoromethoxyphenyl, 4-(2,2,2-trifluoroethan-1-yl)oxyphenyl, 3-methyl-4-(2,2,2- trifluoroethan-1-yl)oxyphenyl, 4-trifluoromethylphenyl, 3-methyl-4-chlorophenyl, 3-methyl- 4-fluorophenyl, 3-methyl-5-fluorophenyl, 3-methyl-4-cyanophenyl, 3-methyl-4- methoxyphenyl, 3-methyl-4-(2,2,2-trifluoroethan-1-yloxy)phenyl, or 3,
  • R 4 is any one of 4-(cyclopropylmethyloxy)phenyl, 4- (morpholin-4-yl)phenyl, 4-(thiomorpholin-4-yl)phenyl, 4-(2-methylpropan-1-yloxy)phenyl, 3- (1-methyl-1H-pyrazol-4-yl)phenyl, 4-(1-methyl-1H-pyrazol-4-yl)phenyl, 3-methyl-4- (morpholin-4-yl)phenyl, 4-(phenyl)phenyl, 3-bromophenyl, 3-cyanophenyl, 4-(2-fluoroethan- 1-yloxy)phenyl, 4-cyanophenyl, , 4-(1,1-dioxothiazinan-4-yl)phenyl, 3-(pyridin-3-yl)phenyl, 3-(pyrimidin-5-yl)phenyl, 3-(1H-imidazol-1-yl)phenyl, 3-(1H-pyr
  • R 4 is some embodi 4 ments, R is . n some aspects of either of these two embodiments, R 5 is hydrogen. In some aspects of either of these embodiments, R 5 is halo. In some aspects of either of these embodiments, R 5 is chloro. In some aspects of either of these embodiments, R 5 is fluoro. In some aspects of either of these embodiments, R 5 is cyano. In some aspects of either of these embodiments, R 5 is C 1 -C 4 alkyl optionally substituted with halo. In some aspects of either of these embodiments, R 5 is C1-C4 alkyl optionally substituted with fluoro. In some aspects of either of these embodiments, R 5 is -O-C 1 -C 4 alkyl.
  • R 5 is methoxy. In some aspects of either of these embodiments, R 5 is methyl. In some aspects of either of these embodiments, R 5 is -CF 3 . In some aspects of either of these embodiments, R 6 is hydrogen. In some aspects of either of these embodiments, R 6 is methyl. In some aspects of either of these embodiments, R 6 is -O-C 1 -C 4 alkyl. In some aspects of either of these embodiments, R 6 is methoxy. In some aspects of either of these embodiments, R 6 is - O-C1-C4 haloalkyl. In some aspects of either of these embodiments, R 6 is 2-fluoroethoxy.
  • R 4 is any one of 2-methylpyridin-4-yl, 6-methylpyridin-3- yl, 2,6-dimethylpyridin-4-yl, 2-trifluoromethyl-6-methylpyridin-4-yl, or 2- trifluoromethylpyridin-4-yl.
  • R 4 is any one of pyridin-3-yl, 2-methylpyridin-5-yl, 2- methoxypyridin-4-yl, 2-methoxypyridin-5-yl, 3-chloropyridin-5-yl, 3-fluoropyridin-5-yl, 3- cyanopyridin-5-yl, 3-methylpyridin-5-yl, or 3-methoxypyridin-5-yl.
  • R 4 is 2-(2-fluoroethoxy)pyridin-5-yl.
  • R 4 is pects of this embodiment, R 7 is C1-C4 alkyl optionally substituted with halo.
  • R 7 is C1-C4 alkyl optionally substituted with fluoro. In some aspects of this embodiment, R 7 is methyl. In some aspects of this embodiment, R 7 is ethyl. In some aspects of this embodiment, R 7 is propyl. In some aspects of this embodiment, R 7 is isopropyl. In some aspects of this embodiment, R 7 is -CH 2 CH(CH 3 ) 2 . In some aspects of this embodiment, R 7 is -CH 2 C(CH 3 ) 2 F. In some aspects of this embodiment, R 7 is -CH 2 CH 2 CF3.
  • R 4 is any one of 1-methyl-1H-pyrazol-4-yl, 1-propyl-1H-pyrazol-4yl, 1-isopropyl-1H-pyrazol-4-yl, 1-(2,2- dimethylethan-1-yl)pyrazol-4-yl, 1-(2-fluoro-2,2-dimethylethan-1-yl)-1H-pyrazol-4-yl, or 1- (3,3,3-trifluoropropan-1-yl)-1H-pyrazol-4-yl.
  • R 4 is ects of this embodiment, each R 6 is hydrogen. In some aspects of this embodiment, R 7 is hydrogen.
  • R 7 is C1-C4 alkyl optionally substituted with halo. In some aspects of this embodiment, R 7 is C 1 -C 4 alkyl optionally substituted with fluoro. In some aspects of this embodiment, R 7 is methyl. In some aspects of this embodiment, R 7 is ethyl. In some aspects of this embodiment, R 7 is propyl. In some aspects of this embodiment, R 7 is isopropyl. In some aspects of this embodiment, R 7 is -CH 2 CH(CH3)2. In some aspects of this embodiment, R 7 is -CH 2 C(CH 3 ) 2 F. In some aspects of this embodiment, R 7 is -CH 2 CH 2 CF 3 .
  • R 7 is cyclopropyl. In some aspects of this embodiment, R 7 is isobutyl. In some aspects of this embodiment, R 7 is 3-fluoropropyl. In some aspects of this embodiment, R 4 is 1-methyl-1H-indazol-5-yl.
  • R 4 is selected from 1H-indazol-5-yl, 1- methyl-1H-indazol-5-yl, 1-ethyl-1H-indazol-5-yl, 1-isopropyl-1H-indazol-5-yl, 1-propyl-1H- indazol-5-yl, 1-(3-fluoropropan-1-yl)-1H-indazol-5-yl, 1-isobutyl-1H-indazol-5-yl, and 1- cyclopropyl-1H-indazol-5-yl.
  • R 4 when R 4 is hydrogen and the other R 6 is methyl or halo.
  • R 4 is 3-chloro-1H-indazol-5-yl, 3- methyl-1H-indazol-5-yl, or 1,3-dimethyl-1H-indazol-5-yl.
  • R 4 is me aspects of this embodiment, R 7 is hydrogen. In some aspects of this embodiment, R 7 is methyl. In some aspects of this embodiment, R 7 is ethyl. In some aspects of this embodiment, R 7 is t-butyl. In some aspects of this embodiment, each R 6 is hydrogen. In some aspects of this embodiment, R 4 is 2-methyl-2H-indazol-5-yl, or 2-ethyl-2H-indazol-5-yl.
  • R 4 is 2-(t-butyl)-2H-indazol-5-yl. [0082] In some embodiments, R 4 is . In some aspects of this embodiment, R 9 is an optionally substituted phenyl. In some aspects of this embodiment, R 9 is unsubstituted phenyl. In some aspects of this embodiment, R 4 is 3-phenylisothiazol-5-yl. [0083] In some embodiments, R 4 is . In some embodiments, R 4 is . In some embodiments, R 4 is . n some aspects of each of these embodiments, each R 6 is hydrogen. In some aspects of each of these embodiments, two R 6 are hydrogen and the other R 6 is other than hydrogen.
  • R 6 is hydrogen and the other two R 6 are other than hydrogen.
  • R 4 is benzofuran-6-yl. In some aspects of this embodiment, R 4 is benzofuran-5-yl.
  • R 4 is . me aspects of this embodiment, R 7 is C 1 -C 4 alkyl. In some aspects of this embodiment, R 7 is methyl. In some aspects of this embodiment, R 7 is -C(O)CH3.
  • each R 6 is hydrogen. In some aspects of this embodiment, two R 6 are hydrogen and the other R 6 is other than hydrogen. In some aspects of this embodiment, one R 6 is hydrogen and the other two R 6 are other than hydrogen.
  • R 4 is 1-methyl-1H-indol-5-yl. In some aspects of this embodiment, R 4 is 1-acetyl-1H-indol-5-yl. [0085] In some embodiments, R 4 is . some aspects of this embodiment, each R 6 is hydrogen. In some aspects of this embodiment, R 4 is 1,2,4-triazolo[1,5- a]pyridin-6-yl. [0086] In some embodiments, R 4 is . ome aspects of this embodiment, each R 6 is hydrogen. In some aspects of this embodiment, R 4 is quinoxalin-6-yl. [0087] In some embodiments, R 4 is selected from any one of , me aspects of this embodiment, each R 6 is hydrogen.
  • R 4 is quinolin-6-yl, quinolin-7-yl, isoquinolin-6-yl, or isoquinolin-7-yl.
  • each R 6 is hydrogen. In some aspects of these embodiments, one R 6 is hydrogen and the other R 6 is methyl. In some aspects of these embodiments, R 4 is 2- methylbenzo[d]oxazol-5-yl or 2-methylbenzo[d]oxazol-6-yl.
  • R 4 is ects of these embodiments, each R 6 is hydrogen. In some aspects of these embodiments, R 4 is benzo[d]isoxazol-6-yl.
  • R 4 is . me aspects of this embodiment, each R 6 is hydrogen. In some aspects of this embodiment, one R 6 is hydrogen and the other R 6 is methyl. In some aspects of this embodiment, R 4 is benzo[d]thiazol-6-yl or 2-methylbenzo[d]thiazol-6-yl. [0091] In some embodiments, R 4 is . ome aspects of this embodiment, each R 6 is hydrogen. In some aspects of this embodiment, R 7 is methyl. In some aspects of this embodiment, R 4 is 1-methyl-1H-benzo[d]imidazol-5-yl. In some aspects of this embodiment, R 7 is isopropyl.
  • R 4 is 1-isopropyl-1H- benzo[d]imidazol-5-yl. [0092] In some embodiments, R 4 is ects of this embodiment, each R 6 is hydrogen. In some aspects of this embodiment, R 4 is benzo[d]thiazol-5-yl. [0093] In some embodiments, R 4 is ects of this embodiment, each R 6 is hydrogen. In some aspects of this embodiment, R 7 is methyl. In some aspects of this embodiment, R 4 is 1-methyl-1H-indazol-6-yl. [0094] In some embodiments, R 4 is e aspects of this embodiment, each R 6 is hydrogen. In some aspects of this embodiment, R 7 is methyl.
  • R 4 is 2-methyl-2H-indazol-6-yl.
  • R 4 is . me aspects of this embodiment, each R 6 is hydrogen.
  • R 4 is imidazo[1,2- a]pyridin-6-yl.
  • R 4 is pects of this embodiment, R 6 is hydrogen.
  • R 7 is C1-C4 alkyl.
  • R 7 is isopropyl.
  • R 4 is 1-isopropyl-1H- pyrazolo[3,4-b]pyridin-5-yl.
  • R 4 is .
  • R 6 is hydrogen.
  • R 7 is C1-C4 alkyl.
  • R 7 is isopropyl.
  • R 4 is 1-isopropyl-1H- benzo[d][1,2,3]triazol-5-yl.
  • R 5 is hydrogen, halo, -CN, - C 1 -C 4 alkyl optionally substituted with halo, or -O-(C 1 -C 4 alkyl) optionally substituted with halo, or R 5 and an R 6 on an adjacent ring atom are taken together to form a 4-7 membered saturated heterocyclic or cycloalkyl ring that is fused to R 4 .
  • R 5 is hydrogen.
  • R 5 is halo.
  • R 5 is chloro.
  • R 5 is fluoro.
  • R 5 is C 1 -C 4 alkyl optionally substituted with halo.
  • R 5 is C1-C4 alkyl optionally substituted with fluoro. In some embodiments, R 5 is methyl. In some embodiments, R 5 is -CF 3 . In some embodiments, R 5 and an R 6 on an adjacent ring atom are taken together to form a 4-7 membered saturated heterocyclic or cycloalkyl ring that is fused to R 4 . In some embodiments, R 5 and an R 6 on an adjacent ring atom are taken together to form a methylenedioxy that is fused to R 4 .
  • each R 6 is independently hydrogen, halo, -CN, -C1-C4 alkyl optionally substituted with halo, or -O-(C1-C4 alkyl) optionally substituted with halo, wherein no more than two R 6 are other than hydrogen. In some embodiments, no more than one R 6 is other than hydrogen. In some embodiments, each R 6 is hydrogen. In some embodiments, one R 6 is halo. In some embodiments, one R 6 is chloro. In some embodiments, one R 6 is fluoro. In some embodiments, one R 6 is bromo. In some embodiments, one R 6 is -C1-C4 alkyl optionally substituted with halo.
  • one R 6 is -C 1 -C 4 alkyl optionally substituted with fluoro. In some embodiments, one R 6 is - CH3. In some embodiments, one R 6 is -CF3. In some aspects of this embodiment, one R 6 is hydrogen and the other R 6 is halo. In some embodiments, one R 6 is -O(C 1 -C 4 alkyl) optionally substituted with halo. In some embodiments, one R 6 is -O(C 1 -C 4 alkyl) optionally substituted with fluoro. In some embodiments, one R 6 is -OCH3. In some embodiments, one R 6 is - OCH 2 CF 3 . In some embodiments, one R 6 is -CN.
  • R 7 is hydrogen, -C1-C4 alkyl, -C 1 -C 4 alkylene-O-C 1 -C 4 alkyl, -C(O)-C 1 -C 4 alkyl, or a C 3 -C 6 cycloalkyl, wherein any C 1 -C 4 alkyl or C1-C4 alkylene portion of R 7 is optionally substituted with one or more substituents independently selected from halo and -CN.
  • R 7 is C 1 -C 4 alkyl optionally substituted with halo.
  • R 7 is C1-C4 alkyl optionally substituted with fluoro.
  • R 7 is methyl. In some embodiments, R 7 is ethyl. In some embodiments, R 7 is propyl. In some embodiments, R 7 is isopropyl. In some embodiments, R 7 is -CH 2 CH(CH3)2. In some embodiments, R 7 is -CH 2 C(CH3)2F. In some embodiments, R 7 is -CH 2 CH 2 CF3. In some embodiments, R 7 is -CH 2 CH 2 CH 2 F3. In some embodiments, R 7 is cyclopropyl. In some embodiments, R 7 is -C(O)CH3.
  • R 8 is hydrogen, -C1-C4 alkyl, or a 4-6 membered saturated heterocycle. In some embodiments, R 8 is hydrogen. In some embodiments, R 8 is methyl. In some embodiments, R 8 is ethyl. In some embodiments, R 8 is propyl. In some embodiments, R 8 is oxetan-3-yl.
  • R 9 is hydrogen, halo, -CN, - C1-C4 alkyl optionally substituted with halo, -O-C1-C4 alkyl optionally substituted with halo, or an optionally substituted phenyl.
  • R 9 is unsubstituted phenyl.
  • that ring may be a spiro-fused bicyclic ring when two R 2 bound to the same carbon atom are taken together to form a C 3 -C 7 cycloalkyl ring.
  • that ring may be a bridged bicyclic ring when two R 2 bound to different carbon atoms are taken together to form a C3-C5 cycloalkyl ring.
  • that ring may be a bridged bicyclic ring when one R 2 and R 8 are taken together to form a 3-5 membered ring. In some embodiments, that ring may be a fused bicyclic ring when two R 2 bound to different carbon atoms are taken together to form a C 3 -C 5 cycloalkyl ring. In some embodiments, that ring may be a fused bicyclic ring when one R 2 and R 8 are taken together to form a 3-5 membered ring. [0104] In some embodiments, the ring represented by the structure: piperidinyl ring.
  • the piperidinyl ring is a piperidin-3- yl ring (i.e., n is 1 and m is 2; or n is 3 and m is 0). In some aspects of these embodiments, the piperidinyl ring is a piperidin-4-yl ring (i.e., n is 2 and m is 1). In some aspects of these embodiments, the R 8 substituent on such piperidinyl ring is hydrogen. In some aspects of these embodiments, the R 8 substituent on such piperidinyl ring is methyl. In some aspects of these embodiments, the R 8 substituent on such piperidinyl ring is trideuteromethyl (methyl-d3).
  • the R 8 substituent on such piperidinyl ring is ethyl. In some aspects of these embodiments, the R 8 substituent on such piperidinyl ring is isopropyl. In some aspects of these embodiments, the R 8 substituent on such piperidinyl ring is propyl. In some aspects of these embodiments, R 2 is absent. In some aspects of these embodiments, one or more R 2 are methyl.
  • the piperidinyl ring is piperidinyl-4-yl, piperidinyl-3-yl, 1-methylpiperidin-4-yl, 1-d3-methylpiperidin-4-yl, 1- methylpiperidin-3-yl, 1-ethylpiperidin-4-yl, 1-propylpiperidin-4-yl, 1,2-dimethylpiperidin-4- yl, 2,2-dimethylpiperidin-4-yl, 2-methylpiperidin-4-yl, 1,2,2-trimethylpiperidin-4-yl, or 1- (oxetan-3-yl)piperindin-4-yl.
  • the ring represented by the structure pyrrolidinyl ring.
  • the pyrrolidinyl ring is a pyrrolidin- 3-yl ring (i.e., n is 1 and m is 1; or n is 2 and m is 0).
  • the R 8 substituent on such pyrrolidinyl ring is hydrogen.
  • the R 8 substituent on such pyrrolidinyl ring is methyl.
  • the R 8 substituent on such pyrrolidinyl ring is ethyl.
  • the R 8 substituent on such pyrrolidinyl ring is isopropyl. In some aspects of these embodiments, R 2 is absent.
  • the R 8 substituent on such quinuclidinyl ring is methyl. In some aspects of these embodiments, the R 8 substituent on such quinuclidinyl ring is ethyl. In some aspects of these embodiments, the R 8 substituent on such quinuclidinyl ring is isopropyl. In some aspects of these embodiments, R 2 is absent. [0107] In some embodiments, the ring represented by the structure: an azabicyclo[3.2.1]octanyl ring.
  • the azabicyclo[3.2.1]octanyl ring is an 8-azabicyclo[3.2.1]octan-3-yl ring (i.e., n is 1 and m is 2; or n is 3 and m is 0; and two R 2 bound to different carbon atoms are taken together to form a bridged C4 cycloalkyl ring).
  • the R 8 substituent on such azabicyclo[3.2.1]octanyl ring is hydrogen.
  • the R 8 substituent on such azabicyclo[3.2.1]octanyl ring is methyl.
  • the R 8 substituent on such azabicyclo[3.2.1]octanyl ring is ethyl. In some aspects of these embodiments, the R 8 substituent on such azabicyclo[3.2.1]octanyl ring is isopropyl. In some aspects of these embodiments, R 2 is absent. [0108] In some embodiments, the ring represented by the structure: an azabicyclo[3.3.1]nonanyl ring.
  • the azabicyclo[3.3.1]nonanyl ring is a 9-azabicyclo[3.3.1]nonan-3-yl ring (i.e., n is 1 and m is 2; or n is 3 and m is 0; and two R 2 bound to different carbon atoms are taken together to form a bridged C 5 cycloalkyl ring).
  • the R 8 substituent on such azabicyclo[3.3.1]nonanyl ring is hydrogen.
  • the R 8 substituent on such azabicyclo[3.3.1]nonanyl ring is methyl.
  • the R 8 substituent on such azabicyclo[3.3.1]nonanyl ring is ethyl. In some aspects of these embodiments, the R 8 substituent on such azabicyclo[3.3.1]nonanyl ring is isopropyl. In some aspects of these embodiments, R 2 is absent. [0109] In some embodiments, the ring represented by the structure: an azabicyclo[2.2.2]octanyl ring.
  • the azabicyclo[2.2.2]octanyl ring is a 2-azabicyclo[2.2.2]octan-5-yl ring (i.e., n is 1 and m is 2; or n is 3 and m is 0; and two R 2 bound to different carbon atoms are taken together to form a bridged C 4 cycloalkyl ring).
  • the R 8 substituent on such azabicyclo[2.2.2]octanyl ring is methyl.
  • the ring represented by the structure: is an azepanyl ring.
  • the azepanyl ring is azepan-4-yl (i.e., n is 2 and m is 2).
  • the R 8 substituent on such azepanyl ring is hydrogen.
  • the R 8 substituent on such azepanyl ring is methyl.
  • the compound of formula I may be a compound, or a pharmaceutically acceptable salt thereof, selected from Table 1, which lists chemical structure for each compound and LCMS and 1 H NMR for those compounds that have therein been so analyzed. Chemical shifts are reported in ppm ( ⁇ ) using the residual solvent as internal standard.
  • Peak multiplicities given in Hz are expressed as follow: s, singlet; d, doublet; dd, doublet of doublets; ddd, doublet of doublet of doublets; t, triplet; dt, doublet of triplets; q, quartet; dq, doublet of quartets; p, pentet; h, heptet; m, multiplet; br s, broad singlet.
  • Table 1 Exemplary Compounds
  • the disclosure provides a composition comprising a compound of this disclosure or a pharmaceutically acceptable derivative thereof and a pharmaceutically acceptable carrier, adjuvant, or vehicle.
  • the amount of compound in compositions of this disclosure is such that is effective to measurably inhibit Wee1A kinase, or a mutant thereof, in a biological sample or in a patient.
  • a composition of this disclosure is formulated for administration to a patient in need of such composition.
  • a composition of this disclosure is formulated for oral administration to a patient.
  • the term “patient”, as used herein, means an animal, preferably a mammal, and most preferably a human.
  • pharmaceutically acceptable carrier, adjuvant, or vehicle refers to a non- toxic carrier, adjuvant, or vehicle that does not destroy the pharmacological activity of the compound with which it is formulated.
  • compositions of this disclosure include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins, such as human serum albumin, buffer substances such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes, such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, cellulose- based substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, polyethylene glycol and wool fat.
  • ion exchangers alumina, aluminum stearate, lecithin
  • serum proteins such as human serum albumin
  • buffer substances such as phosphates, glycine, sorbic acid, potassium sorbate
  • compositions of the present disclosure may be administered orally, parenterally, by inhalation spray, topically, rectally, nasally, buccally, vaginally or via an implanted reservoir.
  • parenteral as used herein includes subcutaneous, intravenous, intramuscular, intra- articular, intra-synovial, intrasternal, intrathecal, intrahepatic, intralesional and intracranial injection or infusion techniques.
  • compositions are administered orally, intraperitoneally or intravenously.
  • Sterile injectable forms of the compositions of this disclosure may be aqueous or oleaginous suspension. These suspensions may be formulated according to techniques known in the art using suitable dispersing or wetting agents and suspending agents.
  • the sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent, for example as a solution in 1,3-butanediol.
  • the acceptable vehicles and solvents that may be employed are water, Ringer’s solution and isotonic sodium chloride solution.
  • sterile, fixed oils are conventionally employed as a solvent or suspending medium.
  • any bland fixed oil may be employed including synthetic mono- or di-glycerides.
  • Fatty acids such as oleic acid and its glyceride derivatives are useful in the preparation of injectables, as are natural pharmaceutically acceptable oils, such as olive oil or castor oil, especially in their polyoxyethylated versions.
  • These oil solutions or suspensions may also contain a long-chain alcohol diluent or dispersant, such as carboxymethyl cellulose or similar dispersing agents that are commonly used in the formulation of pharmaceutically acceptable dosage forms including emulsions and suspensions.
  • compositions of this disclosure are administered orally.
  • Pharmaceutically acceptable compositions of this disclosure may be orally administered in any orally acceptable dosage form including, but not limited to, capsules, tablets, aqueous suspensions or solutions.
  • carriers commonly used include lactose and corn starch.
  • Lubricating agents, such as magnesium stearate, are also typically added.
  • useful diluents include lactose and dried cornstarch.
  • compositions of this disclosure may be administered in the form of suppositories for rectal administration. These can be prepared by mixing the agent with a suitable non-irritating excipient that is solid at room temperature but liquid at rectal temperature and therefore will melt in the rectum to release the drug. Such materials include cocoa butter, beeswax and polyethylene glycols.
  • compositions of this disclosure may also be administered topically, especially when the target of treatment includes areas or organs readily accessible by topical application, including diseases of the eye, the skin, or the lower intestinal tract. Suitable topical formulations are readily prepared for each of these areas or organs.
  • Topical application for the lower intestinal tract can be effected in a rectal suppository formulation (see above) or in a suitable enema formulation. Topically-transdermal patches may also be used.
  • provided pharmaceutically acceptable compositions may be formulated in a suitable ointment containing the active component suspended or dissolved in one or more carriers.
  • Carriers for topical administration of compounds of this disclosure include, but are not limited to, mineral oil, liquid petrolatum, white petrolatum, propylene glycol, polyoxyethylene, polyoxypropylene compound, emulsifying wax and water.
  • provided pharmaceutically acceptable compositions can be formulated in a suitable lotion or cream containing the active components suspended or dissolved in one or more pharmaceutically acceptable carriers.
  • Suitable carriers include, but are not limited to, mineral oil, sorbitan monostearate, polysorbate 60, cetyl esters wax, cetearyl alcohol, 2octyldodecanol, benzyl alcohol and water.
  • compositions may be formulated as micronized suspensions in isotonic, pH adjusted sterile saline, or, preferably, as solutions in isotonic, pH adjusted sterile saline, either with or without a preservative such as benzalkonium chloride.
  • the pharmaceutically acceptable compositions may be formulated in an ointment such as petrolatum.
  • Pharmaceutically acceptable compositions of this disclosure may also be administered by nasal aerosol or inhalation.
  • compositions are prepared according to techniques well-known in the art of pharmaceutical formulation and may be prepared as solutions in saline, employing benzyl alcohol or other suitable preservatives, absorption promoters to enhance bioavailability, fluorocarbons, and/or other conventional solubilizing or dispersing agents.
  • pharmaceutically acceptable compositions of this disclosure are formulated for oral administration.
  • the amount of compounds of the present disclosure that may be combined with the carrier materials to produce a composition in a single dosage form will vary depending upon the host treated, the particular mode of administration.
  • provided compositions should be formulated so that a dosage of between 0.001 - 100 mg/kg body weight/day of the inhibitor can be administered to a patient receiving these compositions.
  • a specific dosage and treatment regimen for any particular patient will depend upon a variety of factors, including the activity of the specific compound employed, the age, body weight, general health, sex, diet, time of administration, rate of excretion, drug combination, and the judgment of the treating physician and the severity of the particular disease being treated.
  • the amount of a compound of the present disclosure in the composition will also depend upon the particular compound in the composition.
  • Uses of Compounds and Pharmaceutically Acceptable Compositions [0128] Compounds and compositions described herein are generally useful for the inhibition of protein kinase activity of one or more enzymes.
  • the activity of a compound utilized in this disclosure as an inhibitor of Wee1A kinase or Myt1 kinase, or a mutant of either of the foregoing, may be assayed in vitro, in vivo or in a cell line.
  • In vitro assays include assays that determine inhibition of either the phosphorylation activity and/or the subsequent functional consequences, or ATPase activity of activated Wee1A kinase, activated Myt1 kinase, or a mutant of either of the foregoing. Alternate in vitro assays quantitate the ability of the inhibitor to bind to Wee1A kinase.
  • DDR DNA damage response
  • Wee1 Kinase Family [0132] The Wee1 kinase family consists of three serine/threonine kinases sharing conserved molecular structures and encoded by the following genes: WEE1 (alternatively, WEE1 G2 checkpoint kinase or Wee1A kinase), PKMYT1 (alternatively MYT1 kinase or membrane-associated tyrosine- and threonine-specific cdc2-inhibitory kinase), and WEE2 (alternatively WEE2 oocyte meiosis inhibiting kinase or Wee1B kinase).
  • WEE1 alternatively, WEE1 G2 checkpoint kinase or Wee1A kinase
  • PKMYT1 alternatively MYT1 kinase or membrane-associated tyrosine- and threonine-specific cdc2-inhibitory kinase
  • Wee1A kinase and Myt1 kinase play a key role in cell cycle regulation, in particular, in the entry into mitosis (Schmidt M, Rohe A, Platzer C, et al. Regulation of G2/M transition by inhibition of Wee1 and PMyt1 Kinases. Molecules.2017;22:2045). Their role as regulators is crucial during normal cell cycle progression and in response to DNA damage as part of the DNA damage response (DDR) pathways.
  • Wee1B kinase regulates cell cycle progression and, in particular, meiosis (Solc P, Schultz RM, Motlik J.
  • Wee1B kinase expression is germ-cell specific and inhibits meiosis by phosphorylating Tyr15 of the CDK1-cyclin B complex (JY Zhu et al., J Med Chem.2017; 60 (18), 7863-7875). Previous and current drug discovery efforts have not been focused on Wee1B kinase due to its characterized role in meiosis.
  • Wee1B kinase plays a dual regulatory role in oocyte meiosis by preventing premature restart prior to ovulation and permitting metaphase II exit at fertilization (Nakanishi M, Ando H, Watanabe N, et al. Identification and characterization of human Wee1B, a new member of the Wee1 family of Cdk-inhibitory kinases. Genes Cells. 2000;5(10):839-47).
  • Myt1 kinase is a multi-functional protein kinase localized to the ER-Golgi complex that is known to play a regulatory role in the cell cycle by inhibiting Cdk1/cyclin B1 mediated mitosis (JY Zhu et al., J Med Chem. 2017; 60 (18), 7863-7875).
  • Myt1 kinase inhibits the Cdk1/cyclin B1 activity through the phosphorylation of Tyr15 and Thr14 of Cdk1 and sequestration of Cdk1 from the nucleus. Additionally, Myt1 kinase has been tied to orchestrating the ER-Golgi complex reassembly during mitotic exit.
  • Wee1A kinase [0135] Wee1A kinase regulates entry into mitosis at the G2/M transition of the S phase by phosphorylating Tyr15 of Cdk1 to inactive the Cdk1/cyclin B complex.
  • Cells with perturbed G1 checkpoint activity e.g., cancer cells
  • Wee1A kinase activity is altered, a perturbed cell may enter mitosis prematurely without having the opportunity to fully replicate the entire DNA content or repair potential DNA lesions that might have occurred during S phase.
  • This characterization of Wee1A kinase’s role in the cell cycle has made it an attractive target for anticancer therapeutics, especially in combination with DNA-damaging agents (JY Zhu et al., J Med Chem.2017; 60 (18), 7863-7875).
  • treatment refers to reversing, alleviating, delaying the onset of, or inhibiting the progress of a disease or disorder, or one or more symptoms thereof, as described herein.
  • treatment may be administered after one or more symptoms have developed.
  • treatment may be administered in the absence of symptoms.
  • treatment may be administered to a susceptible individual prior to the onset of symptoms (e.g., in light of a history of symptoms and/or in light of genetic or other susceptibility factors). Treatment may also be continued after symptoms have resolved, for example to prevent or delay their recurrence.
  • Provided compounds are inhibitors of Wee1A kinase and are therefore useful for treating one or more disorders associated with activity of Wee1A kinase.
  • the present disclosure provides a method for treating a Wee1A kinase-mediated disorder comprising the step of administering to a patient in need thereof a compound of the present disclosure, or pharmaceutically acceptable composition thereof.
  • Some of the provided compounds also demonstrate potent inhibitory activity against Myt1 kinase and therefore are dual inhibitors useful for treating one or more disorders associated with activity of both Wee1A kinase and Myt1 kinase.
  • the present disclosure provides a method for treating a Wee1A kinase/Myt1 kinase-mediated disorder comprising the step of administering to a patient in need thereof a compound of the present disclosure that is a dual inhibitor, or a pharmaceutically acceptable composition thereof.
  • a Wee1A kinase-mediated disorder or condition as used herein means any disease or other deleterious condition in which Wee1A kinase, or a mutant thereof, is known to play a role.
  • another embodiment of the present disclosure relates to treating or lessening the severity of one or more diseases in which Wee1A kinase, or a mutant thereof, is known to play a role.
  • the present disclosure relates to a method of treating or lessening the severity of a disease or condition selected from a proliferative disorder, wherein said method comprises administering to a patient in need thereof a compound or composition according to the present disclosure.
  • a disease or condition selected from a proliferative disorder comprising administering to a patient in need thereof a compound or composition according to the present disclosure.
  • the term “Wee1A kinase/Myt1 kinase-mediated” disorder or condition as used herein means any disease or other deleterious condition in which both Wee1A kinase and Myt1 kinase, or a mutant of either or both of the foregoing, are known to play a role.
  • another embodiment of the present disclosure relates to treating or lessening the severity of one or more diseases in which both Wee1A kinase and Myt1 kinase, or a mutant of either or both of the foregoing, are known to play a role.
  • the present disclosure relates to a method of treating or lessening the severity of a disease or condition selected from a proliferative disorder, wherein said method comprises administering to a patient in need thereof a compound or composition according to the present disclosure.
  • the present disclosure provides a method of inhibiting Wee1A kinase activity in a subject comprising the step of administering to the subject an effective amount of a compound, or a pharmaceutically acceptable composition, of the present disclosure.
  • the present disclosure provides a method of inhibiting both Wee1A kinase and Myt1 kinase activity in a subject comprising the step of administering to the subject an effective amount of a compound of the present disclosure that is a dual inhibitor, or a pharmaceutically acceptable composition.
  • the present disclosure provides a method for treating or lessening the severity of one or more disorders selected from a cancer comprising the step of administering to the subject an effective amount of a compound, or a pharmaceutically acceptable composition thereof, of the present disclosure.
  • the cancer is associated with a solid tumor.
  • the present disclosure provides a method of treating a subject suffering from a cancer or other disordered cell growth characterized by aberrant Wee1A kinase activity comprising the step of administering to the subject an effective amount of a compound, or a pharmaceutically acceptable composition thereof, of the present disclosure.
  • aberrant Wee1A kinase activity includes elevated activity, or overexpression, or undesirable activity as compared to a non-diseased state.
  • aberrant Wee1A kinase activity may include perturbed p53 activity, Cdk1 activity, Cdk2 activity, replication stress, altered mitosis, and DNA damage.
  • the subject is suffering from a cancer associated with inactivation of p53.
  • the present disclosure provides a method of treating a subject suffering from a cancer or other disordered cell growth characterized by both aberrant Wee1A kinase and aberrant Myt1 kinase activity comprising the step of administering to the subject an effective amount of a compound of the present disclosure that is a dual inhibitor, or a pharmaceutically acceptable composition thereof.
  • aberrant Wee1A kinase and aberrant Myt1 kinase activity includes elevated activity, or overexpression, or undesirable activity as compared to a non-diseased state.
  • aberrant Wee1A kinase activity and aberrant Myt1 kinase activity may include perturbed Cdk1 activity, replication stress, altered mitosis, and DNA damage.
  • the subject to be treated has previously been treated with either a mono-specific Wee1A kinase inhibitor or Myt1 kinase inhibitor (neither of which is a dual inhibitor) and has developed resistance to or is refractory to such treatment.
  • a subject could be resistant or refractory to the Myt1 kinase inhibitor RP-6306, or the Wee1A kinase inhibitors AZD1775, Debio0123 or ZnC 3 .
  • the cancer to be treated by a compound disclosed herein is selected from a brain cancer, a cervicocerebral cancer, a cardiac cancer, a gastrointestinal cancer, an esophageal cancer, a thyroid cancer, a small cell cancer, a non-small cell cancer, a breast cancer, a lung cancer, a stomach cancer, a gallbladder/bile duct cancer, a liver cancer, a pancreatic cancer, a colon cancer, a rectal cancer, an ovarian cancer, a choriocarcinoma, an uterus body cancer, an uterocervical cancer, a renal pelvis/ureter cancer, a bladder cancer, a prostate cancer, a penis cancer, a testicular cancer, a fetal cancer, Wilms' cancer, a skin cancer, malignant melanoma, a neuroblastoma, an osteosarcoma, an Ewing's tumor, a soft part sarcoma,
  • the subject is suffering from a cancer selected from a uterine serous carcinoma and a renal cancer.
  • the breast cancer is selected from ductal carcinoma in situ (DCIS), invasive ductal carcinoma (IDC), lobular carcinoma in situ (LCIS), invasive lobular cancer (ILC), triple negative breast cancer (TNBC), inflammatory breast cancer (IBC), metastatic breast cancer (MBC), medullary carcinoma, tubular carcinoma, mucinous carcinoma (colloid), and Paget disease of the breast or nipple (commonly known as Paget disease).
  • the uterine cancer is selected from endometrial cancer and uterine sarcoma.
  • the uterine cancer is endometrial cancer. In some embodiments, the uterine cancer is uterine sarcoma.
  • the ovarian cancer is selected from epithelial ovarian carcinomas, germ cell tumors, and stromal cell tumors.
  • the stomach cancer is selected from adenocarcinoma, lymphoma, gastrointestinal stromal tumors (GISTs), carcinoid tumors, and hereditary (familial) diffuse gastric cancer.
  • the esophageal cancer is selected from squamous cell carcinoma, small cell carcinoma, and adenocarcinoma.
  • the esophageal cancer is selected from squamous cell carcinoma and adenocarcinoma. In some embodiments, the esophageal cancer is squamous cell carcinoma. In some embodiments, the esophageal cancer is adenocarcinoma.
  • the lung cancer is selected from non-small cell lung cancer, lung nodules, small cell lung cancer, and mesothelioma. In some embodiments, the lung cancer is non-small cell lung cancer.
  • the colorectal cancer is selected from adenocarcinoma, gastrointestinal stromal tumors (GIST), lymphoma, carcinoids, Turcot syndrome, Peutz- Jeghers syndrome (PJS), familial colorectal cancer (FCC), and juvenile polyposis coli.
  • the cancer is associated with deregulation of cyclin E1.
  • the cancer associated with deregulation of cyclin E1 is ovarian cancer.
  • the cancer is associated with deregulation of p53.
  • the cancer associated with deregulation of p53 is selected from a brain cancer, a cervicocerebral cancer, a cardiac cancer, a gastrointestinal cancer, an esophageal cancer, a thyroid cancer, a small cell cancer, a non-small cell cancer, a breast cancer, a lung cancer, a stomach cancer, a gallbladder/bile duct cancer, a liver cancer, a pancreatic cancer, a colon cancer, a rectal cancer, an ovarian cancer, a choriocarcinoma, an uterus body cancer, an uterocervical cancer, a renal pelvis/ureter cancer, a bladder cancer, a prostate cancer, a penis cancer, a testicular cancer, a fetal cancer, Wilms' cancer, a skin cancer, malignant melanoma, a neuroblastoma, an osteosarcoma, an Ewing's tumor, a soft part sarcoma, an acute leukemia,
  • the cancer associated with deregulation of p53 is selected from uterine serous carcinoma and a renal cancer.
  • the cancer is associated with deregulation of Cdk1.
  • the cancer associated with deregulation of Cdk1 is selected from a brain cancer, a cervicocerebral cancer, a cardiac cancer, a gastrointestinal cancer, an esophageal cancer, a thyroid cancer, a small cell cancer, a non-small cell cancer, a breast cancer, a lung cancer, a stomach cancer, a gallbladder/bile duct cancer, a liver cancer, a pancreatic cancer, a colon cancer, a rectal cancer, an ovarian cancer, a choriocarcinoma, an uterus body cancer, an uterocervical cancer, a renal pelvis/ureter cancer, a bladder cancer, a prostate cancer, a penis cancer, a testicular cancer, a fetal cancer, Wilms'
  • the cancer associated with deregulation of Cdk1 is selected from uterine serous carcinoma and a renal cancer.
  • the cancer is associated with deregulation of Cdk2.
  • the cancer associated with deregulation of Cdk2 is selected from a brain cancer, a cervicocerebral cancer, a cardiac cancer, a gastrointestinal cancer, an esophageal cancer, a thyroid cancer, a small cell cancer, a non-small cell cancer, a breast cancer, a lung cancer, a stomach cancer, a gallbladder/bile duct cancer, a liver cancer, a pancreatic cancer, a colon cancer, a rectal cancer, an ovarian cancer, a choriocarcinoma, an uterus body cancer, an uterocervical cancer, a renal pelvis/ureter cancer, a bladder cancer, a prostate cancer, a penis cancer, a testicular cancer, a fetal cancer, Wilms
  • the cancer associated with deregulation of Cdk1 is selected from uterine serous carcinoma and a renal cancer.
  • additional therapeutic agents which are normally administered to treat that condition, may also be present in the compositions of this disclosure.
  • additional therapeutic agents that are normally administered to treat a particular disease, or condition are known as “appropriate for the disease, or condition, being treated.”
  • compounds of the present disclosure, or a pharmaceutically acceptable composition thereof are administered in combination with chemotherapeutic agents to treat proliferative diseases and cancer.
  • chemotherapeutic agents include, but are not limited to, Adriamycin, dexamethasone, vincristine, cyclophosphamide, fluorouracil, topotecan, taxol, interferons, platinum derivatives, taxane (e.g., paclitaxel), vinca alkaloids (e.g., vinblastine), anthracyclines (e.g., doxorubicin), epipodophyllotoxins (e.g., etoposide), cisplatin, an mTOR inhibitor (e.g., a rapamycin), methotrexate, actinomycin D, dolastatin 10, colchicine, emetine, trimetrexate, metoprine, cyclosporine, daunorubicin, teniposide, amphotericin, alkylating agents (e.g., chlorambucil), 5-fluorouracil, camptothecin, cisplatin, met
  • a compound of the present disclosure is administered in combination with a biologic agent, such as Avastin or VECTIBIX.
  • a biologic agent such as Avastin or VECTIBIX.
  • compounds of the present disclosure or a pharmaceutically acceptable composition thereof are administered in combination with an agent selected from fasudil, sirolimus, imatinib, gefitinib, erlotinib, sorafenib, sunitinib, dasatinib, lapatinib, nilotinib, temsirolimus, everolimus, pazopanib, ruxolitinib, vandetanib, vemurafenib, crizotinib, icotinib, axitinib, tofacitinib, bosutinib, cabozantinib, ponatinib, regorafenib, afatinib, dabrafeni
  • compounds of the present disclosure, or a pharmaceutically acceptable composition thereof are administered in combination with an antiproliferative or chemotherapeutic agent selected from any one or more of abarelix, aldesleukin, alemtuzumab, alitretinoin, allopurinol, altretamine, amifostine, anastrozole, arsenic trioxide, asparaginase, azacitidine, BCG Live, bevacizumab, fluorouracil, bexarotene, bleomycin, bortezomib, busulfan, calusterone, capecitabine, camptothecin, carboplatin, carmustine, celecoxib, cetuximab, chlorambucil, cladribine, clofarabine, cyclophosphamide, cytarabine, dactinomycin, darbepoetin alfa, daunorubicin, de
  • compounds of the present disclosure, or a pharmaceutically acceptable composition thereof are co-administered with a pharmaceutically acceptable Myt1 kinase inhibitor.
  • the Myt1 kinase inhibitor is RP-6306.
  • compounds of the present disclosure, or a pharmaceutically acceptable composition thereof are co-administered with a pharmaceutically acceptable DNA damaging agent.
  • compounds of the present disclosure, or a pharmaceutically acceptable composition thereof are co-administered with radiation.
  • compounds of the present disclosure, or a pharmaceutically acceptable composition thereof are administered in combination with a monoclonal antibody or an siRNA therapeutic.
  • compounds of the present disclosure are administered in combination with a targeted therapy selected from (i) an inhibitor of a kinase selected from MET, MEK, mTOR, FLT3, BRAF, KIT, PDGFR, FDFR, PI3K, EGFR, AKT, and KRAS, (ii) an inhibitor of a fusion kinase like BCR-ABL, ALK, RET and ROS, JAK, CDK4/6, and KRAS, (iii) epigenetic modulators such as an HDAC inhibitor, (iv) immuno-oncology agents such as those targeting PD1, PDL1, and CTLA4, (v) antibody drug conjugates such as those targeting Her2, CD38, BCMA, CD19, nectin 4 , trop2, CD79, and CD22, (vi) bispecific T cell engagers (BiTEs), (vii) transcription factor modulators such as those targeting IKZF (i.e.,
  • compounds of the present disclosure are administered in combination with an inhibitor of a DNA repair protein other than Wee1A kinase or Myt1 kinase.
  • a DNA repair protein other than Wee1A kinase or Myt1 kinase.
  • Such inhibitors include those that inhibit one or more of the following CHK1, CHK2, ATM, ATR, Pol Theta, CDC7, DNAPK, PLK1, WRN, PARP and Aurora A/B.
  • Those additional agents may be administered separately from an inventive compound-containing composition, as part of a multiple dosage regimen. Alternatively, those agents may be part of a single dosage form, mixed together with a compound of this disclosure in a single composition.
  • the two active agents may be submitted simultaneously, sequentially or within a period of time from one another, for example, within one, two, three, four, five, six, seven, eight, nine, ten, eleven, or twelve hours from one another.
  • compounds of the present disclosure, or a pharmaceutically acceptable salt thereof particularly those compounds that do not have significant Myt1 kinase activity (e.g., compounds not rated A or B in the Myt1 kinase binding assay disclosed in Example 6 herein), are administered as part of a multiple dosage regimen with a pharmaceutically acceptable Myt1 kinase inhibitor.
  • compounds of the present disclosure, or a pharmaceutically acceptable composition thereof are administered as part of a multiple dosage regimen with a Myt1 kinase inhibitor selected from RP-6306.
  • a Myt1 kinase inhibitor selected from RP-6306.
  • compounds of the present disclosure, or a pharmaceutically acceptable salt thereof, particularly those compounds that do not have significant Myt1 kinase activity are administered to a subject wherein a Myt1 kinase inhibitor is used as the first or second line therapy.
  • compounds of the present disclosure, or a pharmaceutically acceptable salt thereof are administered to a subject wherein a Myt1 kinase inhibitor is used as a first line therapy.
  • compounds of the present disclosure, or a pharmaceutically acceptable salt thereof are administered to a subject wherein a Myt1 kinase inhibitor selected from RP-6306, is used as a first line therapy.
  • compounds of the present disclosure, or a pharmaceutically acceptable salt thereof, particularly those compounds that do not have significant Myt1 kinase activity are administered to a subject wherein a Myt1 kinase inhibitor is used as a second line therapy.
  • compounds of the present disclosure, or a pharmaceutically acceptable salt thereof are administered to a subject wherein a Myt1 kinase inhibitor selected from RP-6306, is used as a second line therapy.
  • the term “combination,” “combined,” “co-administered” and related terms refer to the simultaneous or sequential administration of therapeutic agents in accordance with this disclosure.
  • a compound of the present disclosure may be administered with another therapeutic agent simultaneously or sequentially in separate unit dosage forms or together in a single unit dosage form.
  • the present disclosure provides a single unit dosage form comprising a provided compound, an additional therapeutic agent, and a pharmaceutically acceptable carrier, adjuvant, or vehicle.
  • the amount of both, an inventive compound and additional therapeutic agent in those compositions which comprise an additional therapeutic agent as described above) that may be combined with the carrier materials to produce a single dosage form will vary depending upon the host treated and the particular mode of administration.
  • compositions of this disclosure should be formulated so that a dosage of between 0.001 - 100 mg/kg body weight/day of an inventive can be administered.
  • that additional therapeutic agent and the compound of this disclosure may act synergistically. Therefore, the amount of additional therapeutic agent in such compositions will be less than that required in a monotherapy utilizing only that therapeutic agent. In such compositions a dosage of between 0.001 - 1,000 ⁇ g/kg body weight/day of the additional therapeutic agent can be administered.
  • the amount of additional therapeutic agent present in the compositions of this disclosure will be no more than the amount that would normally be administered in a composition comprising that therapeutic agent as the only active agent.
  • the amount of additional therapeutic agent in the presently disclosed compositions will range from about 50% to 100% of the amount normally present in a composition comprising that agent as the only therapeutically active agent.
  • the present disclosure provides a method for inhibiting Wee1A kinase in vitro. In some such embodiments, the amount of Wee1A kinase inhibition is assessed based on a competitive ATP-binding assay. [0177] In some embodiments, the present disclosure provides a method for inhibiting Wee1A kinase in a biological sample. [0178] In some embodiments, the present disclosure provides a method for inhibiting both Wee1A kinase and Myt1 kinase in a biological sample.
  • the present disclosure provides a method for inhibiting both Wee1A kinase and Myt1 kinase in vitro. In some such embodiments, the amount of Wee1A kinase and Myt1 kinase inhibition are both assessed based on a competitive ATP-binding assay. [0180] In some embodiments, the present disclosure provides a method for assessing Cdk1 phosphorylation in a cell, comprising contacting said cell with a compound described herein. In one embodiment, the contacting step comprises incubating a cell with a compound presented herein. In some such embodiments, the cell is incubated for at least 4 hours.
  • the cell may comprise a DAOY medulloblastoma cell.
  • Reagents and solvents were purchased from commercial suppliers and used as received unless otherwise noted. Solvents were dried over molecular sieves 4 ⁇ . Reactions were stirred using magnetic stir bars in glass vials or round bottomed flasks and heated using stirring plates. Solvents were removed on rotary evaporators, vacuum centrifuge, or by freeze-drying. Reaction progress was monitored by LC-MS or thin layer chromatography (TLC). Aluminium-backed TLC plates (60F 254 ) were visualized by UV-light (254 nm).
  • Flash chromatography was performed on silica gel or C18 functionalized silica were performed on an automated flash purification system equipped with a diode array detector (200-400 nm), eluting with gradients of ethyl acetate and petroleum ether or methanol and dichloromethane (containing 0.03 % of ammonia.
  • Purity analyses were performed by HPLC reversed phase C18 columns eluting with acetonitrile and water (containing 0.1% TFA or 0.03% ammonia). UV-traces were recorded at 220 nm. Purifications by preparative HPLC were performed using reversed phase C18 columns eluting with acetonitrile and water (containing 0.1% TFA or 0.03% ammonia).
  • reaction mixture was heated to 90 °C and stirred for several days until LCMS indicated full conversion.
  • the reaction mixture was diluted with brine and aq. NaHCO3 (sat.) and extracted with ethyl acetate ( ⁇ 3).
  • the combined organic layers were dried using a phase-separator and concentrated under reduced pressure giving an oil. This material that was either used without further purification or purified by flash chromatography.
  • CuI (1.2 equivalents) followed by N,N’-dimethylethylenediamine (1 equivalent) was added to a stirred degassed suspension of tert-butyl 4-((6-bromopyridin-2- yl)amino)piperidine-1-carboxylate (1 equivalent), 6-(methylsulfanyl)-2-(prop-2-en-1-yl)- 1H,2H,3H-pyrazolo[3,4-d]pyrimidin-3-one (1 equivalent), and cesium carbonate (3 equivalents) in dioxane (0.3 mol/L) at room temperature.
  • the reaction was heated to 90 °C in a closed vial overnight.
  • the reaction was diluted with water and a few drops of aq. ammonia (28%) then extracted with ethyl acetate ( ⁇ 3).
  • the combined organic layers were dried using a phase-separator and concentrated under reduced pressure giving an oil. This material that was either used without further purification or purified by flash chromatography.
  • tert-butyl 4-((6-bromopyridin-2-yl)oxy)piperidine-1-carboxylate [0202] Sodium hydride was added to a stirred solution of tert-butyl 4-hydroxypiperidine- 1-carboxylate (1 equivalent) in tetrahydrofuran (0.5 mol/mL) at 0 °C. A solution of 2,6- dibromopyridine (1 equivalent) in THF (0.5 mol/mL) was added to the mixture, upon complete addition the mixture was brought to room temperature and stirred until LCMS indicated full conversion, typically overnight. The reaction mixture was concentrated, diluted with brine and aq.
  • CuI (1.2 equivalents) followed by N,N’-dimethylethylenediamine (1 equivalent) was added to a stirred degassed suspension of tert-butyl 4-((6-bromopyridin-2- yl)oxy)piperidine-1-carboxylate (1 equivalent), 6-(methylsulfanyl)-2-(prop-2-en-1-yl)- 1H,2H,3H-pyrazolo[3,4-d]pyrimidin-3-one (1 equivalent), and cesium carbonate (3 equivalents) in dioxane (0.3 mol/L) at room temperature.
  • the reaction was heated to 90 °C in a closed vial overnight.
  • the reaction was diluted with water and a few drops of aq. ammonia (28%) then extracted with ethyl acetate ( ⁇ 3).
  • the combined organic layers were dried using a phase-separator and concentrated under reduced pressure giving an oil. This material that was either used without further purification or purified by flash chromatography.
  • reaction mixture was diluted with brine and aq. NaHCO3 (sat.) and extracted with ethyl acetate ( ⁇ 3).
  • the combined organic layers were dried using a phase-separator and concentrated under reduced pressure giving an oil. This material that was either used without further purification or purified by flash chromatography.
  • the reaction was heated to 90 °C in a closed vial overnight.
  • the reaction was diluted with water and a few drops of aq. ammonia (28%) then extracted with ethyl acetate ( ⁇ 3).
  • the combined organic layers were dried using a phase-separator and concentrated under reduced pressure giving an oil. This material that was either used without further purification or purified by flash chromatography.
  • the reaction was controlled by LCMS. After 15 min, DCM was removed in vacuo, and the crude solubilized in MeCN (5 ml), then p-bromoaniline (69 mg, 0.401 mmol) was added. The reaction mixture stirred at 60 °C in a closed vial. After 96 h, the reaction mixture was allowed to cool to RT, and mCPBA quenched with 1M NaOH (5 mL) which was added dropwise. The aqueous phase was extracted with EtOAc (3x20 mL) then washed with brine (20 mL). The combined organic layers were dried (MgSO4) and concentrated under reduced pressure to give the product as a yellow powder.
  • Example 5 The resulting material was purified by reversed phase chromatography (Gemini NX-C18,21*150 mm, water (0.1% TFA)/acetonitrile, gradient over 12 minutes, 25 ml/min). The pure fractions were pooled and concentrated giving the title compound [50.9 mg, yield 33.2 %]. [0228] Example 5.
  • the reaction mass was degassed for 15 minutes. Iron bis[2-(diphenylphosphino)-2,4-cyclopentadien-1-ide]— dichloro-palladamethane (33 mg, 0.12 eq., 45.1 ⁇ mol) as was added and the screw cap was tightened on the seal tube. The contents were heated to 100 °C and stirred over night. The reaction mass was cooled to RT, diluted with EtOAc, washed with water followed by brine solution. The organic layer was dried over anhydrous Na2SO4 and the solvent was removed under reduced pressure to obtain a crude mass. The collected material (150 mg) was dissolved in DCM:TFA 4:1 v/v .The reaction mixture was stirred at room temperature for 1 h.
  • Example 20 Synthesis of 2-allyl-6-(2-methoxy-4-pyridylamino)-1-[6-(1- methyl-4-piperidyloxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazainden-3-one (Compound 263) [0272] tert-butyl 4-(3-(2-allyl-6-amino-3-oxo-2,3-dihydro-1H-pyrazolo[3,4-d]pyrimidin- 1-yl)phenoxy)piperidine-1-carboxylate [0273] In a flask, was taken tert-butyl 4-(3-(2-allyl-6-(methylsulfonyl)-3-oxo-2,3- dihydro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)phenoxy)piperidine-1-car
  • Example 27 Synthesis of 2-allyl-6-(6-methoxy-3-pyridylamino)-1-[6-(4- piperidyloxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazainden-3-one (Compound 251) [0291] This compound was made using a similar method as described in the synthesis of 2-allyl-6-(2-methoxy-4-pyridylamino)-1-[6-(1-methyl-4-piperidyloxy)-2-pyridyl]-1,2- dihydro-3H-1,2,5,7-tetraazainden-3-one. [0292] Example 28.
  • Example 30 Synthesis of 2-allyl-6-(1-isopropyl-1H-indazol-5-ylamino)-1-[6- (1-methyl-4-piperidyloxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazainden-3-one (Compound 248) [0297] This compound was made using a similar method as described in the synthesis of allyl-6-(p-bromophenylamino)-1-[6-(1-methyl-4-piperidyloxy)-2-pyridyl]-1,2-dihydro-3H- 1,2,5,7-tetraazainden-3-one.
  • Example 31 Synthesis of 2-allyl-6-(1,3,3a-triaza-5-indenylamino)-1-[6-(4- piperidyloxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazainden-3-one (Compound 245) [0299] This compound was made using a similar method as described in the synthesis of allyl-6-(p-bromophenylamino)-1-[6-(1-methyl-4-piperidyloxy)-2-pyridyl]-1,2-dihydro-3H- 1,2,5,7-tetraazainden-3-one.
  • Example 32 Synthesis of 2-allyl-6-(2,1,3-benzothiadiazol-5-ylamino)-1-[6-(1- methyl-4-piperidyloxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazainden-3-one (Compound 208) [0301] This compound was made using a similar method as described in the synthesis of allyl-6-(p-bromophenylamino)-1-[6-(1-methyl-4-piperidyloxy)-2-pyridyl]-1,2-dihydro-3H- 1,2,5,7-tetraazainden-3-one.
  • Example 33 Synthesis of 2-allyl-6-(6-isoquinolylamino)-1-[6-(4-piperidyloxy)- 2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazainden-3-one (Compound 244) [0303] tert-butyl 4- ⁇ 6-[2-allyl-6-(6-isoquinolylamino)-3-oxo-1,2-dihydro-3H-1,2,5,7- tetraazainden-1-yl]-2-pyridyloxy ⁇ -1-piperidinecarboxylate [0304] Tert-butyl 4-[6-(2-allyl-6-amino-3-oxo-1,2-dihydro-3H-1,2,5,7-tetraazainden-1- yl)-2-pyridyloxy]-1-piperidinecarboxylate (
  • Example 40 Synthesis of 2-allyl-6-(5-fluoro-3-pyridylamino)-1-[6-(4- piperidyloxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazainden-3-one (Compound 329) [0320] This compound was made using a similar method as described in the synthesis of 2-allyl-6-(2-methoxy-4-pyridylamino)-1-[6-(1-methyl-4-piperidyloxy)-2-pyridyl]-1,2- dihydro-3H-1,2,5,7-tetraazainden-3-one.
  • Example 41 Synthesis of 6-(1,3a-diaza-5-indenylamino)-2-allyl-1-[6-(1- methyl-4-piperidyloxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazainden-3-one (Compound 328) [0322] This compound was made using a similar method as described in the synthesis of allyl-6-(p-bromophenylamino)-1-[6-(1-methyl-4-piperidyloxy)-2-pyridyl]-1,2-dihydro-3H- 1,2,5,7-tetraazainden-3-one.
  • Example 42 Synthesis of 2-allyl-6-(5-fluoro-3-pyridylamino)-1-[6-(1-methyl- 4-piperidyloxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazainden-3-one (Compound 327) [0324] This compound was made using a similar method as described in the synthesis of 2-allyl-6-(2-methoxy-4-pyridylamino)-1-[6-(1-methyl-4-piperidyloxy)-2-pyridyl]-1,2- dihydro-3H-1,2,5,7-tetraazainden-3-one.
  • Example 43 Synthesis of 2-allyl-6-(7-isoquinolylamino)-1-[6-(1-methyl-4- piperidyloxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazainden-3-one (Compound 326) [0326] This compound was made using a similar method as described in the synthesis of 2-allyl-6-(2-methoxy-4-pyridylamino)-1-[6-(1-methyl-4-piperidyloxy)-2-pyridyl]-1,2- dihydro-3H-1,2,5,7-tetraazainden-3-one.
  • Example 44 Synthesis of 6-(1,3a-diaza-5-indenylamino)-2-allyl-1-[6-(4- piperidyloxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazainden-3-one (Compound 324) [0328] This compound was made using a similar method as described in the synthesis of allyl-6-(p-bromophenylamino)-1-[6-(1-methyl-4-piperidyloxy)-2-pyridyl]-1,2-dihydro-3H- 1,2,5,7-tetraazainden-3-one. Yield: 100 mg TFA salt (42%). [0329] Example 45.
  • Example 54 Synthesis of 2-allyl-6-(2-methyl-4-pyridylamino)-1-[m-(4- piperidyloxy)phenyl]-1,2-dihydro-3H-1,2,5,7-tetraazainden-3-one (Compound 222) [0349] tert-butyl 4-(3-bromophenoxy)piperidine-1-carboxylate [0350] To a stirred solution of tert-butyl 4-hydroxypiperidine-1-carboxylate (7.48 g, 37.1 mmol) in DMF (50 ml) was added NaH (2.285 g, 57.1 mmol) at 0 °C under inert atmosphere and stirred for 1 hour at 50 °C.
  • tert-butyl-4-(3-(2-allyl-6-(methylsulfonyl)-3-oxo-2,3-dihydro-1H-pyrazolo[3,4- d]pyrimidin-1-yl)phenoxy)piperidine-1-carboxylate [0354] To a stirred solution of tert-butyl 4-(3-(2-allyl-6-(methylthio)-3-oxo-2,3-dihydro- 1H-pyrazolo[3,4-d]pyrimidin-1-yl)phenoxy)piperidine-1-carboxylate (60 mg, 0.121 mmol) in DCM (2 ml) was added m-CPBA (41.6 mg, 0.241 mmol).
  • tert-butyl 4-(3-(2-allyl-6-(methylsulfonyl)-3-oxo-2,3-dihydro-1H- pyrazolo[3,4-d]pyrimidin-1-yl)phenoxy)piperidine-1-carboxylate 400mg, 0.755 mmol
  • ammonia in THF (5ml, 5.00 mmol)at rt under inert atmosphere and stirred for 16 h.
  • tert-butyl-4-(3-(2-allyl-6-((2-methylpyridin-4-yl)amino)-3-oxo-2,3-dihydro-1H- pyrazolo[3,4-d]pyrimidin-1-yl)phenoxy)piperidine-1-carboxylate [0358] A stirred solution of tert-butyl 4-(3-(2-allyl-6-amino-3-oxo-2,3-dihydro-1H- pyrazolo[3,4-d]pyrimidin-1-yl)phenoxy)piperidine-1-carboxylate (300mg, 0.643 mmol), 4- bromo-2-methylpyridine (133 mg, 0.772 mmol), K2CO3 (267 mg, 1.929 mmol) and N, N'- dimethylethylenediamine (56.6 mg, 0.643 mmol) in dioxane (4 ml) was degassed with N 2 for 30 minutes.
  • Example 55 Synthesis of 2-allyl-1-[6-(4-piperidyloxy)-2-pyridyl]-6-(3- pyridylamino)-1,2-dihydro-3H-1,2,5,7-tetraazainden-3-one (Compound 230) [0362] This compound was made using a similar method as described in the synthesis of 2-allyl-6-(2-methoxy-4-pyridylamino)-1-[6-(1-methyl-4-piperidyloxy)-2-pyridyl]-1,2- dihydro-3H-1,2,5,7-tetraazainden-3-one. Yield: 93 mg as a white powder. [0363] Example 56.
  • Example 60 Synthesis of 2-allyl-6-(1-methyl-1H-indazol-5-ylamino)-1-[2-(1- methyl-4-piperidylamino)-4-pyrimidinyl]-1,2-dihydro-3H-1,2,5,7-tetraazainden-3-one (Compound 313) [0372] This compound was made using a similar method as described in the synthesis of 2-allyl-6-((4-chlorophenyl)amino)-1-(6-((1-methylpiperidin-4-yl)amino)pyridin-2-yl)-1,2- dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one.
  • Example 61 Synthesis of 2-allyl-6-(p-fluorophenylamino)-1-[2-(1-methyl-4- piperidylamino)-4-pyrimidinyl]-1,2-dihydro-3H-1,2,5,7-tetraazainden-3-one (Compound 312) [0374] This compound was made using a similar method as described in the synthesis of 2-allyl-6-((4-chlorophenyl)amino)-1-(6-((1-methylpiperidin-4-yl)amino)pyridin-2-yl)-1,2- dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one.
  • Example 62 Synthesis of 2-allyl-1-[m-(1-methyl-4-piperidyloxy)phenyl]-6-(1- methyl-4-pyrazolylamino)-1,2-dihydro-3H-1,2,5,7-tetraazainden-3-one (Compound 221) [0376] tert-butyl 4-(3-(2-allyl-6-((1-methyl-1H-pyrazol-4-yl)amino)-3-oxo-2,3-dihydro- 1H-pyrazolo[3,4-d]pyrimidin-1-yl)phenoxy)piperidine-1-carboxylate [0377] To a stirred solution of tert-butyl 4-(3-(2-allyl-6-(methylsulfonyl)-3-oxo-2,3- dihydro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)
  • Example 64 Synthesis of 1- ⁇ m-[N-methyl(1-methyl-4- piperidyl)amino]phenyl ⁇ -2-allyl-6-(1-methyl-1H-indazol-5-ylamino)-1,2-dihydro-3H- 1,2,5,7-tetraazainden-3-one (Compound 225) [0385] 2-allyl-6-(methylthio)-1-(3-nitrophenyl)-1,2-dihydro-3H-pyrazolo[3,4- d]pyrimidin-3-one [0386] To a stirred solution of 2-allyl-6-(methylthio)-1,2-dihydro-3H-pyrazolo[3,4- d]pyrimidin-3-one (1.2 g, 5.40 mmol) in dichloromethane (15 mL) was added 3-nitrophenyl boronic acid (1.352 g, 8.10 mmol), sodium carbonate (1.707 g, 16.20 mmol) and copper (II)
  • the mixture was heated to 70 °C and stirred for 16 h. The progress of the reaction was monitored by TLC and LCMS. After completion of the reaction, the mixture was filtered through celite and washed with 10% MeOH in DCM (2 X 100 mL).
  • the mixture was stirred at under inert atmosphere for 16 h at room temperature.
  • the progress of the reaction was monitored by TLC and UPLC.
  • the reaction mixture was diluted with water (100 mL) and extracted with ethyl acetate (2X 200 mL).
  • Example 65 Synthesis of 2-allyl-6-(1-methyl-1H-indazol-5-ylamino)-1-[m-(1- methyl-4-piperidyloxy)phenyl]-1,2-dihydro-3H-1,2,5,7-tetraazainden-3-one (Compound 220) [0402] To a stirred solution of 2-allyl-6-((1-methyl-1H-indazol-5-yl)amino)-1-(3- (piperidin-4-yloxy)phenyl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (210 mg, 0.423 mmol) in THF (5 mL) was added 37% aq formaldehyde (172 mg, 2.114 mmol) at 25 °C, and resulting mixture was stirred for 10 min.
  • Example 68 Synthesis of 2-allyl-6-(1-benzofuran-6-ylamino)-1-[6-(1-methyl- 4-piperidyloxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazainden-3-one (Compound 308) [0408] This compound was made using a similar method as described in the synthesis of allyl-6-(p-bromophenylamino)-1-[6-(1-methyl-4-piperidyloxy)-2-pyridyl]-1,2-dihydro-3H- 1,2,5,7-tetraazainden-3-one.
  • Example 69 Synthesis of 2-allyl-6-(2H-1,3-benzodioxol-5-ylamino)-1-[6-(1- methyl-4-piperidyloxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazainden-3-one (Compound 307) [0410] This compound was made using a similar method as described in the synthesis of allyl-6-(p-bromophenylamino)-1-[6-(1-methyl-4-piperidyloxy)-2-pyridyl]-1,2-dihydro-3H- 1,2,5,7-tetraazainden-3-one.
  • Example 70 Synthesis of m- ⁇ 2-allyl-1-[6-(1-methyl-4-piperidyloxy)-2- pyridyl]-3-oxo-1,2-dihydro-3H-1,2,5,7-tetraazainden-6-ylamino ⁇ benzonitrile (Compound 305) [0412] This compound was made using a similar method as described in the synthesis of allyl-6-(p-bromophenylamino)-1-[6-(1-methyl-4-piperidyloxy)-2-pyridyl]-1,2-dihydro-3H- 1,2,5,7-tetraazainden-3-one.
  • Example 71 Synthesis of 2-allyl-6-(1,3-benzothiazol-6-ylamino)-1-[6-(1- methyl-4-piperidyloxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazainden-3-one (Compound 303) [0414] This compound was made using a similar method as described in the synthesis of allyl-6-(p-bromophenylamino)-1-[6-(1-methyl-4-piperidyloxy)-2-pyridyl]-1,2-dihydro-3H- 1,2,5,7-tetraazainden-3-one.
  • Example 72 Synthesis of p- ⁇ 2-ethyl-3-oxo-1-[6-(4-piperidyloxy)-2-pyridyl]- 1,2-dihydro-3H-1,2,5,7-tetraazainden-6-ylamino ⁇ benzonitrile (Compound 302) [0416] This compound was made using a similar method as described in the synthesis of 2-allyl-6-(2-methoxy-4-pyridylamino)-1-[6-(1-methyl-4-piperidyloxy)-2-pyridyl]-1,2- dihydro-3H-1,2,5,7-tetraazainden-3-one using p-bromobenzonitrile and tert ⁇ butyl 4 ⁇ [(6 ⁇ 6 ⁇ amino ⁇ 2 ⁇ ethyl ⁇ 3 ⁇ oxo ⁇ 1H,2H,3H ⁇ pyrazolo[3,4 ⁇ d]pyrimi
  • Example 73 Synthesis of p- ⁇ 2-allyl-1-[6-(1-methyl-4-piperidyloxy)-2-pyridyl]- 3-oxo-1,2-dihydro-3H-1,2,5,7-tetraazainden-6-ylamino ⁇ benzonitrile (Compound 301) [0418] This compound was made using a similar method as described in the synthesis of 2-allyl-6-(2-methoxy-4-pyridylamino)-1-[6-(1-methyl-4-piperidyloxy)-2-pyridyl]-1,2- dihydro-3H-1,2,5,7-tetraazainden-3-one using p-bromobenzonitrile.
  • Example 74 p- ⁇ 2-allyl-3-oxo-1-[6-(4-piperidyloxy)-2-pyridyl]-1,2-dihydro- 3H-1,2,5,7-tetraazainden-6-ylamino ⁇ benzonitrile (Compound 300) [0420] 4- ⁇ 6-[2-allyl-6-(p-cyanophenylamino)-3-oxo-1,2-dihydro-3H-1,2,5,7- tetraazainden-1-yl]-2-pyridyloxy ⁇ -1-piperidinecarboxylate [0421] tert-butyl-4-[6-(2-allyl-6-amino-3-oxo-1,2-dihydro-3H-1,2,5,7-tetraazainden-1- yl)-2-pyridyloxy]-1-piperidinecarboxylate (200mg, 0.428 mmol), p-bromobenzonitrile (155.
  • Example 75 2-allyl-6-(1,2-benzisoxazol-6-ylamino)-1-[6-(1-methyl-4- piperidyloxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazainden-3-one (Compound 299) [0425] This compound was made using a similar method as described in the synthesis of allyl-6-(p-bromophenylamino)-1-[6-(1-methyl-4-piperidyloxy)-2-pyridyl]-1,2-dihydro-3H- 1,2,5,7-tetraazainden-3-one. Yield: 55 mg (42%) as a white powder.
  • reaction mixture was cooled to room temperature and a solution of 1-bromo-3-fluorobenzene (5 g, 28.6 mmol) dissolved in DMF (5 mL) was added. After addition, the reaction mixture was stirred at 70 °C for 3 h. The progress of the reaction was monitored by LCMS and TLC. After completion of the reaction, the reaction mixture was quenched with ice cold water and extracted with ethyl acetate (2X 400 mL).
  • tert-butyl-4-(3-(2-allyl-6-(methylthio)-3-oxo-2,3-dihydro-1H-pyrazolo[3,4- d]pyrimidin-1-yl)phenoxy)piperidine-1-carboxylate [0430] A stirred solution of tert-butyl 4-(3-bromophenoxy)piperidine-1-carboxylate (481 mg, 1.350 mmol), 2-allyl-6-(methylthio)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (250 mg, 1.125 mmol), K 2 CO 3 (466 mg, 3.37 mmol) and N,N′-dimethylethylenediamine (99 mg, 1.125 mmol) in dioxane (10 mL) was degassed for 20 min at room temperature under inert atmosphere.
  • tert-butyl-4-(3-(2-allyl-6-(methylsulfonyl)-3-oxo-2,3-dihydro-1H-pyrazolo[3,4- d]pyrimidin-1-yl)phenoxy)piperidine-1-carboxylate [0432] To the stirred solution of tert-butyl 4-(3-(2-allyl-6-(methylthio)-3-oxo-2,3- dihydro-1H-pyrazolo[3,4-d]pyrimidin-1yl)phenoxy)piperidine-1-carboxylate (80 mg, 0.161 mmol) in DCM (5 mL) was added m-CPBA (55.5 mg, 0.322 mmol) at room temperature and the mixture was stirred for 2 h.
  • m-CPBA 55.5 mg, 0.322 mmol
  • tert-butyl 4-(3-(2-allyl-6-(methylsulfonyl)-3-oxo-2,3- dihydro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)phenoxy)piperidine-1-carboxylate (65 mg, 0.123 mmol) in AcOH (3 mL) was added 1-methyl-1H-indazol-5-amine (18.06 mg, 0.123 mmol) at room temperature.
  • reaction mixture was allowed to stir for 16 h. The progress of the reaction was monitored by TLC and LCMS. After completion of the reaction, the reaction mass was quenched with aq. sodium bicarbonate solution and extracted with 10% MeOH in DCM (2X 100 mL).
  • 2-allyl-6-(methylthio)-1-(3-nitrophenyl)-1,2-dihydro-3H-pyrazolo[3,4- d]pyrimidin-3-one [0439] To a stirred solution of 2-allyl-6-(methylthio)-1,2-dihydro-3H-pyrazolo[3,4- d]pyrimidin-3-one (1, 1.2 g, 5.40 mmol) in dichloroethane (15 mL) was added 3-nitrophenyl boronic acid (1.352 g, 8.10 mmol), sodium carbonate (1.707 g, 16.20 mmol) and copper (II) acetate (0.490
  • Example 78 2-ethyl-1-[6-(4-piperidyloxy)-2-pyridyl]-6-m-toluidino-1,2- dihydro-3H-1,2,5,7-tetraazainden-3-one (Compound 298) [0449] This compound was made using a similar method as described in the synthesis of allyl-6-(p-bromophenylamino)-1-[6-(1-methyl-4-piperidyloxy)-2-pyridyl]-1,2-dihydro-3H- 1,2,5,7-tetraazainden-3-one. Yield: 55 mg (42%) as a white powder. Yield: 92.5 mg, 80% as off-white solid.
  • Example 80 2-allyl-6-(1-methyl-4-pyrazolylamino)-1-[m-(4- piperidyloxy)phenyl]-1,2-dihydro-3H-1,2,5,7-tetraazainden-3-one (Compound 224)
  • reaction mixture was allowed to stir at 25 °C for 16 h. The progress of the reaction was monitored by TLC and LCMS. After completion of the reaction. The reaction mixture was concentrated under reduced pressure and then diluted with 10% aq. sodium bicarbonate and then extracted with 10% MeOH in DCM.
  • This compound was made using a similar method as described in the synthesis of 2-allyl-6-(2-methoxy-4-pyridylamino)-1-[6-(1-methyl-4-piperidyloxy)-2-pyridyl]-1,2- dihydro-3H-1,2,5,7-tetraazainden-3-one. Yield: 27 mg, 47%.
  • This compound was made using a similar method as described in the synthesis of allyl-6-(p-bromophenylamino)-1-[6-(1-methyl-4-piperidyloxy)-2-pyridyl]-1,2-dihydro-3H- 1,2,5,7-tetraazainden-3-one. Yield: 55 mg (42%) as a white powder.
  • This compound was made using a similar method as described in the synthesis of allyl-6-(p-bromophenylamino)-1-[6-(1-methyl-4-piperidyloxy)-2-pyridyl]-1,2-dihydro-3H- 1,2,5,7-tetraazainden-3-one. Yield: 55 mg (42%) as a white powder.
  • Example 88 2-allyl-1-[6-(1-methyl-4-piperidyloxy)-2-pyridyl]-6-(6-methyl-3- pyridylamino)-1,2-dihydro-3H-1,2,5,7-tetraazainden-3-one (Compound 197) [0480] This compound was made using a similar method as described in the synthesis of allyl-6-(p-bromophenylamino)-1-[6-(1-methyl-4-piperidyloxy)-2-pyridyl]-1,2-dihydro-3H- 1,2,5,7-tetraazainden-3-one. Yield: 29 mg TFA salt (83%) as a pale-yellow powder.
  • Example 89 2-allyl-6-(2H-1,3-benzodioxol-5-ylamino)-1-[6-(1-methyl-4- piperidylamino)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazainden-3-one (Compound 196) [0482] This compound was made using a similar method as described in the synthesis of2-allyl-6-(1-methyl-1H-indazol-5-ylamino)-1-[m-(1-methyl-4-piperidylamino)phenyl]-1,2- dihydro-3H-1,2,5,7-tetraazainden-3-one.
  • Example 90 2-allyl-6-(1-benzofuran-6-ylamino)-1-[6-(1-methyl-4- piperidylamino)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazainden-3-one (Compound 195)
  • This compound was made using a similar method as described in the synthesis of 2-allyl-6-(1-methyl-1H-indazol-5-ylamino)-1-[m-(1-methyl-4-piperidylamino)phenyl]-1,2- dihydro-3H-1,2,5,7-tetraazainden-3-one.
  • This compound was made using a similar method as described in the synthesis of 2-allyl-6-(1-methyl-1H-indazol-5-ylamino)-1-[m-(1-methyl-4-piperidylamino)phenyl]-1,2- dihydro-3H-1,2,5,7-tetraazainden-3-one starting with tert-butyl (S)-3-amino-1- piperidinecarboxylate.
  • This compound was made using a similar method as described in the synthesis of 2-allyl-6-(1-methyl-1H-indazol-5-ylamino)-1-[m-(1-methyl-4-piperidylamino)phenyl]-1,2- dihydro-3H-1,2,5,7-tetraazainden-3-one starting with tert-butyl (R)-3-amino-1- piperidinecarboxylate.
  • Example 106 (R)-2-allyl-6-((4-chlorophenyl)amino)-1-(6-(methyl(1- methylpiperidin-3-yl)amino)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3- one (Compound 163) [0522] This compound was made using a similar method as described in the synthesis of 2-allyl-6-((4-chlorophenyl)amino)-1-(6-((1-methylpiperidin-4-yl)amino)pyridin-2-yl)-1,2- dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one starting with tert-butyl (R)-3-amino-1- piperidinecarboxylate.
  • This compound was made using a similar method as described in the synthesis of 2-allyl-6-((4-chlorophenyl)amino)-1-(6-((1-methylpiperidin-4-yl)amino)pyridin-2-yl)-1,2- dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one.
  • This compound was made using a similar method as described in the synthesis of 2-allyl-6-((4-chlorophenyl)amino)-1-(6-((1-methylpiperidin-4-yl)amino)pyridin-2-yl)-1,2- dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one.
  • This compound was made using a similar method as described in the synthesis of 2-allyl-6-((4-chlorophenyl)amino)-1-(6-((1-methylpiperidin-4-yl)amino)pyridin-2-yl)-1,2- dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one using 3,4-dicholoroaniline and tert ⁇ butyl 4 ⁇ ( ⁇ 6 ⁇ [6 ⁇ (methylsulfanyl) ⁇ 3 ⁇ oxo ⁇ 2 ⁇ (prop ⁇ 2 ⁇ en ⁇ 1 ⁇ yl) ⁇ 1
  • reaction mixture was concentrated to a yellow oil (175 mg, crude), half of which was purified by prep-HPLC then converted to the free base by solid-phase extraction (1 g SCX-2, MeOH, 1.4 M MeOH/NH3). Yield: 15 mg free base (31%) as a powder.
  • reaction mixture was stirred at room temperature for 16 h. The progress of the reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to afford a residue. The resultant residue was basified with sodium bicarbonate and extracted with ethyl acetate (10x2 mL). The combined organic layer was dried over Na 2 SO 4 , filtered, and concentrated under reduced pressure to afford crude.
  • Example 120 2-allyl-6-(4-fluoro-3-toluidino)-1-[6-(1-methyl-4- piperidylamino)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazainden-3-one (Compound 140) [0557] This compound was made using a similar method as described in the synthesis of This compound was made using a similar method as described in the synthesis of 2-allyl-6- ((4-chlorophenyl)amino)-1-(6-((1-methylpiperidin-4-yl)amino)pyridin-2-yl)-1,2-dihydro-3H- pyrazolo[3,4-d]pyrimidin-3-one.
  • This compound was made using a similar method as described in the synthesis of 2-allyl-6-((4-chlorophenyl)amino)-1-(6-((1-methylpiperidin-4-yl)amino)pyridin-2-yl)-1,2- dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one using 4-chloro-3-methylaniline and tert ⁇ butyl 4 ⁇ ( ⁇ 6 ⁇ [6 ⁇ (methylsulfanyl) ⁇ 3 ⁇ oxo ⁇ 2 ⁇ (prop ⁇ 2 ⁇ en ⁇ 1 ⁇ yl) ⁇ 1H,2H,3H ⁇ pyrazolo[3,4 ⁇ d]pyrimidin ⁇ 1 ⁇
  • This compound was made using a similar method as described in the synthesis of 2-allyl-6-((4-chlorophenyl)amino)-1-(6-((1-methylpiperidin-4-yl)amino)pyridin-2-yl)-1,2- dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one using tert ⁇ butyl 4 ⁇ ( ⁇ 6 ⁇ [6 ⁇ (methylsulfanyl) ⁇ 3 ⁇ oxo ⁇ 2 ⁇ (prop ⁇ 2 ⁇ en ⁇ 1 ⁇ yl) ⁇ 1H,2H,3H ⁇ pyrazolo[3,4 ⁇ d]pyrimidin ⁇ 1 ⁇ y
  • Example 139 2-allyl-6-(2-methyl-4-pyridylamino)-1-[6-(4-piperidyloxy)-2- pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazainden-3-one (Compound 113) [0595] This compound was made using a similar method as described in the synthesis of 2-allyl-6-(2-methoxy-4-pyridylamino)-1-[6-(1-methyl-4-piperidyloxy)-2-pyridyl]-1,2- dihydro-3H-1,2,5,7-tetraazainden-3-one. Yield 2 mg.
  • Example 141 Synthesis of 1- ⁇ 6-[(S)-3-pyrrolidinylamino]-2-pyridyl ⁇ -2-allyl-6- (1-methyl-1H-indazol-5-ylamino)-1,2-dihydro-3H-1,2,5,7-tetraazainden-3-one (Compound 107) [0599] This compound was made using a similar method as described in the synthesis of 2-allyl-6-((4-chlorophenyl)amino)-1-(6-((1-methylpiperidin-4-yl)amino)pyridin-2-yl)-1,2- dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one.
  • Example 142 Synthesis of 2-allyl-6-(1-methyl-1H-indazol-5-ylamino)-1-[6-(4- piperidyloxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazainden-3-one (Compound 106) [0601] This compound was made using a similar method as described in the synthesis of allyl-6-(p-bromophenylamino)-1-[6-(1-methyl-4-piperidyloxy)-2-pyridyl]-1,2-dihydro-3H- 1,2,5,7-tetraazainden-3-one using tert ⁇ butyl 4 ⁇ ( ⁇ 6 ⁇ [6 ⁇ (methylsulfanyl) ⁇ 3 ⁇ oxo ⁇ 2 ⁇ (prop ⁇ 2 ⁇ en ⁇ 1 ⁇ yl) ⁇ 1H,2H,3H ⁇ pyrazolo[3,4 ⁇ d
  • Example 143 Synthesis of 2-allyl-6-(1-methyl-1H-indazol-5-ylamino)-1-[6-(1- methyl-4-piperidyloxy)-2-pyridyl]-1,2-dihydro-3H-1,2,5,7-tetraazainden-3-one (Compound 109) [0603] This compound was made using a similar method as described in the synthesis of allyl-6-(p-bromophenylamino)-1-[6-(1-methyl-4-piperidyloxy)-2-pyridyl]-1,2-dihydro-3H- 1,2,5,7-tetraazainden-3-one using 6 ⁇ [(1 ⁇ methyl ⁇ 1H ⁇ indazol ⁇ 5 ⁇ yl)amino] ⁇ 1 ⁇ [6 ⁇ (piperidin ⁇ 4 ⁇ yloxy)pyridin ⁇ 2 ⁇ yl] ⁇ 2 ⁇ (prop ⁇ 2 ⁇ en ⁇
  • Example 144 Synthesis of 1- ⁇ 6-[(S)-1-methyl-3-pyrrolidinyloxy]-2-pyridyl ⁇ -2- allyl-6-(1-methyl-1H-indazol-5-ylamino)-1,2-dihydro-3H-1,2,5,7-tetraazainden-3-one (Compound 105) [0605] This compound was made using a similar method as described in the synthesis of allyl-6-(p-bromophenylamino)-1-[6-(1-methyl-4-piperidyloxy)-2-pyridyl]-1,2-dihydro-3H- 1,2,5,7-tetraazainden-3-one using tert-butyl (3S)-3-hydroxypyrrolidine-1-carboxylate.
  • Example 145 Synthesis of 1- ⁇ 6-[(S)-3-piperidyloxy]-2-pyridyl ⁇ -2-allyl-6-(1- methyl-1H-indazol-5-ylamino)-1,2-dihydro-3H-1,2,5,7-tetraazainden-3-one (Compound 104) [0607] This compound was made using a similar method as described in the synthesis of allyl-6-(p-bromophenylamino)-1-[6-(1-methyl-4-piperidyloxy)-2-pyridyl]-1,2-dihydro-3H- 1,2,5,7-tetraazainden-3-one using 1 ⁇ methyl ⁇ 1H ⁇ indazol ⁇ 5 ⁇ amine (1 equiv.) and tert ⁇ butyl (3S) ⁇ 3 ⁇ ( ⁇ 6 ⁇ [6 ⁇ (methylsulfanyl) ⁇ 3 ⁇ oxo ⁇ 2 ⁇ (prop ⁇ 2 ⁇ en ⁇ 1 ⁇
  • Example 146 Synthesis of 1- ⁇ 6-[(R)-3-pyrrolidinyloxy]-2-pyridyl ⁇ -2-allyl-6-(1- methyl-1H-indazol-5-ylamino)-1,2-dihydro-3H-1,2,5,7-tetraazainden-3-one (Compound 101) [0609] This compound was made using a similar method as described in the synthesis of allyl-6-(p-bromophenylamino)-1-[6-(1-methyl-4-piperidyloxy)-2-pyridyl]-1,2-dihydro-3H- 1,2,5,7-tetraazainden-3-one using 1-methyl-1H-indazol-5-amine and tert ⁇ butyl (3R) ⁇ 3 ⁇ ( ⁇ 6 ⁇ [6 ⁇ (methylsulfanyl) ⁇ 3 ⁇ oxo ⁇ 2 ⁇ (prop ⁇ 2 ⁇ en ⁇ 1 ⁇
  • Example 147 Synthesis of 1- ⁇ 6-[(S)-3-pyrrolidinyloxy]-2-pyridyl ⁇ -2-allyl-6-(1- methyl-1H-indazol-5-ylamino)-1,2-dihydro-3H-1,2,5,7-tetraazainden-3-one (Compound 100) [0611] This compound was made using a similar method as described in the synthesis of allyl-6-(p-bromophenylamino)-1-[6-(1-methyl-4-piperidyloxy)-2-pyridyl]-1,2-dihydro-3H- 1,2,5,7-tetraazainden-3-one using 1-methyl-1H-indazol-5-amine and tert ⁇ butyl (3S) ⁇ 3 ⁇ ( ⁇ 6 ⁇ [6 ⁇ (methylsulfanyl) ⁇ 3 ⁇ oxo ⁇ 2 ⁇ (prop ⁇ 2 ⁇ en ⁇ 1 ⁇ yl) ⁇ 1H,2
  • Example 150 Synthesis of 1- ⁇ 6-[(R)-3-pyrrolidinylamino]-2-pyridyl ⁇ -2-allyl-6-(1- methyl-1H-indazol-5-ylamino)-1,2-dihydro-3H-1,2,5,7-tetraazainden-3-one (Compound 102) [0617] tert-butyl (3R)-3-[(6-bromopyridin-2-yl)amino]pyrrolidine-1-carboxylate [0618] 2-Bromo-6-fluoropyridine (0.37 g, 2.1 mmol), DIPEA (6.3 mmol, 1.1 ml) and tert- butyl (3R)-3-aminopyrrolidine-1-carboxylate hydrochloride (0.52 g, 2.1 mmol) were mixed in 10 ml of DMSO.
  • reaction mixture was stirred at 100 °C for two days and partitioned between ethyl acetate and water.
  • the organic phase was washed with water, sat. NaHCO 3 and brine, dried over MgSO4, filtered and concentrated.
  • the residue was slurried with a mixture of heptane and ethyl acetate and collected by filtration giving 0.42 g (58%) of tert-butyl (3R)- 3-[(6-bromopyridin-2-yl)amino]pyrrolidine-1-carboxylate.
  • Step 2 6-(Methylsulfanyl)-2-(prop-2-en-1-yl)-1H,2H,3H-pyrazolo[3,4-d]pyrimidin-3-one (56 mg, 0.25 mmol), copper (I) iodide (52 mg, 0.27 mmol), potassium carbonate (104 mg, 0.75 mmol), 1,2-dimethylethylenediamine (44 mg, 0.50 mmol) and tert-butyl (3R)-3-[(6- bromopyridin-2-yl)amino]pyrrolidine-1-carboxylate (86 mg, 0.25 mmol) were mixed in 20 ml of dioxane under nitrogen.
  • Example 152 Synthesis of 6 ⁇ [(1 ⁇ methyl ⁇ 1H ⁇ indazol ⁇ 5 ⁇ yl)amino] ⁇ 1 ⁇ [6 ⁇ (piperidin ⁇ 4 ⁇ yloxy)pyridin ⁇ 2 ⁇ yl] ⁇ 2 ⁇ propyl ⁇ 1H,2H,3H ⁇ pyrazolo[3,4 ⁇ d]pyrimidin ⁇ 3 ⁇ one (Compound 110) [0626] To a solution of 6 ⁇ [(1 ⁇ methyl ⁇ 1H ⁇ indazol ⁇ 5 ⁇ yl)amino] ⁇ 1 ⁇ [6 ⁇ (piperidin ⁇ 4 ⁇ yloxy)pyridin ⁇ 2 ⁇ yl] ⁇ 2 ⁇ (prop ⁇ 2 ⁇ en ⁇ 1 ⁇ yl) ⁇ 1H,2H,3H ⁇ pyrazolo[3,4 ⁇ d]pyrimidin ⁇ 3 ⁇ one; trifluoroacetic acid (60 mg, 0.098 mmol) in ethanol (5 mL) was added 10% palladium on charcoal (4.4 mg) and the mixture was
  • Example 157 Synthesis of 6 ⁇ [(1 ⁇ methyl ⁇ 1H ⁇ indazol ⁇ 5 ⁇ yl)amino] ⁇ 2 ⁇ (prop ⁇ 2 ⁇ en ⁇ 1 ⁇ yl) ⁇ 1 ⁇ (6 ⁇ [(3R) ⁇ 1 ⁇ (propan ⁇ 2 ⁇ yl)pyrrolidin ⁇ 3 ⁇ yl]amino ⁇ pyridin ⁇ 2 ⁇ yl) ⁇ 1H,2H,3H ⁇ pyrazolo[3,4 ⁇ d]pyrimidin ⁇ 3 ⁇ one (Compound 120) [0641] This compound was made using a similar method as described in the synthesis of 2- allyl-6-(p-bromophenylamino)-1-[6-(1-methyl-4-piperidyloxy)-2-pyridyl]-1,2-dihydro-3H- 1,2,5,7-tetraazainden-3-one using 6-[(1-methyl-1H-indazol-5-yl)amin
  • Example 159 Synthesis of 6 ⁇ (phenylamino) ⁇ 1 ⁇ 6 ⁇ [(piperidin ⁇ 4 ⁇ yl)amino]pyridin ⁇ 2 ⁇ yl ⁇ 2 ⁇ (prop ⁇ 2 ⁇ en ⁇ 1 ⁇ yl) ⁇ 1H,2H,3H ⁇ pyrazolo[3,4 ⁇ d]pyrimidin ⁇ 3 ⁇ one (Compound 123) [0645] This compound was made using a similar method as described in the synthesis of 2- allyl-6-((4-chlorophenyl)amino)-1-(6-(piperidin-4-ylamino)pyridin-2-yl)-1,2-dihydro-3H- pyrazolo[3,4-d]pyrimidin-3-one using tert ⁇ butyl 4 ⁇ ( ⁇ 6 ⁇ [6 ⁇ (methylsulfanyl) ⁇ 3 ⁇ oxo ⁇ 2 ⁇ (prop ⁇ 2 ⁇ en ⁇ 1 ⁇ yl) ⁇ 1H
  • Example 160 Synthesis of 6 ⁇ [(4 ⁇ fluorophenyl)amino] ⁇ 1 ⁇ 6 ⁇ [(piperidin ⁇ 4 ⁇ yl)amino]pyridin ⁇ 2 ⁇ yl ⁇ 2 ⁇ (prop ⁇ 2 ⁇ en ⁇ 1 ⁇ yl) ⁇ 1H,2H,3H ⁇ pyrazolo[3,4 ⁇ d]pyrimidin ⁇ 3 ⁇ one (Compound 125) [0647] This compound was made using a similar method as described in the synthesis of 2- allyl-6-((4-chlorophenyl)amino)-1-(6-(piperidin-4-ylamino)pyridin-2-yl)-1,2-dihydro-3H- pyrazolo[3,4-d]pyrimidin-3-one using tert ⁇ butyl 4 ⁇ ( ⁇ 6 ⁇ [6 ⁇ (methylsulfanyl) ⁇ 3 ⁇ oxo ⁇ 2 ⁇ (prop ⁇ 2 ⁇ en ⁇
  • Example 164 Synthesis of 6 ⁇ [3 ⁇ methyl ⁇ 4 ⁇ (trifluoromethoxy)phenyl]amino ⁇ 1 ⁇ 6 ⁇ [(piperidin ⁇ 4 ⁇ yl)amino]pyridin ⁇ 2 ⁇ yl ⁇ 2 ⁇ (prop ⁇ 2 ⁇ en ⁇ 1 ⁇ yl) ⁇ 1H,2H,3H ⁇ pyrazolo[3,4 ⁇ d]pyrimidin ⁇ 3 ⁇ one (Compound 136) [0655] This compound was made using a similar method as described in the synthesis of 2- allyl-6-((4-chlorophenyl)amino)-1-(6-((1-methylpiperidin-4-yl)amino)pyridin-2-yl)-1,2- dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one using tert ⁇ butyl 4 ⁇ ( ⁇ 6 ⁇ [6 ⁇ (methylsulfanyl) ⁇ 3 ⁇ oxo ⁇ 2 ⁇
  • Example 165 Synthesis of 6 ⁇ [(4 ⁇ bromophenyl)amino] ⁇ 1 ⁇ 6 ⁇ [(piperidin ⁇ 4 ⁇ yl)amino]pyridin ⁇ 2 ⁇ yl ⁇ 2 ⁇ (prop ⁇ 2 ⁇ en ⁇ 1 ⁇ yl) ⁇ 1H,2H,3H ⁇ pyrazolo[3,4 ⁇ d]pyrimidin ⁇ 3 ⁇ one (Compound 143) [0657] This compound was made using a similar method as described in the synthesis of 2- allyl-6-((4-chlorophenyl)amino)-1-(6-((1-methylpiperidin-4-yl)amino)pyridin-2-yl)-1,2- dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one using tert ⁇ butyl 4 ⁇ ( ⁇ 6 ⁇ [6 ⁇ (methylsulfanyl) ⁇ 3 ⁇ oxo ⁇ 2 ⁇ (prop ⁇ 2
  • Example 166 Synthesis of 6 ⁇ [(3 ⁇ bromo ⁇ 5 ⁇ chlorophenyl)amino] ⁇ 1 ⁇ 6 ⁇ [(piperidin ⁇ 4 ⁇ yl)amino]pyridin ⁇ 2 ⁇ yl ⁇ 2 ⁇ (prop ⁇ 2 ⁇ en ⁇ 1 ⁇ yl) ⁇ 1H,2H,3H ⁇ pyrazolo[3,4 ⁇ d]pyrimidin ⁇ 3 ⁇ one (Compound 144) [0659] This compound was made using a similar method as described in the synthesis of 2- allyl-6-((4-chlorophenyl)amino)-1-(6-((1-methylpiperidin-4-yl)amino)pyridin-2-yl)-1,2- dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one using tert ⁇ butyl 4 ⁇ ( ⁇ 6 ⁇ [6 ⁇ (methylsulfanyl) ⁇ 3 ⁇ oxo ⁇ 2 ⁇
  • This intermediate was made using a similar method as described in the synthesis of tert-butyl 4-((6-(2-allyl-6-(methylthio)-3-oxo-2,3-dihydro-1H-pyrazolo[3,4-d]pyrimidin-1- yl)pyridin-2-yl)amino)piperidine-1-carboxylate using tert ⁇ butyl 4 ⁇ [(6 ⁇ bromopyridin ⁇ 2 ⁇ yl)(propyl)amino]piperidine ⁇ 1 ⁇ carboxylate (475 mg
  • Example 168 Synthesis of 6 ⁇ [3 ⁇ methyl ⁇ 5 ⁇ (trifluoromethoxy)phenyl]amino ⁇ 1 ⁇ 6 ⁇ [(piperidin ⁇ 4 ⁇ yl)amino]pyridin ⁇ 2 ⁇ yl ⁇ 2 ⁇ (prop ⁇ 2 ⁇ en ⁇ 1 ⁇ yl) ⁇ 1H,2H,3H ⁇ pyrazolo[3,4 ⁇ d]pyrimidin ⁇ 3 ⁇ one (Compound 155) [0668] This compound was made using a similar method as described in the synthesis of 2- allyl-6-((4-chlorophenyl)amino)-1-(6-((1-methylpiperidin-4-yl)amino)pyridin-2-yl)-1,2- dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one using tert ⁇ butyl 4 ⁇ ( ⁇ 6 ⁇ [6 ⁇ (methylsulfanyl) ⁇ 3 ⁇ oxo
  • Example 169 Synthesis of 6 ⁇ [(4 ⁇ chlorophenyl)amino] ⁇ 1 ⁇ 6 ⁇ [methyl(piperidin ⁇ 4 ⁇ yl)amino]pyridin ⁇ 2 ⁇ yl ⁇ 2 ⁇ (prop ⁇ 2 ⁇ en ⁇ 1 ⁇ yl) ⁇ 1H,2H,3H ⁇ pyrazolo[3,4 ⁇ d]pyrimidin ⁇ 3 ⁇ one (Compound 156) [0670] This compound was made using a similar method as described in the synthesis of : 6 ⁇ [(4 ⁇ chlorophenyl)amino] ⁇ 1 ⁇ 6 ⁇ [(1 ⁇ methylpiperidin ⁇ 4 ⁇ yl)(propyl)amino]pyridin ⁇ 2 ⁇ yl ⁇ 2 ⁇ (prop ⁇ 2 ⁇ en ⁇ 1 ⁇ yl) ⁇ 1H,2H,3H ⁇ pyrazolo[3,4 ⁇ d]pyrimidin ⁇ 3 ⁇ one using tert ⁇ butyl 4 ⁇ [methyl( ⁇ 6 ⁇ [
  • Example 170 Synthesis of 6 ⁇ [(4 ⁇ chlorophenyl)amino] ⁇ 1 ⁇ 6 ⁇ [methyl(1 ⁇ methylpiperidin ⁇ 4 ⁇ yl)amino]pyridin ⁇ 2 ⁇ yl ⁇ 2 ⁇ (prop ⁇ 2 ⁇ en ⁇ 1 ⁇ yl) ⁇ 1H,2H,3H ⁇ pyrazolo[3,4 ⁇ d]pyrimidin ⁇ 3 ⁇ one (Compound 157) [0672] This compound was made using a similar method as described in the synthesis of : 6 ⁇ [(4 ⁇ chlorophenyl)amino] ⁇ 1 ⁇ 6 ⁇ [(1 ⁇ methylpiperidin ⁇ 4 ⁇ yl)(propyl)amino]pyridin ⁇ 2 ⁇ yl ⁇ 2 ⁇ (prop ⁇ 2 ⁇ en ⁇ 1 ⁇ yl) ⁇ 1H,2H,3H ⁇ pyrazolo[3,4 ⁇ d]pyrimidin ⁇ 3 ⁇ one using 6 ⁇ [(4 ⁇ chlorophenyl)a
  • Example 172 Synthesis of 6 ⁇ [(4 ⁇ chlorophenyl)amino] ⁇ 1 ⁇ (6 ⁇ [(3S) ⁇ 1 ⁇ methylpiperidin ⁇ 3 ⁇ yl]oxy ⁇ pyridin ⁇ 2 ⁇ yl) ⁇ 2 ⁇ (prop ⁇ 2 ⁇ en ⁇ 1 ⁇ yl) ⁇ 1H,2H,3H ⁇ pyrazolo[3,4 ⁇ d]pyrimidin ⁇ 3 ⁇ one (Compound 162) [0676] This compound was made using a similar method as described in the synthesis of 2- allyl-6-((4-chlorophenyl)amino)-1-(6-((1-methylpiperidin-4-yl)amino)pyridin-2-yl)-1,2- dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one using 6 ⁇ [(4 ⁇ chlorophenyl)amino] ⁇ 1 ⁇ 6 ⁇ [(3R) ⁇ pipe
  • Example 174 Synthesis of 1 ⁇ 6 ⁇ [methyl(piperidin ⁇ 4 ⁇ yl)amino]pyridin ⁇ 2 ⁇ yl ⁇ 6 ⁇ [1 ⁇ (2 ⁇ methylpropyl) ⁇ 1H ⁇ pyrazol ⁇ 4 ⁇ yl]amino ⁇ 2 ⁇ (prop ⁇ 2 ⁇ en ⁇ 1 ⁇ yl) ⁇ 1H,2H,3H ⁇ pyrazolo[3,4 ⁇ d]pyrimidin ⁇ 3 ⁇ one (Compound 170) [0680] This compound was made using a similar method as described in the synthesis of 6 ⁇ [(4 ⁇ chlorophenyl)amino] ⁇ 1 ⁇ 6 ⁇ [(1 ⁇ methylpiperidin ⁇ 4 ⁇ yl)(propyl)amino]pyridin ⁇ 2 ⁇ yl ⁇ 2 ⁇ (prop ⁇ 2 ⁇ en ⁇ 1 ⁇ yl) ⁇ 1H,2H,3H ⁇ pyrazolo[3,4 ⁇ d]pyrimidin ⁇ 3 ⁇ one using tert ⁇ but
  • Example 176 Synthesis of 1 ⁇ 6 ⁇ [methyl(1 ⁇ methylpiperidin ⁇ 4 ⁇ yl)amino]pyridin ⁇ 2 ⁇ yl ⁇ 6 ⁇ [1 ⁇ (2 ⁇ methylpropyl) ⁇ 1H ⁇ pyrazol ⁇ 4 ⁇ yl]amino ⁇ 2 ⁇ (prop ⁇ 2 ⁇ en ⁇ 1 ⁇ yl) ⁇ 1H,2H,3H ⁇ pyrazolo[3,4 ⁇ d]pyrimidin ⁇ 3 ⁇ one (Compound 175) [0684] This compound was made using a similar method as described in the synthesis of 2- allyl-6-((4-chlorophenyl)amino)-1-(6-((1-methylpiperidin-4-yl)amino)pyridin-2-yl)-1,2- dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one using 1 ⁇ 6 ⁇ [methyl(piperidin ⁇ 4 ⁇ yl)amino]
  • Example 177 Synthesis of 1 ⁇ 6 ⁇ [(3R) ⁇ 1 ⁇ azabicyclo[2.2.2]octan ⁇ 3 ⁇ yloxy]pyridin ⁇ 2 ⁇ yl ⁇ 6 ⁇ [(4 ⁇ chlorophenyl)amino] ⁇ 2 ⁇ (prop ⁇ 2 ⁇ en ⁇ 1 ⁇ yl) ⁇ 1H,2H,3H ⁇ pyrazolo[3,4 ⁇ d]pyrimidin ⁇ 3 ⁇ one (Compound 176) [0686] This compound was made using a similar method as described in the synthesis of (R)- 2-allyl-6-((1-methyl-1H-pyrazol-3-yl)amino)-1-(6-(quinuclidin-3-yloxy)pyridin-2-yl)-1,2- dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one using 1 ⁇ 6 ⁇ [(3R) ⁇ 1 ⁇ azabicyclo[2.2.2]oc
  • Example 178 Synthesis of 2-allyl-1-[6-(4-piperidyloxy)-2-pyridyl]-6-(1-propyl-4- pyrazolylamino)-1,2-dihydro-3H-1,2,5,7-tetraazainden-3-one (Compound 179)
  • This compound was made using a similar method as described in the synthesis of 2- allyl-6-((4-chlorophenyl)amino)-1-(6-((1-methylpiperidin-4-yl)amino)pyridin-2-yl)-1,2- dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one using tert-butyl 4- ⁇ 6-[2-allyl-6-(methylthio)-3- oxo-1,2-dihydro-3H-1,2,5,7-tetraazainden-1-yl]-2-pyr
  • Example 180 Synthesis of 1-[6-(N-methyl-N-4-piperidylamino)-2-pyridyl]-2- allyl-6-[1-(2-fluoro-2-methylpropyl)-4-pyrazolylamino]-1,2-dihydro-3H-1,2,5,7- tetraazainden-3-one (Compound 183) [0692]
  • This compound was made using a similar method as described in the synthesis of 6 ⁇ [(4 ⁇ chlorophenyl)amino] ⁇ 1 ⁇ 6 ⁇ [(1 ⁇ methylpiperidin ⁇ 4 ⁇ yl)(propyl)amino]pyridin ⁇ 2 ⁇ yl ⁇ 2 ⁇ (prop ⁇ 2 ⁇ en ⁇ 1 ⁇ yl) ⁇ 1H,2H,3H ⁇ pyrazolo[3,4 ⁇ d]pyrimidin ⁇ 3 ⁇ one using tert-butyl 4-( ⁇ 6-[2-allyl-6- (methylthio)-3
  • Example 183 Synthesis of 1- ⁇ 6-[(R)-1-methyl-3-piperidyloxy]-2-pyridyl ⁇ -2-allyl- 6-(p-chlorophenylamino)-1,2-dihydro-3H-1,2,5,7-tetraazainden-3-one (Compound 187) [0698] This compound was made using a similar method as described in the synthesis of 2- allyl-6-((4-chlorophenyl)amino)-1-(6-((1-methylpiperidin-4-yl)amino)pyridin-2-yl)-1,2- dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one using 1- ⁇ 6-[(R)-3-piperidyloxy]-2-pyridyl ⁇ -2- allyl-6-(p-chlorophenylamino)-1,2-dihydro-3H-1,2,5,
  • Example 184 Synthesis of 6 ⁇ [(1 ⁇ methyl ⁇ 1H ⁇ pyrazol ⁇ 4 ⁇ yl)amino] ⁇ 1 ⁇ (6 ⁇ [(trans) ⁇ 8 ⁇ methyl ⁇ 8 ⁇ azabicyclo[3.2.1]octan ⁇ 3 ⁇ yl]oxy ⁇ pyridin ⁇ 2 ⁇ yl) ⁇ 2 ⁇ (prop ⁇ 2 ⁇ en ⁇ 1 ⁇ yl) ⁇ 1H,2H,3H ⁇ pyrazolo[3,4 ⁇ d]pyrimidin ⁇ 3 ⁇ one (Compound 190) and 6 ⁇ [(1 ⁇ methyl ⁇ 1H ⁇ pyrazol ⁇ 4 ⁇ yl)amino] ⁇ 1 ⁇ (6 ⁇ [(cis) ⁇ 8 ⁇ methyl ⁇ 8 ⁇ azabicyclo[3.2.1]octan ⁇ 3 ⁇ yl]oxy ⁇ pyridin ⁇ 2 ⁇ yl) ⁇ 2 ⁇ (prop ⁇ 2 ⁇ en ⁇ 1 ⁇ yl) ⁇ 1H,2H,3H ⁇ pyrazolo[3,4 ⁇ d]pyrimidin ⁇ 3 ⁇ one
  • Example 185 Synthesis of 2-allyl-6-(1-isopropyl-4-pyrazolylamino)-1- ⁇ 6-(9- methyl-9-azabicyclo[3.3.1]non-3-yloxy)-2-pyridyl ⁇ -1,2-dihydro-3H-1,2,5,7- tetraazainden-3-one (Compound 193) [0709] This compound was made using a similar method as described in the synthesis of allyl-6-(p-bromophenylamino)-1-[6-(1-methyl-4-piperidyloxy)-2-pyridyl]-1,2-dihydro-3H- 1,2,5,7-tetraazainden-3-one.
  • Example 186 Synthesis of 6 ⁇ [(2,6 ⁇ dimethylpyridin ⁇ 4 ⁇ yl)amino] ⁇ 1 ⁇ 6 ⁇ [(1 ⁇ methylpiperidin ⁇ 4 ⁇ yl)oxy]pyridin ⁇ 2 ⁇ yl ⁇ 2 ⁇ (prop ⁇ 2 ⁇ en ⁇ 1 ⁇ yl) ⁇ 1H,2H,3H ⁇ pyrazolo[3,4 ⁇ d]pyrimidin ⁇ 3 ⁇ one (Compound 199) [0711] This compound was made using a similar method as described in the synthesis of 2- allyl-6-(2-methoxy-4-pyridylamino)-1-[6-(1-methyl-4-piperidyloxy)-2-pyridyl]-1,2-dihydro- 3H-1,2,5,7-tetraazainden-3-one using 6 ⁇ [(2,6 ⁇ dimethylpyridin ⁇ 4 ⁇ yl)amino] ⁇ 1 ⁇ [6 ⁇ (piperidin ⁇ 4 ⁇ yloxy)pyridin ⁇
  • Example 188 Synthesis of 2 ⁇ methyl ⁇ 4 ⁇ [(1 ⁇ 6 ⁇ [(1 ⁇ methylpiperidin ⁇ 4 ⁇ yl)oxy]pyridin ⁇ 2 ⁇ yl ⁇ 3 ⁇ oxo ⁇ 2 ⁇ (prop ⁇ 2 ⁇ en ⁇ 1 ⁇ yl) ⁇ 1H,2H,3H ⁇ pyrazolo[3,4 ⁇ d]pyrimidin ⁇ 6 ⁇ yl)amino]benzonitrile (Compound 204) [0715] This compound was made using a similar method as described in the synthesis of 2- allyl-6-((4-chlorophenyl)amino)-1-(6-((1-methylpiperidin-4-yl)oxy)pyridin-2-yl)-1,2- dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one using 2 ⁇ methyl ⁇ 4 ⁇ ( ⁇ 3 ⁇ oxo ⁇ 1 ⁇ [6 ⁇ (piperidin ⁇ 4 ⁇ yloxy)pyridin
  • Example 189 Synthesis of 6 ⁇ [(3 ⁇ fluoro ⁇ 5 ⁇ methylphenyl)amino] ⁇ 1 ⁇ 6 ⁇ [(1 ⁇ methylpiperidin ⁇ 4 ⁇ yl)oxy]pyridin ⁇ 2 ⁇ yl ⁇ 2 ⁇ (prop ⁇ 2 ⁇ en ⁇ 1 ⁇ yl) ⁇ 1H,2H,3H ⁇ pyrazolo[3,4 ⁇ d]pyrimidin ⁇ 3 ⁇ one (Compound 205) [0717] This compound was made using a similar method as described in the synthesis of 2- allyl-6-((4-chlorophenyl)amino)-1-(6-((1-methylpiperidin-4-yl)oxy)pyridin-2-yl)-1,2- dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one using 1 ⁇ 6 ⁇ [(1 ⁇ methylpiperidin ⁇ 4 ⁇ yl)oxy]pyridin ⁇ 2 ⁇ yl ⁇ 6 ⁇ (methylsulf
  • Example 190 Synthesis of 6 ⁇ [(4 ⁇ fluoro ⁇ 3 ⁇ methylphenyl)amino] ⁇ 1 ⁇ 6 ⁇ [(1 ⁇ methylpiperidin ⁇ 4 ⁇ yl)oxy]pyridin ⁇ 2 ⁇ yl ⁇ 2 ⁇ (prop ⁇ 2 ⁇ en ⁇ 1 ⁇ yl) ⁇ 1H,2H,3H ⁇ pyrazolo[3,4 ⁇ d]pyrimidin ⁇ 3 ⁇ one (Compound 206) [0719] This compound was made using a similar method as described in the synthesis of 2- allyl-6-((4-chlorophenyl)amino)-1-(6-((1-methylpiperidin-4-yl)oxy)pyridin-2-yl)-1,2- dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one using 1 ⁇ 6 ⁇ [(1 ⁇ methylpiperidin ⁇ 4 ⁇ yl)oxy]pyridin ⁇ 2 ⁇ yl ⁇ 6 ⁇ (
  • Example 192 Synthesis of 2-allyl-6-(benzo[d]thiazol-5-ylamino)-1-(6-(piperidin- 4-yloxy)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (Compound 246) and 2-allyl-6-(benzo[d]thiazol-5-ylamino)-1-(6-(piperidin-4-yloxy)pyridin-2-yl)-1,2- dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (Compound 209) [0723] tert-butyl 4-((6-(2-allyl-6-(benzo[d]thiazol-5-ylamino)-3-oxo-2,3-dihydro-1H- pyrazolo[3,4-d]pyrimidin-1-yl)pyridin-2-
  • the resultant reaction mixture was subjected to stirring at 25 °C for 16 h. The progress of the reaction was monitored by TLC. After completion of the reaction, the reaction mixture was directly concentrated under reduced pressure, and the residue was diluted with water and extracted with 10% methanol-DCM (50 mL x 2). The combined organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure.
  • Example 195 Synthesis of 2-allyl-1-(3-(methyl(1-methylpiperidin-4- yl)amino)phenyl)-6-((1-methyl-1H-indazol-5-yl)amino)-1,2-dihydro-3H-pyrazolo[3,4- d]pyrimidin-3-one (Compound 217) [0734] 2-allyl-6-(methylthio)-1-(3-nitrophenyl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3- one [0735] To a solution of 2-allyl-6-(methylthio)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3- one (1.2 g, 5.40 mmol) in dichloroethane (15 mL) was added 3-nitrophenyl boronic acid (1.352 g, 8.10 mmol), sodium carbonate (1.707 g, 16.
  • tert-butyl (2-allyl-1-(3-((1-methylpiperidin-4- yl)amino)phenyl)-3-oxo-2,3-dihydro-1H-pyrazolo[3,4-d]pyrimidin-6-yl)(1-methyl-1H- indazol-5-yl)carbamate (0.130 g, 0.213 mmol) in THF (2 mL) was added NaH (0.012 g, 0.533 mmol) and allowed to stir for 30 min at 0 °C under inert atmosphere.
  • Example 196 Synthesis of 2-allyl-6-((1-methyl-1H-indazol-5-yl)amino)-1-(3-((1- methylpiperidin-4-yl)oxy)phenyl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (Compound 218) [0751] To a stirred solution of 2-allyl-6-((1-methyl-1H-indazol-5-yl)amino)-1-(3-(piperidin- 4-yloxy)phenyl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (210 mg, 0.423 mmol) in THF (5 mL) was added 37% aq.
  • reaction mixture was stirred at 25 °C for 1 h. The progress of the reaction was monitored by TLC and LCMS. After completion of the reaction, the reaction mixture was quenched with NH 3 in MeOH followed by TFA at rt and extracted with 10% MeOH in DCM (2 X 100 mL). The combined organic layers were dried over anhydrous Na 2 SO 4 .
  • the resulting reaction mixture was stirred at rt for 16 h. The progress of the reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated under reduced pressure. The residue was basified with sodium bicarbonate and extracted with ethyl acetate (10 mL x 2).
  • the progress of the reaction was monitored by TLC. After completion of the reaction, the reaction mixture was diluted with ice water (150 mL) and extracted with 10% methanol-DCM (2 X 100 mL). The combined organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure. The resulting crude material was purified by prep.
  • reaction mixture was stirred at rt for 2 h. The reaction progress was monitored by TLC. The reaction mixture was quenched with 10% sodium bicarbonate solution in water (50 mL) and extracted with DCM (250 mL). The organic layer was washed with brine water (250 ml ), and dried over Na2SO4, filtered, and concentrated under reduced pressure to give tert-butyl 4-((6-(2-allyl-6-(methylsulfonyl)-3-oxo-2,3-dihydro-1H-pyrazolo[3,4-d]pyrimidin- 1-yl)pyridin-2-yl)oxy)piperidine-1-carboxylate (2.5 g, 3.58 mmol, 66.1 % yield) as a yellow gum.
  • the progress of the reaction was monitored by TLC.
  • the reaction mixture was concentrated under vacuum to afford crude compound.
  • the crude product was purified by flash column chromatography (silica-gel, mesh size 60- 120) using ethyl acetate-hexane (55 to 60%) as an eluent to obtain tert-butyl 4-((6-(2-allyl-6- ((1-methyl-1H-indol-5-yl)amino)-3-oxo-2,3-dihydro-1H-pyrazolo[3,4-d]pyrimidin-1- yl)pyridin-2-yl)oxy)piperidine-1-carboxylate (600 mg, 0.674 mmol) as an brown solid.
  • the progress of the reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated under vacuum to afford crude compound.
  • the crude compound was neutralized by aqueous sodium bicarbonate solution (50 ml) and extracted with 10% methanol-DCM. Organic layers were dried over sodium sulfate and concentrated under vacuum to afford crude compound.
  • the crude compound was purified by prep-HPLC (Ammonium acetate method).
  • Example 201 Synthesis of 6-((1H-indazol-5-yl)amino)-2-allyl-1-(6-((1- methylpiperidin-4-yl)oxy)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (Compound 238) [0785] 2-allyl-1-(6-fluoropyridin-2-yl)-6-(methylsulfonyl)-1,2-dihydro-3H-pyrazolo[3,4- d]pyrimidin-3-one [0786] To a stirred solution of 2-allyl-1-(6-fluoropyridin-2-yl)-6-(methylthio)-1,2-dihydro- 3H-pyrazolo[3,4-d]pyrimidin-3-one (1 g, 3.15 mmol) in DCM (10 ml) was added m-CPBA (1.679 g, 6.62 mmol
  • the resultant reaction mixture was stirred at 25 °C for 16 h. The progress of the reaction was monitored by TLC and LCMS. After completion of the reaction, the reaction mixture was diluted with water and extracted with 10% MeOH in DCM (2 X 50 mL).
  • Example 202 Synthesis of 2-allyl-1-[6-(4-piperidyloxy)-2-pyridyl]-6-(7- quinolylamino)-1,2-dihydro-3H-1,2,5,7-tetraazainden-3-one (Compound 243) [0796] This compound was made using a similar method as described in the synthesis of 2- allyl-6-(2-methoxy-4-pyridylamino)-1-[6-(1-methyl-4-piperidyloxy)-2-pyridyl]-1,2-dihydro- 3H-1,2,5,7-tetraazainden-3-one. Yield 50 mg. [0797] Example 203.
  • tert-butyl 4-((4-iodopyrimidin-2-yl)oxy)piperidine-1-carboxylate To a stirred solution of tert-butyl 4-hydroxypiperidine-1-carboxylate (800 mg, 3.97 mmol) in THF (10 ml) at 0 °C under inert atmosphere was added NaH (477 mg, 11.92 mmol), followed by the addition of 4-iodo-2-(methylsulfonyl)pyrimidine (1.129 g, 3.97 mmol), which was dissolved in THF (5 mL). The mixture was stirred at rt for 20 min. The progress of the reaction was monitored by TLC and LCMS.
  • tert-butyl 4-((4-(2-allyl-6-(methylthio)-3-oxo-2,3-dihydro-1H- pyrazolo[3,4-d]pyrimidin-1-yl)pyrimidin-2-yl)oxy)piperidine-1-carboxylate (450 mg, 0.901 mmol) in DCM (10 ml) was added mCPBA (311 mg, 1.801 mmol) and the mixture was stirred at rt for 2 h.
  • 4-(4-(2-allyl-6-(methylsulfonyl)-3-oxo-2,3-dihydro- 1H-pyrazolo[3,4-d]pyrimidin-1-yl)pyrimidin-2-yl)oxy)piperidine-1-carboxylate 400 mg, 0.752 mmol
  • acetic acid (2 ml) was added 4-(2,2,2-trifluoroethoxy)aniline (216 mg, 1.129 mmol) at rt under inert atmosphere.
  • the resulting reaction mixture was stirred at 25 °C for 16 h. The progress of the reaction was monitored by TLC and LCMS. After completion of the reaction, the reaction mixture was diluted with water and extracted with 10% MeOH-DCM (2 X 50 mL).
  • the resulting reaction mixture was stirred at room temperature for 16 h. The progress of the reaction was monitored by TLC and LCMS. After completion of the reaction, the reaction mixture was concentrated under reduced pressure. The crude residue was washed with water and extracted with 10% MeOH in DCM. The organic layer was dried over anhydrous Na 2 SO 4 , and evaporated under reduced pressure to give the crude compound.
  • reaction mixture was stirred for 16 h. The progress of the reaction was monitored by TLC and LCMS. After completion of the reaction, the reaction mixture was concentrated under reduced pressure, water was added (100 mL). The resulting mixture wasextracted with 10% MeOH in DCM (2 X 100 mL). The combined organic layers was washed with 10% aq.
  • Example 210 Synthesis of rel-(S)-2-allyl-1-(6-(azepan-4-yloxy)pyridin-2-yl)-6- ((4-fluorophenyl)amino)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (Compound 266), rel-(S)-2-allyl-6-((4-fluorophenyl)amino)-1-(6-((1-methylazepan-4-yl)oxy)pyridin- 2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (Compound 351), rel-(R)-2-allyl- 1-(6-(azepan-4-yloxy)pyridin-2-yl)-6-((4-fluorophenyl)amino)-1,2-dihydro-3H- pyrazolo[3,4-d]pyrimidin-3-one (Compound 351)
  • tert-butyl 4-((6-(2-allyl-6-(methylthio)-3-oxo-2,3-dihydro-1H-pyrazolo[3,4- d]pyrimidin-1-yl)pyridin-2-yl)oxy)azepane-1-carboxylate To a stirred solution of tert-butyl 4-((6-bromopyridin-2-yl)oxy)azepane-1-carboxylate (5.01 g, 13.50 mmol) in 1,4-dioxane (15 ml) was added 2-allyl-6-(methylthio)-1,2-dihydro- 3H-pyrazolo[3,4-d]pyrimidin-3-one (2.5 g, 11.25 mmol), N,N-Dimethylethylenediamine (0.992 g, 11.25 mmol) and K2CO3 (4.66 g, 33.7 mmol).
  • tert-butyl 4-((6-(2-allyl-6-(methylsulfonyl)-3-oxo-2,3-dihydro-1H-pyrazolo[3,4- d]pyrimidin-1-yl)pyridin-2-yl)oxy)azepane-1-carboxylate To a stirred solution of tert-butyl 4-((6-(2-allyl-6-(methylthio)-3-oxo-2,3-dihydro-1H- pyrazolo[3,4-d]pyrimidin-1-yl)pyridin-2-yl)oxy)azepane-1-carboxylate (2.0 g, 3.90 mmol), in DCM (20 ml) was added mCPBA (1.885 g, 8.19 mmol) at 0 °C under inert atmosphere, and the resulting mixture was allowed to stir at rt for 3 h.
  • mCPBA 1.885 g, 8.19 mmol
  • Example 212 Synthesis of 2-allyl-6-((2-methyl-2H-indazol-5-yl)amino)-1-(6- (piperidin-4-yloxy)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (Compound 274) and 2-allyl-6-((2-methyl-2H-indazol-5-yl)amino)-1-(6-((1- methylpiperidin-4-yl)oxy)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (Compound 273) [0878] tert-butyl 4-((6-(2-allyl-6-((2-methyl-2H-indazol-5-yl)amino)-3-oxo-2,3-dihydro-1H- pyrazolo[3,4-d]pyrimidin-1-yl)pyri
  • the resultant reaction mixture was subjected to stirring at 70 °C for 16 h. The progress of the reaction was monitored by TLC. After completion of the reaction, the reaction mixture was diluted with water and extracted with 10% methanol-DCM (50 mL x 2). The combined organic layer was dried over Na 2 SO 4 , filtered, and concentrated under reduced pressure.
  • reaction mixture was stirred at 25 °C for 5 min.
  • STAB (575 mg, 2.71 mmol) was added portion-wise. After the complete addition of STAB, the reaction mixture was stirred at 25 °C for 20 min. The progress of the reaction was monitored by TLC and LCMS. The reaction mixture was quenched with TFA, followed by methanolic ammonia 2 molar solution diluted with water and extracted with 10% MeOH in DCM (2 X 100 mL). The combined organic extracts was washed with NaHCO3, dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to afford crude compound, which was purified by prep.
  • reaction mixture was subjected to stirring at 75 °C for 16 h. The progress of reaction was monitored by TLC. After completion of the reaction, the reaction mixture was diluted with water and extracted with 10% methanol-DCM (50 mL x 2). The combined organic layer was dried over Na 2 SO 4 , filtered, and concentrated under reduced pressure.
  • Example 214 Synthesis of Tert-butyl 4-((6-(2-allyl-6-((2-ethyl-2H-indazol-5- yl)amino)-3-oxo-2,3-dihydro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)pyridin-2- yl)oxy)piperidine-1-carboxylate (Compound 276) and 2-allyl-6-((2-ethyl-2H-indazol-5- yl)amino)-1-(6-((1-methylpiperidin-4-yl)oxy)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4- d]pyrimidin-3-one (Compound 282) [0890] tert-butyl 4-((6-(2-allyl-6-((2-ethyl-2H-indazol-5-yl)amino)-3-oxo-2,3-
  • reaction mixture was stirred at 70 °C for 16 h. The progress of the reaction was monitored by TLC. After completion of the reaction, the reaction mixture was diluted with water and extracted with 10% methanol-DCM (50 mL x 2). The combined organic layer was dried over Na 2 SO 4 , filtered, and concentrated under reduced pressure.
  • reaction mixture was stirred at 25 °C for 5 min. Then, STAB (0.829 g, 3.91 mmol) was added portion-wise. After the complete addition of STAB, the reaction mixture was stirred at 25 °C for 20 minutes. The progress of the reaction was monitored by TLC and LCMS. The reaction mixture was quenched with TFA followed by NH 3 in MeOH, diluted with water and extracted with 10% MeOH in DCM (2 X 100 mL). The above combined organic extracts were washed with NaHCO 3 , dried over anhydrous Na 2 SO 4 , filtered, and concentrated under reduced pressure. The above crude was purified by prep.
  • the reaction mixture was stirred at 70 °C for 16 h. The progress of reaction was monitored by LCMS, After the completion of the reaction, the reaction mixture was concentrated under reduced pressure to obtain crude product.
  • the crude product was purified by flash column chromatography (silica gel, 100-200) using ethyl acetate-hexane (50 to 100%) as an eluent to obtain tert-butyl 4-((6-(2-allyl-6-((4-morpholinophenyl)amino)-3-oxo- 2,3-dihydro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)pyridin-2-yl)oxy)piperidine-1-carboxylate (500 mg, 0.445 mmol, 45.8 % yield) as a red colored gum liquid.
  • the resulting reaction mixture was stirred at room temperature for 16 h. The progress of reaction was monitored by LCMS. After completion of the reaction, the reaction mixture was concentrated under reduced pressure. The resultant crude residue was washed with sodium bicarbonate and extracted with DCM. The solvent was removed in vacuo. The crude compound was submitted to prep.
  • Example 217 Synthesis of 2-allyl-6-((1-ethyl-1H-indazol-5-yl)amino)-1-(6- (piperidin-4-yloxy)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (Compound 280) [0911] 1-ethyl-5-nitro-1H-indazole [0912] To a stirred solution of 5-nitro-1H-indazole (1.5 g, 9.19 mmol) in N,N- Dimethylformamide (15 ml) was added sodium hydride (60% in oil, 1.57 g, 10.11 mmol) at 25 °C and the mixture was stirred for 30 min.
  • the resulting reaction mixture was subjected to stirring at 70 °C for 16 h. The progress of the reaction was monitored by TLC. After completion of the reaction, the reaction mixture was diluted with water and extracted with 10% methanol-DCM (50 mL x 2). The combined organic layer was dried over Na 2 SO 4 , filtered, and concentrated under reduced pressure.
  • the crude material was purified by flash column chromatography (silica gel, 100- 200 mesh size) using EtOH-hexane (60-80%) as an eluent to obtain tert-butyl 4-((6-(2-allyl- 6-((1-ethyl-1H-indazol-5-yl)amino)-3-oxo-2,3-dihydro-1H-pyrazolo[3,4-d]pyrimidin-1- yl)pyridin-2-yl)oxy)piperidine-1-carboxylate (350 mg, 0.244 mmol, 41.9 % yield) as a brown solid.
  • reaction mixture was stirred at 25 °C for 5 min. Then, sodium borodeuteride (33.7 mg, 0.804 mmol) was added portion-wise, and the reaction mixture was stirred at 25 °C for 20 min. The progress of the reaction was monitored by TLC and LCMS. After completion of the reaction, the reaction mixture was quenched with sodium hydroxide solution (20 mL) diluted with water and extracted with 10% MeOH in DCM (2 X 100 mL). The combined organic extracts were washed with NaHCO3, dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by prep.
  • Example 220 Synthesis of 2 ⁇ ethyl ⁇ 6 ⁇ [(1 ⁇ methyl ⁇ 1H ⁇ indazol ⁇ 5 ⁇ yl)amino] ⁇ 1 ⁇ 6 ⁇ [(1 ⁇ methylpiperidin ⁇ 4 ⁇ yl)amino]pyridin ⁇ 2 ⁇ yl ⁇ 1H,2H,3H ⁇ pyrazolo[3,4 ⁇ d]pyrimidin ⁇ 3 ⁇ one (Compound 288) [0924] This compound was made using a similar method as described in the synthesis of 2- allyl-6-((4-chlorophenyl)amino)-1-(6-((1-methylpiperidin-4-yl)amino)pyridin-2-yl)-1,2- dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one using 2 ⁇ ethyl ⁇ 6 ⁇ [(1 ⁇ methyl ⁇ 1H ⁇ indazol ⁇ 5 ⁇ yl)amino] ⁇
  • Example 224 Synthesis of 6 ⁇ [(1 ⁇ methyl ⁇ 1H ⁇ pyrazol ⁇ 4 ⁇ yl)amino] ⁇ 1 ⁇ (6 ⁇ ⁇ [(1S,4S,5R) ⁇ 2 ⁇ methyl ⁇ 2 ⁇ azabicyclo[2.2.2]octan ⁇ 5 ⁇ yl]oxy ⁇ pyridin ⁇ 2 ⁇ yl) ⁇ 2 ⁇ (prop ⁇ 2 ⁇ en ⁇ 1 ⁇ yl) ⁇ 1H,2H,3H ⁇ pyrazolo[3,4 ⁇ d]pyrimidin ⁇ 3 ⁇ one (Compound 295) [0937] This compound was made using a similar method as described in the synthesis of 2- allyl-6-((4-chlorophenyl)amino)-1-(6-((1-methylpiperidin-4-yl)amino)pyridin-2-yl)-1,2- dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one using 1 ⁇ 6 ⁇
  • Example 225 Synthesis of 2-allyl-1-(6-(piperidin-4-yloxy)pyridin-2-yl)-6-(m- tolylamino)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (Compound 296) [0939] tert-butyl 4-((6-(2-allyl-6-(methylsulfonyl)-3-oxo-2,3-dihydro-1H-pyrazolo[3,4- d]pyrimidin-1-yl)pyridin-2-yl)oxy)piperidine-1-carboxylate [0940] To a solution of tert-butyl 4-((6-(2-allyl-6-(methylthio)-3-oxo-2,3-dihydro-1H- pyrazolo[3,4-d]pyrimidin-1-yl)pyridin-2-yl)oxy)piperidine-1-carboxylate [0940] To a solution of
  • the progress of the reaction was monitored by TLC.
  • the reaction mixture was concentrated under vacuum.
  • the crude residue was neutralized by aqueous sodium bicarbonate solution (50 ml) and extracted with 10% methanol-DCM. The combined organic layers were dried over sodium sulfate and concentrated under vacuum.
  • the crude product was purified by flash column chromatography (silica-gel, mesh size 60-120) using ethyl acetate (100%) as an eluent to obtain desired product tert-butyl 4-((6-(2-allyl-3-oxo-6-(m-tolylamino)-2,3-dihydro-1H-pyrazolo[3,4- d]pyrimidin-1-yl)pyridin-2-yl)oxy)piperidine-1-carboxylate (400 mg, 0.572 mmol, 50.6 % yield) as a yellow solid.
  • tert-butyl 4-((6-(2-ethyl-6-(methylsulfonyl)-3-oxo-2,3- dihydro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)pyridin-2-yl)oxy)piperidine-1-carboxylate 400 mg, 0.771 mmol
  • acetic acid 5 mL
  • 1-methyl-1H-benzo[d]imidazol-5-amine 114 mg, 0.771 mmol
  • reaction mixture was stirred at rt for 16 h. The progress of the reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated under reduced pressure. The residue was basified with sodium bicarbonate and extracted with ethyl acetate (10 mL x 2). The combined organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure.
  • acetic acid 5 mL
  • reaction mixture was stirred at rt for 16 h. The progress of the reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated under reduced pressure. The residue was basified with sodium bicarbonate and extracted with ethyl acetate (10 mL x 2). The combined organic layer was dried over Na 2 SO 4 , filtered, and concentrated under reduced pressure.
  • Example 230 Synthesis of 2-allyl-1-[6-(1-methyl-4-piperidyloxy)-2-pyridyl]-6-(6- quinolylamino)-1,2-dihydro-3H-1,2,5,7-tetraazainden-3-one (Compound 325) [0964] This compound was made using a similar method as described in the synthesis of 2- allyl-6-(2-methoxy-4-pyridylamino)-1-[6-(1-methyl-4-piperidyloxy)-2-pyridyl]-1,2-dihydro- 3H-1,2,5,7-tetraazainden-3-one using tert-butyl 4-[6-(2-allyl-6-amino-3-oxo-1,2-dihydro-3H- 1,2,5,7-tetraazainden-1-yl)-2-pyridyloxy]-1-piperidinecarboxylate (100 mg) and 6- bromoquinoline (44.5 mg
  • Example 232 Synthesis of 6 ⁇ [(1 ⁇ methyl ⁇ 1H ⁇ indazol ⁇ 5 ⁇ yl)amino] ⁇ 1 ⁇ (6 ⁇ [(3R) ⁇ 1 ⁇ methylpiperidin ⁇ 3 ⁇ yl]oxy ⁇ pyridin ⁇ 2 ⁇ yl) ⁇ 2 ⁇ (prop ⁇ 2 ⁇ en ⁇ 1 ⁇ yl) ⁇ 1H,2H,3H ⁇ pyrazolo[3,4 ⁇ d]pyrimidin ⁇ 3 ⁇ one (Compound 331) [0973] This compound was made using a similar method as described in the synthesis of 2- allyl-6-((4-chlorophenyl)amino)-1-(6-(piperidin-4-yloxy)pyridin-2-yl)-1,2-dihydro-3H- pyrazolo[3,4-d]pyrimidin-3-one using 6 ⁇ [(1 ⁇ methyl ⁇ 1H ⁇ indazol ⁇ 5 ⁇ yl)amin
  • Example 233 Synthesis of 2-allyl-6-((3-(methyl-d3)phenyl)amino)-1-(6- (piperidin-4-yloxy)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (Compound 334) and 2-allyl-6-((3-(methyl-d3)phenyl)amino)-1-(6-((1-methylpiperidin- 4-yl)oxy)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (Compound 333) [0975] 1-bromo-3-(methyl-d3)benzene [0976] To a stirred solution of 1,3-dibromobenzene (4 g, 16.96 mmol) in THF (20 ml) was added n-
  • tert-butyl (3-(methyl-d3)phenyl)carbamate [0978] To a stirred solution of 1-bromo-3-(methyl-d3)benzene (400 mg, 2.298 mmol), tert- butyl carbamate 3538 mg, 4.60 mmol), Cs2CO3 (2246 mg, 6.89 mmol) and Xphos (219 mg, 0.460 mmol) in 1,4-dioxane (40 ml), were added. The mixture was degassed for 20 minutes at 25 °C. Then was added Pd(OAc)2 (103 mg, 0.460 mmol) and the mixture was stirred at 100 °C for 16 h.
  • tert-butyl 4-((6-(2-allyl-6-(methylthio)-3-oxo-2,3-dihydro- 1H-pyrazolo[3,4-d]pyrimidin-1-yl)pyridin-2-yl)oxy)piperidine-1-carboxylate 500 mg, 1.003 mmol
  • DCM 5 ml
  • m-CPBA 346 mg, 2.006 mmol
  • tert-butyl 4-((6-(2-allyl-6-(methylsulfonyl)-3-oxo-2,3-dihydro- 1H-pyrazolo[3,4-d]pyrimidin-1-yl)pyridin-2-yl)oxy)piperidine-1-carboxylate 450 mg, 0.848 mmol
  • acetic acid 3 ml
  • the reaction mixture was allowed to stir at room temperature for 16 h. The progress of the reaction was monitored by TLC and LCMS. After completion of the reaction, the solvent was evaporated under reduced pressure, and the residue was quenched with aq. sodium bicarbonate solution (100 mL) and extracted with 5% MeOH in DCM. The combined organic layers were dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to give a crude compound.
  • mCPBA 0.548 g, 3.17 mmol
  • reaction was allowed to stir at 25 °C for 16 h. The progress of the reaction was monitored by TLC. After completion of the reaction, the reaction mixture was diluted with water and extracted with ethyl acetate (100 mL x 2). The combined organic layer was dried over Na 2 SO 4 , filtered, and concentrated under reduced pressure to afford tert-butyl 4- hydroxy-2,2-dimethylpiperidine-1-carboxylate (500 mg, 2.093 mmol, 95 % yield) as a colorless liquid.
  • tert-butyl 4-((6-bromopyridin-2-yl)oxy)-2,2-dimethylpiperidine-1-carboxylate [1004] To a stirred solution of tert-butyl 4-hydroxy-2,2-dimethylpiperidine-1-carboxylate (500 mg, 2.180 mmol) in THF (10 ml) was added NaH (60% in oil, 157 mg, 6.54 mmol). After 10 min at 25 °C, was added 2,6-dibromopyridine (3, 620 mg, 2.62 mmol). The reaction was allowed to stir at 25 °C for 2 h. The progress of the reaction was monitored by TLC and UPLC.
  • tert-butyl 4-((6-(2-allyl-6-(methylthio)-3-oxo-2,3-dihydro-1H- pyrazolo[3,4-d]pyrimidin-1-yl)pyridin-2-yl)oxy)-2,2-dimethylpiperidine-1-carboxylate 400 mg, 0.760 mmol
  • DCM 5 ml
  • mCPBA 262 mg, 1.519 mmol
  • reaction mixture was subjected to stirring at 70 °C for 16 h. The progress of the reaction was monitored by TLC. After completion of the reaction, the reaction mixture was diluted with water and extracted with 10% methanol-DCM (50 mL x 2). The combined organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure.
  • the crude material was purified by flash column chromatography (silica gel, 100-200 mesh size) using ethyl acetate-hexane (40 % to 50%) as an eluent to obtain tert-butyl 4-((6-(2-allyl-6-((1-methyl-1H-indazol-5-yl)amino)-3-oxo-2,3-dihydro-1H- pyrazolo[3,4-d]pyrimidin-1-yl)pyridin-2-yl)oxy)-2,2-dimethylpiperidine-1-carboxylate (425 mg, 0.506 mmol, 66.9 % yield) as a brown solid.
  • reaction mixture stirred at 25 °C for 5 min.
  • STAB (1089 mg, 5.14 mmol) was added portion-wise.
  • the reaction mixture was stirred at 25 °C for 20 min.
  • the progress of the reaction was monitored by TLC and LCMS.
  • the reaction mixture was quenched with TFA, followed by NH3 in MeOH, diluted with water and extracted with 10% MeOH in DCM (2 X 100 mL).
  • the combined organic extracts was washed with NaHCO3, dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to afford crude compound, which was purified by prep.
  • Example 238 Synthesis of 2-allyl-6-((3-methyl-1H-indazol-5-yl)amino)-1-(6- (piperidin-4-yloxy)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (Compound 345) and 2-allyl-6-((3-methyl-1H-indazol-5-yl)amino)-1-(6-((1- methylpiperidin-4-yl)oxy)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (Compound 355) [1020] tert-butyl 4-((6-(2-allyl-6-((3-methyl-1H-indazol-5-yl)amino)-3-oxo-2,3-dihydro-1H-
  • reaction mixture was subjected to stirring at 70 °C for 16 h. The progress of the reaction was monitored by TLC. After completion of the reaction, the reaction mixture was diluted with water and extracted with 10% methanol-DCM (50 mL x 2). The combined organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure.
  • the resulting reaction mixture was stirred at room temperature for 4 h. The progress of the reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated under reduced pressure. Of the crude material, 800 mg was taken for the next step without any further purification. The remaining 200 mg crude residue was purified by prep.
  • reaction mixture was stirred at 25 °C for 5 min. Then, was added portion-wise STAB (1022 mg, 4.82 mmol). After the complete addition of STAB, the reaction mixture was stirred at 25 °C for 20 min. The reaction was monitored by TLC and LCMS. After completion of the reaction, the reaction mixture was quenched with TFA, followed by NH3 in MeOH diluted with water and extracted with 10% MeOH in DCM (2 X 100 mL). The above combined organic extracts were washed with NaHCO3, dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The resultant crude material was purified by prep.
  • reaction mixture was subjected to stirring at 70 °C for 16 h. The progress of the reaction was monitored by TLC. After completion of the reaction, the reaction mixture was diluted with water and extracted with 10% methanol-DCM (50 mL x 2). The combined organic layer was dried over Na 2 SO 4 , filtered, and concentrated under reduced pressure.
  • the crude material was purified by flash column chromatography (silica gel, 100- 200 mesh size) using ethyl acetate-hexane (40% to 50%) as an eluent to obtain tert-butyl 4- ((6-(2-allyl-6-((1,3-dimethyl-1H-indazol-5-yl)amino)-3-oxo-2,3-dihydro-1H-pyrazolo[3,4- d]pyrimidin-1-yl)pyridin-2-yl)oxy)piperidine-1-carboxylate (1.1 g, 1.068 mmol, 55.0 % yield) as a brown solid.
  • the resulting reaction mixture was stirred at room temperature for 4 h. The progress of the reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated under reduced pressure. Of the resulting crude residue, 200 mg was purified by prep HPLC, and the remaining was taken to the next step without any further purification.
  • reaction mixture was stirred at 25 °C for 5 min. Then, STAB (0.456 g, 2.150 mmol) was added portion-wise. After the complete addition of STAB, the reaction mixture was stirred at 25 °C for 20 min. The progress of the reaction was monitored by TLC and LCMS. The reaction mixture was quenched with TFA, followed by NH 3 in MeOH diluted with water and extracted with 10% MeOH in DCM (2 X 100 mL). The combined organic extracts were washed with NaHCO 3 , dried over anhydrous Na 2 SO 4 , filtered, and concentrated under reduced pressure. The residue was purified by prep.
  • mCPBA 0.479 g, 2.77 mmol
  • reaction mixture was stirred at rt for 2 h. The progress of the reaction was monitored by TLC and LCMS. After completion of the reaction, aq. NaHCO 3 (150 mL) was added and the mixture was extracted with ethyl acetate (2 X 150 mL).
  • reaction mixture was stirred at 70 °C for 16 h.
  • the progress of the reaction was monitored by TLC and LCMS.
  • the reaction mixture was concentrated under reduced pressure. Then was added ice cold water (100 mL). The mixture was extracted with 10% MeOH in DCM (2 X 250 mL).
  • the reaction mixture was subjected to stirring at 70 °C for 16 h. The progress of the reaction was monitored by LCMS. After the completion of the reaction, the reaction mixture was concentrated under reduced pressure.
  • the crude product was purified by flash column chromatography (silica gel, 100-200) using ethyl acetate-hexane (50 to 100%) as an eluent to obtain tert-butyl 4-((6-(2-allyl-6-((1-isobutyl-1H-indazol-5- yl)amino)-3-oxo-2,3-dihydro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)pyridin-2-yl)oxy)piperidine- 1-carboxylate (310 mg, 0.359 mmol, 30.8 % yield) as a red colored solid.
  • the mixture was stirred at rt for 1 h. The progress of the reaction was monitored by TLC and LCMS. After completion of the reaction, the reaction mixture was quenched with methanolic ammonia 2 molar solution, followed by TFA at rt. The mixture was diluted with water and extracted with 10% MeOH in DCM (2 X 50 mL). The combined organic layers were dried over anhydrous Na 2 SO 4 .
  • the mixture was stirred at rt for 16 h. The progress of the reaction was monitored by TLC and LCMS. After completion of the reaction, the reaction mixture was distilled, and residual solvent was neutralized with aq. NaHCO3 (50 mL) and the mixture was extracted with DCM (2 X 250 mL).
  • reaction mixture was stirred at 25 °C for 2 h. The progress of the reaction was monitored by TLC and LCMS. After completion of the reaction, the reaction mixture was quenched with NH3 in MeOH followed by TFA at rt. The mixture was diluted with water and extracted with 10% MeOH in DCM (2 X 100 mL). The combined organic layers was dried over anhydrous Na2SO4.
  • reaction mixture was stirred at 25 °C for 16 h. The progress of the reaction was monitored by TLC and LCMS. After completion of the reaction, the solvent was evaporated under reduced pressure, and then the reaction mixture was quenched with aq. NaHCO3 (50 mL) at 0 °C and extracted with 10% methanol in DCM (2 X 50 mL).
  • the mixture was stirred for 2 h. The progress of the reaction was monitored by TLC and LCMS. After completion of the reaction, the reaction mixture was quenched with NH3 in MeOH followed by TFA at rt and water. The mixture was extracted with 10% MeOH in DCM (2 X 100 mL).
  • the resulting reaction mixture was stirred at 70 °C for 3 h. The progress of the reaction was monitored by TLC and LCMS. After completion of the reaction, the reaction mixture was quenched with water (100 mL) and extracted with ethyl acetate (2 X 100 mL). The combined organic layers were dried over anhydrous Na 2 SO 4 , filtered, and evaporated under reduced pressure to give crude compound which was purified by column chromatography (SiO 2 /100-200 mesh; 40-50% ethyl acetate-hexane) to give 1-cyclopropyl-5- nitro-1H-indazole (3, 700 mg, 3.44 mmol, 37.5 % yield).
  • reaction mixture was stirred at 25 °C for 16 h. The progress of the reaction was monitored by TLC and LCMS. After completion of the reaction, the solvent was evaporated under reduced pressure and the residue was quenched with aq. NaHCO 3 (100 mL) at 0 °C and extracted with 10% MeOH-DCM (2 X 50 mL).
  • reaction mixture was stirred at room temperature for 1 h. The progress of the reaction was monitored by TLC and LCMS. After completion of the reaction, the reaction mixture was quenched with NH3 in MeOH followed by TFA at rt. The mixture was diluted with water and extracted with 10% MeOH in DCM (2 X 100 mL).
  • Example 25 Synthesis of 2-allyl-6-((4-(3-fluoropropoxy)phenyl)amino)-1-(6-((1- methylpiperidin-4-yl)oxy)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (Compound 377) and 2-allyl-6-((4-(3-fluoropropoxy)phenyl)amino)-1-(6-(piperidin-4- yloxy)pyridin-2-yl)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (Compound 378) [1106] (1-(3-fluoropropoxy)-4-nitrobenzene [1107] To a stirred solution of 4-nitrophenol (1 g, 7.19 mmol) in DMF (10 mL) was added potassium carbonate (0.993 g, 7.19 mmol) at 0 °C and
  • reaction mixture was subjected to stirring at 70 °C for 16 h. The progress of the reaction was monitored by TLC. After completion of the reaction, the reaction mixture was diluted with water and extracted with 10% methanol-DCM (50 mL x 2). The combined organic layer was dried over Na 2 SO 4 , filtered, and concentrated under reduced pressure.
  • the crude material was purified by flash column chromatography (silica gel, 100- 200 mesh size) using methanol-DCM (15-20%) as an eluent to obtain tert-butyl 4-((6-(2- allyl-6-((4-(3-fluoropropoxy)phenyl)amino)-3-oxo-2,3-dihydro-1H-pyrazolo[3,4- d]pyrimidin-1-yl)pyridin-2-yl)oxy)piperidine-1-carboxylate (510 mg, 0.757 mmol, 67.0 % yield) as a brown solid.
  • tert-butyl 4-((6-bromopyridin-2-yl)oxy)-2-methylpiperidine-1-carboxylate [1120] To a stirred solution of tert-butyl 4-hydroxy-2-methylpiperidine-1-carboxylate (3 g, 13.93 mmol) in THF (10 ml) was added sodium hydride (60% in oil, 0.669 g, 16.72 mmol). After 10 min at 25 °C, was added 2,6-dibromopyridine (3.14 g, 13.24 mmol) at 0 °C. The reaction was allowed to stir at 25 °C for 2 h. The progress of the reaction was monitored by TLC and UPLC.
  • the crude material was purified by flash column chromatography (silica gel, 100-200 mesh size) using ethyl acetate-petroleum ether (10% - 20%) as an eluent to obtain tert-butyl 4-((6-(2-allyl-6- (methylthio)-3-oxo-2,3-dihydro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)pyridin-2-yl)oxy)-2- methylpiperidine-1-carboxylate (2.2 g, 4.25 mmol, 79 % yield) off white solid.
  • mCPBA (65-70%, 148 mg, 0.858 mmol
  • reaction mixture was quenched with saturated sodium thiosulphate pentahydrate (100 mL) and extracted with DCM (250 mL X 2). The combined organic extract was washed with saturated aqueous sodium bicarbonate solution (100 mL).
  • tert-butyl 4-((6-(2-allyl-6-(methylsulfinyl)-3-oxo-2,3-dihydro- 1H-pyrazolo[3,4-d]pyrimidin-1-yl)pyridin-2-yl)oxy)-2-methylpiperidine-1-carboxylate (1.65 g, 3.12 mmol) in acetonitrile (15 mL) was added 1-methyl-1H-indazol-5-amine (0.551 g, 3.75 mmol) at room temperature.
  • the reaction mixture was subjected to stirring at 70 °C for 16 h. The progress of the reaction was monitored by TLC. After the completion of the reaction, the reaction mixture was concentrated under reduced pressure. The crude product (1.8 g) was purified by achiral SFC purification.
  • reaction mixture was stirred at 25 °C for 5 min. Then sodium triacetoxyborohydride (331 mg, 1.564 mmol) was added portion-wise. After the complete addition of STAB, the reaction mixture was stirred at 25 °C for 20 min. The progress of the reaction was monitored by TLC and LCMS. The reaction mixture was quenched with TFA, followed by NH3 in MeOH, then diluted with water and extracted with 10% MeOH in DCM (2 X 100 mL). The combined organic extract was washed with NaHCO 3 , dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to afford crude compound which was purified by prep.
  • reaction mixture was stirred at 25 °C for 5 min.
  • sodium triacetoxyborohydride (249 mg, 1.173 mmol) was added portion-wise.
  • the reaction mixture was stirred at 25 °C for 20 min.
  • the progress of the reaction was monitored by TLC and LCMS.
  • the reaction mixture was quenched with TFA followed by NH3 in MeOH, then diluted with water and extracted with 10% MeOH in DCM (2 X 100 mL). The combined organic extract was washed with NaHCO3, dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to afford crude compound which was purified by prep.
  • reaction mixture was subjected to stirring at 70 °C for 16 h. The progress of the reaction was monitored by TLC. After completion of the reaction, the reaction mixture was diluted with water and extracted with 10% methanol-DCM (50 mL x 2). The combined organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure.
  • the crude material was purified by flash column chromatography (silica gel, 100-200 mesh size) using ethyl acetate-hexane (40 % to 50%) as an eluent to obtain rel-tert-butyl(2S,4S)-4-((6-(2-allyl-6-((1-methyl-1H-indazol-5- yl)amino)-3-oxo-2,3-dihydro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)pyridin-2-yl)oxy)-2- methylpiperidine-1-carboxylate (1.03 g, 1.566 mmol, 59.1 % yield) as a brown solid.
  • reaction mixture was stirred at 25 °C for 5 min.
  • sodium triacetoxyborohydride (621 mg, 2.93 mmol) was added portion-wise.
  • the reaction mixture was stirred at 25 °C for 20 min.
  • the progress of the reaction was monitored by TLC and LCMS.
  • the reaction mixture was quenched with TFA, followed by NH3 in MeOH, diluted with water and extracted with 10% MeOH in DCM (2 X 100 mL). The combined organic extracts was washed with NaHCO 3 , dried over anhydrous Na 2 SO 4 , filtered, and concentrated under reduced pressure to afford crude compound which was purified by prep.

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Abstract

L'invention concerne des composés pyrazolo-pyrimidinone représentés par la formule I :, destinés à être utilisés dans des méthodes d'inhibition de la kinase Wee1A ou bien de la kinase Wee1A et de la kinase Myt1.
PCT/US2024/033065 2023-06-09 2024-06-07 Composés pyrazolo-pyrimidinone destinés à être utilisés dans des méthodes d'inhibition de la kinase wee1 Ceased WO2024254490A1 (fr)

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IL325136A IL325136A (en) 2023-06-09 2024-06-07 Pyrazolo-pyrimidinone compounds for use in methods for inhibiting kinase week1 a
AU2024284667A AU2024284667A1 (en) 2023-06-09 2024-06-07 Pyrazolo-pyrimidinone compounds for use in methods of inhibiting wee1 a kinase
CN202480051896.8A CN121752567A (zh) 2023-06-09 2024-06-07 用于抑制wee1 a激酶方法中的吡唑并嘧啶酮化合物
KR1020267000376A KR20260039689A (ko) 2023-06-09 2024-06-07 Wee1 a 인산화효소 억제 방법에 사용하기 위한 피라졸로-피리미딘온 화합물
MX2025014761A MX2025014761A (es) 2023-06-09 2025-12-08 Compuestos de pirazolo-pirimidinona para uso en metodos de inhibicion de la cinasa wee1 a
CONC2026/0000188A CO2026000188A2 (es) 2023-06-09 2026-01-08 Compuestos de pirazolo-pirimidinona para uso en métodos de inhibición de la cinasa wee1 a

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WO2019074979A1 (fr) * 2017-10-09 2019-04-18 Girafpharma, Llc Composés hétérocycliques et leurs utilisations
EP3572413A1 (fr) * 2017-01-23 2019-11-27 Shijiazhuang Sagacity New Drug Development Co., Ltd. Dérivé de 1,2-dihydro-3h-pyrazolo[3,4-d]pyrimidin-3-one utilisé en tant qu'inhibiteur de wee1

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EP3572413A1 (fr) * 2017-01-23 2019-11-27 Shijiazhuang Sagacity New Drug Development Co., Ltd. Dérivé de 1,2-dihydro-3h-pyrazolo[3,4-d]pyrimidin-3-one utilisé en tant qu'inhibiteur de wee1
WO2019074979A1 (fr) * 2017-10-09 2019-04-18 Girafpharma, Llc Composés hétérocycliques et leurs utilisations

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