Wee1 degrading compounds and uses thereof

Abstract

Provided herein are compounds and compositions thereof that reduce WEE1 kinase protein levels. In some embodiments, the compounds and compositions are provided for treatment WEE1 associated diseases such as cancer.

Description

[0001] WEE1 DEGRADING COMPOUNDS AND USES THEREOF

[0002] FIELD

[0003] The present disclosure relates generally to compounds, compositions, and methods for their preparation and use of the compounds and compositions for treating diseases and conditions associated with the WEE1 protein.

[0004] CROSS-REFERENCE TO RELATED APPLICATIONS

[0005] This application claims the benefit under 35 U.S.C. §119(e) of US Provisional Application Ser. No. 63 / 678,740, filed August 02, 2024; the disclosure of which is incorporated herein by reference.

[0006] BACKGROUND

[0007] Targeted protein degradation is a therapeutic modality whereby small molecules induce novel protein-protein interactions and enable destruction of target proteins that drive disease. CRBN E3 ligase modulator (CELMoD) small molecules are a class of targeted protein therapeutics that co-opts the CRL4-Cereblon E3 ubiquitin ligase complex, generating a new molecular “glue” interface on the surface of Cereblon that recruits and polyubiquitinates target proteins that are not normally ubiquitinated by Cereblon. The ubiquitin tagged proteins are then trafficked to and subsequently degraded by the 26S proteasome. The selection of target proteins recruited by Cereblon is determined by the specific molecular structure of the CELMoD compound.

[0008] WEE1 is a tyrosine kinase that phosphorylates the CDK1 and CDK2 cyclin-dependent kinases at their tyrosine- 15 residue consequently inhibiting kinase activity and halting the cell cycle at the intra-S and G2 / M cell cycle checkpoints. While halted at these checkpoints, cells repair stalled replication forks and DNA damage before entering mitosis; if the DNA damage cannot be repaired or rises above acceptable thresholds, cells undergo programmed apoptosis or mitotic catastrophe. Cancer cells frequently exhibit excessive replication stress, mutagenesis, and genomic instability prompting a reliance on cell cycle checkpoints to maintain DNA damage below apoptotic thresholds. Compared to normal cells, cancer cells frequently rely solely upon the intra-S and G2 / M checkpoints because their G1 cell cycle checkpoint is disabled by a various mechanisms, for instance, by restricting p53 and pRB activation, or, by hyper-activating replication-promoting factors such as Cyclin D and Cyclin E. Tumors that rely heavily upon the WEE 1 -mediated intra-S and G2 / M checkpoints are hypothesized to be exceptionally sensitive to WEE1 loss and therapeutics that target WEE1 are expected to exhibit antitumor activity with a favorable therapeutic window.

[0009] Certain cancer therapeutics function by inducing replication stress and DNA damage, including, for example, carboplatin, cisplatin, gemcitabine, pemetrexed, topotecan, doxorubicin, decitabine, and methotrexate. By inducing DNA damage and replication stress, these agents enhance the reliance of dividing tumor cells upon the WEE 1 -mediated cell cycle checkpoints. Therefore, CELMoD compounds that degrade WEE1 are proposed to synergistically combine with DNA damaging agents.

[0010] Accordingly, in one aspect, provided herein are CELMoD compounds that induce the Cereblon-mediated ubiquitination and degradation of WEE 1 to elicit broad antitumor activity as a single agent or in combination with sensitizing therapeutics.

[0011] SUMMARY

[0012] Described herein, in certain embodiments, are compounds and compositions thereof that degrade WEE1. In various embodiments, the compounds and compositions thereof may be used for treatment of diseases associated with WEE1, for example, cancer.

[0013] The present embodiments can be understood more fully by reference to the detailed description and examples, which are intended to exemplify non-limiting embodiments.

[0014] In some embodiments, provided herein are compounds selected from:

[0015] or a pharmaceutically acceptable salt, tautomer, isotopologue, or stereoisomer thereof.

[0016] In some embodiments, provided herein are methods for reducing WEE1 kinase protein levels, the method comprising contacting a cell with an effective amount of a compound of the present disclosure or a pharmaceutically acceptable salt, tautomer, isotopologue, or stereoisomer thereof. In some embodiments, provided herein are uses of a compound of the present disclosure or a pharmaceutically acceptable salt, tautomer, isotopologue, or stereoisomer thereof, in the manufacture of a medicament for reducing WEE1 kinase protein levels.

[0017] In some embodiments, provided herein are methods of preventing or treating cancer in a subject comprising administering to a subject in need thereof an effective amount of a compound of the present disclosure or a pharmaceutically acceptable salt, tautomer, isotopologue, or stereoisomer thereof. In some embodiments, provided herein are uses of a compound of the present disclosure or a pharmaceutically acceptable salt, tautomer, isotopologue, or stereoisomer thereof, in the manufacture of a medicament for the prevention or treatment of cancer. DETAILED DESCRIPTION

[0018] Considerable opportunities to target the "undruggable" utilizing targeted protein degradation have been realized over the last decade. Molecular glue molecules co-opting cereblon to degrade transcriptional regulators are clinically approved, providing meaningful benefit to patients. Several clinical Cereblon E3 Ligase Modulators (CELMoDs) for heme indications and more recently, for the potential treatment of solid tumors have been enrolled in clinical studies. The degradation of WEE1, an essential tyrosine kinase whose primary function is inhibiting cell cycle progression, leads to cell death and significant activity in select solid tumor oncology preclinical models.

[0019] Definitions

[0020] As used herein, the terms “comprising’’ and “including” can be used interchangeably. The terms “comprising” and “including” are to be interpreted as specifying the presence of the stated features or components as referred to, but does not preclude the presence or addition of one or more features, or components, or groups thereof. Additionally, the terms “comprising” and “including” are intended to include examples encompassed by the term “consisting of’. Consequently, the term “consisting of’ can be used in place of the terms “comprising” and “including” to provide for more specific embodiments of the invention.

[0021] The term “consisting of' means that a subject-matter has at least 90%, 95%, 97%, 98% or 99% of the stated features or components of which it consists. In another embodiment the term “consisting of’ excludes from the scope of any succeeding recitation any other features or components, excepting those that are not essential to the technical effect to be achieved.

[0022] As used herein, the term “or” is to be interpreted as an inclusive “or” meaning any one or any combination. Therefore, “A. B or C” means any of the following: “A; B; C; A and B; A and C; B and C; A, B and C”. An exception to this definition will occur only when a combination of elements, functions, steps or acts are in some way inherently mutually exclusive.

[0023] In the present description, any concentration range, percentage range, ratio range, or integer range is to be understood to include the value of any integer within the recited range and, when appropriate, fractions thereof (such as one tenth and one hundredth of an integer), unless otherwise indicated. Also, any number range recited herein relating to any physical feature, such as polymer subunits, size, or thickness, are to be understood to include any integer within the recited range, unless otherwise indicated. As used herein, the terms “about” and “approximately” mean ± 20%. ± 10%. ± 5%, or ± 1% of the indicated range, value, or structure, unless otherwise indicated. An “alkyl” group is a saturated, partially saturated, or unsaturated straight chain or branched non-cyclic hydrocarbon having from 1 to 10 carbon atoms (C1-C10 alkyl), typically from 1 to 8 carbons (Ci-Cs alkyl) or, in some embodiments, from 1 to 6 (Ci-Ce alkyl), 1 to 4 (C1-C4 alkyl), 1 to 3 (C1-C3 alkyl), or 2 to 6 (C2-C6 alkyl) carbon atoms. In some embodiments, the alky l group is a saturated alkyl group. Representative saturated alkyl groups include -methyl, -ethyl, -n-propyl, -n-butyl, -n-penlyl and -n-hexyl; while saturated branched alkyls include -isopropyl, -sec-butyl, -isobutyl, -tert-butyl, -isopentyl, -neopentyl, tert-pentyl. -2- methylpentyl, -3 -methylpentyl, -4-methylpentyl, -2,3-dimethylbutyl and the like. In some embodiments, an alkyd group is an unsaturated alkyd group, also termed an alkenyl or alkynyl group. An “alkenyl” group is an alkyl group that contains one or more carbon-carbon double bonds. An “alkynyl” group is an alkyl group that contains one or more carbon-carbon triple bonds. Examples of unsaturated alkyl groups include, but are not limited to, vinyl, allyl, -CH=CH(CH3), -CH=C(CH3)2, -C(CH3)=CH2, -C(CH3)=CH(CH3), -C(CH2CH3)=CH2, -C=CH. -C=C(CH3). -C=C(CH2CH3). -CH2C-CH. -CH2C=C(CH3) and -CH2C=C(CH2CH3). among others. An alkyd group can be substituted or unsubstituted. When the alkyl groups described herein are said to be “substituted,” they may be substituted with any substituent or substituents as those found in the exemplary' compounds and embodiments disclosed herein, as well as halogen; hydroxy; alkoxy; cycloalkydoxy, aryloxy, heterocyclyloxy, heteroarydoxy, cy cloalky dalkyloxy, ar dalkyloxy, heterocyclylalkyloxy, heteroarydalkyloxy; oxo (=0); amino, alkylamino, cycloalkylamino, arylamino, heterocyclylamino, heteroarylamino, cycloalkylalkylamino, arylalkydamino, heterocyclylalkylamino, heteroarylalkylamino; imino; imido; amidino; guanidino; enamino; acylamino; sulfonylamino; urea, nitrourea; oxime; hydroxylamino; alkoxyamino; aralkoxy amino; hydrazino; hydrazido; hydrazono; azido; nitro; thio (-SH), alkylthio; =S; sulfinyl; sulfonyl; aminosulfonyl; phosphonate: phosphinyl; acyl; formyl; carboxy; ester; carbamate; amido; cyano; isocyanato; isothiocyanate; cyanato; thiocyanato; or -B(OH)2. In certain embodiments, when the alkyl groups described herein are said to be “substituted,” they may be substituted with any substituent or substituents as those found in the exemplary’ compounds and embodiments disclosed herein, as well as halogen (chloro, iodo, bromo, or fluoro); alkyl; hydroxyl; alkoxy; alkoxyalkyl; amino; alkylamino; carboxy; nitro; cyano; thiol; thioether; imine; imide; amidine; guanidine; enamine; aminocarbonyl; acylamino; phosphonate; phosphine; thiocarbonyl; sulfinyl; sulfone; sulfonamide; ketone: aldehyde; ester; urea; urethane; oxime; hydroxyl amine; alkoxyamine; aralkoxyamine; N-oxide; hydrazine; hydrazide; hydrazone; azide; isocyanate; isothiocyanate; cyanate; thiocyanate; B(0H)2, or -O(alkyl)aminocarbonyl. A “cycloalkyf' group is a saturated, or partially saturated cyclic alkyl group of from 3 to 10 carbon atoms (C3-C10 cycloalkyd) having a single cyclic ring or multiple condensed or bridged rings that can be optionally substituted. In some embodiments, the cycloalkyl group has 3 to 8 ring carbon atoms (C3-C8 cycloalkyd), whereas in other embodiments the number of ring carbon atoms ranges from 3 to 5 (C3-C5 cycloalkyd), 3 to 6 (C3-C6 cycloalkyl), or 3 to 7 (C3-C7 cycloalkyl). In some embodiments, the cycloalkyl groups are saturated cycloalkyd groups. Such saturated cycloalkyl groups include, by way of example, single ring structures such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, 1 -methylcyclopropyl, 2-methylcyclopentyl, 2-methylcyclooctyl, and the like, or multiple or bridged ring structures such as l-bicyclo[I.I.I]pentyl, bicyclo[2. l.l]hexyl, bicyclo[2.2.1]heptyl, bicyclo[2.2.2]octyl, adamantyl and the like. In other embodiments, the cycloalkyl groups are unsaturated cycloalkyl groups. Examples of unsaturated cycloalkyl groups include cyclohexenyl, cyclopentenyl, cyclohexadienyl, butadienyl, pentadienyl, hexadienyl, among others. A cycloalkyl group can be substituted or unsubstituted. Such substituted cycloalk d groups include, by way of example, cyclohexanol and the like.

[0024] A “heterocyclyl” is a non-aromatic cycloalkyl in which one to four of the ring carbon atoms are independently replaced with a heteroatom selected from O, S and N. In some embodiments, heterocyclyl groups include 3 tolO ring members, whereas other such groups have 3 to 5, 3 to 6, or 3 to 8 ring members. Heterocyclyls can also be bonded to other groups at any ring atom (i.e.. at any carbon atom or heteroatom of the heterocyclic ring). A heterocycloalkyl group can be substituted or unsubstituted. Heterocyclyl groups encompass saturated and partially saturated ring systems. Further, the term heterocyclyl is intended to encompass any non-aromatic ring containing at least one heteroatom, which ring may be fused to an aryl or heteroaryl ring, regardless of the attachment to the remainder of the molecule. The phrase also includes bridged polycyclic ring systems containing a heteroatom. Representative examples of a heterocyclyl group include, but are not limited to, aziridinyl, azetidinyl, azepanyl, pyrrolidyl, imidazolidinyl (e.g., imidazolidin-4-onyl or imidazolidin-2,4-dionyl), pyrazolidinyl, thiazolidinyl. tetrahydrothiophenyl, tetrahydrofuranyl, piperidyl, piperazinyl (e.g., piperazin-2- onyl), morpholinyl, thiomorpholinyl, tetrahydropyranyl (e.g.. tetrahydro-2H-pyranyl), tetrahydrothiopyranyl, oxathianyl, dithianyl, l,4-dioxaspiro[4.5]decanyl, homopiperazinyl, quinuclidyl, or tetrahydropyrimidin-2(lH)-one. Representative substituted heterocyclyl groups may be mono-substituted or substituted more than once, such as, but not limited to, pyridyl or morpholinyl groups, which are 2-. 3-, 4-, 5-. or 6-substituted, or disubstituted with various substituents such as those listed below'. An “aryl” group is an aromatic carbocyclic group of from 6 to 14 carbon atoms (C6-C14 aryl) having a single ring (e.g., phenyl) or multiple condensed rings (e.g.. naphthyl or anthryl). In some embodiments, aryl groups contain 6-14 carbons (C6-C14 aryl), and in others from 6 to 12 (C6-C 12 aryl) or even 6 to 10 carbon atoms (Ce-Cio aryl) in the ring portions of the groups. Particular ary ls include phenyl, biphenyl, naphthyl and the like. An aryl group can be substituted or unsubstituted. The phrase “aryl groups” also includes groups containing fused rings, such as fused aromatic-aliphatic ring systems (e.g.. indanyl, tetrahydronaphthyl, and the like).

[0025] A “heteroaryl” group is an aromatic ring system having one to four heteroatoms as ring atoms in a heteroaromatic ring system, wherein the remainder of the atoms are carbon atoms. In some embodiments, heteroaryl groups contain 3 to 6 ring atoms, and in others from 6 to 9 or even 6 to 10 atoms in the ring portions of the groups. Suitable heteroatoms include oxygen, sulfur and nitrogen. In certain embodiments, the heteroaryl ring system is monocyclic or bicyclic. Non-limiting examples include but are not limited to, groups such as pyrrolyl, pyrazolyl, imidazolyl, triazolyl, tetrazolyl. oxazolyl, isoxazolyl. benzisoxazolyl (e.g.. benzo|d]isoxazolyl). thiazolyl, pyrolyl, pyndazinyl. pyrimidyl, pyrazinyl. thiophenyl, benzothiophenyl, furanyl, benzofuranyl, indolyl (e g., indolyl-2-onyl or isoindolin-l-onyl), azaindolyl (pyrrolopyridyl or lH-pyrrolo[2,3-b]pyridyl), indazolyl, benzimidazolyl (e.g., lH-benzo[d]imidazolyl). imidazopyridyl (e.g., azabenzimidazolyl or lH-imidazo[4,5-b]pyridyl), pyrazolopyridyl, triazol opyridyl, benzotriazolyl (e.g., lH-benzo[d][l ,2,3]triazolyl), benzoxazolyl (e g., benzo[d]oxazolyl), benzothiazolyl, benzothiadiazolyl, isoxazolopyridyl, thianaphthalenyl, purinyl, xanthinyl, adeninyl, guaninyl, quinolinyl, isoquinolinyl (e.g., 3,4-dihydroisoquinolin-l(2H)-onyl). tetrahydroquinolinyl, quinoxalinyl. and quinazolinyl groups. A heteroaryl group can be substituted or unsubstituted.

[0026] A “halogen” or “halo” is fluorine, chlorine, bromine or iodine.

[0027] An “alkoxy” group is -O-(alkyl), wherein alky l is defined above.

[0028] An “oxo” group is a “=O” group bonded to a carbon.

[0029] An “amino” group is -NH2, wherein one or both of the hydrogen atoms may be substituted with alkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl.

[0030] An “amido” group is an amide group with the formula -NHC(O)-, wherein the hydrogen atom may be substituted with alkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl.

[0031] A “heteroaryl-oxy” group is -O-(heteroaryl), wherein the heteroaryl is defined above. A “heterocyclyl-oxy” group is -O-(heterocyclyl), wherein the heterocyclyl is defined above. A “cycloalkyl-oxy” group is -O-(cycloalkyl), wherein the cycloalkyl is defined above When the groups described herein, with the exception of alkyl group, amino group, and amido group, are said to be ' substituted." they may be substituted with any appropriate substituent or substituents. Illustrative examples of substituents are those found in the exemplary compounds and embodiments disclosed herein, as well as halogen (chloro, iodo, bromo, or fluoro); alkyl; hydroxyl; alkoxy; alkoxyalkyl; amino; alkylamino; carboxy; nitro; cyano; thiol; thioether; imine; imide; amidine; guanidine; enamine; aminocarbonyl; acylamino; phosphonate; phosphine; thiocarbonyl; sulfinyl; sulfone; sulfonamide; ketone; aldehyde; ester; urea; urethane; oxime; hydroxyl amine; alkoxyamine; aralkoxyamine; N-oxide; hydrazine; hydrazide; hydrazone; azide; isocyanate; isothiocyanate; cyanate; thiocyanate; oxo (=0); B(0H)2, -O(alkyl)aminocarbonyl; cycloalkyl, which may be monocyclic or fused or non-fused polycyclic (e.g., cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl), or a heterocyclyl, which may be monocyclic or fused or non-fused polycyclic (e.g., pyrrolidyl, piperidyl, piperazinyl, morpholinyl, or thiazinyl); monocyclic or fused or non-fused polycyclic aryl or heteroaryl (e.g., phenyl, naphthyl, pyrrolyl, indolyl, furanyl, thiophenyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, triazolyl, tetrazolyl, pyrazolyl. pyridyl, quinolinyl, isoquinolinyl, acridinyl. pyrazinyl, pyridazinyl, pyrimidyl, benzimidazolyl, benzothiophenyl, or benzofuranyl); cycloalkyloxy, aryloxy, heterocyclyloxy, heteroaryloxy, cycloalkylalkyloxy, arylalkyloxy, heterocyclylalkyloxy, and heteroarylalkyloxy.

[0032] Embodiments of the disclosure are meant to encompass pharmaceutically acceptable salts, tautomers, isotopologues, and stereoisomers of the compounds provided herein, such as the compounds of the present disclosure.

[0033] As used herein, the term “pharmaceutically acceptable salt(s)” refers to a salt prepared from a pharmaceutically acceptable non-toxic acid or base including an inorganic acid and base and an organic acid and base. Suitable pharmaceutically acceptable base addition salts of the compounds of the present disclosure include, but are not limited to metallic salts made from aluminum, calcium, lithium, magnesium, potassium, sodium and zinc or organic salts made from lysine, N,N’ -dibenzylethylenediamine, chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine (N-methyl-glucamine) and procaine. Suitable non-toxic acids include, but are not limited to. inorganic and organic acids such as acetic, alginic, anthranilic, benzenesulfonic, benzoic, camphorsulfonic, citric, ethenesulfonic, formic, fumaric, furoic, galacturonic, gluconic, glucuronic, glutamic, glycolic, hydrobromic, hydrochloric, isethionic, lactic, maleic, malic, mandelic, methanesulfonic, mucic, nitric, pamoic. pantothenic, phenylacetic, phosphoric, propionic, salicylic, stearic, succinic, sulfanilic, sulfuric, tartaric acid, and p-toluenesulfonic acid. Specific non-toxic acids include hydrochloric, hydrobromic, maleic, phosphoric, sulfuric, and methanesulfonic acids. Examples of specific salts thus include hydrochloride, formic, and mesylate salts. Others are well-known in the art, see for example, Remington 's Pharmaceutical Sciences, 18theds.. Mack Publishing, Easton PA (1990) or Remington: The Science and Practice of Pharmacy, 19theds., Mack Publishing, Easton PA (1995).

[0034] As used herein and unless otherwise indicated, the term “stereoisomer” or “stereoisomerically pure” means one stereoisomer of a particular compound that is substantially free of other stereoisomers of that compound. For example, a stereoisomerically pure compound having one chiral center will be substantially free of the opposite enantiomer of the compound. A stereoisomerically pure compound having two chiral centers will be substantially free of other diastereomers of the compound. A typical stereoisomerically pure compound comprises greater than about 80% by weight of one stereoisomer of the compound and less than about 20% by weight of other stereoisomers of the compound, greater than about 90% by weight of one stereoisomer of the compound and less than about 10% by weight of the other stereoisomers of the compound, greater than about 95% by weight of one stereoisomer of the compound and less than about 5% by weight of the other stereoisomers of the compound, or greater than about 97% by weight of one stereoisomer of the compound and less than about 3% by weight of the other stereoisomers of the compound. The compounds disclosed herein can have chiral centers and can occur as racemates, individual enantiomers or diastereomers, and mixtures thereof. All such isomeric forms are included within the embodiments disclosed herein, including mixtures thereof.

[0035] The use of stereoisomerically pure forms of the compounds disclosed herein, as well as the use of mixtures of those forms, are encompassed by the embodiments disclosed herein. For example, mixtures comprising equal or unequal amounts of the enantiomers of a particular compound may be used in methods and compositions disclosed herein. These isomers may be asymmetrically synthesized or resolved using standard techniques such as chiral columns or chiral resolving agents. See, e.g., Jacques, J., et al., Enantiomers, Racemates and Resolutions (Wiley-Interscience, New York, 1981); Wilen, S. H., et al., Tetrahedron 33:2725 (1977); Eliel, E. L.. Stereochemistry of Carbon Compounds (McGraw-Hill. NY, 1962); Wilen. S. H., Tables of Resolving Agents and Optical Resolutions p. 268 (E.L. Eliel, Ed., Univ, of Notre Dame Press, Notre Dame, IN, 1972); Todd, M., Separation Of Enantiomers : Synthetic Methods (Wiley-VCH Verlag GmbH & Co. KgaA, Weinheim, Germany, 2014); Toda, F., Enantiomer Separation: Fundamentals and Practical Methods (Springer Science & Business Media, 2007); Subramanian, G. Chiral Separation Techniques: A Practical Approach (John Wiley & Sons, 2008); Ahuja, S.. Chiral Separation Methods for Pharmaceutical and Biotechnological Products (John Wiley & Sons. 2011).

[0036] ■‘Tautomers” refers to isomeric forms of a compound that are in equilibrium based on proton transfers. The concentrations of the isomeric forms will depend on the environment the compound is found in and may be different depending upon, for example, whether the compound is a solid or is in an organic or aqueous solution. For example, in aqueous solution, pyrazoles may exhibit the following isomeric forms, which are referred to as tautomers of each other:

[0037] As readily understood by one skilled in the art, a wide variety of functional groups and other structures may exhibit tautomerism and all tautomers of compounds of the present disclosure are within the scope of the present disclosure.

[0038] It should also be noted the compounds disclosed herein can contain unnatural proportions of atomic isotopes at one or more of the atoms. For example, the compounds may be radiolabeled with radioactive isotopes, such as for example tritium (3H), iodine- 125 (125I), sulfur-355S). or carbon- 14 (14C), or may be isotopically enriched, such as with deuterium (2H), carbon- 13 (13C), or nitrogen- 15 (15N). As used herein, an “isotopologue” is an isotopically enriched compound. The term “isotopically enriched” refers to an atom having an isotopic composition other than the natural isotopic composition of that atom. “Isotopically enriched” may also refer to a compound containing at least one atom having an isotopic composition other than the natural isotopic composition of that atom. The term “isotopic composition” refers to the amount of each isotope present for a given atom. Radiolabeled and isotopically enriched compounds are useful as therapeutic agents, e.g., cancer therapeutic agents, research reagents, e.g., binding assay reagents, and diagnostic agents, e.g., in vivo imaging agents. All isotopic variations of the compounds as described herein, whether radioactive or not, are intended to be encompassed within the scope of the embodiments provided herein. In some embodiments, there are provided isotopologues of the compounds disclosed herein, for example, the isotopologues are deuterium, carbon-13, and / or nitrogen- 15 enriched compounds. As used herein, “deuterated”, means a compound wherein at least one hydrogen (H) has been replaced by deuterium (indicated by D or2H), that is, the compound is enriched in deuterium in at least one position. It is understood that, independently of stereoisomerical or isotopic composition, each compound disclosed herein can be provided in the form of any of the pharmaceutically acceptable salts discussed herein. Equally, it is understood that the isotopic composition may vary independently from the stereoisomerical composition of each compound referred to herein. Further, the isotopic composition, while being restricted to those elements present in the respective compound or salt thereof disclosed herein, may otherwise vary independently from the selection of the pharmaceutically acceptable salt of the respective compound.

[0039] It should be noted that if there is a discrepancy between a depicted structure and a name for that structure, the depicted structure is to be accorded more weight.

[0040] “Treating” as used herein, means an alleviation, in whole or in part, of a disorder, disease or condition, or one or more of the symptoms associated with a disorder, disease, or condition, or slowing or halting of further progression or worsening of those symptoms, or alleviating or eradicating the cause(s) of the disorder, disease, or condition itself. In one embodiment, the disorder is a neurodegenerative disease, as described herein, or a symptom thereof.

[0041] “Preventing” as used herein, means a method of delaying and / or precluding the onset, recurrence or spread, in whole or in part, of a disorder, disease or condition; barring a subject from acquiring a disorder, disease, or condition; or reducing a subject’s risk of acquiring a disorder, disease, or condition. In one embodiment, the disorder is a neurodegenerative disease, as described herein, or symptoms thereof.

[0042] The term “effective amount” in connection with a compound disclosed herein means an amount capable of treating or preventing a disorder, disease or condition, or symptoms thereof, disclosed herein.

[0043] The term “subject” or “patient” as used herein include an animal, including, but not limited to, an animal such a cow, monkey, horse, sheep, pig, chicken, turkey, quail, cat, dog. mouse, rat, rabbit or guinea pig, in one embodiment a mammal, in another embodiment a human. In one embodiment, a subject is a human having or at risk for having an WEE1 mediated disease, or a symptom thereof.

[0044] Although various features of the invention may be described in the context of a single embodiment, the features may also be provided separately or in any suitable combination. Conversely, although the invention may be described herein in the context of separate embodiments for clarity, the invention may also be implemented in a single embodiment. Compounds

[0045] In one aspect, provided herein are compounds selected from:

[0046] or a pharmaceutically acceptable salt, tautomer, isotopologue, or stereoisomer thereof.

[0047] In the descriptions herein, it is understood that every description, variation, embodiment, or aspect of a moiety may be combined with every7description, variation, embodiment, or aspect of other moieties the same as if each and every7combination of descriptions is specifically and individually listed. It is also understood that all descriptions, variations, embodiments, or aspects of the compounds, where applicable, apply equally to other formulae detailed herein, and are equally described, the same as if each and every7description, variation, embodiment, or aspect were separately and individually listed for all formulae. In some embodiments, provided is a compound selected from the compounds in Table 1 or a pharmaceutically acceptable salt thereof. Although certain compounds described in the present disclosure, including in Table 1, may be presented as specific stereoisomers and / or in a non-stereochemical form, it is understood that any or all stereochemical forms, including any enantiomeric or diastereomeric forms, and any tautomers or other forms of the compounds of the present disclosure, including in Table 1, are herein described.

[0048] Table 1.

[0049] It is understood that in the present description, combinations of substituents and / or variables of the depicted formulae are permissible only if such contributions result in stable compounds.

[0050] Furthermore, all compounds of the present disclosure that exist in free base or acid form can be converted to their pharmaceutically acceptable salts by treatment with the appropriate inorganic or organic base or acid by methods known to one skilled in the art. Salts of the compounds of the present disclosure can be converted to their free base or acid form by standard techniques.

[0051] Methods of Synthesis

[0052] The compounds described herein can be made using conventional organic syntheses and commercially available starting materials, or the methods provided herein. By way of example and not limitation, compounds of the present disclosure can be prepared as outlined in Scheme 1, as well as in the examples set forth herein. It should be noted that one skilled in the art would know how to modify the procedures set forth in the illustrative schemes and examples to arrive at the desired products, including, for example, selecting starting materials having different stereochemistry' (or racemic starting materials) to arrive at desired products having different stereochemistry.

[0053] In general, the compounds of the present disclosure can be obtained by the following general Scheme I:

[0054]

[0055] In all embodiments, the reaction of the aldehyde is the last step in the reaction sequence.

[0056] Methods of Use

[0057] Embodiments of the present disclosure provide a method for degrading WEE1, a method for reducing WEE1 proteins levels, and a method of preventing or treating diseases such as cancer in a subject in need thereof.

[0058] In one aspect, provided herein is a method for degrading WEE1 in a subject in need thereof, the method comprising contacting a cell with an effective amount of a compound of the present disclosure. Degradation of WEE1 can be assessed and demonstrated by a wide variety’ of methods known in the art. Kits and commercially available assays, including cell-based assays, can be utilized for determining whether and to what degree WEE1 has been degraded. In some embodiments, the compound of the present disclosure partially degrades WEE1. In some embodiments, the compound of the present disclosure fully degrades WEE1.

[0059] In some embodiments, a compound of the present disclosure degrades WEE1 by about 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%. 90%. 95%. or 100%. In some embodiments, a compound of the present disclosure degrades WEE1 by about 1-100%, 5-100%, 10-100%, 15-100%, 20-100%, 25-100%, 30-100%, 35-100%, 40-100%, 45-100%, 50-100%, 55-100%, 60-100%, 65-100%, 70-100%, 75-100%, 80-100%, 85-100%, 90-100%, 95-100%, 5-95%, 5-90%, 5-85%, 5-80%, 5-75%, 5-70%, 5-65%, 5-60%. 5-55%. 5-50%, 5-45%, 5-40%, 5-35%, 5-30%, 5-25%, 5-20%, 5-15%, 5-10%, 10-90%, 20-80%, 30-70%, or 40-60%.

[0060] In some embodiments, provided herein is a method for reducing WEE1 kinase protein levels, the method comprising contacting a cell with an effective amount of a compound of the present disclosure. Reduction of WEE1 kinase protein levels can be assessed and demonstrated by a wide variety of methods known in the art. Kits and commercially available assays, including cell-based assays, can be utilized for determining whether and to what degree kinase protein levels have been reduced.

[0061] In some embodiments, a compound of the present disclosure reduces WEE1 kinase protein levels by about 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%. In some embodiments, a compound of the present disclosure reduces WEE1 kinase protein levels by about 1-100%. 5-100%, 10-100%. 15- 100%, 20-100%, 25-100%, 30-100%, 35-100%, 40-100%, 45-100%, 50-100%, 55-100%, 60- 100%, 65-100%, 70-100%, 75-100%, 80-100%, 85-100%, 90-100%, 95-100%, 5-95%, 5-90%, 5-85%, 5-80%. 5-75%, 5-70%, 5-65%, 5-60%, 5-55%, 5-50%, 5-45%, 5-40%, 5-35%, 5-30%, 5-25%. 5-20%. 5-15%, 5-10%, 10-90%. 20-80%, 30-70%, or 40-60%.

[0062] In some embodiments, a compound of the present disclosure has an EC so value as measured in a WEE1 degradation assay of from about 0.0003 pM to about 1 pM or from about 0.0003 pM to about 0.2 pM or from about 0.0003 pM to about 0.05 pM. In some embodiments, a compound of the present disclosure has an ECso of from about 0.05 pM to about 0.2 pM. In some embodiments, a compound of the present disclosure has an ECso of from about 0.2 pM to about 1 pM. In some embodiments, a compound of the present disclosure has an EC50 of less than about 1 pM. In some embodiments, a compound of the present disclosure has an EC50 value of less than 0.2 pM, less than 0.05 pM, less than 0.001 pM, or less than about 0.0003 pM.

[0063] In another aspect, provided herein is a method for treating cancer in a subject in need thereof, comprising administering to the subject an effective amount of a compound of the present disclosure. In some embodiments, provided herein is a method for preventing cancer in a subject in need thereof, comprising administering to the subject an effective amount of a compound of the present disclosure. In some embodiments, the cancer is selected from a brain cancer, a cervi cocerebral cancer, an esophageal cancer, a thyroid cancer, small cell cancer, a non-small cell cancer, a breast cancer, a lung cancer , a stomach cancer, 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, a chronic lymphatic leukemia, a chronic myelocytic leukemia, polycy themia vera, a malignant lymphoma, multiple myeloma, a Hodgkin's lymphoma, and a non-Hodgkin?s lymphoma. In some embodiments, the cancer is selected from gastric, lung, pancreatic, ovarian, breast, skin, colon, neuroblastoma, osteosarcoma, uterine, rectal, and kidney cancer. In some embodiments, the cancer is selected from pancreatic ductal adenocarcinoma (PDAC), small cell lung cancer, non-small cell lung cancer (NSCLC), high grade serous ovarian cancer, triple negative breast cancer, uterine serous carcinoma, Ewing’s sarcoma, melanoma, colon, and clear cell renal cell carcinoma (ccRCC).

[0064] In some embodiments, administering a compound of the present disclosure to a subject that is predisposed to cancer prevents the subject from developing any symptoms of the cancer (such as tumor growth or metastasis). In some embodiments, administering a compound of the present disclosure to a subject that does not yet display symptoms of cancer prevents the subject from developing any symptoms of the cancer. In some embodiments, administering a compound of the present disclosure to a subject in need thereof diminishes the extent of the cancer in the subject. In some embodiments, administering a compound of the present disclosure to a subject in need thereof stabilizes the cancer (prevents or delays the worsening of the cancer). In some embodiments, administering a compound of the present disclosure to a subject in need thereof delays the occurrence or recurrence of the cancer. In some embodiments, administering a compound of The present invention to a subject in need thereof slow s the progression of the cancer. In some embodiments, administering a compound of The present invention to a subject in need thereof provides a partial remission of the cancer. In some embodiments, administering a compound of The present invention to a subject in need thereof provides a total remission of the cancer. In some embodiments, administering a compound of The present invention to a subject in need thereof decreases the dose of one or more other medications required to treat the cancer. In some embodiments, administering a compound of The present invention to a subject in need thereof enhances the effect of another medication used to treat the cancer. In some embodiments, administering a compound of the present disclosure to a subject in need thereof delays the progression of the cancer. In some embodiments, administering a compound of the present disclosure to a subject in need thereof increases the quality of life of the subject having cancer. In some embodiments, administering a compound of the present disclosure to a subject in need thereof prolongs survival of a subject having cancer.

[0065] In one aspect, provided herein is method of preventing a subj ect that is predisposed to cancer from developing cancer, the method comprising administering a compound of the present disclosure to the subject.

[0066] In some aspects, provided herein is a method of diminishing the extent of cancer in a subject, the method comprising administering a compound of the present disclosure to the subject. In some embodiments, provided herein is a method of stabilizing cancer in a subject, the method comprising administering a compound of the present disclosure to the subject. In some embodiments, the method prevents the worsening of the cancer.

[0067] In another aspect, provided herein is a method of delaying the occurrence or recurrence of cancer in a subject, the method comprising administering a compound of the present disclosure to the subject.

[0068] In some embodiments, provided herein is a method of slowing the progression of cancer in a subject, the method comprising administering a compound of the present disclosure to the subject. In some embodiments, the method provides a partial remission of the cancer. In some embodiments, the method provides a total remission of the cancer.

[0069] In further aspects, provided herein is a method of decreasing the dose of one or more other medications required to treat cancer in a subject, the method comprising administering a compound of the present disclosure to the subject. In some embodiments, provided herein is a method of enhancing the effect of another medication used to treat cancer in a subject, the method comprising administering a compound of the present disclosure to the subject.

[0070] Also provided here is a method of delaying the progression of cancer in a subject, the method comprising administering a compound of the present disclosure to the subject. In some embodiments, the method increases the quality of life of the subject having cancer. In some embodiments, the method prolongs survival of the subject having cancer.

[0071] In some embodiments, compounds of the present disclosure are useful in the manufacture of a medicament for reducing WEE1 kinase protein levels. In some embodiments, compounds of the present disclosure are useful in the manufacture of a medicament for the prevention or treatment of a disease associates with WEE1. In some embodiments, compounds of the present disclosure are useful in the manufacture of a medicament for the prevention or treatment of cancer. In some embodiments, the cancer is selected from gastric, lung, pancreatic, ovarian, breast, skin, colon, neuroblastoma, osteosarcoma, uterine, rectal, and kidney cancer. In some embodiments, the cancer is selected from pancreatic ductal adenocarcinoma (PDAC), small cell lung cancer, non-small cell lung cancer (NSCLC), high grade serous ovarian cancer, triple negative breast cancer, uterine serous carcinoma, Ewing’s sarcoma, melanoma, colon, and clear cell renal cell carcinoma (ccRCC).

[0072] The methods and uses of the present disclosure may include a compound of the present disclosure used alone or in combination with one or more additional therapies (e.g., non-drug treatments or therapeutic agents).

[0073] A compound of the present disclosure may be administered before, after, or concurrently with one or more of such additional therapies. When combined, dosages of the compound of the present disclosure and dosages of the one or more additional therapies (e.g., non-drug treatment or therapeutic agent) may provide a therapeutic effect (e.g., synergistic or additive therapeutic effect). A compound of the present disclosure and an additional therapy, such as an anti-cancer agent, may be administered together, such as in a unitary7pharmaceutical composition, or separately and, when administered separately, this may occur simultaneously or sequentially. Such sequential administration may be close or remote in time.

[0074] In some embodiments, the additional therapy is the administration of side-effect limiting agents (e.g., agents intended to lessen the occurrence or severity of side effects of treatment). For example, in some embodiments, the compounds of the present disclosure can be used in combination with a therapeutic agent that treats nausea. Examples of agents that can be used to treat nausea include, but are not limited to, dronabinol, granisetron, metoclopramide, ondansetron, prochlorperazine, and pharmaceutically acceptable salts thereof.

[0075] In some embodiments, one or more additional therapies includes a non-drug treatment (e.g., surgery or radiation therapy). In some embodiments, one or more additional therapies includes a therapeutic agent (e.g., a compound or biologic that is an antiproliferative agent). In some embodiments, one or more additional therapies includes a non-drug treatment and a therapeutic agent. In other embodiments, one or more additional therapies includes two therapeutic agents. In still other embodiments, one or more additional therapies includes three therapeutic agents. In some embodiments, one or more additional therapies includes four or more therapeutic agents.

[0076] Pharmaceutical Compositions and Routes of Administration

[0077] The compounds provided herein can be administered to a subject orally, topically or parenterally in the conventional form of preparations, such as capsules, microcapsules, tablets, granules, powder, troches, pills, suppositories, injections, suspensions, syrups, patches, creams, lotions, ointments, gels, sprays, solutions and emulsions.

[0078] The compounds disclosed herein can be administered to a subject orally, topically or parenterally in the conventional form of preparations, such as capsules, microcapsules, tablets, granules, powder, troches, pills, suppositories, injections, suspensions, syrups, patches, creams, lotions, ointments, gels, sprays, solutions and emulsions. Suitable formulations can be prepared by methods commonly employed using conventional, organic or inorganic additives, such as an excipient (e.g., sucrose, starch, mannitol, sorbitol, lactose, glucose, cellulose, talc, calcium phosphate or calcium carbonate), a binder (e.g., cellulose, methylcellulose, hydroxymethylcellulose, polypropylpyrrolidone, polyvinylpyrrolidone, gelatin, gum arabic, polyethyleneglycol, sucrose or starch), a disintegrator (e.g, starch, carboxymethylcellulose, hydroxypropylstarch, low substituted hydroxypropylcellulose, sodium bicarbonate, calcium phosphate or calcium citrate), a lubricant (e.g., magnesium stearate, light anhydrous silicic acid, talc or sodium laury l sulfate), a flavoring agent (e.g., citric acid, menthol, glycine or orange powder), a preservative (e.g, sodium benzoate, sodium bisulfite, methylparaben or propylparaben), a stabilizer (e.g., citric acid, sodium citrate or acetic acid), a suspending agent (e.g., methylcellulose, polyvinyl pyrrolidone or aluminum stearate), a dispersing agent (e.g., hydroxypropylmethylcellulose), a diluent (e.g., water), and base wax (e.g., cocoa butter, white petrolatum or polyethylene glycol). The effective amount of the compounds of the present disclosure in the pharmaceutical composition may be at a level that will exercise the desired effect; for example, about 0.005 mg / kg of a subject’s body weight to about 10 mg / kg of a subject’s body weight in unit dosage for both oral and parenteral administration.

[0079] The dose of a compound of the present disclosure to be administered to a subject is rather widely variable and can be subject to the judgment of a health-care practitioner. In general, the compounds disclosed herein can be administered one to four times a day in a dose of about 0.001 mg / kg of a subject’s body weight to about 10 mg / kg of a subject’s body weight, but the above dosage may be properly varied depending on the age, body weight and medical condition of the subject and the type of administration. In one embodiment, the dose is about 0.001 mg / kg of a subject’s body weight to about 5 mg / kg of a subject’s body weight, about 0.01 mg / kg of a subject’s body weight to about 5 mg / kg of a subject’s body weight, about 0.05 mg / kg of a subject’s body weight to about 1 mg / kg of a subject’s body weight, about 0. 1 mg / kg of a subject’s body weight to about 0.75 mg / kg of a subject’s body weight or about 0.25 mg / kg of a subject’s body weight to about 0.5 mg / kg of a subject’s body weight. In one embodiment, one dose is given per day. In any given case, the amount of the compound of the present disclosure administered will depend on such factors as the solubility of the active component, the formulation used, and the route of administration.

[0080] In some embodiments, a compound of the present disclosure is administered to a subject at a dose of about 0.01 mg / day to about 750 mg / day, about 0. 1 mg / day to about 375 mg / day. about 0. 1 mg / day to about 150 mg / day, about 0. 1 mg / day to about 75 mg / day, about 0. 1 mg / day to about 50 mg / day, about 0.1 mg / day to about 25 mg / day, or about 0.1 mg / day to about 10 mg / day.

[0081] In another embodiment, provided herein are unit dosage formulations that comprise between about 0. 1 mg and 500 mg, about 1 mg and 250 mg, about 1 mg and about 100 mg. about 1 mg and about 50 mg, about 1 mg and about 25 mg, or between about 1 mg and about 10 mg of a compound of the present disclosure.

[0082] In a particular embodiment, provided herein are unit dosage formulations comprising about 0. 1 mg or 100 mg of a compound of the present disclosure.

[0083] In another embodiment, provided herein are unit dosage formulations that comprise 0.5 mg, 1 mg, 5 mg, 10 mg, 15 mg, 20 mg, 30 mg, 35 mg, 50 mg, 70 mg. 100 mg, 125 mg, 140 mg, 175 mg, 200 mg. 250 mg, 280 mg. 350 mg, 500 mg, 560 mg. 700 mg, 750 mg. 1000 mg. or 1400 mg of a compound of the present disclosure.

[0084] A compound of the present disclosure can be administered once, twice, three, four or more times daily. As a nonlimiting example, doses of 100 mg or less are administered as a once daily dose and doses of more than 100 mg are administered twice daily in an amount equal to one half of the total daily dose.

[0085] A compound of the present disclosure can be administered orally for reasons of convenience. In one embodiment, when administered orally, a compound of the present disclosure is administered with a meal and water. In another embodiment, the compound of the present disclosure is dispersed in water or juice (e.g, apple juice or orange juice) or any other liquid and administered orally as a solution or a suspension.

[0086] The compounds disclosed herein can also be administered intradermally, intramuscularly, intraperitoneally, percutaneously, intravenously, subcutaneously, intranasally, epidurally, sublingually, intracerebrally, intravaginally, transdermally, rectally, mucosally. by inhalation, or topically to the ears, nose, eyes, or skin. The mode of administration is left to the discretion of the health-care practitioner, and can depend in-part upon the site of the medical condition.

[0087] In one embodiment, provided herein are capsules containing a compound of the present disclosure without an additional earner, excipient or vehicle.

[0088] In another embodiment, provided herein are compositions comprising an effective amount of a compound of the present disclosure and a pharmaceutically acceptable carrier or vehicle, wherein a pharmaceutically acceptable carrier or vehicle can comprise an excipient, diluent, or a mixture thereof. In one embodiment, the composition is a pharmaceutical composition.

[0089] The compositions can be in the form of tablets, chewable tablets, capsules, solutions, parenteral solutions, troches, suppositories, spray dried dispersions, and suspensions and the like. Compositions can be formulated to contain a daily dose, or a convenient fraction of a daily dose, in a dosage unit, which may be a single tablet or capsule or convenient volume of a liquid. In one embodiment, the solutions are prepared from water-soluble salts, such as the hydrochloride salt. In general, all of the compositions are prepared according to known methods in pharmaceutical chemistry. Capsules can be prepared by mixing a compound of the present disclosure with a suitable carrier or diluent and filling the proper amount of the mixture in capsules. The usual carriers and diluents include, but are not limited to, inert powdered substances such as starch of many different kinds, powdered cellulose, especially crystalline and microcrystalline cellulose, sugars such as fructose, mannitol and sucrose, grain flours and similar edible powders.

[0090] Tablets can be prepared by direct compression, by wet granulation, or by dry granulation. Their formulations usually incorporate diluents, binders, lubricants, and disintegrators as well as the compound. Typical diluents include, for example, various types of starch, lactose, mannitol, kaolin, calcium phosphate or sulfate, inorganic salts such as sodium chloride, and powdered sugar. Powdered cellulose derivatives are also useful. Typical tablet binders are substances such as starch, gelatin and sugars such as lactose, fructose, glucose and the like. Natural and synthetic gums are also convenient, including acacia, alginates, methylcellulose, polyvinylpyrrolidine and the like. Polyethylene glycol, ethylcellulose, and waxes can also serve as binders.

[0091] A lubricant might be necessary in a tablet formulation to prevent the tablet and punches from sticking in the dye. The lubricant can be selected from such slippery solids as talc, magnesium and calcium stearate, stearic acid, and hydrogenated vegetable oils. Tablet disintegrators are substances that swell when wetted to break up the tablet and release the compound. They include starches, clays, celluloses, algins, and gums. More particularly, com and potato starches, methylcellulose, agar, bentonite, wood cellulose, powdered natural sponge, cation-exchange resins, alginic acid, guar gum, citrus pulp, and carboxymethyl cellulose, for example, can be used as well as sodium lauryl sulfate. Tablets can be coated with sugar as a flavor and sealant, or with film-forming protecting agents to modify the dissolution properties of the tablet. The compositions can also be formulated as chewable tablets, for example, by using substances such as mannitol in the formulation.

[0092] When it is desired to administer a compound of the present disclosure as a suppository, ty pical bases can be used. Cocoa butter is a traditional suppository base, which can be modified by addition of waxes to raise its melting point slightly. Water-miscible suppository' bases comprising, particularly, polyethylene glycols of various molecular weights are in wide use.

[0093] The effect of the compound of the present disclosure can be delayed or prolonged by proper formulation. For example, a slowly soluble pellet of the compound of the present disclosure can be prepared and incorporated in a tablet or capsule, or as a slow-release implantable device. The technique also includes making pellets of several different dissolution rates and filling capsules with a mixture of the pellets. Tablets or capsules can be coated with a film that resists dissolution for a predictable period of time. Even the parenteral preparations can be made long-acting, by dissolving or suspending the compound of the present disclosure in oily or emulsified vehicles that allow it to disperse slowly in the serum.

[0094] EXAMPLES

[0095] The following Examples are presented by way of illustration, not limitation. One skilled in the art can modify the procedures set forth in the illustrative examples to arrive at the desired products.

[0096] Salts of the compounds described herein can be prepared by standard methods, such as inclusion of an acid (for example TFA, formic acid, or HC1) in the mobile phases during chromatography purification, or stirring of the products after chromatography purification, with a solution of an acid (for example, aqueous HC1). All reactions are at room temperature unless noted otherwise.

[0097] The following abbreviations may be relevant for the application.

[0098] Abbreviations

[0099] Synthetic Examples

[0100] As will be apparent to one skilled in the art, the compounds disclosed below and in Table 1 can exist in various stereochemical forms. Unless otherwise stated, where stereochemistry is shown for the ether linkage between the pyrrolidine and isoindolinone as well as stereochemistry shown between the piperidine-dione to the isoindolinone in the compounds in the Examples and Table 1, the stereochemistry is absolute. Other stereocenters in which stereochemistry is shown may be relative stereochemistry.

[0101] Intermediate A: 3-(l-oxo-5-(((S)-pyrrolidin-3-yl)oxy)isoindolin-2-yl)piperidine-2,6- dione hydrogen chloride

[0102] 2 tert-butyl (7?)-3-((methyIsulfonyl)oxy)pyrrolidine-l -carboxylate. To a solution of tert-butyl (J?)-3-hydroxypyrrolidine-l -carboxylate (75 g, 401 mmol) in PhMe (1000 mL) was added EtaN (65.7 g, 649 mmol). The mixture was stirred and cooled to 3 °C. Methanesulfonyl chloride (66.0 g, 576 mmol) was slowly added with continuous ice bath cooling. The resulting viscous mixture was stirred and gradually warmed to rt over 3 h. The mixture was filtered, and the filtrate was washed with 5% aqueous NaHCCh (2 x 1000 mL), using 15 min of stirring and 15 min of settling time for each wash. The resulting organic layer was dried over anhydrous Na2SO4, filtered, and concentrated to give crude tert-butyl (7 )-3- ((methylsulfonyl)oxy)pyrrolidine-l -carboxylate (106 g) as an orange oil. which was used in the next step without further purification. Step A2:

[0103] 4 tert-butyl (5)-3-(4-bromo-3-formylphenoxy)pyrrolidine-l-carboxylate. To a solution of tert-butyl (A)-3-((methylsulfonyl)oxy)pyrrolidine- 1 -carboxylate (106 g, 398 mmol) in DMF (1500 mL) was added 2-bromo-5-hydroxy -benzaldehyde (65 g, 323 mmol) and K2CO3 (67.8 g, 491 mmol). The reaction mixture was stirred at 90 °C for 1 h. The reaction mixture was diluted with water (1000 mL) and extracted with EtOAc (1000 mL). The organic layer was washed with brine (1000 mL), dried over anhydrous Na2SO4, filtered, and concentrated by rotary evaporation. The residue was purified by silica gel chromatography to give tert-butyl ( )- 3-(4-bromo-3-formylphenoxy)pyrrolidine-l-carboxylate. (109 g, 91% yield) as an off-white solid. 400 MHz, DMSO-cL 5: 10.16 (s, 1H), 7.70 (d, J= 8.8 Hz, 1H), 7.34 (d, J= 2.8 Hz, 1H), 7.27-7.24 (m, 1H), 5.11 (s. 1H), 3.57-3.39 (m. 4H), 2.25-2.05 (m. 2H), 1.41 (t, J= 4.0 Hz, 1H). tert-butyl (3A)-3-(4-bromo-3-(((2,6-dioxopiperidin-3-yl)amino)methyl) phenoxy)pyrrolidine-l-carboxylate. To a stirred solution of 3-aminopiperidine-2, 6-dione hydrochloride (53.3 g, 324 mmol) in MeOH (1000 mL) was added NH3 in MeOH until the pH = 7. The reaction was stirred at rt, and AcOH was added dropwise until the pH = 6. tert-butyl (5)- 3-(4-bromo-3-formylphenoxy)pyrrolidine-l -carboxylate (109 g, 294mmol) in MeOH (80 mL) was added, and the mixture was stirred at rt for 10 min. Sodium cyanoborohydride (56.5 g, 883 mmol) was then added at 0 °C in portions. The reaction mixture was stirred at rt for 16 h. The reaction mixture was concentrated by rotary evaporation. The residue was diluted with water (1000 mL) and extracted with EtOAc (1000 mL). The organic layer was washed with brine (600 mL), dried over anhydrous Na2SO4, filtered, and concentrated. The residue w as purified by silica gel chromatography (petroleum ether / EtOAc, 5 : 1 to 0: 1) to give impure tert-butyl (3S)-3- (4-bromo-3-(((2,6-dioxopiperidin-3-yl)amino)methyl)phenoxy)pyrrolidine-l -carboxylate (120 g) as a light yellow- oil, which was used into next step without any further purification. Step A4:

[0104] 5 tert-butyl (3S)-3-((2-(2,6-dioxopiperidin-3-yl)-l-oxoisoindolin-5-yl)oxy)pyrrolidine- 1-carboxylate. The following procedure was performed in 3 batches that were combined for purification. To a solution of tert-butyl (3S)-3-(4-bromo-3-(((2,6-dioxopiperidin-3- yl)amino)methyl)phenoxy)pyrrolidine-l -carboxylate (40 g, 82.9 mmol) and dccp*HBF4 (7.59 g, 12.4 mmol) in DMF (500 mL) was added Pd(OAc)2 (2.79 g, 12.4 mmol) and K2CO3 (17.4 g, 126 mmol) under nitrogen. The suspension was degassed under vacuum and purged with carbon monoxide gas several times. The mixture was stirred under carbon monoxide (50 psi) at 80 °C for 48 hrs. The reaction mixture was cooled to rt, filtered, and concentrated by rotary evaporation. The residue was purified by preparative reverse phase HPLC to give tert-butyl (35)-3-((2-(2,6-dioxopiperidin-3-yl)-l-oxoisoindolin-5-yl)oxy)pyrrohdine-l -carboxylate (48 g, yield 39%, 2 steps) as an off-white solid. 400 MHz, DMSO-r : 10.97 (s, 1H), 7.64 (d, J = 8.4 Hz, 1H), 7.19 (s, 1H), 7.06 (d, J= 8.8 Hz, 1H), 5.12-5.06 (m, 2H), 4.42-4.26 (m, 2H), 3.59-3.57 (m, 1H), 3.47-3.42 (m, 3H), 2.91-2.85 (m, 1H), 2.62-2.57 (s, 1H), 2.40-2.38 (m, 1H), 2.08-1.98 (m. 3H), 1.40 (d, J= 5.2 Hz, 9H). MS(ESI) m / z 374.0 [M-56]+.

[0105] Step A5:

[0106] 6

[0107] 3-(l-oxo-5-(((A)-pyrrolidin-3-yl)oxy)isoindolin-2-yl)piperidine- 2, 6-dione hydrogen chloride. To a solution of tert-butyl (3S)-3-((2-(2,6-dioxopiperidin-3-yl)-l-oxoisoindolin-5- yl)oxy)pyrrolidine-l -carboxylate (51 g, 119 mmol) in 1,4-di oxane (460 mL) was added HCI in dioxane(230 mL, 4M). The reaction mixture was stirred at 20 °C for 16 h. The reaction mixture was concentrated by rotary evaporation, diluted wi th MTBE (500 mL), and stirred at 20 °C for 30 mins. The mixture was filtered and the solid was dried under vacuum at 50 °C for 16 h to give 3-(l-oxo-5-(((S)-pyrrolidin-3-yl)oxy)isoindolin-2-yl)piperidine-2, 6-dione hydrogen chloride (44 g. 120 mmol. 100%) as an off-white solid. 400 MHz, DMSO-rfc 6: 10.98 (s, 1H), 9.74 (s. 1H), 9.54 (s. 1H), 7.66 (d. J = 8.4 Hz. 1H), 7.23 (s. 1H), 7.09 (d, J = 8.4 Hz, 1H), 5.26 (s, 1H). 5.11-5.06 (m, 1H), 4.44-4.27 (m, 2H). 3.57-3.50 (m, 1H), 3.39-3.34 (m, 3H). 2.91-2.85 (m. 1H), 2.62-2.57 (s, 1H), 2.41-2.38 (m, 1H), 2.03-1.97 (m, 2H), 1.99-1.97 (m, 1H). MS(ESI) m / z 330.0 [M+l]+.

[0108] Example 1

[0109] 3-(5-(((S)- l-(furo [3,2-b] pyridin-2-ylmethyl)pyrrolidin-3-yl)oxy)- l-oxoisoindolin-2- yl)piperidine-2, 6-dione

[0110] A. 3-(5-(((5)-l-(furo[3,2-Z>]pyridin-2-ylmethyl)pyrrolidin-3-yl)oxy)-l- oxoisoindolin-2-yl)piperidine-2, 6-dione A vial containing furo[3,2-b]pyridine-2-carbaldehyde (44.7 mg, 0.304 mmol), 3-(l-oxo-5-(((S)-pyrrolidin-3-yl)oxy)isoindolin-2-yl)piperidine-2,6- dione (50 mg, 0. 152 mmol) was charged with MeCN (1 mL). The mixture was stirred, and NaBH(OAc)3 (129 mg, 0.607 mmol) was added in a single portion. Stirring was continued overnight. The reaction mixture was diluted with MeOH, filtered, and purified by reverse-phase HPLC (5% to 95% MeCN in w ater. 10 mM ammonium acetate modifier). The obtained material was redissolved in 1:1 water / MeCN. frozen, and lyophilized. The material was redissolved in 1: 1 water / MeCN, frozen, and lyophilized again to afford 3-(5-(((<S’)-l-(furo[3,2- >]pyridin-2- ylmethyl)pyrrolidin-3-yl)oxy)-l-oxoisoindolin-2-yl)piperidine-2, 6-dione (38.8 mg, 0.084 mmol, 55.4 % yield). 'H NMR (400 MHz. DMSO-d6) 5 = 10.96 (s, 1H), 8.51 (br s, 1H), 8.01 (br d, J= 4.9 Hz, 1H), 7.62 (br d, J= 8.1 Hz, 1H), 7.39 - 7.30 (m, 1H), 7.15 - 6.97 (m, 3H), 5.07 (br dd, J = 5.1, 13.3 Hz, 2H), 4.41 - 4.35 (m, 1H), 4.28 - 4.23 (m, 1H), 2.94 - 2.85 (m, 2H), 2.68 - 2.55 (m. 5H), 2.42 - 2.30 (m, 3H), 2.02 - 1.93 (m, 2H): MS (ESI) m / z 461.2 [M+H]+.

[0111] Example 10

[0112] 3-(5-(((A)-l-((l-methyl-5-(3-(tetrahydro-2 / / -pyran-4-yl)phenyl)-lH-imidazol-2- yl)methyl)pyrrolidin-3-yl)oxy)-l-oxoisoindolin-2-yl)piperidine-2, 6-dione A. Ethyl l-methyl-5-(3-(tetrahydro-2 / / -pyran-4-yl)phenyl)-l / / -imidazole-2-ca- rboxylate. A mixture of ethyl 5-bromo-l -methyl- l / 7-imidazole-2-carboxylate (600 mg, 2.57 mmol), (3-(tetrahydro-277-pyran-4-yl)phenyl)boronic acid (586 mg, 2.84 mmol), Pd(dppf)Ch-DCM (211 mg, 0.26 mmol) and Na2CCh (824 mg, 7.77 mmol) in 1,4-dioxane (10 ml) and water (1 mL) was stirred overnight at 100 °C under nitrogen. The mixture was fdtered, and the filtrate was concentrated. The crude product was purified by reversed phase flash column chromatography (0-100% MeCN + 0.05% ammonium bicarbonate in water, over 20 min) to afford the title compound (400 mg, 1.27 mmol, 49% yield) as a yellow solid. MS (ESI) m'z 315.1 [M+l]+.

[0113] B. (l-Methyl-5-(3-(tetrahydro-2 / 7-pyran-4-yl)phenyl)-l / / -iinidazol-2-yl)meth- anol. To a solution of ethyl l-methyl-5-(3-(tetrahydro-277-pyran-4-yl)phenyl)-177- imidazole-2-carboxylate (200 mg, 0.63 mmol) in THF (3 mL) was added LAH (2.5 M in THF, 0.5 mL, 1.25 mmol) at 0 °C under nitrogen, and the reaction solution was stirred at this temperature for 0.5 h. The resulting solution was quenched with EtOH and concentrated. The crude product was purified by reversed phase flash (0-100% MeCN + 0.05% ammonium bicarbonate in water, over 20 min) to afford the title compound (110 mg, 0.40 mmol, 63% yield) as a yellow solid. MS (ESI) mfz 273.2 [M+l]+.

[0114] C. l-Metliyl-5-(3-(tetrahydr()-2 / / -pyraii-4-yl)phenyl)-l / / -iniidazole-2-carbald- ehyde. To a solution of (1 -methyl-5-(3-(tetrahydro-277-pyran-4-yl)phenyl)-l 77-imi- dazol-2-yl)methanol (100 mg, 0.37 mmol) in DCM (2 mL) was added Dess-Martin periodinane (234 mg, 0.55 mmol) in several portions at 0 °C under nitrogen, and the reaction mixture was stirred overnight at room temperature. The resulting mixture was filtered, and the filtrate was concentrated. The crude product was purified by flash column chromatography (0 to 50% EtOAc in petroleum ether) to afford the title compound (75 mg, 0.28 mmol, 76% yield) as a yellow solid. MS (ESI) m / z 271. 1 [M+l]+.

[0115] D. 3-(5-((G$)-l-(( l-nietliyl-5-(3-(tetraliy(lro-2 / / -pyran-4-yl)plieiiyl)-l / / -iniidaz- ol-2-yI)methyl)pyrrolidin-3-yl)oxy)-l-oxoisoindolin-2-yI)piperidine-2, 6-dione. A mixture of

[0116] 1-methyl-5-(3-(tetrahydro-2H-pyran-4-yl)phenyl)-lE7-imidazole-2- carbaldehyde (50 mg, 0.18 mmol), 3-(l-oxo-5-(((S)-pyrrolidin-3-yl)oxy)isoindohn-

[0117] 2-yl)piperidine-2,6-dione;TFA (80 mg, 0. 18 mmol) and EtsN (0.05 mL, 0.38 mmol) in DCM (5 mL) pre-stirred for 15 min was added NaBH(OAc)? (157 mg, 0.74 mmol), and the reaction mixture was stirred overnight at room temperature under nitrogen. The resulting mixture was concentrated. The residue was purified by flash column chromatography (0-5% MeOH in DCM) and further purified by preparative HPLC with the following conditions Sunfire preparative Cl 8 column, 30*150 mm, 5 pm; Mobile Phase A: water (0.1%FA), Mobile Phase B: acetonitrile; Flow rate; 60 mL / min; Gradient: 5% B to 35% B in 7 min; Wave Length: 254 / 210 nm; RT1 (min): 5.92. Pure fractions were evaporated to afford the title compound (39.9 mg. 0.068 mmol. 38% yield) as a white solid. 'H NMR (300 MHz, DMSO-ds) 5 10.96 (s, 1H). 7.62 (m, 1H), 7.41 (m, 1H), 7.34-7.23 (m, 3H), 7.13 (d, J= 2.2 Hz, 1H), 7.02 (dd, J= 8.5, 2.2 Hz, 1H), 6.93 (d, J= 0.9 Hz, 1H), 5.14-4.96 (m, 2H), 4.40 (d, J= 17.3 Hz, 1H), 4.26 (d, J= 16.8 Hz, 1H), 4.01-3.92 (m, 2H), 3.76 (s, 2H), 3.66 (s, 3H), 3.50-3.41 (m, 2H), 3.04-2.88 (m, 2H), 2.80 (m. 3H), 2.59 (m, 2H). 2.36 (m, 2H), 2.05-1.92 (m, 1H), 1.85 (m. 1H), 1.78-1.66 (m. 4H); MS (ESI) m / z 584.4 [M+l]+.

[0118] Example 14 3-(l-Oxo-5-(((5)-l-((5-(4-(tetrahydro-2H-pyran-4-yl)phenyl)-l,3,4-oxadiazol-2- yl)methyl)pyrrolidin-3-yl)oxy)isoindolin-2-yl)piperidine-2, 6-dione

[0119] A. Methyl 4-(3,6-dihydro-2 / / -pyran-4-yl)benzoate. To a stirred mixture of methyl 4- iodobenzoate (1.5 g, 5.72 mmol), 2-(3,6-dihydro-27f-pyran-4-yl)-4.4.5.5- tetramethyl-L3,2-dioxaborolane (1.8 g, 8.59 mmol) and sodium carbonate (1.8 g, 16.98 mmol) in 1,4-di oxane (20 rnL) and water (4 mL) was added l,l’-bis(diphenylphosphino)ferrocene- palladium(II)dichloride dichloromethane complex (0.84 g, 1.14 mmol) and the above mixture was stirred for 2 h at 90 °C under nitrogen. The resulting mixture was filtered and the filtrate was concentrated. The residue was purified by flash column chromatography (0 to 30% EtOAc in petroleum ether) to afford the title compound (1.16 g, 5.32 mmol, 93% yield) as a yellow solid.

[0120] B. Methyl 4-(tetrahydro-2 / / -pyran-4-yl)benzoate. To a stirred solution of methyl 4- (3,6-dihydro-2f / -pyran-4-yl)benzoate (1.16 g, 5.32 mmol) in MeOH (15 mL) was added 10% palladium on carbon (0.75 g. wetted with ca. 50% water) in portions at room temperature under nitrogen. The resulting mixture was stirred for 24 h at 30 °C under hydrogen (~ 2.5 bar). The resulting mixture was filtered, and the filter cake was washed with MeOH. The filtrate was concentrated under reduced pressure to afford the title compound (1.1 g, 4.98 mmol, 93% yield) as a white solid. MS (ESI) m / z 221.1 [M+l]+.

[0121] C. 4-(Tetrahydro-2H-pyran-4-yl)benzohydrazide. A solution of methyl 4- (tetrahydro-27 / -pyran-4-y I (benzoate (600 mg, 2.72 mmol) in MeOH (7 mL) was added hydrazine hydrate (7 mL) and the above solution was stirred for 2 h at 80 °C under nitrogen. The solution was concentrated and the residue was purified by reverse phase Cl 8 chromatography (20-60% acetonitrile + 0.05% ammonium bicarbonate in water, over 25 min) to afford the title compound (400 mg, 1.82 mmol, 67% yield) as a white solid. MS (ESI) m / z 221.1 [M+l]+.

[0122] D. Ethyl 5-(4-(tetrahydro-2 / / -pyran-4-yl)phenyl)-l .3.4-oxadiazole-2-carboxy- late. To a stirred mixture of 4-(tetrahydro-2 / 7-pyran-4-yl)benzohydrazide (400 mg, 1.82 mmol) and EtsN (0.76 mL, 5.45 mmol) in DCM (5 mL) was added ethyl 2-chloro-2-oxoacetate (248 mg, 1.82 mmol) at 0 °C and the above mixture was stirred for 2 h at room temperature under nitrogen. The resulting mixture was added EtsN (0.76 mL, 5.45 mmol) and 4-methylbenzene-I- sulfonyl chloride (346 mg. 1.82 mmol) at 0 °C. The resulting mixture was stirred for extra 2 h at room temperature under nitrogen. The resulting mixture was concentrated and the residue was purified by silica gel chromatography (0 to 50% EtOAc in petroleum ether) to afford the title compound (240 mg, 0.79 mmol, 43% yield) as a white solid. MS (ESI) m / z 303. 1 [M+l]+.

[0123] E. (5-(4-(Teti ahydro-2 / / -pyran-4-yl)plienyl)- 1.3.4-oxadiazol-2-y I (methanol. To a stirred mixture of ethyl 5-(4-(tetrahydro-2 -pyran-4-yl)phenyl)-l,3,4-oxadiazol-e-2-carboxylate (230 mg, 0.76 mmol) and calcium chloride (168 mg, 1.52 mmol) in ethanol (5 mL) was added sodium borohydride (57 mg. 1.52 mmol) in portions at 0 °C. The mixture was stirred for 2 h at room temperature. The resulting mixture was concentrated and the residue was purified by silica gel chromatography (0 to 10% MeOH in DCM) to afford the title compound (100 mg, 0.38 mmol, 51% yield) as a white solid. MS (ESI) m / z 261.1 [M+l]+. F. 2-(Cliloromethyl)-5-(4-(tetrahydro-2 / / -pyran-4-yl)phenyl)-l,3,4-oxadiazole. To a stirred mixture of (5-(4-(tetrahydro-2 / f-pyran-4-yl)phenyl)-L3.4-oxadiazol-2-yl)methanol (60 mg, 0.23 mmol) in DCM was added thionyl chloride (57 mg, 0.69 mmol) and DMF (0.02 mL) and the above mixture was stirred for 2 h at room temperature. The resulting mixture was concentrated under vacuum. The residue was basified to pH 8 with saturated sodium bicarbonate and extracted with EtOAc. The combined extracts were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to afford the title compound (60 mg, 0.22 mmol, 96% yield) as a yellow oil. MS (ESI) mJz 279.1 [M+l]+.

[0124] G. 3-(l-Oxo-5-(((5)-l-((5-(4-(tetrahydro-2Ef-pyran-4-yl)phenyl)-l,3,4-oxadiazo- l-2-yl)methyl)pyrrolidin-3-yl)oxy)isoindolin-2-yl)piperidine-2, 6-dione. To a stirred mixture of 2-(chloromelhyl)-5-(4-(letrahydro-2H-pyran-4-yl)phenyl)-1.3.4-oxadi azole (60 mg, 0.22 mmol) in DMF (4 mL) were added 3-(l-oxo-5-(((S)-pyrrolidin-3-yl)oxy)isoindolin-2- yl)piperidine-2,6-dione;2,2,2-trifluoroacetic acid (95 mg, 0.21 mmol) and potassium carbonate (90 mg, 0.65 mmol) at room temperature. The resulting mixture was stirred for 2 h at 30 °C under nitrogen. The resulting mixture was filtered and the filtrate was purified by preparative reverse phase HPLC with the following conditions: Column: Sunfire prep C18 column, 30x150 mm, 5 pm; Mobile Phase A: Water (0.1% formic acid), Mobile Phase B: acetonitrile; Flow rate: 60 mL / min; Gradient: 5% B to 25% B in 10 min, 25% B; Wave Length: 254 / 210 nm; RT1 (min): 11.28. The fractions containing the desired product were collected and evaporated under reduced pressure to afford the title compound (20.8 mg, 0.036 mmol, 16% yield) as a white solid. 'H NMR (400 MHz, Methanol-d-i) 5 8.01-7.97 (m, 2H), 7.71-7.68 (m, 1H), 7.49 (m, 2H), 7.10 (s, 1H), 7.05-7.02 (m, 1H), 5.15-5.10 (m. 2H), 4.50-4.39 (m, 2H), 4.27 (s, 2H), 4.09-4.07 (m. 2H), 3.62-3.57 (m. 2H), 3.25-3.19 (m. 3H), 3.01-2.88 (m. 3H), 2.82-2.77 (m. 1H), 2.54-2.43 (m, 2H), 2.20-2.05 (m, 2H), 1.90-1.81 (m, 4H); MS (ESI) m / z 572.2 [M+l]+.

[0125] Example 16

[0126] 3-(l-Oxo-5-(((5)-l-((2-(4-(tetrahydro-2 / f-pyran-4-yl)phenyl)oxazol-5-yl)methyl)- pyrrolidin-3-yl)oxy)isoindolin-2-yl)piperidine-2, 6-dione A. Ethyl 2-(4-(tetrahydro-2 / / -pyran-4-yl)phenyl)oxazole-5-carboxylate. To a stirred mixture of ethyl 2-bromooxazole-5 -carboxylate (300 mg, 1.36 mmol), (4-(tetrahydro-277-pyran- 4-yl)phenyl)boronic acid (421 mg, 2.04 mmol) and sodium carbonate (433 mg, 4.09 mmol) in 1,4-dioxane (5 mL) and water (1 mL) was added l,l ’-bis(diphenylphosphino)ferrocene- palladium(II)di chloride di chloromethane complex (222 mg, 0.27 mmol) and the above mixture was stirred for 2 h at 100 °C under nitrogen. The resulting mixture was diluted with water and extracted with EtOAc. The extracts combined was washed with brine, dried over anhydrous sodium sulfate. After filtrate, the filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography (0 to 60% EtOAc in petroleum ether) to afford the title compound (338 mg, 1.12 mmol, 82% yield) as an off-white solid. MS (ESI) m / z 302.2 [M+l]+.

[0127] B. (2-(4-(Tetrahydro-2 / / -pyran-4-yl)phenyl)oxazol-5-yl)methanol. To a stirred solution of ethyl 2-(4-(tetrahydro-27f-pyran-4-yl)phenyl)oxazole-5-carboxylate (320 mg. 1.06 mmol) in THF (5 mL) was added lithium aluminum hydride (2.5 M in THF, 0.45 mL, 1.13 mmol) dropwise at 0 °C and the above mixture was stirred for 1 h at 0 °C under nitrogen. The reaction was quenched by the addition of sodium hydroxide (2 M in water). The resulting mixture was filtered and the filter cake was washed with THF. The filtrate was concentrated under reduced pressure to afford the title compound (270 mg, 1.04 mmol, 98% yield) as a white solid. MS (ESI) mz 260.2 [M+l] ' .

[0128] C. 2-(4-(Tetrahydro-2 / / -pyran-4-yl)phenyl)oxazole-5-carbaldehyde. To a stirred mixture of (2-(4-(tetrahydro-27 / -pyran-4-yl)phenyl)oxazol-5-yl)methanol (150 mg, 0.58 mmol) in DCM (5 mL) was added Dess-Martin periodinane (300 mg, 0.71 mmol) 0 °C and the above mixture was stirred for 1 h at room temperature under nitrogen. The resulting mixture was concentrated and the residue was purified by flash column chromatography (0 to 50% EtOAc in petroleum ether) to afford the title compound (95 mg, 0.37 mmol, 64% yield) as a white solid. MS (ESI) m / z 256.1 [M-l]’.

[0129] D. 3-(l-Oxo-5-(((A)-l-((2-(4-(tetrahydro-2H-pyran-4-yl)phenyI)oxazoI-5-yl)methyl) pyrrolidin-3-yl)oxy)isoindolin-2-yl)piperidine-2, 6-dione. To a stirred mixture of 2-(4- (tetrahydro-27 / -pyran-4-yl)phenyl )oxazole-5-carbaldehyde (90 mg, 0.35 mmol), 3-(l-oxo-5- (((S)-pyrrolidin-3-yl)oxy)isoindolin-2-yl)piperidine-2,6-dione;hydrochloride (120 mg, 0.33 mmol) and EtsN (0.15 mL, 1.07 mmol) in DCM (5 mL) was added NaBH(OAc)? (160 mg, 0.75 mmol) in portions and the above mixture was stirred for 1 h at 30 °C under nitrogen. The resulting mixture was concentrated. The residue was firstly purified by flash column chromatography (0 to 50% EtOAc in petroleum ether) and purified further by preparative HPLC with following conditions: Column: Sunfire prep C18 column, 30x 150 mm, 5 um; Mobile Phase A: Water (0.1% formic acid), Mobile Phase B: acetonitrile; Flow rate:60 mL / min; Gradients B to 35 B in 7 min; 254 / 220 nm; RT1 :5.35; RT2. The fractions containing the desired product were collected and evaporated under reduced pressure to afford the title compound (30.8 mg, 0.054 mmol, 16% yield) as a white solid.XH NMR (400 MHz, Methanol-d4) 5 7.97-7.93 (m, 2H), 7.71-7.68 (m, 1H), 7.42 (d, J=8.20 Hz, 2H), 7.24 (s, 1H), 7.09-7.02 (m, 2H), 5.16-5.10 (m, 2H), 4.45-4.39 (m, 2H), 4.09-4.07 (m, 4H), 3.63-3.50 (m, 2H), 3.27-3.12 (m, 3H), 2.95-2.77 (m, 4H), 2.54-2.43 (m. 2H), 2.19-2.09 (m. 2H), 1.89-1.80 (m. 4H); MS (ESI) m / z 571.1 [M+l]+.

[0130] Example 36 3-(l-Oxo-5-(((A)-l-((5-phenylisoxazol-3-yl)methyl)pyi olidiii-3-yl)oxy)isoindolin-2- yl)piperidine-2, 6-dione

[0131] A. To a stirred solution of 5-phenylisoxazole-3-carbaldehyde (57 mg, 0.33 mmol, 1.22 equiv.) in DCM (10 mL) were added 3-(l-oxo-5-(((S)-pyrrolidin-3-yl)oxy) isoindolm-2- yl)piperidine-2,6-dione; hydrochloride (100 mg, 0.27 mmol, 1.00 equiv.), EtsN (83 mg, 0.82 mmol, 3.04 equiv.) and NaBH(OAc)s (232 mg, 1.09 mmol, 4.05 equiv.) sequentially at room temperature and the above mixture was stirred at room temperature for 2 h under nitrogen. The resulting mixture was purified firstly by normal flash (0-5% MeOH in DCM) and further purified by Prep-HPLC. Pure fractions were evaporated to afford the title compound (26 mg, 0.05 mmol, 20% yield). 'H NMR (400 MHz, Methanol-d4) 5 7.89-7.82 (m, 2H), 7.73 (d, J= 8.4 Hz, 1H), 7.58-7.46 (m. 3H), 7.13-7.03 (m, 2H), 6.87 (s, 1H), 5.13 (m, 2H), 4.52-4.37 (m, 2H), 3.96 (s, 2H), 3.17 (m, 1H). 3.11 (m, 2H), 2.92 (m. 1H), 2.88-2.75 (m. 2H), 2.48 (m, 2H). 2.21- 2.08 (m, 2H). ESI (MS) m / z 487.1 [M+l]+.

[0132] Example 56

[0133] 3-(l-Oxo-5-(((A)-l-((2-phenylthiazol-5-yl)methyl)pyrrolidin-3-yl)oxy)isoindolin- 2-yl)piperidine-2, 6-dione

[0134] A. To a stirred solution of 2-phenylthiazole-5-carbaldehyde (63 mg, 0.33 mmol, 1.22 equiv.), 3-(l-oxo-5-(((S)-pyrrolidin-3-yl)oxy)isoindolin-2-yl)piperidine-2, 6-dione; hydrochloride (100 mg, 0.27 mmol, 1.00 equiv.) and Et3N (83 mg, 0.82 mmol. 3.04 equiv.) in DCM (10 mL) was added NaBH(OAc)3 (252 mg. 1.19 mmol, 4.40 equiv.) in several portions at room temperature and the above mixture was stirred at this temperature for 2 h under nitrogen. The resulting solution was concentrated. The residue was purified firstly by normal flash (0-5% MeOH in DCM) and further purified by Prep-HPLC. Pure fractions were evaporated to afford the title compound (15.3 mg, 0.030 mmol. 11% yield). 1H NMR (400 MHz. DMSO-d6) 5 10.98 (s, 1H), 8.15 (s, 1H), 7.93 (dd, J = 7.6, 1.9 Hz, 2H), 7.78 (s, 1H), 7.62 (d, J = 8.3 Hz, 1H), 7.55- 7.47 (m, 3H), 7.13 (s, 1H), 7.02 (dd, J = 8.4, 2.1 Hz, 1H), 5.08 (m, 2H), 4.43-4.26 (m, 2H), 3.93 (s, 2H), 3.01-2.62 (m, 4H), 2.42-2.30 (m, 4H), 2.02-1.86 (m, 2H). MS (ESI) m / z 503.1 [M+l]+.

[0135] Example 61

[0136] 3-(5-(((5)-l-((l-Methyl-3-phenyl-l / 7-pyrazol-5-yl)methyl)pyrrolidin-3-yl)oxy)-l- oxoisoindolin-2-yl)piperidine-2, 6-dione

[0137] A. To a solution of 3-(l-oxo-5-(((<S)-pyrrolidin-3-yl)oxy)isoindolin-2-yl)piperidine-2,6- dione; hydrochloride (100 mg, 0.27 mmol, 1.00 equiv.), l-methyl-3 -phenyl- 177-pyraz- ole-5-carbaldehyde (61 mg, 0.33 mmol 1.21 equiv) and EtsN (33 mg, 0.33 mmol, 1.21 equiv.) in DCM (10 mL) was added NaBH(OAc)s (232 mg, 1.09 mmol. 4.05 equiv.) in several portions at room temperature and the above mixture was stirred for 2 h at this temperature under nitrogen. The resulting mixture was concentrated under reduced pressure. The crude was dissolved into DMSO and purified directly by Prep-HPLC. Pure fractions were evaporated to afford the title compound (37.4 mg. 0.075 mmol, 27% yield).rH NMR (400 MHz, Methanol-dr) 5 8.18 (s, 1H), 7.74 (m, 3H). 7.39 (dd. J = 8.3. 6.9 Hz. 2H), 7.35-7.27 (m. 1H). 7. 13-7.03 (m. 2H). 6.66 (d. J= 2.1 Hz, 1H), 5.17-5.08 (m, 2H), 4.43 (s, 2H), 3.96 (d, J= 1.1 Hz, 5H), 3.18 (s, 1H), 3.07 (s, 2H), 2.93-2.81 (s, 3H), 2.47 (m, 2H), 2.19-2.03 (m, 2H). MS(ESI) m / z 500.2 [M+l]+. Example 62

[0138] 3-(4-(((35)-3-((2-(2,6-Dioxopiperidin-3-yl)-l-oxoisoindolin-5-yl)oxy)pyrrolidin-l- yl)methyl)-lH-pyrazol-l-yl)-5-(trifluoromethoxy)benzonitrile

[0139] A. 3-(4-Formyl-l / / -pyrazol-l-yl)-5-(trifluoromethoxy)benzonitrile To a solution of 3-bromo-5-(trifluoromethoxy)benzonitrile (300 mg, 1.13 mmol), 177-pyrazole-4-carbaldehyde (102 mg, 1.06 mmol), rac-(17?,27?)-A1,A2-dimethylcyclohexane-l,2-diamine (64 mg, 0.45 mmol) and K3PO4 (717 mg, 3.38 mmol) in DMF (5 rnL) was added Cui (42 mg, 0.22 mmol), and the mixture was stirred at 100 °C overnight under nitrogen. The resulting mixture was diluted with water and extracted with EtOAc. The extracts were washed with brine, dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (0 to 35% EtOAc in petroleum ether) to afford the title compound (45 mg, 0. 16 mmol, 14% yield) as an off-white solid. MS (ESI) m / z 280.2 [M-l]’.

[0140] B. 3-(4-(((35)-3-((2-(2,6-Dioxopiperidin-3-yl)-l-oxoisoindolin-5-yl)oxy)pyrrolidin-l- yl)mcthyl)-lH-pyrazol-l-yl)-5-(trifluoromethoxy (benzonitrile To a solution of 3-(4-formyl- l / 7-pyrazol-l-yl)-5-(trifluoromethoxy)benzonitrile (45 mg, 0.16 mmol), 3-(l-oxo-5-(((5)- pyrrolidin-3-yl)oxy)isoindolin-2-yl)piperidine-2,6-dione;TFA (90 mg, 0.20 mmol) and Et?N (45 mg, 0.45 mmol) in DCM (6 rnL) was added NaBH(OAc)3 (135 mg, 0.64 mmol) in several portions. The mixture was stirred 12 h at room temperature under nitrogen. The resulting mixture was concentrated. The residue was purified by flash column chromatography (0-10% MeOH in DCM) and further purified by preparative HPLC with the following conditions: Column: Sunfire prep C18 column, 30*150 mm, 5 pm; Mobile Phase A: water (0.1% formic acid), Mobile Phase B: acetonitrile; Flow rate: 60 mL / min; Gradient: 7% B to 37% B in 7 min, 37% B; Wave Length: 254 / 220 nm; RTi (min): 6.8; Number of Runs: 0. Pure fractions were evaporated to afford the title compound (26.7 mg. 0.045 mmol, 28% yield) as a white solid.XH NMR (400 MHz, Methanol-d4) 5 8.53 (d, J = 1.9 Hz, 1H), 8.24-8.21 (m, 1H), 8.13 (s, 1H), 7.90 (s, 1H), 7.76-7.68 (m, 2H), 7.14-7.04 (m, 2H), 5.22-5.09 (m, 2H), 4.52-4.38 (m, 2H), 4.10 (s, 2H), 3.36 (m, 3H), 3.09 (m, 1H), 2.92-2.88 (m, 1H), 2.82-2.74 (m, 1H), 2.58-2.42 (m, 2H), 2.27- 2.12 (m, 2H); MS (ESI) rn / z 595.1 [M+l]+.

[0141] Example 76

[0142] 3-(l-Oxo-5-(((A)-l-((l-phenyl-lH-l,2,3-triazol-4-yl)methyl)pyrrolidin-3-yl)oxy)- isoindolin-2-yl)piperidine-2, 6-dione

[0143] A. To a solution of l-phenyl-lE7-l,2,3-triazole-4-carbaldehyde (50 mg, 0.29 mmol, 1.11 equiv.), 3-(l-oxo-5-(((S)-pyrrolidin-3-yl)oxy)isoindolin-2-yl)piperidine-2, 6-dione; hydrochloride (95 mg, 0.26 mmol. 1.00 equiv.) and EtsN (58 mg. 0.57 mmol, 2.19 equiv.) in DCM (5 mL) was added NaBH(OAc)s (246 mg, 1.16 mmol, 4.46 equiv.) in several portions at room temperature and the above mixture was stirred 2 h at this temperature under nitrogen. The resulting mixture was concentrated under reduced pressure. The residue was purified firstly by normal flash (0-10% MeOH in DCM) and further purified by Prep-HPLC. Pure fractions were evaporated to afford the title compound (37.6 mg, 0.077 mmol, 30% yield). (400 MHz, Methanol-d4) 5 8.54 (s, 1H), 7.89-7.83 (m, 2H), 7.72-7.70 (m, 1H), 7.61-7.56 (m, 2H), 7.55-7.50 (m, 1H), 7.13-7.03 (m, 2H), 5.17-5.08 (m, 2H), 4.44 (d, J= 9.4 Hz, 2H), 4.13 (s, 2H), 3.26 (m, 1H), 3.21 (m, 2H), 2.98-2.77 (m, 3H), 2.53-2.43 (m, 2H), 2.22-2.06 (m, 2H). ESI (MS) m / z 487.1 [M+l]1.

[0144] Example 86 3-(l-Oxo-5-(((A)-l-((5-(4-(tetrahydro-2 / 7-pyran-4-yl)phenyl)oxazol-2-yl)methyl)- pyrrolidin-3-yl)oxy)isoindolin-2-yl)piperidine-2, 6-dione

[0145] A. (5-(4-(Tetrahydro-2 / / -pyran-4-yl)phenyl)oxazol-2-yl)methanol. To a stirred mixture of (5-bromooxazol-2-yl)methanol (200 mg, 1.12 mmol), (4-(tetrahydro-2H-pyran-4- yl)phenyl)boronic acid (231 mg, 1.12 mmol) and sodium carbonate (357 mg, 3.37 mmol) in 1,4- dioxane (5 mL) and water (1 mL) was added l,l ’-bis(diphenylphosphino)ferrocene- palladium(II)di chloride di chloromethane complex (164 mg. 0.22 mmol) and the above mixture was stirred for 2 h at 90 °C under nitrogen. The resulting mixture was fdtered and the filtrate was concentrated. The residue was purified by silica gel chromatography (0 to 10% MeOH in DCM) to afford the title compound (254 mg, 0.98 mmol, 88% yield) as a brown solid. MS (ESI) m / z 260.1 [M+l]+.

[0146] B. 5-(4-(Tetrahydro-2 / 7-pyran-4-yl)phenyl)oxazole-2-carbaldehyde. To a stirred mixture of (5-(4-(tetrahydro-2E7-pyran-4-yl)phenyl)oxazol-2-yl)methanol (250 mg, 0.96 mmol) in DCM (5 mL) was added Dess-Martin periodinane (500 mg, 1.18 mmol) in portions at 0 °C. The above mixture was stirred for 1 h at room temperature under nitrogen. The resulting mixture was concentrated and the residue was purified by silica gel chromatography (0 to 10% MeOH in DCM) to afford the title compound (128 mg, 0.50 mmol. 52% yield) as a white solid. MS (ESI) m / z 256.1 [M-l]’.

[0147] C. 3-(l-Oxo-5-(((5)-l-((5-(4-(tetrahydro-2 / 7-pyran-4-yl)phenyl)oxazol-2-yl)met- hyl)pyrrolidin-3-yl)oxy)isoindolin-2-yl)piperidine-2, 6-dione. To a stirred mixture of

[0148] 5-(4-(tetrahydro-27f-pyran-4-yl)phenyl)oxazole-2-carbaldehyde (70 mg, 0.27 mmol), 3-(l-oxo- 5-(((.S)-pyrrolidin-3-yl)oxy)isoindolin-2-yl)piperidine- 2,6-dione hydrochloride (90 mg, 0.25 mmol) and EtsN (79 mg, 0.79 mmol) in DCM (5 mL) was added NaBH(OAc)s (160 mg, 0.75 mmol) in portions at room temperature and the above mixture was stirred for 1 h at 30 °C under nitrogen. The resulting mixture was concentrated. The residue was firstly purified by flash column chromatography (0 to 10% MeOH in DCM) and purified further by preparative HPLC with following conditions: Column: Sunfire prep C18 column, 30x 150 mm, 5 um; Mobile Phase A: Water (0.1% formic acid), Mobile Phase B: acetonitrile; Flow rate:60 rnL / min; Gradients B to 28 B in 10 min; 254 / 220 nm; RT1 : 10.38. The fractions containing the desired product were collected and evaporated under reduced pressure to afford the title compound (45.6 mg, 0.080 mmol, 32% yield) as a white solid. 'H NMR (400 MHz, Methanol-dr) 5 7.71-7.68 (m, 1H), 7.66-7.63 (m, 2H), 7.42-7.34 (m, 3H), 7.09-7.01 (m, 2H), 5.15-5.05 (m, 2H), 4.44-4.39 (m, 2H),

[0149] 4 09-4.04 (m, 2H), 4.02-3.94 (m, 2H), 3.62-3.55 (m, 2H), 3.20-3.15 (m, 1H), 3.09-3.02 (m, 2H),

[0150] 2.96-2.76 (m, 4H). 2.52-2.35 (m, 2H). 2.19-2.14 (m, 1H). 2.10-2.03 (m, 1H). 1.80 (m, 4H); MS

[0151] (ESI) m / z 571.3 [M+l]+.

[0152] Example 87

[0153] 3-(5-(((5)-l-((2-(2-Methoxypyridin-4-yl)-l-methyl-17 / -imidazol-5-yl)methyl)pyrr- olidin-3-yl)oxy)-l-oxoisoindolin-2-yl)piperidine-2, 6-dione

[0154] A. 3-(5-(((5)-l-((2-Bromo-l-methyl-lff-imidazol-5-yl)methyl)pyrrolidin-3- yl)oxy)-l-oxoisoindolin-2-yl)piperidine-2, 6-dione. To a solution of 2-bromo-l -methyl- \H- imidazole-5-carbaldehyde (300 mg, 1.59 mmol, 1.00 equiv.), 3-(l-oxo-5-(((S)-pyrrolidin-3- yl)oxy)isoindolin-2-yl)piperidine-2, 6-dione; hydrochloride (581 mg, 1.59 mmol, 1.00 equiv.) and EtsN (241 mg, 2.39 mmol, 1.50 equiv.) in DCM (20 mb) was added NaBH(OAc)3 (1.4 g, 6.60 mmol, 4. 15 equiv.) in several portions at room temperature and the above mixture was stirred for 2 h at this temperature under nitrogen. The resulting mixture was concentrated under reduced pressure. The crude product was purified by flash column chromatography (silica, 0- 5% MeOH in DCM). Pure fractions were evaporated to afford the title compound (750 mg, 1.49 mmol, 94% yield) as a colorless oil. MS(ESI) m / z 502. 1 [M+l]+.

[0155] B. 3-(5-(((A)-l-((2-(2-Methoxypyridin-4-yl)-l-methyl-lH-imidazol-5-yl)methyl) pyrr-olidin-3-yl)oxy)-l-oxoisoindolin-2-yl)piperidine-2, 6-dione. To a solution of 3-(5-(((S - l-((2-bromo-l-methyl-177-imidazol-5-yl)methyl)pyrrolidin-3-yl)oxy)-l-oxoisoindolin-2- yl)piperidine-2, 6-dione (450 mg, 0.90 mmol, 1.00 equiv.), (2-methoxypyridin-4-yl)boronic acid (411 mg, 2.69 mmol, 2.99 equiv.) and Pd(dppf)2.CH2Ch (73 mg, 0.089 mmol, 0.10 equiv.) in 1,4-dioxane (10 mb) were added K2CO3 (248 mg, 1.79 mmol, 1.99 equiv.) and water (81 mg, 4.48 mmol, 4.98 equiv.). The flask was evacuated and flushed three times with nitrogen. The mixture was stirred at 90 °C for 2 h under nitrogen. The resulting mixture was added water and extracted with EtOAc. The extracts combined were washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by flash column chromatography (0 to 10% MeOH in DCM) and further purified by Prep-HPLC. Pure fractions were evaporated to afford the title compound (128.3 mg, 0.24 mmol, 27% yield). ’H NMR (400 MHz, Methanol-d4) 5 8.21 (m. 1H), 8.18 (s, 1H), 7.72 (d, J= 8.4 Hz, 1H), 7.22 (d, J = 1.1 Hz, 1H), 7.12-7.01 (m, 3H), 6.93-6.88 (m, 1H), 5.13 (m, 1H), 5.06 (m, 1H), 4.48 (d, J= 17.0 Hz, 1H), 4.42 (d, J= 17.0 Hz, 1H), 3.96 (m, 5H), 3.83 (s, 3H), 3.15-3.07 (m, 1H), 3.05-2.85 (m, 3H), 2.84-2.69 (m, 2H), 2.56-2.38 (m, 2H), 2.18 (m, 1H), 2.05 (m, 1H). MS(ESI) m / z 531.1 [M+l]+. Biological Examples

[0156] Example Bl. WEEl-ePL degradation assay

[0157] To generate LentiX-WEEl-ePL-GSPTl-ND cells for the WEE1 degradation assay, LentiX-HEK293T (Invitrogen) were tranduced by lentivirus with a C-terminal ePL-tagged (DiscoverX, Fremont, CA) full-length WEE1 expression construct. A separate lentiviral construct was introduced to over-express a mutated form of GSPT1 (NP_002085), termed GSPT1-GN, which contains a G575N mutation and is further truncated in the first 158 amino acids. GSPT1-GN is functional but not degradable by Cereblon. Cells were selected and maintained in complete DMEM media (DMEM, 10% Heat Inactivated FBS, 400pg / mL Geneticin, and 1 pg / mL puromycin at 37C and 5% CO2). Cells were passaged every 3-4 days by reseeding 1x106cells in a T150 flask.

[0158] Cells were dispensed into 384-well white plates (Coming #3570, NY) that were prespotted with compounds using an acoustic dispenser (Echo acoustic transfer system, Beckman Coulter Life Sciences, Carlsbad, CA) as a 10-point dose-response curve with a 3-fold dilution starting at either luM or lOuM and including a DMSO control. Twenty -five microliters of media containing 2500 cells for LentiX-oeWEEl-ePL was dispensed per well. Assay plates were incubated at 37°C with 5% CO2 for four hours (WEE1) After incubation, 25 pL of the InCELL Hunter™ Detection Reagent Working Solution (DiscoverX, Fremont, CA) was added to each well and incubated at room temperature for 30 minutes protected from light. After 30 minutes, luminescence was read on a PHERAstar luminometer (Cam, NC).

[0159] To determine the half-maximal effective concentration (EC50) values for WEEl -ePL degradation, a four parameter logistic model was used: (Sigmoidal Dose-Response Model) (FIT= (A+((B-A) / l+((C / x)AD)))) where C is the inflection point (EC50), D is the Hill slope, and A and B are the low and high limits of the fit respectively). The lower limit of the fit (value A) is referred to as Ymin. The Ymin was constrained by using a luciferase inhibitor at a concentration of 20 pM to fully inhibit the lucifi erase signal and was used as the Ymin= 0 constraint value within each assay. The maximum limit constraint, Ymax, is similarly derived using DMSO control. The curves were processed and evaluated using Activity Base (IDBS, Alameda, CA) or using Dotmatics (Boston, MA). The results are shown in Table 2 below.

[0160] Example B2. MKN45 and MDA-MB-231 cell proliferation assays

[0161] MKN45 cells were obtained from Celgene internal cell banks and cultured in RPMI complete media (RPMI- 1640, IX Penicillin / Streptomycin, IX glutamine, IX Sodium Pyruvate, Non-essential Amino Acids. 10% Fetal Bovine serum, all component from Thermo-Fisher). MDA-MB-231 cells were obtained from the ATCC (HTB-26) and cultured in RPMI- 1640 and 10% Fetal Bovine serum; MDA-MB-231 cells were expanded and stored as aliquots in liquid nitrogen in Celgene internal cell banks. Compounds were dispensed into 384-well plates using an HP DE300 compound printer in triplicate with 8 compounds on each plate. Compounds were dispensed into 384-well black clear bottom plates (Coming 3764BC) using an EDC acoustic dispenser at BMS. For each compound, a 10-point dose response was printed at half-log dilutions starting at either luM or lOuM in triplicate for MKN45 cells and in duplicate for MDA-MB-231 cells. The DMSO concentration was kept constant for a final assay concentration of 0.1% per well assuming 50ul final media volume. Compound plates were sealed and frozen at -20C until use.

[0162] For each assay batch, compound plates were thawed and allowed to reach room temperature, and cells were seeded at 400 MKN45 cells per well, or 800 MDA-MB-231 cells per well, in 50ul media listed above for each cell line. After 120 hours of incubation at 37C and 5% CO2, cells were lysed by addition of 25 pL of Cell-Titer-Glo Reagent (Promega Corporation, Madison, WI), as per manufacturer’s instructions, protected from light, shaken for 45 minutes, and total luminescence read by a Perkin Elmer Envision plate reader. Total luminescence signal is linearly correlated with cell number remaining in the well.

[0163] Luminescence data was processed per plate by subtracting reagent-only “blank” well luminescence from all measured values and calculating a percent of DMSO control value for each treatment well. Then, to determine the half-maximal effective concentration (EC50) values for cell growth inhibition, a four parameter logistic model was fit to the data for each DMSO- normalized compound: Sigmoidal Dose-Response Model: (FIT= (A+((B-A) / l+((C / x)AD)))) where C is the inflection point (EC50), D is the Hill slope, and A and B are the low and high limits of the fit respectively. The lower limit of the fit (value A) is referred to as Ymin- calculated. The minimal percent of DMSO control that is observed in the concentrations tested for each compound is labeled as “Ymin-obs”, and was recorded and reported in the tables. Sigmoidal fit curves were processed and evaluated using Activity Base (IDBS, Alameda, CA) or using Dotmatics (Boston, MA). The results are shown in Table 2 below. N / A = not available at the present time.

[0164] Table 2. MKN45 cell proliferation and WEEl-ePL degradation.

[0165]

[0166] “ Denotes not available

[0167] Although the present invention has been described in some detail by way of illustration and example for purposes of clarity of understanding, the descriptions and examples should not be construed as limiting the scope of the invention. The disclosures of all patent and scientific literature cited herein are expressly incorporated herein in their entirety' by reference.

Claims

CLAIMS1. A compound, selected from:or a pharmaceutically acceptable salt, tautomer, isotopologue. or stereoisomer thereof.

2. A method for reducing WEE1 kinase protein levels, the method comprising contacting a cell with an effective amount of a compound of claim 1 or a pharmaceutically acceptable salt, tautomer, isotopologue, or stereoisomer thereof.

3. The method of claim 2, wherein the cell is in a subject.

4. A method of preventing or treating cancer in a subject comprising administering to a subject in need thereof an effective amount of a compound of claim 1 or a pharmaceutically acceptable salt, tautomer, isotopologue, or stereoisomer thereof.

5. The method according to claim 4, wherein the cancer is selected from gastric, lung, pancreatic, ovarian, breast, skin, colon, neuroblastoma, osteosarcoma, uterine, rectal, and kidney.

6. The method according to claim 5, wherein the cancer is selected from pancreatic ductal adenocarcinoma (PDAC), small cell lung cancer, non-small cell lung cancer (NSCLC), high grade serous ovarian cancer, triple negative breast cancer, uterine serous carcinoma, Ewing's sarcoma, melanoma, colon, and clear cell renal cell carcinoma (ccRCC).

7. The use of a compound of claim 1 or a pharmaceutically acceptable salt, tautomer, isotopologue, or stereoisomer thereof, in the manufacture of a medicament for reducing WEE1 kinase protein levels.

8. The use of a compound of claim 1 or a pharmaceutically acceptable salt, tautomer, isotopologue, or stereoisomer thereof, in the manufacture of a medicament for the prevention or treatment of cancer.

9. The use according to claim 8, wherein the cancer is selected from gastric, lung, pancreatic, ovarian, breast, skin, colon, neuroblastoma, osteosarcoma, uterine, rectal, and kidney.

10. The use according to claim 9, wherein the cancer is selected from pancreatic ductal adenocarcinoma (PDAC), small cell lung cancer, non-small cell lung cancer (NSCLC), high grade serous ovarian cancer, triple negative breast cancer, uterine serous carcinoma, Ewing’s sarcoma, melanoma, colon, and clear cell renal cell carcinoma (ccRCC).

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