WEE1 INHIBITORS AND METHODS FOR TREATING CANCER

INCORPORATION BY REFERENCE TO PRIORITY APPLICATIONS

[0001] Any and all applications for which a foreign or domestic priority claim is identified in the Application Data Sheet as filed with the present application are hereby expressly incorporated by reference under 37 CFR 1.57, and Rules 4.18 and 20.6, including U.S. Provisional Application No. 63/376,024, filed September 16, 2022, which is hereby incorporated by reference in its entirety.

BACKGROUND

Field

[0002] The present application relates generally to compounds that are WEE1 inhibitors and/or degraders thereof, and methods of using them to treat conditions characterized by excessive cellular proliferation, such as cancer.

Description

[0003] DNA is constantly damaged from the environment. Light, chemicals, stress, and cellular replication lead to single- or double- stranded breakage along DNA’s backbone. Typically, organisms defend against DNA damage by repair proteins that either reconnect, or re-synthesize damaged DNA. The correct functioning of these proteins are essential for life. The incorrect replacement of nucleotides into DNA can cause mutations (and other genetic alterations including but not limited to insertions, deletions, and frameshifts), genetic disease, and loss of protein function. The altogether loss of DNA repair can cause cell death, tumor progression, and cancer.

[0004] Cell cycle checkpoints are important for proper DNA repair, ensuring that cells do not progress with cellular- replication until their genomic integrity is restored. WEE1 is a nuclear kinase involved in the G2-M cell-cycle checkpoint arrest for DNA repair before mitotic entry. Normal cells repair damaged DNA during G1 arrest. Cancer cells often have a deficient Gl-S checkpoint and depend on a functional G2-M checkpoint for DNA repair. WEE1 is overexpressed in various cancer types. [0005] Various inhibitors and/or degraders of WEE 1 are known to those skilled in the art. Sec, c.g., WO 2019/173082 and WO 2020/069105. However, there remains an urgent need for inhibitors and/or degraders of WEE1 that are useful for the treatment of conditions characterized by excessive cellular proliferation, such as cancer.

SUMMARY

[0006] Various embodiments provide a compound of Formula (I), or a pharmaceutically acceptable salt thereof, having the structure: wherein:

R ¹ can be absent, a substituted or an unsubstituted 6 to 10 membered aryl, -NH-(a substituted or an unsubstituted 6 to 10 membered aryl)-, a substituted or an unsubstituted 5 to 10 membered heteroaryl, -NH-(a substituted or an unsubstituted 5 to 10 membered heteroaryl)- or -NH-(a substituted or an unsubstituted 6 to 10 membered cycloalkyl)-;

R ² can be absent, a substituted or an unsubstituted 4 to 10 membered heterocyclyl or -O-(a substituted or an unsubstituted 4 to 10 membered heterocyclyl)-;

R ³ can be absent, a substituted or an unsubstituted Ci-Ce alkylene, -NH-, -NH-(a substituted or an unsubstituted Ci-Ce alkylene)-, -O-(a substituted or an unsubstituted Ci-Ce alkylene)-NH- or -O-(a substituted or an unsubstituted Ci-Ce alkylene)-NH-(a substituted or an unsubstituted Ci-Ce alkylene)-;

R ⁴ can be absent or a substituted or an unsubstituted 4 to 10 membered heterocyclyl; with the proviso that at least two of R ¹ , R ² , R ³ and R ⁴ are present;

R ^D can be a substituted or an unsubstituted Ci-Cb alkyl; and Y is -CH or N (nitrogen).

[0007] Another embodiment provides a compound, or a pharmaceutically acceptable salt thereof, wherein the compound can have a structure selected from those of Compound Nos. 1 to 18 as described herein.

[0008] Another embodiment provides a pharmaceutical composition that can include an effective amount of a compound of the Formula (I) as described herein, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier, diluent, excipient or combination thereof.

[0009] Another embodiment provides a use of an effective amount of a compound of the Formula (I) as described herein, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition that can include such a compound as described herein, in the manufacture of a medicament for ameliorating or treating a cancer, wherein the cancer can be selected from a brain cancer, a cervicocerebral cancer, an esophageal cancer, a thyroid cancer, a small cell cancer, a non- small cell cancer, a breast cancer, a lung cancer, a stomach cancer, a gallbladder/bile duct cancer, a liver cancer, a pancreatic cancer, a colon cancer, a rectal cancer, an ovarian cancer, a choriocarcinoma, an uterus body cancer, an uterocervical cancer, a renal pelvis/ureter cancer, a bladder cancer, a prostate cancer, a penis cancer, a testicular cancer, a fetal cancer, Wilms’ cancer, a skin cancer, malignant melanoma, a neuroblastoma, an osteosarcoma, an Ewing’s tumor, a soft pail sarcoma, an acute leukemia, a chronic lymphatic leukemia, a chronic myelocytic leukemia, polycythemia vera, a malignant lymphoma, multiple myeloma, a Hodgkin’s lymphoma and a non-Hodgkin’ s lymphoma.

[0010] Another embodiment provides a use of an effective amount of a compound of the Formula (I) as described herein, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition that can include such a compound as described above, in the manufacture of a medicament for inhibiting replication of a malignant growth or a tumor, wherein the malignant growth or tumor can be due to a cancer selected from a brain cancer, a cervicocerebral cancer, an esophageal cancer, a thyroid cancer, a small cell cancer, a nonsmall cell cancer, a breast cancer, a lung cancer, a stomach cancer, a gallbladder/bile duct cancer, a liver cancer, a pancreatic cancer, a colon cancer, a rectal cancer, an ovarian cancer, a choriocarcinoma, an uterus body cancer, an uterocervical cancer, a renal pelvis/ureter cancer, a bladder cancer, a prostate cancer, a penis cancer, a testicular cancer, a fetal cancer, Wilms’ cancer, a skin cancer, malignant melanoma, a neuroblastoma, an osteosarcoma, an Ewing’s tumor, a soft part sarcoma, an acute leukemia, a chronic lymphatic leukemia, a chronic myelocytic leukemia, polycythemia vera, a malignant lymphoma, multiple myeloma, a Hodgkin’s lymphoma and a non-Hodgkin’s lymphoma.

[0011] Another embodiment provides a use of an effective amount of a compound of the Formula (I) as described herein, or a pharmaceutical composition that can include such a compound as described above, in the manufacture of a medicament for ameliorating or treating a malignant growth or tumor, wherein the malignant growth or tumor can be due to a cancer selected from a brain cancer, a cervicocerebral cancer, an esophageal cancer, a thyroid cancer, a small cell cancer, a non-small cell cancer, a breast cancer, a lung cancer , a stomach cancer, a gallbladder/bile duct cancer, a liver cancer, a pancreatic cancer, a colon cancer, a rectal cancer, an ovarian cancer, a choriocarcinoma, an uterus body cancer, an uterocervical cancer, a renal pelvis/ureter cancer, a bladder cancer, a prostate cancer, a penis cancer, a testicular cancer, a fetal cancer, Wilms’ cancer, a skin cancer, malignant melanoma, a neuroblastoma, an osteosarcoma, an Ewing’s tumor, a soft part sarcoma, an acute leukemia, a chronic lymphatic leukemia, a chronic myelocytic leukemia, polycythemia vera, a malignant lymphoma, multiple myeloma, a Hodgkin’s lymphoma and a non-Hodgkin’s lymphoma.

[0012] These and other embodiments are described in greater detail below.

BRIEF DESCRIPTION OF THE DRAWINGS

[0013] FIG. 1 illustrates a general synthetic scheme for preparing compounds of the Formula (I).

[0014] FIG. 2 illustrates a general synthetic scheme for preparing embodiments of compounds of the Formula (I).

[0015] FIG. 3 illustrates a general synthetic scheme for preparing embodiments of compounds of the Formula (I).

[0016] FIG. 4 illustrates a synthetic scheme for preparing Intermediate 1.

[0017] FIG. 5 illustrates a synthetic scheme for preparing Intermediate 2.

[0018] FIG. 6 illustrates a synthetic scheme for preparing Intermediate 3.

[0019] FIG. 7 illustrates a synthetic scheme for preparing Intermediate 4.

[0020] FIG. 8 illustrates a synthetic scheme for preparing Intermediate 5. [0021] FIG. 9 dlustrates a synthetic scheme for preparing Intermediate 6.

[0022] FIG. 10 illustrates a synthetic schcmc for preparing Intermediate 7.

[0023] FIG. 11 illustrates a synthetic scheme for preparing Intermediate 8.

[0024] FIG. 12 illustrates a synthetic scheme for preparing Intermediate 9.

[0025] FIG. 13 illustrates a synthetic scheme for preparing Intermediate 10.

[0026] FIG. 14 illustrates a synthetic scheme for preparing Intermediate 12.

[0027] FIG. 15 illustrates a synthetic scheme for preparing Intermediate 13.

[0028] FIG. 16 illustrates a synthetic scheme for preparing Intermediate 14.

[0029] FIG. 17 illustrates a synthetic scheme for preparing Intermediate 15.

[0030] FIG. 18 illustrates a synthetic scheme for preparing Intermediate 16.

[0031] FIG. 19 illustrates a synthetic scheme for preparing Intermediate 17.

[0032] FIG. 20 illustrates a synthetic scheme for preparing Intermediate 18.

[0033] FIG. 21 illustrates a synthetic scheme for preparing Intermediate 19.

[0034] FIG. 22 illustrates a synthetic scheme for preparing Intermediate 21.

[0035] FIG. 23 illustrates a synthetic scheme for preparing Intermediate 22.

[0036] FIG. 24 illustrates a synthetic scheme for preparing Intermediate 23.

[0037] FIG. 25 illustrates a synthetic scheme for preparing Intermediate 24.

[0038] FIG. 26 illustrates a synthetic scheme for preparing Intermediate 25.

[0039] FIG. 27 illustrates a synthetic scheme for preparing Intermediate 26.

[0040] FIG. 28 illustrates a synthetic scheme for preparing Intermediate 27.

[0041] FIG. 29 illustrates a synthetic scheme for preparing Intermediate 28.

[0042] FIG. 30 illustrates a synthetic scheme for preparing Intermediate 29.

[0043] FIG. 31 illustrates a synthetic scheme for preparing Intermediate 30.

[0044] FIG. 32 illustrates a synthetic scheme for preparing Intermediate 31.

[0045] FIG. 33 illustrates a synthetic scheme for preparing Intermediate 33.

[0046] FIG. 34 illustrates a synthetic scheme for preparing Intermediate 34.

[0047] FIG. 35 illustrates a synthetic scheme for preparing Intermediate 35.

[0048] FIG. 36 illustrates a synthetic scheme for preparing Intermediate 36.

[0049] FIG. 37 illustrates a synthetic scheme for preparing Intermediate 37.

[0050] FIG. 38 illustrates a synthetic scheme for preparing Intermediate 38.

[0051] FIG. 39 illustrates a synthetic scheme for preparing Intermediate 40. [0052] FIG. 40 illustrates a synthetic scheme for preparing Intermediate 43.

[0053] FIG. 41 illustrates a synthetic scheme for preparing Intermediate 44.

[0054] FIG. 42 illustrates a synthetic scheme for preparing Intermediate 47 and Intermediate 48.

[0055] FIG. 43 illustrates a synthetic scheme for preparing Intermediate 50.

[0056] FIG. 44 illustrates a synthetic scheme for preparing Intermediate 51.

[0057] FIG. 45 illustrates a synthetic scheme for preparing Intermediate 52.

[0058] FIG. 46 illustrates a synthetic scheme for preparing Intermediate 53.

[0059] FIG. 47 illustrates a synthetic scheme for preparing Intermediate 54.

[0060] FIG. 48 illustrates a synthetic scheme for preparing Intermediate 55.

[0061] FIG. 49 illustrates a synthetic scheme for preparing Intermediate 56.

[0062] FIG. 50 illustrates a synthetic scheme for preparing Intermediate 57.

[0063] FIG. 51 illustrates a synthetic scheme for preparing Intermediate 58.

[0064] FIG. 52 illustrates a synthetic scheme for preparing Intermediate 60.

DETAILED DESCRIPTION

[0065] WEE1 is a tyrosine kinase that is a critical component of the ATR- mediated G2 cell cycle checkpoint control that prevents entry into mitosis in response to cellular DNA damage. ATR phosphorylates and activates CHK1, which in turn activates WEE1, leading to the selective phosphorylation of cyclin-dependent kinase 1 (CDK1) at Tyrl5, thereby stabilizing the CDKl-cyclin B complex and halting cell-cycle progression. This process confers a survival advantage by allowing tumor cells time to repair damaged DNA prior to entering mitosis. Inhibition of WEE 1 abrogates the G2 checkpoint, promoting cancer cells with DNA damage to enter into unscheduled mitosis and undergo cell death via mitotic catastrophe. Therefore, WEE1 inhibition and/or degradation has the potential to sensitize tumors to DNA-damaging agents, such as cisplatin, and to induce tumor cell death.

Definitions

[0066] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as is commonly understood by one of ordinary skill in the art. All patents, applications, published applications and other publications referenced herein are incorporated by reference in their entirety unless stated otherwise. In the event that there are a plurality of definitions for a term herein, those in this section prevail unless stated otherwise.

[0067] Whenever a group is described as being “optionally substituted” that group may be unsubstituted or substituted with one or more of the indicated substituents. Likewise, when a group is described as being “unsubstituted or substituted” if substituted, the substituent(s) may be selected from one or more the indicated substituents. If no substituents are indicated, it is meant that the indicated “optionally substituted” or “substituted” group may be substituted with one or more group(s) individually and independently selected from alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl, heteroaryl, heterocyclyl, aryl(alkyl), cycloalkyl(alkyl), heteroaryl(alkyl), heterocyclyl(alkyl), hydroxy, alkoxy, acyl, cyano, halogen, thiocarbonyl, O-carbamyl, N-carbamyl, O-thiocarbamyl, N-thiocarbamyl, C-amido, N-amido, S- sulfonamide, N-sulfonamido, C-carboxy, O-carboxy, nitro, sulfenyl, sulfinyl, sulfonyl, haloalkyl, hydroxyalkyl, haloalkoxy, an amino, a mono-substituted amine group, a di-substituted amine group and an amine(Ci-C6 alkyl).

[0068] As used herein, “C _a to Cb” in which “a” and “b” are integers refer to the number of carbon atoms in a group. The indicated group can contain from “a” to “b”, inclusive, carbon atoms. Thus, for example, a “Ci to C4 alkyl” group refers to all alkyl groups having from 1 to 4 carbons, that is, CH3-, CH3CH2-, CH3CH2CH2-, (CH ₃ ) ₂ CH-, CH3CH2CH2CH2-. CH ₃ CH2CH(CH ₃ )- and (CH ₃ ) ₃ C-. If no “a” and “b” are designated, the broadest range described in these definitions is to be assumed.

[0069] If two “R” groups are described as being “taken together” the R groups and the atoms they are attached to can form a cycloalkyl, cycloalkenyl, aryl, heteroaryl or heterocycle. For example, without limitation, if R ^a and R ^b of an NR ^a R ^b group are indicated to be “taken together,” it means that they are covalently bonded to one another to form a ring:

.R ^a

— N ' R ^b

[0070] As used herein, the term “alkyl” refers to a fully saturated aliphatic hydrocarbon group. The alkyl moiety may be branched or straight chain. Examples of branched alkyl groups include, but are not limited to, iso-propyl, sec-butyl, t-butyl and the like. Examples of straight chain alkyl groups include, but are not limited to, methyl, ethyl, n- propyl, n-butyl, n-pentyl, n-hexyl, n-heptyl and the like. The alkyl group may have 1 to 30 carbon atoms (whenever it appears herein, a numerical range such as “1 to 30” refers to each integer in the given range; e.g., “1 to 30 carbon atoms” means that the alkyl group may consist of 1 carbon atom, 2 carbon atoms, 3 carbon atoms, etc., up to and including 30 carbon atoms, although the present definition also covers the occurrence of the term “alkyl” where no numerical range is designated). The alkyl group may also be a medium size alkyl having 1 to 12 carbon atoms. The alkyl group could also be a lower alkyl having 1 to 6 carbon atoms. An alkyl group may be substituted or unsubstituted.

[0071] The term “alkenyl” used herein refers to a monovalent straight or branched chain radical of from two to twenty carbon atoms containing a carbon double bond(s) including, but not limited to, 1 -propenyl, 2-propenyl, 2-methyl-l -propenyl, 1-butenyl, 2- butenyl and the like. An alkenyl group may be unsubstituted or substituted.

[0072] The term “alkynyl” used herein refers to a monovalent straight or branched chain radical of from two to twenty carbon atoms containing a carbon triple bond(s) including, but not limited to, 1-propynyl, 1-butynyl, 2-butynyl and the like. An alkynyl group may be unsubstituted or substituted.

[0073] As used herein, “cycloalkyl” refers to a completely saturated (no double or triple bonds) mono- or multi- cyclic hydrocarbon ring system. When composed of two or more rings, the rings may be joined together in a fused, bridged or spiro fashion. As used herein, the term “fused” refers to two rings which have two atoms and one bond in common. As used herein, the term “bridged cycloalkyl” refers to compounds wherein the cycloalkyl contains a linkage of one or more atoms connecting non-adjacent atoms. As used herein, the term “spiro” refers to two rings which have one atom in common and the two rings are not linked by a bridge. Cycloalkyl groups can contain 3 to 30 atoms in the ring(s), 3 to 20 atoms in the ring(s), 3 to 10 atoms in the ring(s), 3 to 8 atoms in the ring(s) or 3 to 6 atoms in the ring(s). A cycloalkyl group may be unsubstituted or substituted. Examples of monocycloalkyl groups include, but are in no way limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and cyclooctyl. Examples of fused cycloalkyl groups are decahydronaphthalenyl, dodecahydro- IH-phenalenyl and tetradecahydroanthracenyl; examples of bridged cycloalkyl groups are bicyclo[l.l.l]pentyl, adamantanyl and norbornanyl; and examples of spiro cycloalkyl groups include spiro[3.3]heptane and spiro[4.5]decane. [0074] As used herein, “cycloalkenyl” refers to a mono- or multi- cyclic hydrocarbon ring system that contains one or more double bonds in at least one ring; although, if there is more than one, the double bonds cannot form a fully delocalized pi-electron system throughout all the rings (otherwise the group would be “aryl,” as defined herein). Cycloalkenyl groups can contain 3 to 10 atoms in the ring(s), 3 to 8 atoms in the ring(s) or 3 to 6 atoms in the ring(s). When composed of two or more rings, the rings may be connected together in a fused, bridged or spiro fashion. A cycloalkenyl group may be unsubstituted or substituted.

[0075] As used herein, “carbocyclyl” refers to a non-aromatic a mono- or multi- cyclic hydrocarbon ring system. When composed of two or more rings, the rings may be joined together in a fused, bridged or spiro fashion, as described herein. Carbocyclyl groups can contain 3 to 30 atoms in the ring(s), 3 to 20 atoms in the ring(s), 3 to 10 atoms in the ring(s), 3 to 8 atoms in the ring(s) or 3 to 6 atoms in the ring(s). A carbocyclyl group may be unsubstituted or substituted. Examples of carbocyclyl groups include, but are in no way limited to, cycloalkyl groups and cycloalkenyl groups, as defined herein, and the non-aromatic portions of 1,2,3 ,4-tetrahydronaphthalene, 2,3-dihydro-lH-indene, 5, 6,7,8- tetrahydroquinoline and 6,7-dihydro-5H-cyclopenta[b]pyridine.

[0076] As used herein, “aryl” refers to a carbocyclic (all carbon) monocyclic or multicyclic aromatic ring system (including fused ring systems where two carbocyclic rings share a chemical bond) that has a fully delocalized pi-electron system throughout all the rings. The number of carbon atoms in an aryl group can vary. For example, the aryl group can be a Ce-Cu aryl group, a Ce-Cio aryl group or a Ce aryl group. Examples of aryl groups include, but are not limited to, benzene, naphthalene and azulene. An aryl group may be substituted or unsubstituted.

[0077] As used herein, “heteroaryl” refers to a monocyclic or multicyclic aromatic ring system (a ring system with fully delocalized pi-electron system) that contain(s) one or more heteroatoms (for example, 1, 2 or 3 heteroatoms), that is, an element other than carbon, including but not limited to, nitrogen, oxygen and sulfur. The number of atoms in the ring(s) of a heteroaryl group can vary. For example, the heteroaryl group can contain 4 to 14 atoms in the ring(s), 5 to 10 atoms in the ring(s) or 5 to 6 atoms in the ring(s), such as nine carbon atoms and one heteroatom; eight carbon atoms and two heteroatoms; seven carbon atoms and three heteroatoms; eight carhon atoms and one heteroatom; seven carhon atoms and two hctcroatoms; six carbon atoms and three hctcroatoms; five carbon atoms and four heteroatoms; five carbon atoms and one heteroatom; four carbon atoms and two heteroatoms; three carbon atoms and three heteroatoms; four carbon atoms and one heteroatom; three carbon atoms and two heteroatoms; or two carbon atoms and three heteroatoms. Furthermore, the term “heteroaryl” includes fused ring systems where two rings, such as at least one aryl ring and at least one heteroaryl ring or at least two heteroaryl rings, share at least one chemical bond. Examples of heteroaryl rings include, but are not limited to, furan, furazan, thiophene, benzothiophene, phthalazine, pyrrole, oxazole, benzoxazole, 1,2,3-oxadiazole, 1,2,4- oxadiazole, thiazole, 1,2,3-thiadiazole, 1,2,4-thiadiazole, benzothiazole, imidazole, benzimidazole, indole, indazole, pyrazole, benzopyrazole, isoxazole, benzoisoxazole, isothiazole, triazole, benzotriazole, thiadiazole, tetrazole, pyridine, pyridazine, pyrimidine, pyrazine, purine, pteridine, quinoline, isoquinoline, quinazoline, quinoxaline, cinnoline and triazine. A heteroaryl group may be substituted or unsubstituted.

[0078] As used herein, “heterocyclyl” or “heteroalicyclyl” refers to three-, four-, five-, six-, seven-, eight-, nine-, ten-, up to 18-membered monocyclic, bicyclic and tricyclic ring system wherein carbon atoms together with from 1 to 5 heteroatoms constitute said ring system. A heterocycle may optionally contain one or more unsaturated bonds situated in such a way, however, that a fully delocalized pi-electron system does not occur throughout all the rings. The heteroatom(s) is an element other than carbon including, but not limited to, oxygen, sulfur and nitrogen. A heterocycle may further contain one or more carbonyl or thiocarbonyl functionalities, so as to make the definition include oxo-systems and thio-systems such as lactams, lactones, cyclic imides, cyclic thioimides and cyclic carbamates. When composed of two or more rings, the rings may be joined together in a fused, bridged or spiro fashion. As used herein, the term “fused” refers to two rings which have two atoms and one bond in common. As used herein, the term “bridged heterocyclyl” or “bridged heteroalicyclyl” refers to compounds wherein the heterocyclyl or heteroalicyclyl contains a linkage of one or more atoms connecting non-adjacent atoms. As used herein, the term “spiro” refers to two rings which have one atom in common and the two rings are not linked by a bridge. Heterocyclyl and heteroalicyclyl groups can contain 3 to 30 atoms in the ring(s), 3 to 20 atoms in the ring(s), 3 to 10 atoms in the ring(s), 3 to 8 atoms in the ring(s) or 3 to 6 atoms in the ring(s). For example, five carbon atoms and one heteroatom; four carbon atoms and two heteroatoms; three carbon atoms and three hctcroatoms; four carbon atoms and one hctcroatom; three carbon atoms and two heteroatoms; two carbon atoms and three heteroatoms; one carbon atom and four heteroatoms; three carbon atoms and one heteroatom; or two carbon atoms and one heteroatom. Additionally, any nitrogens in a heteroalicyclic may be quaternized. Heterocyclyl or heteroalicyclic groups may be unsubstituted or substituted. Examples of such “heterocyclyl” or “heteroalicyclyl” groups include but are not limited to, 1,3-dioxin, 1,3- dioxane, 1,4-dioxane, 1,2-dioxolane, 1,3-dioxolane, 1,4-dioxolane, 1,3-oxathiane, 1,4- oxathiin, 1,3 -oxathiolane, 1,3-dithiole, 1,3-dithiolane, 1,4-oxathiane, tetrahydro- 1,4-thiazine, 2H-l,2-oxazine, maleimide, succinimide, barbituric acid, thiobarbituric acid, dioxopiperazine, hydantoin, dihydrouracil, trioxane, hexahydro-1, 3, 5-triazine, imidazoline, imidazolidine, isoxazoline, isoxazolidine, oxazoline, oxazolidine, oxazolidinone, thiazoline, thiazolidine, morpholine, oxirane, piperidine N-Oxide, piperidine, piperazine, pyrrolidine, azepane, pyrrolidone, pyrrolidione, 4-piperidone, pyrazoline, pyrazolidine, 2-oxopyrrolidine, tetrahydropyran, 4H-pyran, tetrahydrothiopyran, thiamorpholine, thiamorpholine sulfoxide, thiamorpholine sulfone and their benzo-fused analogs (e.g., benzimidazolidinone, tetrahydroquinoline and/or 3,4-methylenedioxyphenyl). Examples of spiro heterocyclyl groups include 2-azaspiro[3.3]heptane, 2-oxaspiro[3.3]heptane, 2-oxa-6- azaspiro[3.3]heptane, 2,6-diazaspiro[3.3]heptane, 2-oxaspiro[3.4]octane and 2- azaspiro[3.4]octane.

[0079] As used herein, “aralkyl” and “aryl(alkyl)” refer to an aryl group connected, as a substituent, via a lower alkylene group. The lower alkylene and aryl group of an aralkyl may be substituted or unsubstituted. Examples include but are not limited to benzyl, 2-phenylalkyl, 3 -phenylalkyl and naphthylalkyl.

[0080] As used herein, “heteroaralkyl” and “heteroaryl(alkyl)” refer to a heteroaryl group connected, as a substituent, via a lower alkylene group. The lower alkylene and heteroaryl group of heteroaralkyl may be substituted or unsubstituted. Examples include but are not limited to 2-thienylalkyl, 3-thienylalkyl, furylalkyl, thienylalkyl, pyrrolylalkyl, pyridylalkyl, isoxazolylalkyl and imidazolylalkyl and their benzo-fused analogs.

[0081] A “heteroalicyclyl(alkyl)” and “heterocyclyl(alkyl)” refer to a heterocyclic or a heteroalicyclic group connected, as a substituent, via a lower alkylene group. The lower alkylene and heterocyclyl of a (heteroalicyclyl)alkyl may be substituted or unsubstituted. Examples include but arc not limited tctrahydro-2H-pyran-4-yl(mcthyl), pipcridin-4- yl(ethyl), piperidin-4-yl(propyl), tetrahydro-2H-thiopyran-4-yl(methyl) and l,3-thiazinan-4- yl(methyl).

[0082] As used herein, “alkylene groups” are straight-chained alkyl groups that are -CH2- tethering, forming bonds to connect molecular fragments via their terminal carbon atoms. Embodiments of alkylene groups may be represented herein by -(CH2) _n - where n is an integer in the range of 1 to 30. “Lower” alkylene groups are alkylene groups that contain 1 to 6 carbon atoms. Examples of lower alkylene groups include but are not limited to methylene (-CH2-), ethylene (-CH2CH2-), propylene (-CH2CH2CH2-) and butylene (- CH2CH2CH2CH2-). An alkylene group can be substituted by replacing one or more hydrogen of the alkylene group and/or by substituting both hydrogens on the same carbon with a cycloalkyl group (e.g., -C- ).

[0083] As used herein, the term “hydroxy” refers to a -OH group.

[0084] As used herein, “alkoxy” refers to the Formula -OR wherein R is an alkyl, an alkenyl, an alkynyl, a cycloalkyl, a cycloalkenyl, aryl, heteroaryl, heterocyclyl, cycloalkyl(alkyl), aryl(alkyl), heteroaryl(alkyl) or heterocyclyl(alkyl) is defined herein. A non-limiting list of alkoxys are methoxy, ethoxy, n-propoxy, 1 -methylethoxy (iso-propoxy), n-butoxy, iso-butoxy, sec -butoxy, tert-butoxy, phenoxy and benzoxy. An alkoxy may be substituted or unsubstituted.

[0085] As used herein, “acyl” refers to a hydrogen, alkyl, alkenyl, alkynyl, aryl, hctcroaryl, heterocyclyl, aryl(alkyl), hctcroaryl(alkyl) and hctcrocyclyl(alkyl) connected, as substituents, via a carbonyl group. Examples include formyl, acetyl, propanoyl, benzoyl and acryl. An acyl may be substituted or unsubstituted.

[0086] A “cyano” group refers to a “-CN” group.

[0087] The term “halogen atom” or “halogen” as used herein, means any one of the radio-stable atoms of column 7 of the Periodic Table of the Elements, such as, fluorine, chlorine, bromine and iodine. [0088] A “thiocarbonyl” group refers to a “-C(=S)R” group in which R can be the same as defined with respect to O-carboxy. A thiocarbonyl may be substituted or unsubstituted.

[0089] An “O-carbamyl” group refers to a “-0C(=0)N(RARB)” group in which RA and RB can be independently hydrogen, an alkyl, an alkenyl, an alkynyl, a cycloalkyl, a cycloalkenyl, aryl, heteroaryl, heterocyclyl, cycloalkyl(alkyl), aryl(alkyl), heteroaryl(alkyl) or heterocyclyl(alkyl). An O-carbamyl may be substituted or unsubstituted.

[0090] An “N-carbamyl” group refers to an “R0C(=0)N(RA)-” group in which R and RA can be independently hydrogen, an alkyl, an alkenyl, an alkynyl, a cycloalkyl, a cycloalkenyl, aryl, heteroaryl, heterocyclyl, cycloalkyl(alkyl), aryl(alkyl), heteroaryl(alkyl) or heterocyclyl(alkyl). An N-carbamyl may be substituted or unsubstituted.

[0091] An “O-thiocarbamyl” group refers to a “-OC(=S)-N(RARB)” group in which RA and RB can be independently hydrogen, an alkyl, an alkenyl, an alkynyl, a cycloalkyl, a cycloalkenyl, aryl, heteroaryl, heterocyclyl, cycloalkyl(alkyl), aryl(alkyl), heteroaryl(alkyl) or heterocyclyl(alkyl). An O-thiocarbamyl may be substituted or unsubstituted.

[0092] An “N-thiocarbamyl” group refers to an “ROC(=S)N(RA)-” group in which R and RA can be independently hydrogen, an alkyl, an alkenyl, an alkynyl, a cycloalkyl, a cycloalkenyl, aryl, heteroaryl, heterocyclyl, cycloalkyl(alkyl), aryl(alkyl), heteroaryl(alkyl) or heterocyclyl(alkyl). An N-thiocarbamyl may be substituted or unsubstituted.

[0093] A “C-amido” group refers to a “-C(=0)N(RARB)” group in which RA and RB can be independently hydrogen, an alkyl, an alkenyl, an alkynyl, a cycloalkyl, a cycloalkenyl, aryl, heteroaryl, heterocyclyl, cycloalky l(alkyl), aryl(alkyl), heteroaryl(alkyl) or heterocyclyl(alkyl). A C-amido may be substituted or unsubstituted.

[0094] An “N-amido” group refers to a “RC(=O)N(RA)-” group in which R and RA can be independently hydrogen, an alkyl, an alkenyl, an alkynyl, a cycloalkyl, a cycloalkenyl, aryl, heteroaryl, heterocyclyl, cycloalkyl(alkyl), aryl(alkyl), heteroaryl(alkyl) or heterocyclyl(alkyl). An N-amido may be substituted or unsubstituted.

[0095] An “S-sulfonamido” group refers to a “-SO2N(RARB)” group in which RA and RB can be independently hydrogen, an alkyl, an alkenyl, an alkynyl, a cycloalkyl, a cycloalkenyl, aryl, heteroaryl, heterocyclyl, cycloalkyl(alkyl), aryl(alkyl), heteroaryl(alkyl) or hctcrocyclyl(alkyl). An S-sulfonamido may be substituted or unsubstituted.

[0096] An “N-sulfonamido” group refers to a “RSO2N(RA)-” group in which R and RA can be independently hydrogen, an alkyl, an alkenyl, an alkynyl, a cycloalkyl, a cycloalkenyl, aryl, heteroaryl, heterocyclyl, cycloalkyl(alkyl), aryl(alkyl), heteroaryl(alkyl) or heterocyclyl(alkyl). An N-sulfonamido may be substituted or unsubstituted.

[0097] An “O-carboxy” group refers to a “RC(=O)O-” group in which R can be hydrogen, an alkyl, an alkenyl, an alkynyl, a cycloalkyl, a cycloalkenyl, aryl, heteroaryl, heterocyclyl, cycloalkyl(alkyl), aryl(alkyl), heteroaryl(alkyl) or heterocyclyl(alkyl), as defined herein. An O-carboxy may be substituted or unsubstituted.

[0098] The terms “ester” and “C-carboxy” refer to a “-C(=O)OR” group in which R can be the same as defined with respect to O-carboxy. An ester and C-carboxy may be substituted or unsubstituted.

[0099] A “nitro” group refers to an “-NO2” group.

[0100] A “sulfenyl” group refers to an “-SR” group in which R can be hydrogen, an alkyl, an alkenyl, an alkynyl, a cycloalkyl, a cycloalkenyl, aryl, heteroaryl, heterocyclyl, cycloalkyl(alkyl), aryl(alkyl), heteroaryl(alkyl) or heterocyclyl(alkyl). A sulfenyl may be substituted or unsubstituted.

[0101] A “sulfinyl” group refers to an “-S(=O)-R” group in which R can be the same as defined with respect to sulfenyl. A sulfinyl may be substituted or unsubstituted.

[0102] A “sulfonyl” group refers to an “SO2R” group in which R can be the same as defined with respect to sulfenyl. A sulfonyl may be substituted or unsubstituted.

[0103] As used herein, “haloalkyl” refers to an alkyl group in which one or more of the hydrogen atoms are replaced by a halogen (e.g., mono-haloalkyl, di-haloalkyl, tri- haloalkyl and polyhaloalkyl). Such groups include but are not limited to, chloromethyl, fluoromethyl, difluoromethyl, trifluoromethyl, l-chloro-2-fluoromethyl, 2-fluoroisobutyl and pentafluoroethyl. A haloalkyl may be substituted or unsubstituted.

[0104] As used herein, “haloalkoxy” refers to an alkoxy group in which one or more of the hydrogen atoms are replaced by a halogen (e.g., mono-haloalkoxy, di- haloalkoxy and tri- haloalkoxy). Such groups include but are not limited to, chloromethoxy, fluoromethoxy, difluoromethoxy, trifluoromethoxy, 1 -chloro-2-fluoromethoxy and 2- fluoroisobutoxy. A haloalkoxy may be substituted or unsubstituted.

[0105] The term “amino” as used herein refers to a -NH2 group.

[0106] A “mono-substituted amine” group refers to a “-NHRA” group in which RA can be an alkyl, an alkenyl, an alkynyl, a cycloalkyl, a cycloalkenyl, aryl, heteroaryl, heterocyclyl, cycloalkyl(alkyl), aryl(alkyl), heteroaryl(alkyl) or heterocyclyl(alkyl), as defined herein. The RA may be substituted or unsubstituted. Examples of mono-substituted amino groups include, but are not limited to, -NH(methyl), -NH(phenyl) and the like.

[0107] A “di- substituted amine” group refers to a “-NRARB” group in which RA and RB can be independently an alkyl, an alkenyl, an alkynyl, a cycloalkyl, a cycloalkenyl, aryl, heteroaryl, heterocyclyl, cycloalkyl(alkyl), aryl(alkyl), heteroaryl(alkyl) or heterocyclyl(alkyl), as defined herein. RA and RB can independently be substituted or unsubstituted. Examples of di-substituted amino groups include, but are not limited to, -N(methyl)2, -N(phenyl)(methyl), -N(ethyl)(methyl) and the like.

[0108] As used herein, “amine(alkyl)” group refers to an -(alkylene)-NR’R” radical where R’ and R” are independently hydrogen or alkyl as defined herein. An amine(alkyl) may be substituted or unsubstituted. Examples of amine(alkyl) groups include, but are not limited to, -CH ₂ NH(methyl), -CH2NH(phenyl), -CH2CH2NH(methyl), -CH2CH2NH(phenyl), -CH2N(methyl)2, -CH2N(phenyl)(methyl), -NCH2(ethyl)(methyl), -CH ₂ CH2N(methyl) ₂ , -CH ₂ CH ₂ N(phenyl)(methyl), -NCH ₂ CH ₂ (ethyl)(methyl) and the like.

[0109] Where the number of substituents is not specified (e.g. haloalkyl), there may be one or more substituents present. For example, “haloalkyl” may include one or more of the same or different halogens. As another example, “C1-C3 alkoxyphenyl” may include one or more of the same or different alkoxy groups containing one, two or three atoms.

[0110] As used herein, a radical indicates species with a single, unpaired electron such that the species containing the radical can be covalently bonded to another species. Hence, in this context, a radical is not necessarily a free radical. Rather, a radical indicates a specific portion of a larger molecule. The term “radical” can be used interchangeably with the term “group.”

[0111] The term “pharmaceutically acceptable salt” refers to a salt of a compound that does not cause significant irritation to an organism to which it is administered and does not abrogate the biological activity and properties of the compound. Tn some embodiments, the salt is an acid addition salt of the compound. Pharmaceutical salts can be obtained by reacting a compound with inorganic acids such as hydrohalic acid (e.g., hydrochloric acid or hydrobromic acid), a sulfuric acid, a nitric acid and a phosphoric acid (such as 2,3- dihydroxypropyl dihydrogen phosphate). Pharmaceutical salts can also be obtained by reacting a compound with an organic acid such as aliphatic or aromatic carboxylic or sulfonic acids, for example formic, acetic, succinic, lactic, malic, tartaric, citric, ascorbic, nicotinic, methanesulfonic, ethanesulfonic, p-toluenesulfonic, trifluoroacetic, benzoic, salicylic, 2- oxopentanedioic or naphthalenesulfonic acid. Pharmaceutical salts can also be obtained by reacting a compound with a base to form a salt such as an ammonium salt, an alkali metal salt, such as a sodium, a potassium or a lithium salt, an alkaline earth metal salt, such as a calcium or a magnesium salt, a salt of a carbonate, a salt of a bicarbonate, a salt of organic bases such as dicyclohexylamine, N-methyl-D-glucamine, tris(hydroxymethyl)methylamine, C1-C7 alkylamine, cyclohexylamine, triethanolamine, ethylenediamine and salts with amino acids such as arginine and lysine. For compounds of Formula (I), those skilled in the art understand that when a salt is formed by protonation of a nitrogen-based group (for example, NH2), the nitrogen-based group can be associated with a positive charge (for example, NH2 can become NH ₃ ⁺ ) and the positive charge can be balanced by a negatively charged counterion (such as CF).

[0112] The terms “WEE1 inhibition”, “WEE1 inhibitor” and similar terms as used herein refer to inhibiting the activity or function of a WEE 1 tyrosine kinase, e.g., by degrading WEE1 tyrosine kinase and/or by reducing the activity of WEE 1 tyrosine kinase with regard to mediating phosphorylation of CDK1. A WEE1 inhibitor that functions by degrading WEE1 tyrosine kinase may be referred to herein as a WEE1 degrader.

[0113] It is understood that, in any compound described herein having one or more chiral centers, if an absolute stereochemistry is not expressly indicated, then each center may independently be of R-configuration or S -configuration or a mixture thereof. Thus, the compounds provided herein may be enantiomerically pure, enantiomerically enriched, racemic mixture, diastereomerically pure, diastereomerically enriched or a stereoisomeric mixture. In addition, it is understood that, in any compound described herein having one or more double bond(s) generating geometrical isomers that can be defined as E or Z, each double bond may independently be E or Z a mixture thereof. Likewise, it is understood that, in any compound described, all tautomeric forms arc also intended to be included.

[0114] It is to be understood that where compounds disclosed herein have unfilled valencies, then the valencies are to be filled with hydrogens or isotopes thereof, e.g., hydrogen- 1 (protium) and hydrogen-2 (deuterium). Compounds described herein can also include all isotopes of atoms occurring in the intermediates or final compounds. Isotopes include those atoms having the same atomic number but different mass numbers. For example, isotopes of hydrogen include tritium and deuterium.

[0115] It is understood that the compounds described herein can be labeled isotopically. Substitution with isotopes such as deuterium may afford certain therapeutic advantages resulting from greater metabolic stability, such as, for example, increased in vivo half-life or reduced dosage requirements. Each chemical element as represented in a compound structure may include any isotope of said element. For example, in a compound structure a hydrogen atom may be explicitly disclosed or understood to be present in the compound. At any position of the compound that a hydrogen atom may be present, the hydrogen atom can be any isotope of hydrogen, including but not limited to hydrogen- 1 (protium), hydrogen-2 (deuterium) and hydrogen-3 (tritium). Thus, reference herein to a compound encompasses all potential isotopic forms unless the context clearly dictates otherwise.

[0116] It is understood that the methods and combinations described herein include crystalline forms (also known as polymorphs, which include the different crystal packing arrangements of the same elemental composition of a compound), amorphous phases, salts, solvates and hydrates. In some embodiments, the compounds described herein exist in solvated forms with pharmaceutically acceptable solvents such as water, ethanol or the like. In other embodiments, the compounds described herein exist in unsolvated form. Solvates contain either stoichiometric or non- stoichiometric amounts of a solvent, and may be formed during the process of crystallization with pharmaceutically acceptable solvents such as water, ethanol or the like. Hydrates are formed when the solvent is water or alcoholates are formed when the solvent is alcohol. In addition, the compounds provided herein can exist in unsolvated as well as solvated forms. In general, the solvated forms are considered equivalent to the unsolvated forms for the purposes of the compounds and methods provided herein. [0117] Where a range of values is provided, it is understood that the upper and lower limit, and each intervening value between the upper and lower limit of the range is encompassed within the embodiments.

[0118] Terms and phrases used in this application, and variations thereof, especially in the appended claims, unless otherwise expressly stated, should be construed as open ended as opposed to limiting. As examples of the foregoing, the term ‘including’ should be read to mean ‘including, without limitation,’ ‘including but not limited to,’ or the like; the term ‘comprising’ as used herein is synonymous with ‘including,’ ‘containing,’ or ‘characterized by,’ and is inclusive or open-ended and does not exclude additional, unrecited elements or method steps; the term ‘having’ should be interpreted as ‘having at least;’ the term ‘includes’ should be interpreted as ‘includes but is not limited to;’ the term ‘example’ is used to provide exemplary instances of the item in discussion, not an exhaustive or limiting list thereof; and use of terms like ‘preferably,’ ‘preferred,’ ‘desired,’ or ‘desirable,’ and words of similar- meaning should not be understood as implying that certain features are critical, essential, or even important to the structure or function, but instead as merely intended to highlight alternative or additional features that may or may not be utilized in a particular embodiment. In addition, the term “comprising” is to be interpreted synonymously with the phrases “having at least” or “including at least”. When used in the context of a compound, composition or device, the term “comprising” means that the compound, composition or device includes at least the recited features or components, but may also include additional features or components.

[0119] With respect to the use of substantially any plural and/or singular terms herein, those having skill in the art can translate from the plural to the singular and/or from the singular to the plural as is appropriate to the context and/or application. The various singular/plural permutations may be expressly set forth herein for sake of clarity. The indefinite article “a” or “an” does not exclude a plurality. The mere fact that certain measures are recited in mutually different dependent claims does not indicate that a combination of these measures cannot be used to advantage. Any reference signs in the claims should not be construed as limiting the scope. Compounds

[0120] Some embodiments disclosed herein relate to a compound of Formula (I), or a pharmaceutically acceptable salt thereof, having the structure: wherein R ¹ , R ² , R ³ , R ⁴ , R ^c and R ^D are as described below.

[0121] In various embodiments, R ¹ in Formula (I) can be absent, a substituted or an unsubstituted 6 to 10 membered aryl, -NH-(a substituted or an unsubstituted 6 to 10 membered aryl)-, a substituted or an unsubstituted 5 to 10 membered heteroaryl, -NH-(a substituted or an unsubstituted 5 to 10 membered heteroaryl)- or -NH-(a substituted or an unsubstituted 6 to 10 membered cycloalkyl)-. In an embodiment, R ¹ can be absent. In another embodiment, R ¹ can be a substituted or an unsubstituted 6 to 10 membered aryl. For example, in such an embodiment R ¹ can be an unsubstituted phenyl. In another embodiment, R ¹ can be -NH-(a substituted or an unsubstituted 6 to 10 membered aryl)-. For example, in such an embodiment R ¹ can substituted or an unsubstituted 5 to 10 membered hctcroaryl. In another embodiment, R ¹ can be -NH-(a substituted or an unsubstituted 5 to 10 membered heteroaryl)-. In another embodiment, R ¹ can be a -NH-(a substituted or an unsubstituted 6 to 10 membered cycloalkyl)-. For example, in such an embodiment R ¹ can be

[0122] In various embodiments, R ² in Formula (I) can be absent, a substituted or an unsubstituted 4 to 10 membered heterocyclyl or -O-(a substituted or an unsubstituted 4 to 10 membered heterocyclyl)-. In an embodiment, R ² can be absent. In another embodiment, R ² can be a substituted or an unsubstituted 4 to 10 membered heterocyclyl. For example, in another embodiment, R ² can be In another embodiment, R ² can be a -0-(a substituted or an unsubstituted 4 to 10 membered heterocyclyl)-. For example, in such an embodiment R can

[0123] In various embodiments, R ³ in Formula (I) can be absent, a substituted or an unsubstituted Ci-Ce alkylene, -NH-, -NH-(a substituted or an unsubstituted Ci-Ce alkylene)-, -O-(a substituted or an unsubstituted Ci-Ce alkylene)-NH- or -O-(a substituted or an unsubstituted Ci-Ce alkylene)-NH-(a substituted or an unsubstituted Ci-Ce alkylene)-. In an embodiment, R ³ can be absent. In another embodiment, R ³ can be a substituted or an unsubstituted Ci-Ce alkylene. For example, in such an embodiment R ³ can be -(CH2)-, - (CH2CH2)- or -(CH2CH2CH2)-. In another embodiment, R ³ can be -(CH2CHOH)-. In another embodiment, R ³ is -NH-. In another embodiment, R ³ can be -NH-(a substituted or an unsubstituted Ci-Ce alkylene)-. For example, in such an embodiment R ³ can be -NH- (CH ₂ )-. In another embodiment, R ³ can be -NH-(CH ₂ CH ₂ )-, -NH-(CH ₂ CH ₂ CH ₂ )- or -NH- (CH ₂ CH ₂ CH ₂ CH ₂ )-. In another embodiment, R ³ can be a -O-(a substituted or an unsubstituted Ci-Ce alkylcnc)-NH-. In another embodiment, R ³ can be -O-(CH ₂ )4-NH- In another embodiment, R ³ can be a -O-(a substituted or an unsubstituted C1-C6 alkylene)-NH- (a substituted or an unsubstituted Ci-Ce alkylene)-. For example, in such an embodiment R ³ can be -O-(CH ₂ ) ₂ -NH-(CH ₂ )-.

[0124] In various embodiments, R ⁴ in Formula (I) can be absent or a substituted or an unsubstituted 4 to 10 membered heterocyclyl. In an embodiment, R ⁴ can be absent. In another embodiment, R ⁴ can be an unsubstituted 4 to 10 membered heterocyclyl. For example, in such an embodiment R ⁴ can be

[0125] In various embodiments, at least two of R ¹ , R ² , R ³ and R ⁴ in Formula (I) are present. In some embodiments, at least three of R ¹ , R ² , R ³ and R ⁴ are present. In some embodiments, all four of R ¹ , R ² , R ³ and R ⁴ are present. [0126] In various embodiments, R ^c in Formula (I) can be , wherein Y is -CH or N (nitrogen). In some embodiments R ^c in Formula (I) can be and Y is -CH. In some embodiments R ^c in Formula (I) can be and Y is N (nitrogen). In some embodiments R ^c in Formula (I) can

-CH. In some embodiments R ^c in Formula (I) can (nitrogen).

[0127] In various embodiments, R ^D can be a substituted or an unsubstituted C1-C6 alkyl. In various embodiments, R ^D can be a substituted Ci-Ce alkyl. In various embodiments, R ^D can be an unsubstituted Ci-Ce alkyl. In various embodiments, R ^D can be a halogensubstituted Ci-C ₆ alkyl. In various embodiments, R ^D can be an ethyl. In various embodiments, R ^D can be a methyl. In various embodiments, R ^D can be a halogen-substituted ethyl. In various embodiments, R ^D can be a halogen-substituted methyl.

[0128] Various embodiments provide a compound (e.g., a compound of Formula (I)), or a pharmaceutically acceptable salt thereof, wherein the compound has a structure selected from those of Compound Nos. 1 to 42 as listed in the following Table A:

Table A Synthcsis

[0129] Compounds of the Formula (I), or pharmaceutically acceptable salts thereof, can be made in various ways by those skilled using known techniques as guided by the detailed teachings provided herein, including the Examples provided below. For example, in an embodiment, compounds of the Formula (I) are prepared in accordance with the general schemes illustrated in FIGS. 1-52. Any preliminary reaction steps required to form starting compounds or other precursors, can be carried out by those skilled in the art, for example by appropriate adjustment of the reagents and conditions described in the Examples. In FIGS. 1-52, the variables including R ¹ , R ² , R ³ , R ⁴ , R ^c and R ^D can be as described elsewhere herein, taking into consideration the synthetic conversions involved as understood by those of skill in the art. R ⁵ , R ⁶ , R ⁷ and R ⁸ are understood by those of skill in the art to be synthetic precursors of R ³ and R ⁴ , as further illustrated in the Examples below.

Pharmaceutical Compositions

[0130] Some embodiments described herein relate to a pharmaceutical composition, which can include an effective amount of one or more compounds described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) and a pharmaceutically acceptable carrier, diluent, excipient or combination thereof.

[0131] The term “pharmaceutical composition” refers to a mixture of one or more compounds and/or salts disclosed herein with other chemical components, such as diluents or carriers. The pharmaceutical composition facilitates administration of the compound to an organism. Pharmaceutical compositions can also be obtained by reacting compounds with inorganic or organic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, and salicylic acid. Pharmaceutical compositions will generally be tailored to the specific intended route of administration.

[0132] The term “physiologically acceptable” defines a carrier, diluent or excipient that does not abrogate the biological activity and properties of the compound nor cause appreciable damage or injury to an animal to which delivery of the composition is intended. [0133] As used herein, a “carrier” refers to a compound that facilitates the incorporation of a compound into cells or tissues. For example, without limitation, dimethyl sulfoxide (DMSO) is a commonly utilized carrier that facilitates the uptake of many organic compounds into cells or tissues of a subject.

[0134] As used herein, a “diluent” refers to an ingredient in a pharmaceutical composition that lacks appreciable pharmacological activity but may be pharmaceutically necessary or desirable. For example, a diluent may be used to increase the bulk of a potent drug whose mass is too small for manufacture and/or administration. It may also be a liquid for the dissolution of a drug to be administered by injection, ingestion or inhalation. A common form of diluent in the art is a buffered aqueous solution such as, without limitation, phosphate buffered saline that mimics the pH and isotonicity of human blood.

[0135] As used herein, an “excipient” refers to an essentially inert substance that is added to a pharmaceutical composition to provide, without limitation, bulk, consistency, stability, binding ability, lubrication, disintegrating ability etc., to the composition. For example, stabilizers such as anti-oxidants and metal-chelating agents are excipients. In an embodiment, the pharmaceutical composition comprises an anti-oxidant and/or a metalchelating agent. A “diluent” is a type of excipient.

[0136] The pharmaceutical compositions described herein can be administered to a human patient per se, or in pharmaceutical compositions where they are mixed with other active ingredients, as in combination therapy, or carriers, diluents, excipients or combinations thereof. Proper formulation is dependent upon the route of administration chosen. Techniques for formulation and administration of the compounds described herein are known to those skilled in the art.

[0137] The pharmaceutical compositions disclosed herein may be manufactured in a manner that is itself known, e.g., by means of conventional mixing, dissolving, granulating, dragee-making, levigating, emulsifying, encapsulating, entrapping or tableting processes. Additionally, the active ingredients are contained in an amount effective to achieve its intended purpose. Many of the compounds used in the pharmaceutical combinations disclosed herein may be provided as salts with pharmaceutically compatible counterions.

[0138] Multiple techniques of administering a compound, salt and/or composition exist in the art including, but not limited to, oral, rectal, pulmonary, topical, aerosol, injection, infusion and parenteral delivery, including intramuscular, subcutaneous, intravenous, intramedullary injections, intrathecal, direct intraventricular, intraperitoneal, intranasal and intraocular injections. In some embodiments, a compound of Formula (I), or a pharmaceutically acceptable salt thereof, can be administered orally.

[0139] One may also administer the compound, salt and/or composition in a local rather than systemic manner, for example, via injection or implantation of the compound directly into the affected area, often in a depot or sustained release formulation. Furthermore, one may administer the compound in a targeted drug delivery system, for example, in a liposome coated with a tissue-specific antibody. The liposomes will be targeted to and taken up selectively by the organ. For example, intranasal or pulmonary delivery to target a respiratory disease or condition may be desirable.

[0140] The compositions may, if desired, be presented in a pack or dispenser device which may contain one or more unit dosage forms containing the active ingredient. The pack may for example comprise metal or plastic foil, such as a blister pack. The pack or dispenser device may be accompanied by instructions for administration. The pack or dispenser may also be accompanied with a notice associated with the container in form prescribed by a governmental agency regulating the manufacture, use, or sale of pharmaceuticals, which notice is reflective of approval by the agency of the form of the drug for human or veterinary administration. Such notice, for example, may be the labeling approved by the U.S. Food and Drug Administration for prescription drugs, or the approved product insert. Compositions that can include a compound and/or salt described herein formulated in a compatible pharmaceutical carrier may also be prepared, placed in an appropriate container, and labeled for treatment of an indicated condition.

Uses and Methods of Treatment

[0141] Some embodiments described herein relate to a method for ameliorating and/or treating a cancer described herein that can include administering an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) to a subject having a cancer described herein. Other embodiments described herein relate to the use of an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) in the manufacture of a medicament for ameliorating and/or treating a cancer described herein. Still other embodiments described herein relate to an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) for ameliorating and/or treating a cancer described herein.

[0142] Some embodiments described herein relate to a method for inhibiting replication of a malignant growth or a tumor that can include contacting the growth or the tumor with an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof), wherein the malignant growth or tumor is due to a cancer described herein. Other embodiments described herein relate to the use of an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) in the manufacture of a medicament for inhibiting replication of a malignant growth or a tumor, wherein the malignant growth or tumor is due to a cancer described herein. Still other embodiments described herein relate to an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) for inhibiting replication of a malignant growth or a tumor, wherein the malignant growth or tumor is due to a cancer described herein.

[0143] Some embodiments described herein relate to a method for ameliorating or treating a cancer described herein that can include contacting a malignant growth or a tumor with an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) to a subject having a cancer described herein. Other embodiments described herein relate to the use of an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) in the manufacture of a medicament for ameliorating or treating a cancer that can include contacting a malignant growth or a tumor, wherein the malignant growth or tumor is due to a cancer described herein. Still other embodiments described herein relate to an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) for ameliorating or treating a cancer that can include contacting a malignant growth or a tumor, wherein the malignant growth or tumor is due to a cancer described herein.

[0144] Some embodiments described herein relate to a method for inhibiting the activity of WEE1 (for example, inhibiting the activity of WEE1 in TP53-mutated cells, inhibiting the activity of WEE1 in TP53 wild-type cells, inhibiting the activity in WEE1 p53- deficient cells and/or decreasing the overexpression of WEE1 in cells) that can include providing an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) to a cancer cell from a cancer described herein. Other embodiments described herein relate to the use of an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) in the manufacture of a medicament for inhibiting the activity of WEE1 (for example, inhibiting the activity of WEE1 in TP53- mutated cells, inhibiting the activity of WEE1 in TP53 wild-type cells, inhibiting the activity in WEE1 p53-dcficicnt cells and/or decreasing the overexpression of WEE1 in cells). Still other embodiments described herein relate to an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) for inhibiting the activity of WEE 1 (for example, inhibiting the activity of WEE 1 in TP53-mutated cells, inhibiting the activity of WEE1 in TP53 wild-type cells, inhibiting the activity in WEE1 p53-deficient cells and/or decreasing the overexpression of WEE 1 in cells). Some embodiments described herein relate to a method for inhibiting the activity of WEE 1 (for example, inhibiting the activity of WEE 1 in TP53-mutated cells, inhibiting the activity of WEE1 in TP53 wild-type cells, inhibiting the activity in WEE1 p53-deficient cells and/or decreasing the overexpression of WEE1 in cells) that can include providing an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) to a cancer cell from a cancer described herein. Other embodiments described herein relate to a method for inhibiting the activity of WEE 1 (for example, inhibiting the activity of WEE 1 in TP53-mutated cells, inhibiting the activity of WEE1 in TP53 wild-type cells, inhibiting the activity in WEE1 p53-deficient cells and/or decreasing the overexpression of WEE1 in cells) that can include contacting a cancer cell from a cancer described herein with an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof), and thereby inhibiting the activity of WEE E

[0145] Some embodiments described herein relate to a method for ameliorating or treating a cancer described herein that can include inhibiting the activity of WEE1 (for example, inhibiting the activity of WEE 1 in TP53-mutated cells, inhibiting the activity of WEE1 in TP53 wild-type cells, inhibiting the activity in WEE1 p53-deficient cells and/or decreasing the overexpression of WEE1 in cells) using an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof). Other embodiments described herein relate to the use of an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) in the manufacture of a medicament for ameliorating or treating a cancer described herein by inhibiting the activity of WEE1 (for example, inhibiting the activity of WEE1 in TP53-mutated cells, inhibiting the activity of WEE1 in TP53 wild-type cells, inhibiting the activity in WEE1 p53-deficient cells and/or decreasing the overexpression of WEE 1 in cells). Still other embodiments described herein relate to an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) for ameliorating or treating a cancer described herein by inhibiting the activity of WEE1 (for example, inhibiting the activity of WEE1 in TP53-mutated cells, inhibiting the activity of WEE1 in TP53 wild-type cells, inhibiting the activity in WEE1 p53- deficient cells and/or decreasing the overexpression of WEE 1 in cells). Some embodiments described herein relate to a method for ameliorating or treating a cancer described herein that can include contacting a cancer cell with an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof), wherein the compound inhibits the activity of WEE1 (for example, inhibiting the activity of WEE1 in TP53-mutated cells, inhibiting the activity of WEE1 in TP53 wild-type cells, inhibiting the activity in WEE1 p53-deficient cells and/or decreasing the overexpression of WEE 1 in cells).

[0146] Some embodiments disclosed herein relate to a method for inhibiting the activity of WEE1 that can include providing an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) to a subject having a cancer described herein or a cancer cell from a cancer described herein. Other embodiments disclosed herein relate to the use of an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) in the manufacture of a medicament for inhibiting the activity of WEE1. Still other embodiments disclosed herein relate to a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) for inhibiting the activity of WEE 1.

[0147] Examples of suitable cancers include, but are not limited to: brain cancers, cervicocerebral cancers, esophageal cancers, thyroid cancers, small cell cancers, non-small cell cancers, breast cancers, lung cancers (for example non-small cell lung cancer and small cell lung cancer), stomach cancers, gallbladder/bile duct cancers, liver cancers, pancreatic cancers, colon cancers, rectal cancers, ovarian cancers, choriocarcinomas, uterus body cancers, uterocervical cancers, renal pelvis/ureter cancers, bladder cancers, prostate cancers, penis cancers, testicular cancers, fetal cancers, Wilms’ cancer, skin cancers, malignant melanoma, neuroblastomas, osteosarcomas, Ewing's tumors, soft part sarcomas, acute leukemia, chronic lymphatic leukemias, chronic myelocytic leukemias, polycythemia vera, malignant lymphomas, multiple myeloma, Hodgkin's lymphomas and non-Hodgkin’s lymphomas.

[0148] As described herein, a cancer can become resistant to one or more anticancer agents. In some embodiments, a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) can be used to treat and/or ameliorate a cancer that has become resistant to one or more anti-cancer agents (such as one or more WEE1 inhibitors). Examples of anti-cancer agents that a subject may have developed resistance to include, but are not limited to, WEE1 inhibitors (such as AZDI 775). In some embodiments, the cancer that has become resistant to one or more anti-canccr agents can be a cancer described herein.

[0149] Several known WEE1 inhibitors can cause one or more undesirable side effects in the subject being treated. Examples of undesirable side effects include, but are not limited to, thrombocytopenia, neutropenia, anemia, diarrhea, vomiting, nausea, abdominal pain, and constipation. In some embodiments, a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) can decrease the number and/or severity of one or more side effects associated with a known WEE1 inhibitor. In some embodiments, a compound of Formula (I), or a pharmaceutically acceptable salt thereof, can result in a severity of a side effect (such as one of those described herein) that is 25% less than compared to the severity of the same side effect experienced by a subject receiving a known WEE1 inhibitor (such as AZD1775, formally known as MK1775 (CAS No.: 955365-80-7, 2-allyl-l-(6-(2-hydroxypropan-2-yl)pyridin-2-yl)-6-(4-(4- methylpiperazin-l-yl)phenylamino)-l,2-dihydropyrazolo[3,4-d] pyrimidin-3-one)). In some embodiments, a compound of Formula (I), or a pharmaceutically acceptable salt thereof, results in a number of side effects that is 25% less than compared to the number of side effects experienced by a subject receiving a known WEE1 inhibitor (for example, AZD1775). In some embodiments, a compound of Formula (I), or a pharmaceutically acceptable salt thereof, results in a severity of a side effect (such as one of those described herein) that is less in the range of about 10% to about 30% compared to the severity of the same side effect experienced by a subject receiving a known WEE1 inhibitor (such as AZD1775). In some embodiments, a compound of Formula (I), or a pharmaceutically acceptable salt thereof, results in a number of side effects that is in the range of about 10% to about 30% less than compared to the number of side effects experienced by a subject receiving a known WEE1 inhibitor (for example, AZD1775).

[0150] The one or more compounds of Formula (I), or a pharmaceutically acceptable salt thereof, that can be used to treat, ameliorate and/or inhibit the growth of a cancer wherein inhibiting the activity of WEE1 is beneficial is provided in any of the embodiments described under the heading titled “Compounds” above. [0151] As used herein, a “subject” refers to an animal that is the object of treatment, observation or experiment. “Animal” includes cold- and warm-blooded vertebrates and invertebrates such as fish, shellfish, reptiles and, in particular, mammals. “Mammal” includes, without limitation, mice, rats, rabbits, guinea pigs, dogs, cats, sheep, goats, cows, horses, primates, such as monkeys, chimpanzees, and apes, and, in particular, humans. In some embodiments, the subject can be human. In some embodiments, the subject can be a child and/or an infant, for example, a child or infant with a fever. In other embodiments, the subject can be an adult.

[0152] As used herein, the terms “treat,” “treating,” “treatment,” “therapeutic,” and “therapy” do not necessarily mean total cure or abolition of the disease or condition. Any alleviation of any undesired signs or symptoms of the disease or condition, to any extent can be considered treatment and/or therapy. Furthermore, treatment may include acts that may worsen the subject’s overall feeling of well-being or appearance.

[0153] The terms “therapeutically effective amount” and “effective amount” are used to indicate an amount of an active compound, or pharmaceutical agent, which elicits the biological or medicinal response indicated. For example, a therapeutically effective amount of compound, salt or composition can be the amount needed to prevent, alleviate or ameliorate symptoms of the disease or condition, or prolong the survival of the subject being treated. This response may occur in a tissue, system, animal or human and includes alleviation of the signs or symptoms of the disease or condition being treated. Determination of an effective amount is well within the capability of those skilled in the art, in view of the disclosure provided herein. The therapeutically effective amount of the compounds disclosed herein required as a dose will depend on the route of administration, the type of animal, including human, being treated and the physical characteristics of the specific animal under consideration. The dose can be tailored to achieve a desired effect, but will depend on such factors as weight, diet, concurrent medication and other factors which those skilled in the medical ai ts will recognize.

[0154] For example, an effective amount of a compound, or radiation, is the amount that results in: (a) the reduction, alleviation or disappearance of one or more symptoms caused by the cancer, (b) the reduction of tumor size, (c) the elimination of the tumor, and/or (d) long-term disease stabilization (growth arrest) of the tumor. In the treatment of lung cancer (such as non-small cell lung cancer) a therapeutically effective amount is that amount that alleviates or eliminates cough, shortness of breath and/or pain. As another example, an effective amount, or a therapeutically effective amount of an WEE1 inhibitor and/or degrader is the amount which results in the reduction in WEE1 activity and/or phosphorylation (such as phosphorylation of CDC2). The reduction in WEE1 activity is known to those skilled in the art and can be determined by the analysis of WEE1 intrinsic kinase activity and downstream substrate phosphorylation.

[0155] The amount of the compound of Formula (I), or a pharmaceutically acceptable salt thereof, required for use in treatment will vary not only with the particular compound or salt selected but also with the route of administration, the nature and/or symptoms of the disease or condition being treated and the age and condition of the patient and will be ultimately at the discretion of the attendant physician or clinician. In cases of administration of a pharmaceutically acceptable salt, dosages may be calculated as the free base. As will be understood by those of skill in the ail, in certain situations it may be necessary to administer the compounds disclosed herein in amounts that exceed, or even far exceed, the dosage ranges described herein in order to effectively and aggressively treat particularly aggressive diseases or conditions.

[0156] In general, however, a suitable dose will often be in the range of from about 0.05 mg/kg to about 10 mg/kg. For example, a suitable dose may be in the range from about 0.10 mg/kg to about 7.5 mg/kg of body weight per day, such as about 0.15 mg/kg to about 5.0 mg/kg of body weight of the recipient per day, about 0.2 mg/kg to 4.0 mg/kg of body weight of the recipient per day, or any amount in between. The compound may be administered in unit dosage form; for example, containing 1 to 500 mg, 10 to 100 mg, 5 to 50 mg or any amount in between, of active ingredient per unit dosage form.

[0157] The desired dose may conveniently be presented in a single dose or as divided doses administered at appropriate intervals, for example, as two, three, four or more sub-doses per day. The sub-dose itself may be further divided, e.g., into a number of discrete loosely spaced administrations.

[0158] As will be readily apparent to one skilled in the art, the useful in vivo dosage to be administered and the particular mode of administration will vary depending upon the age, weight, the severity of the affliction, the mammalian species treated, the particular compounds employed and the specific use for which these compounds are employed. The determination of effective dosage levels, which is the dosage levels necessary to achieve the desired result, can be accomplished by one skilled in the art using routine methods, for example, human clinical trials, in vivo studies and in vitro studies. For example, useful dosages of a compound of Formula (I), or pharmaceutically acceptable salts thereof, can be determined by comparing their in vitro activity, and in vivo activity in animal models. Such comparison can be done by comparison against an established drug, such as cisplatin and/or gemcitabine)

[0159] Dosage amount and interval may be adjusted individually to provide plasma levels of the active moiety which are sufficient to maintain the modulating effects, or minimal effective concentration (MEC). The MEC will vary for each compound but can be estimated from in vivo and/or in vitro data. Dosages necessary to achieve the MEC will depend on individual characteristics and route of administration. However, HPLC assays or bioassays can be used to determine plasma concentrations. Dosage intervals can also be determined using MEC value. Compositions should be administered using a regimen which maintains plasma levels above the MEC for 10-90% of the time, preferably between 30-90% and most preferably between 50-90%. In cases of local administration or selective uptake, the effective local concentration of the drug may not be related to plasma concentration.

[0160] It should be noted that the attending physician would know how to and when to terminate, interrupt or adjust administration due to toxicity or organ dysfunctions. Conversely, the attending physician would also know to adjust treatment to higher levels if the clinical response were not adequate (precluding toxicity). The magnitude of an administrated dose in the management of the disorder of interest will vary with the severity of the disease or condition to be treated and to the route of administration. The severity of the disease or condition may, for example, be evaluated, in part, by standard prognostic evaluation methods. Further, the dose and perhaps dose frequency, will also vary according to the age, body weight and response of the individual patient. A program comparable to that discussed above may be used in veterinary medicine.

[0161] Compounds, salts and compositions disclosed herein can be evaluated for efficacy and toxicity using known methods. For example, the toxicology of a particular compound, or of a subset of the compounds, sharing certain chemical moieties, may be established by determining in vitro toxicity towards a cell line, such as a mammalian, and preferably human, cell line. The results of such studies arc often predictive of toxicity in animals, such as mammals, or more specifically, humans. Alternatively, the toxicity of particular compounds in an animal model, such as mice, rats, rabbits, dogs or monkeys, may be determined using known methods. The efficacy of a particular compound may be established using several recognized methods, such as in vitro methods, animal models, or human clinical trials. When selecting a model to determine efficacy, the skilled artisan can be guided by the state of the ait to choose an appropriate model, dose, route of administration and/or regime.

EXAMPLES

[0162] Additional embodiments are disclosed in further detail in the following examples, which are not in any way intended to limit the scope of the claims.

Intermediate 1

3-((4-(4-(4-aminobenzyl)piperazin- l-yl)phenyl)amino)piperidine-2, 6-dione (FIG. 4)

[0163] Step 1: To a mixture of 3-(4-piperazin-l-ylanilino)piperidine-2, 6-dione (Reference: WO 2021/083949) (0.200 g, 497 pmol, TFA salt) in DCM (2 mL) were added tert-butyl (4-formylphenyl) carbamate (132 mg, 596 pmol), NaBH(OAc)3 (211mg, 994 pmol) and DIEA (321 mg, 2.49 mmol) at 0 °C. The mixture was stirred at 20 °C for 12 h under N2. LCMS showed the reaction was completed and the desired product was detected. The mixture was poured into H2O (10 mL). The mixture was extracted with DCM (3 x 10 mL). The organic phase was washed with brine (10 mL), dried over anhydrous Na S04, filtrated, and the filtrate was concentrated under reduced pressure to give the crude product. The crude product was purified by prep-TLC (MeOH:EA = 1: 10, Rf = 0.5) to afford tert-butyl A-[4-[[4-[4-[(2,6-dioxo-3-piperidyl)amino] phenyl]piperazin- l-yl]methyl]phenyl] carbamate (Boc protected Intermediate 1) (0.120 g, 48% yield, 98% purity). LCMS (ESI ⁺ ) m/z 494.2 [M+H] ⁺ , RT: 0.481 min. NMR (400 MHz, DMSO-d ₆ ) 5 10.75 (br s, 1 H, NH), 9.29 (s, 1 H, NH), 7.39 (br d, J = 8.4 Hz, 2H), 7.18 (d, J = 8.4 Hz, 2H), 6.73 (d, J = 8.9 Hz, 2H), 6.60 (d, J = 8.9 Hz. 2H), 5.36 (br d, J = 7.1 Hz, 1H, NH), 4.23 - 4.14 (m, 1H). 3.41 (s, 2H), 2.90 - 2.97 (m, 4H), 2.75 - 2.65 (m, 1H), 2.63 - 2.53 (m, 1H), 2.50 - 2.43 (m, 4H), 2.16 - 2.04 (m, 1H), 1.80 - 1.65 (m, 1H), 1.47 (s, 9H).

[0164] Step 2: A mixture of Boc protected Intermediate 1 (0.120 g, 238 pmol, 98% purity) in TFA (0.2 mL) and DCM (2 mL) was stirred at 20 °C for 12 h under N2. LCMS showed the reaction was completed and the desired product was detected. The mixture was poured into NaHCCh solution (10 mL). The mixture was extracted with DCM (3 x 10 mL). The organic phase was washed with brine (10 mL), dried over anhydrous Na SCL and filtrated. The filtrate was concentrated under reduced pressure to give Intermediate 1, 3-[4-[4-[(4- aminophenyl)methyl]piperazin-l-yl]anilino]piperidine-2, 6-dione (80 mg, 71% yield, 84% purity). LCMS (ESI ⁺ ) m/z 394.2 [M+H] ⁺ , RT: 0.688 min.

Intermediate 2 3-((4-(4-(4-aminophenethyl)piperazin- 1 -yl)phenyl)amino)piperidine-2, 6-dione (FIG. 5)

[0165] Step 1: Boc protected Intermediate 2 was prepared following the procedure described in Step 1 for Intermediate 1 using tert-butyl (4-(2- oxoethyl)phenyl)carbamate in place of tert-butyl (4-formylphenyl)carbamate in yield 57% with purity 95%. LCMS (ESI ⁺ ) m/z 508.3 [M+H] ⁺ , RT: 0.554 min. ’ H NMR (400 MHz, DMSO-d ₆ ) 5 10.75 (s, 1H, NH), 9.21 (s, 1H, NH). 7.34 (br d. J= 8.4 Hz, 2H), 7.10 (d, J= 8.4 Hz, 2H), 6.75 (d, J = 8.9 Hz, 2H), 6.61 (d, J = 8.9 Hz, 2H), 5.37 (d, J = 7.3 Hz, 1H, NH), 4.19 (ddd, J= 11.2, 6.7, 4.9 Hz, 1H), 2.98 - 2.89 (m, 4H), 2.78 - 2.71 (m, 1H), 2.70 - 2.64 (m, 2H), 2.62 - 2.57 (m, 1H), 2.57 - 2.53 (m. 4H). 2.54 - 2.50 (m, 2H), 2.15 - 2.06 (m, 1H), 1.88 - 1.77 (m, 1H), 1.46 (s, 9H). [0166] Step 2: Intermediate 2 was prepared following the procedure described in Step 2 for Intermediate 1. The isolated yield 73% with 95% purity was obtained. LCMS (ESI ⁺ ) m/z 408.2 [M-H] ⁺ . RT: 0.149 min. ’ H NMR (400 MHz, DMSO-d ₆ ) 8 10.74 (s, 1H, NH), 6.87 (d, J = 8.3 Hz, 2H), 6.75 (d, J= 8.9 Hz, 2H), 6.61 (d, J = 8.9 Hz, 2H), 6.48 (d, J = 8.3 Hz, 2H), 5.36 (d, J = 7.3 Hz, 1H), 4.80 (br s, 2H), 4.19 (ddd, J =11.3, 7.0, 4.8 Hz, 1H), 2.99 - 2.88 (m. 4H), 2.78 - 2.68 (m. 1H). 2.62 - 2.52 (m, 8H), 2.47 - 2.44 (m, 1H), 2.15 - 2.07 (m, 1H), 1.89 - 1.77 (m, 1H).

Intermediate 3 3-((4-(4-(4-aminophenyl)piperazin-l-yl)phenyl)amino)piperidi ne-2, 6-dione (FIG. 6)

[0167] Step 1: To a solution of 3-(4-piperazin-l-ylanilino)piperidine-2, 6-dione (300 mg, 1.04 mmol) in DMF (6 mL) were added l-fluoro-4-nitrobenzene (117 mg, 832 pmol) and DIEA (672 mg, 5.20 mmol) at 20 °C. The reaction was stirred at 80 °C for 12 h. LCMS showed the starting material was consumed and the desired product was detected. The mixture was diluted with water (10 mL) and extracted with ethyl acetate (EA) (3 x 10 mL). The organic combined organic layers were washed with brine (3 x 10 mL) and dried over NaiSCU. After filtered, the filtrate was concentrated under reduced pressure to give 3-[4-[4- (4-nitrophenyl)piperazin-l-yl]anilino]piperidine-2, 6-dione (140 mg, 31% yield, 95% purity). LCMS (ESI ⁺ ) m/z 410.1 [M+H] ⁺ , RT: 0.483 min. ¹ H NMR (400 MHz, DMSO-d ₆ ) 8 10.76 (s, 1H, NH). 8.07 (d, 7= 9.4 Hz. 2H), 7.09 (d, 7 = 9.4 Hz, 2H), 6.82 (d. 7 = 8.8 Hz, 2H), 6.64 (d, 7 = 8.8 Hz, 2H), 5.45 (d, 7 = 7.3 Hz, 1H, NH), 4.21 (ddd, 7 = 11.2, 6.8, 4.8 Hz, 1H), 3.64 - 3.55 (m, 4H), 3.10 - 3.02 (m, 4H), 2.73 (ddd, 7 = 17.4, 11.8, 5.6 Hz, 1H), 2.57 (ddd, 7 = 17.6, 4.4, 4.0 Hz, 1H), 2.11 (ddd, 7 = 12.9, 8.8, 4.4 Hz, 1H), 1.85 (ddd, 7 = 24.1, 11.5, 4.3 Hz, 1H).

[0168] Step 2: To a solution of 3-[4-[4-(4-nitrophenyl)piperazin-l- yl]anilino]piperidine-2, 6-dione (from Step 1) (140 mg, 324 pmol, 95% purity) in TFE (3 mL) was added Pd/C (40.2 mg, 10% on carbon) at 25 °C. The reaction was stirred at 25 °C under Hi atmosphere (15 psi) for 12 h. LCMS showed the reaction was completed and the desired compound was detected. The mixture was filtered and the filter cake was washed with TFE (20 mL). The collected filtrate was concentrated to give Intermediate 3, 3-[4-[4-(4- aminophenyl) piperazin-l-yl]anilino]piperidine-2, 6-dione (70.0 mg, 54% yield, 94.91% purity). LCMS (ESI ⁺ ) ni/z 380.2 [M+H] ⁺ , RT: 0.559 min. NMR (400 MHz, DMSO-d ₆ ) 8 = 10.76 (br s, 1H, NH). 6.80 (br d. J = 8.8 Hz, 2H), 6.73 (d. J = 8.8 Hz, 2H), 6.63 (br d, J = 8.8 Hz, 2H), 6.51 (d, J - 8.8 Hz, 2H), 5.40 (d, J - 7.3 Hz, 1H, NH), 4.58 (br s, 2H, NH ₂ ), 4.20 (ddd, J = 11.2, 6.8, 5.0 Hz, 1H), 3.10 - 3.00 (m, 8H). 2.73 (ddd. J = 17.4, 11.8, 5.6 Hz, 1H), 2.58 (ddd, J = 17.4, 4.4, 4.4 Hz, 1H), 2.11 (ddd, J = 12.9, 8.8 Hz, 1H), 1.84 (ddd, J = 24.1, 12.0, 4.5 Hz, 1H).

Intermediate 4 3-((4-(4-(3-(4-aminophenyl)propyl)piperazin-l-yl)phenyl)amin o)piperidine-2, 6-dione (FIG. 7)

[0169] Step 1 : Boc protected Intermediate 4 was prepared following the procedure described in Step 1 for Intermediate 1 using tert-butyl (4-(3- oxopropyl)phenyl)carbamate (Reference: WO 2012/177061, CN 103649199 A) in place of tert-butyl (4-formylphenyl)carbamate. The reaction was directly purified by prep-TLC (EA: MeOH = 10:1, Rf = 0.4) to give Boc-protected Intermediate 4 (0.18 g, 60% yield, 95% purity). LCMS (ESI ⁺ ) m/z 522.3 [M+H] ⁺ . RT: 0.578 min. ¹ H NMR (400 MHz, DMSO-d ₆ ) 8 10.75 (s, 1H, NH), 9.22 (br s, 1H, NH), 7.35 (br d, J - 8.1 Hz, 2H), 7.09 (br d, J - 8.1 Hz, 2H), 6.76 (br d, J = 8.6 Hz. 2H), 6.61 (br d. J = 8.6 Hz, 2H), 5.41 (br s, 1H), 4.19 (ddd, J = 10.2, 6.8, 4.8 Hz, 1H), 3.04 - 2.82 (m, 4H), 2.79 - 2.65 (m, 3H), 2.63 - 2.57 (m, 1H), 2.57 - 2.48 (m, 6H), 2.15 - 2.05 (m, 1H), 1.89 - 1.64 (m, 3H), 1.46 (s, 9H).

[0170] Step 2: Intermediate 4 was prepared following the procedure described in Step 2 for Intermediate 1. The mixture was concentrated under reduced pressure at 40 °C to give a product. The product was used to next step without further purification. Intermediate 4, 3-((4-(4-(3-(4-aminophenyl)propyl)piperazin-l-yl)phenyl)amin o)piperidine-2, 6-dione, was obtained (81.0 mg, 192 pmol, 56% yield, 95% purity). LCMS (ESI ⁺ ) m/z 422.2 [M+H] ⁺ , RT: 0.152 min. ’ H NMR (400 MHz, DMSO-d ₆ ) 8 10.75 (s, 1H. NH), 6.84 (d. J = 8.2 Hz, 2H), 6.74 (d, J = 8.9 Hz, 2H), 6.60 (d, J = 8.9 Hz, 2H), 6.47 (d, J = 8.2 Hz, 2H), 5.36 (d, J = 7.2 Hz, 1H), 4.79 (br s, 2H, NH ₂ ), 4.18 (ddd, J = 11.3, 6.9, 4.9 Hz, 1H), 3.00 - 2.86 (m, 4H), 2.72 (ddd, J = 17.5, 11.8. 5.6 Hz, 1H). 2.57 (ddd. J = 17.5, 4.2, 4.2 Hz. 1H). 2.47 (br s. 4H). 2.42 (br t, J - 7.6 Hz, 2H), 2.35 - 2.24 (m, 2H), 2.10 (ddd, J = 13.3, 8.7, 4.4 Hz, 1H), 1.83 (ddd, J = 24.2, 11.5, 4.4 Hz, 1H), 1.66 (ddd, J = 14.5, 7.4, 7.4 Hz, 2H).

Intermediate 5 tert-Butyl (4-(4-(3-aminophenyl)piperazin-l-yl)phenyl)carbamate (FIG. 8)

[0171] Step 1: To a mixture of l-bromo-3-nitrobenzene (0.500 g, 2.48 mmol) in toluene (8 mL) were added tert-butyl /V-(4-pipcrazin- l-ylphcnyl)carbamatc (Reference: WO 2001/97810, page 41, 824 mg, 2.97 mmol), BINAP (154 mg, 248 pmol), Pd(OAc) ₂ (55.6 mg, 248 pmol) and Cs ₂ COs (1.61 g, 4.95 mmol) at 20 °C. The reaction was stirred at 100 °C for 12 h under N ₂ . LCMS showed that the starting material was consumed and the desired product was detected. The mixture was poured into H ₂ O (10 mL). The mixture was extracted with DCM (3 x 10 mL). The organic phase was washed with brine (5 x 10 mL), dried over anhydrous Na ₂ SO4 and filtrated. The filtrate was concentrated under reduced pressure to give crude product. The crude product was purified by prep-TLC (PE: EA = 0: 1, Rf = 0.5) to give tert-butyl A-[4-[4-(3-nitrophenyl)piperazin-l-yl]phenyl]carbamate, the precursor of Intermediate 5 (0.700 g. 54% yield, 76% purity). LCMS (ESI ⁺ ) m/z 399.2 [M+H] ⁺ , RT: 0.672 min. ’H NMR (400 MHz, DMSO-d ₆ ) 8 9.07 (br s, 1H, NH), 7.73 - 7.68 (m, 1H), 7.61 (ddd, J = 7.2, 2.0, 2.0 Hz. 1H), 7.54 - 7.43 (m, 2H), 7.32 (br d, J = 8.8 Hz. 2H), 6.92 (d, J = 8.8 Hz, 2H), 3.45 - 3.38 (m, 4H), 3.23 - 3.16 (m, 4H), 1.46 (s, 9H). [0172] Step 2: To a solution of the product of Step 1 (0.700 g, 1.34 mmol 76% purity) in THF (10 mL) was added Pd/C (285 mg, 268 pmol, 10% purity) under N2. The suspension was degassed under vacuum and purged with H2 several times. The mixture was stirred under H2 (15 psi) at 25 °C for 12 h. LCMS showed the reaction was completed the starting material consumed and the desired product was detected. The mixture was filtered and the filtrate was concentrated to give Intermediate 5, tert-butyl A-[4-[4-(3- aminophenyl)piperazin-l-yl] phenyl]carbamate (0.500 g, 91% yield, 90.15% purity). LCMS (ESI ⁺ ) m/z 369.3 [M+H] ⁺ , RT: 0.863 min.

Intermediate 6 3-((3-(4-(4-aminophenyl)piperazin- l-yl)phenyl)amino)piperidine-2, 6-dione (FIG. 9)

[0173] Step 1: To a mixture of Intermediate 5 (0.500 g, 1.36 mmol, 90.15% purity) in dioxane (6 mL) were added 3-bromopiperidine-2, 6-dione (313 mg, 1.63 mmol), DIEA (351 mg, 2.71 mmol) and Nal (40.7 mg, 271 pmol). The mixture was stirred at 100 °C for 12 h under N2. LCMS showed the reaction was completed the starting material consumed and the desired product was detected. The mixture was poured into H2O (30 mL). The mixture was extracted with DCM (3 x 30 mL). The organic phase was washed with brine (3 x 30 mL), dried over anhydrous Na2SO4 and filtrated. The filtrate was concentrated under reduced pressure to give Boc-protected Intermediate 6 (0.30 g, 43.54% yield, 85.15% purity). LCMS (ESI ⁺ ) m/z 480.3 [M+H] ⁺ , RT: 0.853 min.

[0174] Step 2: A mixture of Boc-protected Intermediate 6 (0.500 g, 888 pmol, 85.15% purity) in DCM (10 mL) and TFA (1 mL) was stirred at 20 °C for 12 h under N2. LCMS showed the reaction was completed and the desired product was detected. The mixture was poured into NaHCCL solution (10 mL). The mixture was extracted with DCM (3 x 10 mL). The organic phase was washed with brine (5 x 10 mL), dried over anhydrous IShrtSCL and filtrated. The filtrate was concentrated under reduced pressure to give Intermediate 6, 3-((3-(4-(4-aminophenyl)piperazin-l-yl)phenyl)amino)piperidi ne-2, 6-dione (0.25 g, 37% yield, 50.47% purity) and used for next step reaction without further purification. LCMS (ESI ⁺ ) m/z 380.2 [M+H] ⁺ , RT: 0.685 min.

Intermediate 7 l-(4-((2,6-dioxopiperidin-3-yl)amino)phenyl)piperidine-4-car baldehyde (FIG. 10)

[0175] Step 1: To a solution of [l-(4-aminophenyl)-4-piperidyl]methanol (Reference: WO 2021/127561, 1.00 g, 4.65 mmol, 95.97% purity) and 3-bromopiperidine- 2, 6-dione (893 mg, 4.65 mmol) in DMF (15.0 mL) was added NaHCOs (782 mg, 9.30 mmol) at 25 °C. The reaction was stirred at 60 °C under N2 for 2 h. LCMS showed the reaction was completed and the desired product was detected. The mixture was concentrated under reduced pressure at 40 °C to give the crude product. The crude product was purified by column chromatography on silica gel (eluted with PE:EA = 3: 1 to 0: 1) to give 3-[4-[4- (hydroxymethyl)-l-piperidyl]anilino]piperidine-2, 6-dione (0.978 g, 3.02 mmol, 65% yield, 98.16% purity). LCMS (ESI ⁺ ) m/z 318.2 [M+H] ⁺ , RT: 0.617 min. NMR (400 MHz, DMSO-d ₆ ) 5 10.75 (s, 1H, NH), 6.75 (d, J = 8.9 Hz, 2H), 6.59 (d, J = 8.9 Hz, 2H), 5.35 (d, J = 7.2 Hz, 1H), 4.44 (t, J = 5.3 Hz, 1H), 4.18 (ddd, J = 11.3, 7.0, 4.8 Hz, 1H), 3.38 (br d, J = 11.9 Hz, 2H), 3.28 (t, J = 5.8 Hz, 2H), 2.79 - 2.66 (m, 1H), 2.62 - 2.53 (m, 1H), 2.48 - 2.40 (m, 2H), 2.15 - 2.05 (m, 1H), 1.89 - 1.77 (m, 1H), 1.72 (br d, J = 10.8 Hz, 2H), 1.48 - 1.34 (m, 1H), 1.23 (dq, J = 12.0, 4.1 Hz, 2H).

[0176] Step 2: To a mixture of 3-[4-[4-(hydroxymethyl)-l- piperidyl]anilino]piperidine-2, 6-dione (0.700 g, 2.16 mmol, 98.16% purity) in DCM (12.0 mL) were added TEA (2.19 g, 21.65 mmol,), pyridine; sulfur trioxide (2.21 g, 13.9 mmol) and DMSO (1.69 g, 21.7 mmol). The reaction was stirred at 25 °C for 12 h under N2. LCMS showed the reaction was completed and the desired product was detected. The reaction was extracted with DCM (3 x 60 mL). The organic layer was dried over NaiSCL. After filtered, the filtrate was concentrated under reduced pressure to give Intermediate 7, l-(4-((2,6- dioxopiperidin-3-yl)amino)phenyl)piperidine-4-carbaldehyde (0.35 g, 31 % yield, 60.12% purity). LCMS (ESI ⁺ ) m/z 316.2 [M+H] ⁺ , RT: 0.677 min.

Intermediate 8

(7?)-2-allyl-l-(7-ethyl-7-hydroxy-6,7-dihydro-5/7-cyclope nta[Z>]pyridin-2-yl)-6-((4-

(piperazin-l-yl)phenyl)amino)-l,2-dihydro-3//-pyrazolo[3, 4-<7]pyrimidin-3-one (FIG. 11)

[0177] Step 1: In a 125 mL round bottom flask, (R)-2-allyl-l-(7-ethyl-7 -hydroxy - 6,7-dihydro-5H-cyclopenta[Z>]pyridin-2-yl)-6-(methylthio) -l/7-pyrazolo[3,4-d]pyrimidin- 3(2H)-one (Reference: WO 2019/173082 Al) and cooled to 0 °C. m-CPBA (616 mg, 2.75 mmol) was slowly added to the flask. The reaction was continued at room temperature (rt) for 1 h, then cooled back to 0 °C. tert-Butyl 4-(4-aminophenyl)piperazine-l -carboxylate (832 mg, 3.00 mmol) and DIEA (2.183 mL, 12.50 mmol) were added to the mixture. The reaction was continued at rt for 16 h. Water (30 mL) was added to the mixture and extracted with EtOAc (2 x 50 mL). The combined organic layer washed with saturated aq. NaHCCh solution and brine, dried over sodium sulfate and evaporated to obtain a black residue, which was then purified by silica gel column chromatography using Combi-Flash eluting with DCM-MeOH (0-10%). Pure product fractions combined and evaporated to give (R)-tert-butyl 4-(4-((2- allyl-l-(7-cthyl-7 -hydroxy-6, 7-dihydro-5//-cyclopcnta[Z?]pyridin-2-yl)-3-oxo-2,3-dihydro- l/7-pyrazolo[3,4-<7]pyrimidin-6yl) amino )phenyl)piperazine-l -carboxylate (1.15 g, 1.88 mmol, 75 % yield). LC/MS (ESI ⁺ ) mA 613.4 [M+H] ⁺ .

[0178] Step 2: In a 25 mL round bottom flask, the product of Step 1 (225 mg, 0.367 mmol) was dissolved in dichloromethane (3 mL) and cooled to 0 °C. 2N HC1 in ether ( 1 mL) was added to the mixture and the reaction was continued at this temperature for 3 h. The reaction was quenched with cold sat. NaHCO aqueous solution and extracted with dichloromethane (3 15 mL). The combined organic layer was washed with brine, dried over sodium sulfate and evaporated. The resulting residue was purified on silica gel column chromatography using dichloromethane-MeOH (0-100%). The product fractions were combined and evaporated to give Intermediate 8, (R)-2-allyl-l-(7-cthyl-7-hydroxy-6,7- dihydro-5H-cyclopenta[b]pyridin-2-yl)-6-((4-(piperazin-l-yl) phenyl)amino)-lH- pyrazolo[3,4-d]pyrimidin-3(2H)-one, (150 mg, 0.293 mmol, 80 % yield). LC/MS (ESI ⁺ ) m/z 513.41 [M+H] ⁺ .

Intermediate 9 l-(4-nitrophenyl)-N-(piperidin-4-yl)piperidin-4-amine (FIG. 12)

[0179] Step 1: To a stirred solution of tert-butyl 4-aminopiperidine-l -carboxylate (1.09 g, 5.45 mmol) and l-(4-nitrophenyl)piperidin-4-one (1.00 g, 4.54 mmol) in 1,2- Dichloroethane (10 mL) at 0 °C, acetic acid (2.5 mL) was added dropwise, and the reaction was stirred at rt for 2 h. To this mixture at 0 °C, sodium triacetoxyborohydride (2.41 g, 11.4 mmol) was added portionwise and the mixture was allowed to stir at rt for an additional 16 h. After completion, the mixture was diluted with DCM (100 mL) and washed with water (2 x 50 mL) and brine (50 mL). The organic phase was dried over anhydrous sodium sulfate and concentrated to get the crude compound. The crude was purified by combi-flash chromatography (silica gel) using 0 to 5% methanol in DCM as eluting solvent to get tertbutyl 4-((l-(4-nitrophenyl)-2V-piperidin-4-yl)amino)piperidine-l -carboxylate (1.05 g, 57%). LC/MS (ESI ⁺ ) m/z 405.42 [M+H] ⁺ . ^} H NMR (400 MHz. CDCL) 5 8.11 (d. J = 9.5, Hz. 2H), 6.82 (d, J = 9.5 Hz, 2H), 5.65 (br s, 1H), 4.14 (br s, 2H), 3.98 (br d, J = 13.2 Hz, 2H), 3.18 - 3.08 (m, 1H), 3.07 - 2.94 (m, 3H), 2.84 - 2.70 (m, 2H), 2.20 - 2.09 (m, 2H). 2.07 - 1.94 (m, 2H), 1.82 - 1.68 (m, 2H), 1.67 - 1.50 (m, 2H), 1.45 (s, 9H).

[0180] Step 2 : To a stirred solution of tert-butyl 4-((l-(4-nitrophenyl)-2V- piperidin-4-yl)amino)piperidine-l -carboxylate (1.30 g, 3.21 mmol) in DCM (13 mL) at 0 °C, trifluoroacetic acid (5 mL) was added drop wise and the mixture was stirred at rt for 2 h. After completion, the mixture was concentrated under vacuum to get the residue, which was washed with diethyl ether (10 mL). The resulting product was dissolved in 20% MeOH in DCM (100 mL) and washed with saturated NaHCCh solution (2 x 100 mL) and brine solution (100 mL). The organic phase was dried over anhydrous sodium sulfate and concentrated to get l-(4-nitrophenyl)-N-(piperidin-4-yl)piperidin-4-amine (0.900 g, 92%). LC/MS (ESI ⁺ ) m/z 305.15 [M+H] ⁺ . ’ H NMR (400 MHz, CDC1 ₃ ) 5 ’ H NMR (400 MHz, CDCI3) 88.10 (d, J = 9.6 Hz, 2H), 6.81 (d, J = 9.6 Hz, 2H), 3.92 (br d, J = 13.2 Hz, 2H), 3.16 (br d, J = 12.8 Hz. 2H). 3.09 - 2.97 (m, 2H), 2.97 - 2.86 (m. 1H), 2.79 - 2.62 (m, 3H). 2.02 - 1.86 (m, 4H), 1.48 - 1.35 (m, 2H), 1.35 - 1.22 (m, 2H).

Intermediate 10

3-(4-(4-((L(4-aminophenyl)piperidin-4-yl)amino)piperidin- l-yl)phenyl)piperidine-2,6-

[0181] Step 1: To a stirred solution of Intermediate 9 (800 mg, 2.63 mmol), methyl 2-(4-bromophenyl)acetate (783 mg, 3.42 mmol) in 1,4-Dioxane (10 mL) at rt, tripotassium phosphate (1.67 g, 7.87 mmol) was added and the mixture degassed with nitrogen gas for 15 minutes. To this mixture, tris(dibenzylideneacetone)dipalladium(0) (241 mg, 0.263 mmol) and X-Phos (125 mg, 0.263 mmol) were added. The mixture was stirred at 120 °C for 16 h. After completion, the mixture was diluted with ice cold 10% aq. AcOH (50 mL) and extracted with 10% MeOH in DCM (2 x 50 mL). The organic layer was dried over sodium sulphate, filtered and concentrated under reduced pressure to get the crude product . The crude was purified by combi-flash chromatography (silica gel, column 40 g) using 0 to 7% MeOH in DCM as an eluent to get methyl 2-(4-(4-((l-(4-nitrophenyl)piperidin-4- yl)amino)piperidin-l-yl)phenyl)acetate (705 mg, 59%). LC/MS (ESI ⁺ ) m/z 453.67 [M+H] ⁺ . ¹ H NMR (400 MHz, CDCh) 8 8.11 (d, J - 9.2 Hz, 2H), 7.16 (d, J = 8.8 Hz, 2H), 6.88 (d, J - 8.4 Hz, 2H), 6.81 (d, J = 9.6 Hz, 2H), 3.67 (s, 3H), 3.66 (d. J = 13.2 Hz, 2H), 3.54 (s, 2H), 3.07-2.88 (m, 5H), 2.77-2.70 (m, 2H), 2.10-2.04 (m, 4H), 1.67-1.62 (m, 4H). [0182] Step 2: To a stirred solution of methyl 2-(4-(4-((l-(4- nitrophcnyl)pipcridin-4-yl)amino)pipcridin-l-yl)phcnyl)accta tc from Step 1 (0.6 g, 1.326 mmol) in DMF (6 mL) at -10 °C, acrylamide (0.104 g, 1.46 mmol) and Potassium tert- butoxide (IM solution in THF, 1.46 mL, 1.46 mmol) were added. The mixture was mixture at -10 °C to 0 °C for 2 h. The progress of reaction was monitored by TLC. After completion, the reaction was quenched with water (100 mL) and extracted with 10% MeOH in DCM (2 x 50 mL). The organic layer was washed with brine (50 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to get the crude product. The crude product was purified by silica gel flash chromatography using 0 to 5% MeOH in DCM as eluent to get 3-(4-(4-((l-(4-nitrophenyl)piperidin-4-yl)amino)piperidin-l- yl)phenyl)piperidine-2,6- dione (0.355 g, 54%). LC/MS (ESI ⁺ ) m/z 492.75 [M+H] ⁺ .

[0183] Step 3: To a stirred solution of 3-(4-(4-((l-(4-nitrophenyl)piperidin-4- yl)amino)piperidin-l-yl)phenyl)piperidine-2, 6-dione from Step 2 (0.250 g, 0.509 mmol) in MeOH (3 mL) under nitrogen atmosphere in a 100 mL round bottom flask, 10% Palladium on activated carbon (50% wet, 0.250 g) was added and the mixture was hydrogenated at rt for 16 h using hydrogen balloon. Upon completion, the reaction mixture was filtered through Celite pad and the filtrate was concentrated under reduced pressure to give Intermediate 10, 3-(4-(4-((l-(4-aminophenyl)piperidin-4-yl)amino)piperidin-l- yl)phenyl)piperidine-2,6- dione (0.160 g, 68%). LC/MS (ESI ⁺ ) m/z 462.33 8 10.77 (s, 1H), 8.82 (br s, 2H), 7.06 (d, J = 8.4 Hz, 2H), 6.93 (d, J = 8.4 Hz, 2H), 6.76 (d, J = 8.4 Hz, 2H), 6.58 (d, J = 8.4 Hz, 2H), 5.59 (br s, 1H), 3.85 - 3.70 (m, 3H), 3.69 - 3.55 (m, 1H), 3.44 (br d, J = 11.6 Hz, 2H), 3.19 - 3.09 (m, 1H). 2.76 (t, J = 12.0 Hz, 2H), 2.70 - 2.55 (m, 3H), 2.19 - 1.95 (m, 5H), 1.79 - 1.60 (m, 4H), 1.30 - 1.18 (m, 2H).

Intermediate 11 l-((l-(4-nitrophenyl)piperidin-4-yl)methyl)piperazine

Intermediate 11 N0 ₂ [0184] To a stirred solution of tert-butyl 4-((l-(4-nitrophenyl)piperidin-4- yl)mcthyl)pipcrazinc- 1 -carboxylate (Reference: US 2018/0099940, 3.10 g, 7.66 mmol) in DCM (30 mL) at 0 °C, trifluoroacetic acid (6 mL) was added dropwise and stirred the mixture at rt for 2 h. After completion, the mixture was concentrated under vacuum to get the crude product, which was washed with diethyl ether (25 mL). The resulting product was redissolved in 20% MeOH in DCM (200 mL) and washed with saturated NaHCCL solution (2 x 100 mL) and brine solution (100 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to get Intermediate 11, l-((l-(4-nitrophenyl)piperidin-4- yl)methyl)piperazine (2.05 g, 87%). LCMS (ESI ⁺ ) m z 305.31 [M+H] ⁺ . 'H NMR (400 MHz, CDC1 ₃ ) 88.17 (d, J = 9.4 Hz, 2H), 6.86 (d, J = 9.4 Hz, 2H), 4.02 (br d, J = 12.8 Hz, 2H), 3.06 - 2.96 (m, 6H). 2.47 (br s. 4H), 2.26 (d, J = 6.8 Hz. 2H), 2.03 - 1.93 (m, 3H), 1.38 - 1.27 (m, 2H).

Intermediate 12 3-(4-(4-((l-(4-aminophenyl)piperidin-4-yl)methyl)piperazin-l -yl)phenyl)piperidine-2,6- dione (FIG. 14)

[0185] Intermediate 12 was prepared by following the similar- procedure described in Intermediate 10 using Intermediate 11 (1.000 g, 3.285 mmol) in place of Intermediate 9, afforded Intermediate 12, 3-(4-(4-((l-(4-aminophenyl)piperidin-4- yl)methyl)piperazin-l-yl)phenyl)piperidine-2,6-dione (51%, 46% and 53% for three steps). LCMS (ESI ⁺ ) m/z 462.57 [M+H] ⁺ . Intermediate 13

3-(4-(4-aminopipcridin-l-yl)phcnyl)pipcridinc-2,6-dionc (FIG. 15)

[0186] Intermediate 13 was prepared by following the similar procedure described in Intermediate 12 Step 2 using methyl 2-(4-(4-((terr- butoxycarbonyl)amino)piperidin-l-yl)phenyl)acetate (Reference: US 2016/0068512, 1.00 g, 2.87 mmol) to afford the Boc-protcctcd Intermediate 13. The Boc group was removed by standard TFA/DCM procedure to give Intermediate 13, TFA salt of 3-(4-(4-aminopiperidin- l-yl)phenyl)piperidine-2, 6-dione (49% and 99% for two steps). LCMS (ESI ⁺ ) mJz 288.47 [M+H] ⁺ .

Intermediate 14

3-(4-(4-(((l-(4-aminophenyl)piperidin-4-yl)methyl)amino)p iperidin-l-yl)phenyl)piperidine- 2,6-dione (FIG. 16)

[0187] Step 1: To a stirred solution of Intermediate 13 TFA salt (540 mg, 1.40 mmol) and l-(4-nitrophenyl)piperidine-4-carbaldehyde (Reference: US 2021/0087170 P54 for similar procedure, 328 mg, 1.40 mmol) in DCE (10 mL) at rt, acetic acid (0.1 mL) was added. The mixture was stirred at rt for 1 h. To this mixture at 0 °C, NaBH(OAc)3 (595 mg, 2.80 mmol) was added and the resulting mixture was stirred at rt for 16 h. The reaction was quenched with water (35 mL) and extracted with dichloromethane (2 x 50 mL). The organic layer was washed with water (20 mL) and saturated sodium bicarbonate solution (20 mL), dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude product was purified by silica gel flash chromatography using 0 to 60% MeOH in DCM as eluent to get 3-(4-(4-(((l-(4-nitrophenyl)piperidin-4-yl)methyl)amino)pipe ridin-l-yl)phenyl) piperidine-2, 6-dione (460 mg, 65%). LCMS (ESI ⁺ ) m z 506.59 [M+H] ⁺ . ’H NMR (400 MHz, DMSO-d ₆ ) 5 10.77 (s. 1H, NH), 8.32 (br s. 1H, NH), 8.05 (d. J = 9.6 Hz, 2H), 7.06 (d, J = 8.8 Hz, 2H), 7.04 (d, 7 - 9.6 Hz, 2H), 6.92 (d, J= 8.8 Hz, 2H), 4.08 (br d, 7 - 10.6 Hz, 2H), 3.81 (br d, J = 12.0 Hz. 2H), 3.75 - 3.60 (m, 1H), 3.30 - 3.15 (m, 1H), 3.00 (t, J = 11.6 Hz, 2H), 2.89 (br s, 2H), 2.74 (t, 7= 12.0 Hz, 2H), 2.70 - 2.58 (m, 1H), 2.54 - 2.45 (m, 1H), 2.20 - 2.05 (m, 1H), 2.05 - 1.88 (m, 4H), 1.84 (br d, 7 = 13.2 Hz, 2H), 1.67 - 1.50 (m, 2H), 1.30 - 1.20 (m, 2H).

[0188] Step 2: To a stirred solution of 3-(4-(4-(((l-(4-nitrophenyl)piperidin-4- yl)methyl)amino)piperidin-l-yl)phenyl)piperidine-2, 6-dione (0.202 g, 0.4 mmol) in MeOH (10.1 mL) under nitrogen atmosphere, 10% palladium on carbon (50% wet, 0.170 g) was added at rt and the mixture was hydrogenated using hydrogen balloon at rt for 2 h. Progress of the reaction was monitored by TLC. After completion of the reaction, the mixture was filtered through a celite bed and washed with MeOH (10 mL). The filtrate was concentrated under reduced pressure to afford Intermediate 14, 3-(4-(4-(((l-(4- aminophenyl)piperidin-4-yl)methyl)amino)piperidin-l-yl)pheny l)piperidine-2,6- dione (190 mg, 99%). LCMS (ESI ⁺ ) m/z 476.63 [M+H] ⁺ .

Intermediate 15

3-(4-(2-aminoethoxy)phenyl)piperidine-2, 6-dione (FIG. 17)

[0189] Step 1: Intermediate 15 was prepared by following the similar procedure described in Intermediate 13 using methyl 2-(4-(2-((tert- butoxycarbonyl)amino)ethoxy)phenyl)acetate (Reference: WO 2019/243576, 1.20 g, 3.88 mmol) in place of methyl 2-(4-(4-((fert-butoxycarbonyl)amino)piperidin-l- yl)phenyl)acetate to afford Boc-protected Intermediate 15, tert-butyl (2-(4-(2,6- dioxopipcridin-3-yl)phcnoxy)cthyl)carbamatc (0.740 g, 55%). LCMS (ESI ⁺ ) m/z 347.07 [M- H] . ’H NMR (400 MHz. CDC1 ₃ ) 5 10.78 (s, 1H. NH). 7.12 (d, J = 8.4 Hz. 2H). 6.99 (br t, J = 6.2 Hz, 1H), 6.88 (d, J = 8.4 Hz, 2H), 3.94 (t, J = 6.0 Hz, 2H), 3.78 (dd, J = 11.4, 5.0 Hz, 1H), 3.34 - 3.24 (m, 2H), 2.72 - 2.58 (m, 1H). 2.52 - 2.45 (m, 1H), 2.23 - 2.10 (m, 1H), 2.07 - 1.98 (m, 1H). 1.38 (s, 9H).

[0190] Step 2: The Boc group was taken off by standard TFA/DCM procedure to give Intermediate 15, TFA salt of 3-(4-(2-aminoethoxy)phenyl)piperidine-2, 6-dione (97%). LCMS (ESI ⁺ ) m/z 249.24 [M+H] ⁺ .

Intermediate 16

3-(4-(2-(((l-(4-aminophenyl)piperidin-4-yl)methyl)amino)e thoxy)phenyl)piperidine-2,6-

[0191] Intermediate 16 was prepared by following the same procedure described in Intermediate 14 using Intermediate 15 (0.500 g, 2.01 mmol) in place of Intermediate 13, afforded Intermediate 16, 3-(4-(2-(((l-(4-aminophenyl)piperidin-4-yl)methyl)amino) ethoxy )phenyl)piperidine-2, 6-dione, (0.21 g, 42% and 56% for two steps). LCMS (ESI ) m/z 435.45 [M-H]-.

Intermediate 17

3-(4-((l-((l-(4-aminophenyl)piperidin-4-yl)methyl)piperid in-4-yl)oxy)phenyl)piperidine-

2,6-dione (FIG. 19) [0192] Intermediate 17 was prepared by following the same procedure described in Intermediate 14 using 3-(4-(pipcridin-4-yloxy)phcnyl)pipcridinc-2,6-dionc (0.250 g, 0.867 mmol) in place of Intermediate 13, afforded Intermediate 17, 3-(4-((l-((l-(4- aminophenyl)piperidin-4-yl)methyl)piperidin-4-yl)oxy)phenyl) piperidine-2, 6-dione, (0.18 g, 57% and 74% for two steps). LCMS (ESI ⁺ ) m z 477.65 [M+H] ⁺ .

Intermediate 18

2',6'-dioxo-[l,3'-bipiperidine]-4-carbaldehyde (FIG. 20)

[0193] Step 1 : To a mixture of 3 -bromopiperidine-2, 6-dione (0.500 g, 2.60 mmol) in DMF (5 mL) were added 4-piperidylmethanol (330 mg, 2.86 mmol) and DIEA (673 mg, 5.21 mmol) at 20 °C. Then the mixture was stirred at 80 °C for 12 h under N2. LCMS showed the starting material consumed and the desired product was detected. The mixture was filtered and the filtrate was purified by prep-HPLC (NH4HCO3 Condition) to afford 3-[4- (hydroxymethyl)-l-piperidyl]piperidine-2, 6-dione (0.250 g, 37% yield, 87.62% purity). LCMS (ESI ⁺ ) m/z 227.2 [M+H] ⁺ , RT: 0.496 min. ’H NMR (400 MHz, DMSO-d ₆ ) 8 10.54 (br s, 1H, NH), 4.38 (br s, 1H, OH), 3.40 - 3.34 (m, 2H), 3.26 - 3.18 (m, 2H), 2.82 - 2.70 (m, 2H), 2.59 - 2.50 (m, 2H), 2.40 - 2.30 (m, 1H). 2.06 - 1.93 (m, 1H), 1.88 - 1.79 (m, 1H), 1.66 - 1.55 (m, 2H), 1.39 - 1.25 (m, 1H), 1.14 - 1.01 (m, 2H).

[0194] Step 2: To a mixture of 3-[4-(hydroxymethyl)-l-piperidyl]piperidine-2,6- dione from Step 1 (0.250 g, 968 pmol, 87.62% purity) in DCM (3 mL) were added TEA (980 mg, 9.68 mmol), pyridine- sulfur trioxide (986 mg, 6.20 mmol) and DMSO (756 mg, 9.68 mmol) at 0 °C. The mixture was stirred at 40 °C for 1 h under N2. LCMS showed the starting material consumed and the desired product was detected. The mixture was poured into H2O (10 mL). The mixture was extracted with DCM (3 x 10 mL). The organic phase was washed with brine (5 mL), dried over anhydrous Na2SO4, concentrated in vacuum to give Intermediate 18, l-(2,6-dioxo-3-piperidyl)piperidine-4-carbaldehyde (0.150 g, 68% yield, 98.30% purity). LCMS (ESI ⁺ ) m/z 225.2 [M+H] ⁺ , RT: 0.513 min. Intermediate 19

4-((4-(4-aminophenyl)piperazin-l-yl)methyl)-[l,3'-bipiper idine]-2',6'-dione (FIG. 21)

[0195] Step 1: To a mixture of Intermediate 18 (0.150 g, 658 pmol, 98.30% purity) in DCM (5 mL) were added tert-butyl A-(4-piperazin-l-ylphenyl)carbamate (182 mg. 658 pmol) and NaBH(OAc)3 (279 mg, 1.32 mmol) at 0 °C. The mixture was stirred at 20 °C for 12 h under N2. LCMS showed the reaction was completed and the desired product was detected. The mixture was poured into H2O (10 mL). The mixture was extracted with DCM (3 x 10 mL). The organic phase was washed with brine (10 mL), dried over anhydrous NaiSCL, concentrated in vacuum to give Boc-protected Intermediate 19, rert-butyl (4-(4- ((2',6'-dioxo-[l,3'-bipiperidin]-4-yl)methyl) piperazin- l-yl)phenyl)carbamate (0.400 g, 44% yield). The crude product was used in the next step without purification. LCMS (ESI ⁺ ) m/z. 486.3 [M+H] ⁺ , RT: 1.739 min.

[0196] Step 2: Standard TFA/DCM Boc deprotection was used. The residue was purified by Prep-TLC (EA: MeOH = 3: 1, Rf = 0.4) to afford pure Intermediate 19, 3-((4-(4- aminophenyl)piperazin-l-yl)methyl)-[l,3'-bipiperidine]-2',6' -dione (0.120 g, 71% yield). LCMS (ESI ⁺ ) m/z 386.2 [M+H] ⁺ , RT: 1.134 min.

Intermediate 21

(l-(3-aminophenyl)piperidin-4-yl)methanol (FIG. 22)

[0197] Step 1: To a solution of l-bromo-3 -nitrobenzene (250 mg, 1.24 mmol) in toluene (8.0 mL) was added 4-piperidylmethanol (171. mg, 1.49 mmol), CS2CO3 (807 mg, 2.48 mmol), BINAP (77.1 mg, 124 pmol) and Pd(OAc)2 (27.8 mg, 124 pmol) at 25 °C in the glove box. The mixture was stirred at 100 °C for 12 h. LCMS showed the starting material was consumed and main product with desired MS was detected. Seven additional reactions were set up as detailed above. After cooling to rt, all eight reaction mixtures were combined. The reaction solution was filtered through celite and the filtrate was concentrated under reduced pressure to give a residue. The residue was purified by MPLC (Biotage, SiO ₂ ): (eluting with PE:EA = 1:0 to 2:1) to give (l-(3-nitrophenyl)piperidin-4-yl)methanol (0.900 g, 3.51 mmol. 35% yield, 92.14% purity). LCMS (ESI ⁺ ) m/z 237.1 [M+H] ⁺ . RT: 0.587 min. ’H NMR (400 MHz, CDC1 ₃ ) 87.73 (t, 7 - 2.2 Hz, 1H), 7.63 (dd, J= 8.2, 1.6 Hz, 1H), 7.36 (t, J = 8.2 Hz, 1H), 7.21 (dd, J = 8.2, 2.0 Hz, 1H). 3.87 - 3.78 (m, 2H), 3.58 (d, J = 6.3 Hz, 2H), 2.85 (td, J = 12.4, 2.4 Hz, 2H), 1.90 (br d, 7 = 13.4 Hz, 2H), 1.80 - 1.67 (m, 1H), 1.41 (qd, J = 12.2, 4.4 Hz, 2H).

[0198] Step 2: To a solution of (l-(3-nitrophenyl)piperidin-4-yl)methanol from Step 1 (0.900 g, 3.51 mmol, 92.14% purity) in THF (30.0 mL) was added Pd/C (369 mg, 351 pmol, 10% purity, 10% Pd on charcoal, wet, containing 50% water) at 25 °C. The mixture was degassed with hydrogen for three times and then stirred at 25 °C for 12 h under H2 (15 psi). LCMS showed the starting material was consumed and main product with desired MS was detected. The reaction solution was filtered through celite and the filtrate was concentrated under reduced pressure to give Intermediate 21, (l-(3-aminophenyl)piperidin- 4-yl)methanol, (0.500 g, 2.09 mmol, 59% yield, 86.03% purity). The crude product was used for the next step directly without purification. LCMS (ESI ⁺ ) m/z 207.1 [M+H] ⁺ , RT: 0.477 min. ^X H NMR (400 MHz, DMSO-d ₆ ) 8 6.81 (t, 7= 7.9 Hz, 1H), 6.14 (d, 7= 1.6Hz, 1H), 6.1O (dd, 7 = 7.9, 1.6 Hz, 1H), 5.99 (dd, 7 = 7.9, 1.4 Hz, 1H), 4.69 (br s, 2H, NH ₂ ), 3.56 (br d, 7 = 12.3 Hz, 2H), 3.27 (d. 7 = 6.2 Hz, 2H), 2.54 (td, 7 = 12.0. 2.0 Hz, 2H), 1.70 (br d, 7 = 12.1 Hz, 2H), 1.54 - 1.40 (m, 1H), 1.19 (qd, 7 = 12.2, 3.8 Hz, 2H).

Intermediate 22 l-(3-((2,6-dioxopiperidin-3-yl)amino)phenyl)piperidine-4-car baldehyde (FIG. 23)

[0199] Intermediate 22 was prepared by following the similar procedure described in Intermediate 7 using Intermediate 21 (0.500 g, 2.09 mmol, 86.03% purity) in place of [l-(4-aminophenyl)-4-piperidyl]methanol, to afford 1 -(3-((2,6-dioxopiperidin-3- yl)amino)phcnyl)pipcridinc-4-carbaldchydc, with yields at 46% and 26% for Step 1 and Step 2 respectively.

[0200] Analytical data for Step 1 product: LCMS (ESI ⁺ ) m/z 318.2 [M+H] ⁺ , RT: 0.315 min. ’H NMR (400 MHz, CDC1 ₃ ) 8 8.08 (br s. 1H), 7.11 (t, J = 8.1 Hz, 1H), 6.45 (dd, J = 8.4. 2.0 Hz, 1H), 6.29 (t, J = 2.1 Hz. 1H). 6.17 (dd, J = 8.0. 2.0 Hz. 1H). 4.67 (br s. 1H). 4.09 (dd, J = 12.0, 4.4 Hz, 1H), 3.69 (br d, J - 12.3 Hz, 2H), 3.55 (d, J = 6.4 Hz, 2H), 2.91 - 2.83 (m, 1H), 2.81 - 2.68 (m, 3H), 2.62 - 2.53 (m, 1H), 1.90 (qd, J = 13.0, 4.8 Hz. 1H), 1.85 (br d, J = 12.3 Hz, 2H), 1.69 - 1.62 (m, 1H), 1.40 (qd, J = 12.2, 4.0 Hz, 2H).

[0201] Analytical data for Intermediate 22: LCMS (ESI ⁺ ) m/z 316.2 [M+H] ⁺ , RT: 1.357 min.

Intermediate 23 te/7-butyl 6-(4-(2,6-dioxopiperidin-3-yl)phenyl)-2,6-diazaspiro[3.3]hep tane-2-carboxylate

(FIG. 24)

[0202] Step 1: To a solution of 2,6-bisbenzyloxy-3-bromo-pyridine (5.000 g, 13.50 mmol) in dioxane (100 mL) were added (4-boronophenyl)boronic acid (4.500 g, 27.01 mmol), Pd(dppf)Cl ₂ (1.50 g, 2.03 mmol), H ₂ O (20 mL), K2CO3 (5.700 g, 27.01 mmol) at 25 °C. The reaction was stirred at 80 °C for 12 h. LCMS showed the starting material was consumed and the desired product was detected. The mixture was diluted with water (10 mL) and extracted with EA (3 x 10 mL). The organic combined organic layers were washed with brine (3 x 10 mL) and dried over NaiSCL. The crude product was purified by column chromatography on silica gel (eluted with PE:EA = 100:0 to 35: 1) to give [4-(2,6- bisbenzyloxy-3-pyridyl)phenyl]boronic acid (2.0 g, 26% yield, 71.76% purity). LCMS (ESI ⁺ ) m/z 412.2 [M+H] ⁺ , RT: 0.977 min. 'H NMR (400 MHz, DMSO-d ₆ ) 8 8.03 (s, 2H), 7.80 (d, J = 8.2 Hz, 2H), 7.76 (d, J = 8.0 Hz, 2H), 7.52 (d, J = 8.2 Hz, 2H), 7.46 - 7.42 (m, 2H), 7.41 - 7.29 (m, 8H), 6.56 (d, J= 8.1 Hz, 1H), 5.48 (s, 2H), 5.39 (s, 2H).

[0203] Step 2: To a solution of the boronic acid from Step 1 (2.00 g, 3.49 mmol, 71.76% purity) in DCM (40 mL) were added tert-butyl 2,6-diazaspiro[3.3]heptane-2- carboxylate (346 mg, 1.74 mmol) and TEA (1.40 g, 13.8 mmol), Cu(OAc)z (634 mg, 3.49 mmol,) at 25 °C. The reaction was stirred at 25 °C for 12 h. LCMS showed the reaction was completed and the desired compound was detected. The mixture was diluted with water (10 mL) and extracted with EA (3 x 10 mL). The organic combined organic layers were washed with brine (3 x 10 mL) and dried over Na2SO4. The crude product was purified by column chromatography on silica gel (eluted with PE:EA = 100:0 to 20: 1) to give tert-butyl 6-[4-(2,6- bisbenzyloxy-3-pyridyl)phenyl]-2,6-diazaspiro[3.3]heptane-2- carboxylate (400 mg, 19% yield, 93.02% purity). LCMS (ESI ⁺ ) m/z 564.3 [M+H] ⁺ , RT: 1.110 min. 'H NMR (400 MHz, DMSO-d ₆ ) 87.64 (d, J = 8.1 Hz, 1H), 7.45 - 7.26 (m, 12H), 6.50 (d, J= 8.1 Hz, 1H), 6.44 (d, J = 8.7 Hz. 2H), 5.38 (s. 2H). 5.35 (s, 2H). 4.02 (br s. 4H). 3.93 (s, 4H). 1.38 (s, 9H).

[0204] Step 3: To a solution of the product from Step 2 (400.0 mg, 660 pmol, 93.02% purity) in THF (10 mL) were added Pd/C (10%, 66 mg, 62 pmol) at 25 °C. The reaction was stirred at 25 °C under H2 atmosphere (15 psi) for 12 h. LCMS showed the reaction was completed and the desired compound was detected. The mixture was filtered and the filter cake was washed with THF (20mL), the collected filtrate was concentrated to give Intermediate 23, tert-butyl 6-(4-(2,6-dioxopiperidin-3-yl)phenyl)-2,6- diazaspiro[3.3]heptane-2-carboxylate (250.0 mg, 84% yield, 85.44% purity). LCMS (ESI ⁺ ) m/z 386.2 [M+H] ⁺ , RT: 0.793 min.

Intermediate 24

3-(4-(6-((l-(4-aminophenyl)piperidin-4-yl)methyl)-2,6-dia zaspiro[3.3]heptan-2- yl)phenyl)piperidine-2, 6-dione (FIG. 25) [0205] Step 1 for the preparation of Intermediate 24 was Boc deprotection of Intermediate 23 under standard TFA/DCM condition with yield 98% and 86.25% purity. LCMS (ESI ⁺ ) m/z 286.2 [M+H] ⁺ , RT: 0.516 min.

[0206] Step 2 for the preparation of Intermediate 24 was followed the same procedure described in Intermediate 14 using de-Boc Intermediate 23 from Step 1 (180 mg, 544 pmol, 86.25% purity in place of Intermediate 13, to react with tert-butyl (4-(4- formylpiperidin-l-yl)phenyl)carbamate (Reference: WO 2007/075783) and afforded Boc- protected Intermediate 24 (200 mg, 46% yield, 71.01% purity). LCMS (ESI ⁺ ) m/z 574.4 [M+H] ⁺ , RT: 0.865 min.

[0207] Step 3 for the preparation of Intermediate 24 was under standard TFA/DCM condition. The reaction was stirred at 25 °C for 12 h. LCMS showed the starting material consumed and the desired product was detected. The reaction was concentrated under reduced pressure to give Intermediate 24, 3-(4-(6-((l-(4-aminophenyl)piperidin-4- yl)methyl)-2,6-diazaspiro[3.3]heptan-2-yl)phenyl)piperidine- 2, 6-dione, (100 mg, 51% yield, 59.48% purity). LCMS (ESI ⁺ ) m/z 474.3 [M+H] ⁺ , RT: 0.686 min.

Intermediate 25

3-(4-(4-aminobutoxy)phenyl)piperidine-2, 6-dione (FIG. 26)

[0208] Step 1: To a stirred solution of 4-(2,6-bis(benzyloxy)pyridin-3-yl)phenol (Reference: WO 2022/012622, 1 -30 g, 3.39 mmol), tert-butyl (4- hydroxybutyl)carbamate (0.962 g, 5.09 mmol) and triphenylphosphine (1.334 g, 5.085 mmol) in THF (10 mL) at 0 °C, Diisopropyl azodicarboxylate (1.028 g, 5.085 mmol) was slowly added and the mixture was stirred at rt for 16 h. After completion of reaction, water (10 mL) was added to quench the reaction. The mixture was concentrated under reduced pressure. The residue was diluted with water (50 mL) and DCM (100 mL). After separating the phases, the aqueous phase was further extracted with DCM (100 mL) and the combined organic phase was concentrated under reduced pressure to get residue. The crude product was purified by silica gel chromatography using 0 to 30% EtOAc in petroleum ether (PE) to obtain tert-butyl (4-(4-(2,6-bis(bcnzyloxy)pyridin-3- yl)phenoxy)butyl)carbamate (1.75 g, 93%). LCMS (ESI ) m/z 553.37 [M-H] .

[0209] Step 2: To a stirred solution of tert-butyl (4-(4-(2,6- bis(benzyloxy)pyridin-3-yl)phenoxy)butyl)carbamate from Step 1 (1.70 g, 3.07 mmol) in methanol (20 mL) under nitrogen atmosphere, 10% Palladium on activated carbon (50% wet, 0.5 g) was added and the resulting mixture was hydrogenated using hydrogen balloon for 16 h. The mixture was filtered through celite bed and celite bed was washed with EA (40 mL). The organic layer was concentrated to give crude compound. The crude compound was purified by silica gel flash chromatography using 0 to 50% EA in PE to get tert-butyl (4-(4- (2,6-dioxopiperidin-3-yl)phenoxy)butyl)carbamate (605 mg, 52%). LCMS (EST) m/z 375.19 [M-H]“. ’ H NMR (400 MHz, DMSO-d ₆ ) 5 10.78 (s, 1H), 7.11 (d, J = 8.4 Hz, 2H), 6.87 (d, J = 8.4 Hz, 2H), 6.84 (t, J = 5.2 Hz, 1H), 3.94 (t, J = 6.4 Hz, 2H), 3.78 (dd, J = 11.4, 5.0 Hz. 1H), 2.98 (q, J = 6.4 Hz, 2H). 2.71 - 2.58 (m, 1H), 2.52 - 2.43 (m, 1H), 2.22 - 2.10 (m, 1H), 2.06 - 1.96 (m, 1H), 1.73 - 1.63 (m, 2H), 1.57 - 1.47 (m, 2H), 1.37 (s, 9H).

[0210] Step 3: To a stirred solution of tert-butyl (4-(4-(2,6-dioxopiperidin-3- yl)phenoxy)butyl)carbamate (500 mg) in DCM from Step 2 at 0 °C, trifluoroacetic acid ( 1 mL) was added and the resulting mixture was stirred at it for 1 h. After completion of reaction, the mixture was concentrated under reduced pressure to give residue. The residue was triturated with EA (5 mL) to get TFA salt of Intermediate 25, 3-(4-(4- aminobutoxy)phenyl)piperidine-2, 6-dione (0.475 g, 95%). LCMS (ESI ⁺ ) m/z 277.48 [M+H] ⁺ .

Intermediate 26

3-(4-(4-((l-(4-aminophcnyl)pipcridin-4-yl)amino)butoxy)ph cnyl)pipcridinc-2,6-dionc

[0211] Intermediate 26 was prepared by following the same procedure described in Intermediate 14 using Intermediate 25 (TFA salt, 550 mg, 1.463 mmol) in place of Intermediate 13 to react with l-(4-nitrophenyl)piperidin-4-one, afforded Intermediate 26, 3-(4-(4-((l-(4-aminophenyl)piperidin-4-yl)amino)butoxy)pheny l)piperidine-2, 6-dione, (0.400 g, 57% and 88% for two steps).

[0212] Analytical data for Step 1 product: LCMS (ESI ⁺ ) m/z 481.67 [M+H] ⁺ . ^! H NMR (400 MHz, DMSO-d ₆ ) 5 10.79 (s. 1H, NH), 8.05 (d, J = 9.4 Hz. 2H), 7.13 (d, J = 8.6 Hz, 2H), 7.06 (d, J = 9.4 Hz, 2H), 6.89 (d, J = 8.6 Hz, 2H), 4.15 - 4.02 (m, 2H), 3.98 (t, J = 6.2 Hz, 2H), 3.78 (dd, J = 11.2, 4.8 Hz, 1H), 3.14 - 3.02 (m, 3H), 2.91 - 2.79 (m, 2H). 2.70 - 2.60 (m, 1H), 2.55 - 2.44 (m, 1H), 2.22 - 2.10 (m, 1H), 2.04 - 1.94 (m, 3H), 1.82 - 1.72 (m, 2H), 1.72 - 1.59 (m, 2H), 1.49 - 1.35 (m, 2H).

[0213] Analytical data for Intermediate 26: LCMS (ESI ) m/z 449.48 [M-H]’.

Intermediate 27

3-[4-[4-(4-aminophenyl)piperazin-l-yl]phenyl] piperidine-2,6-dione (FIG. 28) [0214] Step 1 : To a solution of [4-(2,6-dibenzyloxy-3-pyridyl)phenyl]boronic acid (1 g, 2.43 mmol) in DCM (20 mL) were added tert-butyl N-(4-pipcrazin-l- ylphenyl)carbamate (404.6 mg, 1.46 mmol), Cu(OAc)2 (441.6 mg, 2.43 mmol) and TEA (984.2 mg, 9.73 mmol) at 25 °C. The reaction was stirred at 25 °C for 12 h. LCMS showed the starting material was consumed and the desired product was detected. The mixture was diluted with water (10 mL) and extracted with DCM (3 x 10 mL). The organic combined organic layers were washed with brine (3 xx 10 mL) and dried over NaoSCL. The crude product was purified by column chromatography on silica gel (eluted with PE:EA = 100:0 to 30: 1) to give tert-butyl N-[4-[4-[4-(2,6-dibenzyloxy-3-pyridyl)phenyl]piperazin-l- yl]phenyl] carbamate (400.3, mg, 12.19% yield, 47.62% purity) as a yellow solid. LCMS (ESI ⁺ ) m/z 643.4 [M+H] ⁺ .

[0215] Step 2: To a solution of tert-butyl N-[4-[4-[4-(2,6-dibenzyloxy-3- pyridyl)phenyl]piperazin-l-yl] phenyl] carbamate (400.3 mg, 296.3 pmol) in THF (10 mL) were added Pd/C (6.28 mg, 29.634 pmol, 50% purity) at 25 °C. The reaction was stirred at 25 °C under H2 atmosphere (15 psi) for 12 h. LCMS showed the reaction was completed and the desired compound was detected. The mixture was filtered and the filter cake was washed with THF (30 mL), the collected filtrate was concentrated to give tert-butyl N-[4-[4-[4-(2,6- dioxo-3-piperidyl)phenyl]piperazin-l-yl] phenyl] carbamate (160.0 mg, 99.43% yield, 67.11% purity) as a yellow solid. LCMS (ESI ⁺ ) m/z 465.3 [M+H] ⁺ .

[0216] Step 3: To a solution of tert-butyl N-[4-[4-[4-(2,6-dioxo-3- piperidyl)phenyl]piperazin-l-yl] phenyl]carbamate (160.0 mg, 231.14 pmol) in DCM (4 mL) were added TFA (0.4 mL) at 25 °C. The reaction was stirred at 25 °C for 12 h. LCMS showed the reaction was completed and the desired compound was detected. The reaction was concentrated under reduced pressure to give 3-[4-[4-(4-aminophenyl)piperazin-l- yl]phenyl]piperidine-2, 6-dione (Intermediate 27) (100.0 mg, 97.18% yield, 81.86% purity)as yellow oil. LCMS (ESI ⁺ ) m/z 365.2 [M+H] ⁺ . Intermediate 28

3-[4-[4-[2-(4-aminophcnyl)cthyl]pipcrazin-l-yl]phcnyl]pip cridinc-2,6-dionc

(FIG. 29)

[0217] Step 1: To a solution of [4-(2,6-dibenzyloxy-3-pyridyl)phenyl]boronic acid (3.0 g, 7.29 mmol) in DCM (30 mL) were added tert-butyl piperazine- 1 -carboxylate (1.4 g, 7.29 mmol). Cu(OAc) ₂ (1.32 g, 7.29 mmol) and TEA (2.9 g. 29.18 mmol) at 20 °C. The reaction was stirred at 20 °C for 12 h. LCMS showed the starting material was consumed and the desired product was detected. The mixture was diluted with water (30 mL) and extracted with DCM (3 x 30 mL). The combined organic layers were washed with brine (3 xx 20 mL) and dried over Na ₂ SO4. After filtered, the filtrate was concentrated to give the crude product. The crude product was purified by column chromatography on silica gel (eluted with PE:EA = 100:0 to 20: 1) to give tert-butyl 4-[4-(2,6-dibenzyloxy-3-pyridyl)phenyl]piperazine-l- carboxylate (0.7 g, 10.9% yield, 63.17% purity) as a yellow solid. LCMS (ESI ⁺ ) m/z 552.3 [M+H] ⁺ .

[0218] Step 2: To a solution of tert-butyl 4-[4-(2,6-dibenzyloxy-3- pyridyl)phenyl]piperazine- 1 -carboxylate (0.7 g, 801.54 pmol) in THF (20 mL) was added Pd/C (85.3 mg, 80.15 pmol, 10% purity) at 20 °C. The reaction was stirred at 20 °C under H ₂ atmosphere (15 psi) for 12 h. LCMS showed the reaction was completed and the desired compound was detected. The mixture was filtered and the filter cake was washed with THF (30 mL), the collected filtrate was concentrated to give tert-butyl 4-[4-(2,6-dioxo-3- piperidyl)phenyl]piperazine- 1 -carboxylate (0.4 g, 95.5% yield, 71.53% purity) as a yellow solid. LCMS (ESI ⁺ ) /n/z 374.2 [M+H] ⁺ . 'H NMR (400 MHz, DMSO-d ₆ ) 5 10.77 (s, 1H), 7.07 (d, J = 8.7 Hz, 2H), 6.91 (d, J = 8.7 Hz, 2H), 3.73 (dd, J = 11.1, 4.9 Hz, 1H), 3.50 - 3.41 (m, 4H), 3.12 - 3.02 (m, 4H), 2.63 (ddd, J = 17.1, 11.4, 5.2 Hz, 1H), 2.48 (br s, 1H), 2.20 - 2.08 (m, 1H), 2.05 - 1.96 (m, 1H), 1.42 (s. 9H).

[0219] Step 3: A solution of tert-butyl 4-[4-(2,6-dioxo-3- piperidyl)phenyl]piperazine- 1 -carboxylate (0.4 g, 766.16 pmol) in DCM (10 mL) were added TFA (1 mL) at 0 °C. The reaction was stirred at 20 °C for 12 h. LCMS showed the reaction was completed and the desired compound was detected. The reaction was concentrated under reduced pressure to give 3-(4-piperazin-l-ylphenyl)piperidine-2.6-dione (0.25 g, 96.6% yield, 80.94% purity) as a yellow solid. LCMS (ESI ⁺ ) m/z 274.2 [M+H] ⁺ . ¹ H NMR (400 MHz, DMSO-d ₆ ) 8 10.79 (s, 1H), 8.78 (br s, 1H), 7.11 (d, J= 8.5 Hz, 2H), 6.95 (d, J= 8.6 Hz, 2H), 3.76 (dd, J = 11.2. 4.9 Hz, 1H). 3.38 - 3.28 (m, 4H). 3.24 (br s. 4H), 2.64 (ddd, J = 17.2, 11.6. 5.3 Hz, 1H), 2.49 - 2.41 (m, 1H), 2.22 - 2.09 (m, 1H), 2.05 - 1.95 (m, 1H).

[0220] Step 4: To a solution of 3-(4-piperazin-l-ylphenyl)piperidine-2, 6-dione (0.25 g, 740.32 pmol) and tert-butyl N-[4-(2-oxoethyl)phenyl]carbamate (209.02 mg, 888.38 pmol) in DCM (6 mL) was added DIEA (191.4 mg, 1.48 mmol) and AcOH (44.5 mg, 740.32 pmol, 42.34 pL) at 25 °C. The reaction was stirred at 25 °C for 2 h. NaBH(OAc)s (313.8 mg,

1.48 mmol) was added to the mixture and the mixture was stirred at 20 °C for 12 h. LCMS showed the starting material was consumed and the desired product was detected. The mixture was diluted with water (10 mL) and extracted with DCM (3 x 10 mL). The combined organic layers were washed with brine (3 x 10 mL) and dried over NaiSO4. After filtered, the filtrate was concentrated under reduced pressure to give tert-butyl N-[4-[2-[4-[4-(2,6-dioxo- 3-piperidyl)phenyl]piperazin-l-yl]ethyl]phenyl]carbamate (0.3 g, 44.0% yield, 53.52% purity) as a yellow solid. LCMS (ESI ⁺ ) m/z 493.3 [M+H] ⁺ .

[0221] Step 5: A solution of tert-butyl N-[4-[2-[4-[4-(2,6-dioxo-3- piperidyl)phenyl]piperazin-l-yl]ethyl]phenyl]carbamate (0.3 g, 325.94 pmol) in DCM (6 mL) were added TFA (0.6 mL) at 0 °C. The reaction was stirred at 20 °C for 12 h. LCMS showed the reaction was completed and the desired compound was detected. The reaction was concentrated under reduced pressure to give a residue. The residue was purified by prep- HPLC (NH4HCO3 Condition) to afford 3-[4-[4-[2-(4-aminophenyl)ethyl]piperazin-l- yl]phenyl]piperidine-2, 6-dione (Intermediate 28) (0.1 g, 74.0% yield, 94.69% purity) as a yellow solid. LCMS (ESI ⁺ ) m/z 393.2 [M+H] ⁺ . ¹ H NMR (400 MHz, DMSO-d ₆ ) 8 10.77 (s, 1H), 7.04 (d, J = 8.8 Hz. 2H), 6.88 (dd, J = 8.4, 6.9 Hz, 4H), 6.52 - 6.44 (m, 2H), 4.81 (s, 2H), 3.72 (dd, J = 10.9, 4.9 Hz, 1H), 3.15 - 3.06 (m, 4H), 2.68 - 2.58 (m, 2H), 2.57 - 2.53 (m, 4H),

2.49 - 2.41 (m, 4H), 2.19 - 2.07 (m, 1H), 2.05 - 1.96 (m, 1H). Intermediate 29

3-[[4-[4-(4-aminophcnyl)pipcrazin-l-yl]cyclohcxyl] amino]pipcridinc-2,6-dionc (FIG. 30)

[0222] Step 1: To a solution of tert-butyl N-(4-oxocyclohexyl)carbamate (500.0 mg, 2.34 mmol) in DCM (10 mL) were added l-(4-nitrophenyl)piperazine (485.83 mg, 2.34 mmol) and NaBH(OAc)3 (993.7 mg, 4.69 mmol) at 25 °C. The reaction was stirred at 25 °C for 12 h. LCMS showed the starting material was consumed and the desired product was detected. The mixture was diluted with water (10 mL) and extracted with DCM (3 x 10 mL). The combined organic layers were washed with brine (3 x 10 mL) and dried over NaiSCL. The crude product was purified by column chromatography on silica gel (eluted with PE:EA = 100:0 to 3: 1) to give tert-butyl N-[4-[4-(4-nitrophenyl)piperazin-l-yl] cyclohexyl] carbamate (400.0 mg, 42.18% yield, 100.00% purity) as a white solid. LCMS (ESI ⁺ ) m/z 405.2 [M+H] ⁺ . 'H NMR (400 MHz, DMSO-d ₆ ) 8 8.10 - 8.00 (m, 2H), 7.06 - 6.98 (m, 2H), 3.42 (br d, J = 4.4 Hz. 4H), 3.32 (br d. J = 2.1 Hz, 1H), 2.59 (br s, 4H), 2.27 - 2.21 (m, 1H), 1.75 - 1.58 (m. 4H), 1.50 - 1.40 (m, 4H). 1.38 (t, J = 4.9 Hz, 9H).

[0223] Step 2: A solution of tert-butyl N-[4-[4-(4-nitrophenyl)piperazin-l- yl]cyclohexyl]carbamate (400.0 mg, 988.87 pmol, 100.00% purity) in HCl/dioxane (10 mL) at 25 °C. The reaction was stirred at 25° C for 12 h. LCMS showed the starting material was consumed and main product with desired MS was detected. The mixture was concentrated under reduced pressure to give 4-[4-(4-nitrophenyl)piperazin-l-yl]cyclohexanamine (300.2 mg, 99.67% yield, 84.78% purity) as a yellow solid. LCMS (ESI ⁺ ) m/z 305.2 [M+H] ⁺ . ‘H NMR (400 MHz, DMSO-d ₆ ) 8 8.11 (d, J = 9.3 Hz, 2H), 7.21 - 7.04 (m, 2H), 4.26 - 4.13 (m, 2H), 3.60 (br s, 1H). 3.53 (br s, 2H), 3.37 (br s, 2H), 3.29 - 3.10 (m, 2H), 2.62 - 2.53 (m. 2H), 2.29 - 2.05 (m, 2H), 2.04 - 1.94 (m, 4H), 1.77 - 1.56 (m, 2H), 1.49 - 1.36 (m, 1H).

[0224] Step 3: To a solution of 4-[4-(4-nitrophenyl)piperazin-l- yl]cyclohexanamine (300.2 mg, 835.58 pmol, 84.78% purity) in DMF (8 mL) were added 3- bromopiperidine-2, 6-dione (240.6 mg, 1.25 mmol) and DIEA (539.9 mg, 4.18 mmol) at 25 °C. The reaction was stirred at 80 °C for 12 h. LCMS showed the reaction was completed and the desired compound was detected. The mixture was diluted with water (10 mL) and extracted with DCM (3 x 10 mL). The organic combined organic layers were washed with brine (3 x 10 mL) and dried over NaiSCL- After filtered, the filtrate was concentrated under reduced pressure to give the residue. The residue was purified by prep-HPLC (NH4HCO3 Condition) to afford 3-[[4-[4-(4-nitrophenyl)piperazin-l-yl]cyclohexyl]amino]pipe ridine- 2,6-dione (150.3 mg, 41.02% yield, 94.94% purity) as a yellow solid. LCMS (ESI ⁺ ) m z 416.2 [M+H] ⁺ . ¹ H NMR (400 MHz, DMSO-d ₆ ) 8 10.85 - 10.48 (m, 1H), 8.05 (d, J = 9.5 Hz, 2H), 7.08 - 6.96 (m, 2H), 3.43 (br d, J = 3.9 Hz, 4H), 3.40 (br d, J = 4.9 Hz. 2H), 2.81 (br s, 1H), 2.58 (br d, J = 4.6 Hz, 4H), 2.54 (br d, J = 4.9 Hz, 2H), 2.22 (br s, 1H), 2.04 - 1.95 (m, 1H), 1.75 - 1.68 (m, 1H), 1.65 (br d, J = 9.0 Hz, 4H), 1.53 - 1.39 (m, 4H).

[0225] Step 4: To a solution of 3-[[4-[4-(4-nitrophenyl)piperazin-l- yl]cyclohexyl]amino]piperidine-2, 6-dione (150.3 mg, 342.76 pmol, 94.94% purity) in THF (2 mL) was added Pd/C (5.7 mg, 34.28 pmol, 30% purity) at 25 °C. The reaction was stirred at 25 °C for 12 h. LCMS showed the stalling material consumed and the desired product was detected. The mixture was filtered and the filter cake was washed with THF (10 mL). The collected filtrate was concentrated to give 3-[[4-[4-(4-aminophenyl)piperazin-l- yl]cyclohexyl] amino] piperidine-2, 6-dione (Intermediate 29) (100.0 mg, 53.13% yield, 70.20% purity) as a yellow solid. LCMS (ESI ⁺ ) m/z 386.2 [M+H] ⁺ .

Intermediate 30

4-[4-(4-oxo-l-piperidyl)phenyl]piperidine-2, 6-dione (FIG. 31)

[0226] Step 1: To a solution of 4-bromobenzaldehyde (5 g, 27.02 mmol) in ethyl 3-oxobutanoate (EAA) (7.74 g, 59.45 mmol) was added piperidine (1 mL) at 0 °C dropwise. The mixture was then stirred at 20 °C for 3 h. The precipitated solid was filtered out and triturated by cold EtOH to give diethyl 2,4-diacetyl-3-(4-bromophenyl)pentanedioate (5 g, 11.7 mmol. 43.2 %) as white solid. The crude solid was then added into aqueous solution of KOH (20 M, 50 mL). The mixture was stirred at 80 °C for 3 h. LCMS showed desired product was formed. After cooling to rt, cone. HC1 was added slowly to the mixture to adjust pH to 3. The formed solid was filtered out, washed with water to give 3-(4-bromophcnyl) pentanedioic acid as a white solid (3.9 g, 13.58 mmol, 50.26% yield). ’H NMR (400 MHz, DMSO-d ₆ ) 8 12.10 (br s, 2H), 7.47 (d, J = 8.4 Hz, 2H), 7.27 - 7.20 (m, 2H), 3.52 - 3.19 (m, 1H), 2.73 - 2.58 (m, 2H), 2.57 - 2.52 (m, 1H), 2.50 - 2.47 (m, 1H).

[0227] Step 2: 3-(4-bromophenyl)pentanedioic acid (2.8 g, 9.75 mmol) and urea (1.46 g, 24.38 mmol) was mixed together and was stirred at 160 °C for 3 h. LCMS showed desired product was formed. The mixture was then cooled to rt. The obtained solid was dissolved in EA and washed with water (3x). The organic layer was concentrated with reduced pressure to give 4-(4-bromophenyl)piperidine-2, 6-dione (1 g, 3.73 mmol, 38.25% yield) as a brown solid. LCMS (ESI ⁺ ) m/z 268.0 (M+H) ⁺ .

[0228] Step 3: To a solution of 4-(4-bromophenyl)piperidine-2, 6-dione (500 mg, 1.31 mmol) 1,4-dioxa- 8-azaspiro[4.5]decane (373.84 mg, 2.61 mmol, 334.68 pL) in dioxane (10 mL) was added CS2CO3 (1.28 g, 3.92 mmol) l,3-bis[2,6-bis(l-propylbutyl)phenyl]-4,5- dichloro-2H-imidazol-l-ium-2-ide;3-chloropyridine;dichloropa lladium (126.99 mg, 130.55 pmol) . The mixture was then stirred at 100 °C for 5 h under the protection of N2. Another batch was set up with the same procedure. LCMS showed desired product was formed. The mixture was extract with EA (3 x 100 mL) and washed with saturated NH4CI aqueous solution. The organic layer was then dried over Na2SO4, filtered and concentrated by reduced pressure to give the crude residue, which was further purified by silica column with eluent of EA:PE = 1: 1. 4-[4-(l,4-dioxa-8-azaspiro[4.5]decan-8-yl)phenyl]piperidine- 2, 6-dione (230 mg, 696.17 pmol, 53.33% yield) was obtained as a white solid. LCMS (ESI ⁺ ) 331 [M+H] ⁺ . 'H NMR (400 MHz, CDCI3) 87.81 (br s, 1H), 7.01 (d, J = 8.7 Hz, 2H), 6.85 (d, J = 8.8 Hz, 2H), 3.92 (s, 4H), 3.33 - 3.21 (m, 5H), 2.89 - 2.76 (m. 2H), 2.70 - 2.55 (m, 2H), 1.86 - 1.69 (m, 4H).

[0229] Step 4: To a solution of 4-[4-(l,4-dioxa-8-azaspiro[4.5]decan-8- yl)phenyl]piperidine-2, 6-dione (230 mg, 302.68 pmol) in formic acid (2.3 mL) was stirred at 90 °C. LCMS showed desired product was formed. The reaction was washed with saturated NH4CI aqueous solution and extracted by EA (2 x 20 mL). The organic layer was combined and concentrated by reduced pressure. The crude reside was purified by prep-TLC with pure EA. 4- [4-(4-oxo- 1 -piperidyl)phenyl]piperidine-2 ,6-dione (80 mg, 139.70 pmol, 46.15% yield) was formed as a white solid. 'H NMR (400 MHz, CDCh) 87.75 (br s, 1H), 7.07 (d, J = 8.7 Hz, 2H), 6.89 (d, 7 = 8.7 Hz, 2H), 3.54 (t, J = 6.1 Hz, 4H), 3.37 - 3.23 (m, 1H), 2.93 - 2.76 (m, 2H), 2.66 (dd, J = 11.4, 17.4 Hz, 2H), 2.49 (t, J = 6.1 Hz, 4H).

Intermediate 31

2- ally 1 - 6- [4- [4-(aminomethy 1)- 1 -piperidyl] anilino] - 1 - [(7R)-7-ethy 1-7 -hydroxy-5 ,6- dihydrocyclopenta[b]pyridin-2-yl]pyrazolo[3,4-d]pyrimidin-3- one (FIG. 32)

[0230] Step 1: To a solution of tert-butyl (piperidin-4-ylmethyl)carbamate (10 g, 70.87 mmol) and l-fluoro-4-nitrobenzene (18.23 g, 85.05 mmol) in DMF (20 mL) was added DIEA (18.32 g, 141.74 mmol). The mixture was stirred at 60 °C for 12 h. LCMS showed one major peak with desired MS was detected. The reaction was added to water (50 mL) and the mixture was filtered. The filter cake was collected by suction filtration and dried in vacuum to give tert-butyl ((l-(4-nitrophenyl)piperidin-4-yl)methyl)carbamate (23 g, 68.58 mmol, 96.76% yield) as a yellow solid. LCMS (ESI ⁺ ) m/z. 336.2 [M+H] ⁺ . 'H NMR (400 MHz. DMSO-d ₆ ) 8 8.03 (d, J = 9.1 Hz, 2H), 7.00 (br d. J = 9.3 Hz, 2H), 6.91 (br t, J = 5.4 Hz, 1H), 4.04 (br d, J = 13.3 Hz, 2H), 2.95 (br t, J = 12.4 Hz, 2H), 2.84 (br t, J = 5.9 Hz, 2H), 1.76 - 1.62 (m, 3H), 1.38 (s, 9H), 1.13 (q, J = 11.3 Hz, 2H).

[0231] Step 2: A solution of tert-butyl ((l-(4-nitrophenyl)piperidin-4- yl)methyl)carbamate (23 g, 68.58 mmol) in HCl/EtOAc (4 M, 30 mL) was stirred at 25 °C for 2 h. LCMS showed the starting material consumed and a new spot was detected. The mixture was concentrated under reduced pressure at 40 °C to give crude l-(4-nitrophenyl)piperidin-4- yl)methanamine (16 g, 99.17% yield) as a yellow solid, which was used to next step without further purification. 'H NMR (400 MHz, DMSO-d ₆ ) 6 8.25 (br s, 3H), 8.06 - 7.99 (m, 2H), 7.72 (br s, 2H), 7.07 - 7.00 (m, 2H), 4.07 (br d, J = 13.4 Hz, 2H), 3.03 - 2.90 (m, 2H), 2.71 (quin, J = 5.9 Hz, 2H), 2.00 - 1.90 (m, 1H), 1.85 (br d, J = 13.1 Hz, 2H), 1.22 (dq, J = 3.8, 12.2 Hz, 2H). [0232] Step 3: To a solution of (l-(4-nitrophenyl)piperidin-4-yl)methanamine (16 g, 68.00 mmol) and TFAA (21.42 g, 102.01 mmol) in DCM (200 mL) was added TEA (20.64 g, 204.01 mmol). The reaction was stirred at 25 °C for 12 h. LCMS showed the reaction was completed and the desired compound was detected. The reaction was purified directly without work-up. The crude product was purified by column chromatography on silica gel (eluted with PE:EA = 100: 1 to 2: 1) to give 2,2,2-trifluoro-N-((l-(4-nitrophenyl)piperidin-4-yl) methyl)acetamide (22 g, 89.48% yield, 91.633% purity) as a white solid. LCMS (ESI ⁺ ) m/z 332.1 [M+H] ⁺ .

[0233] Step 4: To a solution of 2,2,2-trifluoro-N-((l-(4-nitrophenyl)piperidin-4- yl)methyl) acetamide (22 g, 66.41 mmol) in THF (220 mL) was added Pd/C (141.34 g, 132.81 mmol, 10% purity) at 2 0°C. The mixture was stirred at 20 °C for 12 h. LCMS showed one major peak with desired MS was detected. The reaction was filtered and the filter cake was washed with THF (200 mL). The combined filtration was concentrated to give the crude product, which was purified by column chromatography on silica gel (eluted with THF:EA = 1: 100 to 3:1) to give N-((l-(4-aminophenyl)piperidin-4-yl)methyl)-2,2,2-trifluoroa cetamide (13 g, 35.34% yield, 54.4% purity) as a brown solid. LCMS (ESI ⁺ ) m/z 302.1 [M+H] ⁺ . 'H NMR (400 MHz, DMSO-d ₆ ) 59.50 (br t, J = 5.4 Hz, 1H), 7.05 - 6.73 (m, 2H), 6.71 - 6.39 (m, 2H), 6.36 - 5.80 (m, 2H), 3.41 (br d, J = 4.6 Hz, 2H), 3.14 (br t, J = 6.1 Hz, 2H), 2.62 (br s, 2H), 1.70 (br s. 3H), 1.30 (br d. J = 11.7 Hz. 2H).

[0234] Step 5: To a solution of (R)-2-allyl-l-(7-ethyl-7-hydroxy-6,7-dihydro-5H- cyclopenta[b]pyridine-2-yl)-6-(methylthio)-l,2-dihydro-3H-py razolo[3,4-d]pyrimidin-3-one (5 g, 13.04 mmol) in toluene (50 mL) was added m-CPBA (5.29 g, 26.08 mmol, 85% purity) at 20 °C. The reaction was stirred at 25 °C for 2 h, then a solution of N-((l-(4- aminophenyl)piperidin-4-yl)methyl)-2,2,2-trifluoroacetamide (4.71 g, 15.65 mmol) and DIEA (8.43 g, 65.19 mmol) in DMF (50 mL) was added to the reaction dropwise at 0 °C. The reaction was stirred at 25 °C for 12 h. Additional one reaction was set up as described as above and the reactions were combined for work-up. The reaction was poured into water (100 mL) and extracted with EA (200 mL). The organic layer was dried over NaiSCk and filtered. The filtrate was concentrated to give the crude product, which was purified by column chromatography on silica gel (eluted with EA) to give (R)-N-((l-(4-((2-allyl-l-(7-ethyl-7- hydroxy-6,7-dihydro-5H-cyclopenta[b]pyridin-2-yl)-3-oxo-2,3- dihydro-lH-pyrazolo[3,4- d]pyrimidin-6-yl)amino)phenyl)piperidin-4-yl)methyl)-2,2,2-t rifluoroacetamide (11 g, 17.28 mmol, 66.25% yield) as a yellow solid. ’H NMR (400 MHz, CDC1 ₃ ) 8 8.81 (s, 1H), 7.73 - 7.67 (m, 2H), 7.46 (br d, J = 8.4 Hz, 2H), 6.92 (br d, J = 8.9 Hz, 2H). 6.49 (br s. 1H). 5.70 (tdd, J = 6.2, 10.4, 16.8 Hz, 1H), 5.03 (d, J = 10.3 Hz, 1H), 4.93 (d, 7 = 17.1 Hz, 1H), 4.90 - 4.80 (m, 1H), 4.78 - 4.64 (m, 1H), 3.68 (br d, 7 = 12.3 Hz, 2H), 3.37 (t, 7 = 6.4 Hz, 2H), 3.14 - 2.98 (m, 1H). 2.95 - 2.82 (m, 1H), 2.72 (br t, 7 = 11.4 Hz, 2H), 2.40 (ddd, 7 = 4.5, 8.6. 13.4 Hz, 2H), 2.29 - 2.20 (m, 1H), 2.05 - 1.97 (m, 1H), 1.90 - 1.81 (m, 3H), 1.80 - 1.71 (m, 1H), 1.70 - 1.59 (m, 1H), 1.48 (dq, 7 = 3.4, 12.0 Hz, 2H), 1.01 (t, 7 = 7.4 Hz, 3H).

[0235] Step 6: To a solution of N-[[l-[4-[[2-allyl-l-[(7R)-7-ethyl-7-hydroxy-5,6- dihydrocyclopentafb] pyridin-2-yl] -3-oxo-pyrazolo [3 ,4-d]pyrimidin-6-yl] amino]phenyl] -4- piperidyl]methyl]-2,2,2-trifluoro-acetamide (10 g, 15.71 mmol) in MeOH (300 mL) was added K2CO3 (21.71 g, 157.07 mmol). The reaction was stirred at 30 °C for 48 h. LCMS showed the reaction was completed. The reaction was diluted with water (300 mL) and extracted with DCM (5 x 300 mL). The organic layer was dried over NaiSCL and filtered. The filtrate was concentrated to give 2-ally 1-6- [4- [4-(aminomethyl)-l -piperidyl] anilino]-l - [(7R)-7-ethyl-7-hydroxy-5,6-dihydrocyclopenta[b]pyridin-2-yl ]pyrazolo[3,4-d]pyrimidin-3- one (Intermediate 31) (9 g, 14.98 mmol, 95.38% yield, 90% purity) as a yellow solid. LCMS (ESI ⁺ ) zn/z 541.3 [M+H] ⁺ . ’ H NMR (400 MHz, DMSO-d ₆ ) 8 10.29 - 9.94 (m, 1H), 8.82 (s, 1H), 7.94 (br d, 7 = 7.1 Hz, 1H), 7.70 (d, 7 = 8.3 Hz, 1H), 7.57 (br s, 2H), 6.92 (br d, 7 = 9.0 Hz, 2H), 5.67 (tdd, 7 = 6.0, 10.6, 16.8 Hz, 1H), 5.08 (br s, 1H), 5.OO (d, 7= 10.3 Hz, 1H), 4.86 (br d, 7 = 17.0 Hz, 1H), 4.80 - 4.71 (m, 1H), 4.57 (br dd, 7 = 5.8, 16.0 Hz, 1H), 3.65 (br d, 7 = 12.0 Hz, 2H), 3.07 - 2.92 (m, 1H), 2.90 - 2.66 (m. 2H), 2.62 - 2.54 (m, 2H), 2.45 (d, 7 = 6.1 Hz, 2H), 2.21 (ddd, 7 = 5.8, 8.2, 13.4 Hz, 1H), 2.08 - 1.97 (m, 1H), 1.90 (br dd, 7 = 7.3, 13.6 Hz, 1H), 1.85 - 1.67 (m, 3H). 1.59 - 1.39 (m, 1H), 1.38 - 1.28 (m, 1H), 1.28 - 1.13 (m, 2H), 0.88 (t, 7 = 7.4 Hz, 3H).

Intermediate 33

3-((3-(4-aminopiperidin-l-yl)phenyl)amino)piperidine-2, 6-dione (FIG. 33) [0236] Step 1 : To a solution of l-bromo-3-nitrobenzene (10.0 g, 40.16 mmol) in toluene (200 mL) were added BINAP (2.5 g, 4.02 mmol), Pd(OAc)i (901.6 mg, 4.02 mmol), tert-butyl piperidin-4-ylcarbamate (8.1 g, 40.16 mmol) and CS2CO3 (26.17 g, 80.32 mmol) at 25 °C. The reaction was stirred at 100 °C for 12 h. LCMS showed the starting material was consumed and the desired product was detected. The reaction was concentrated under reduced pressure to give the product. The crude product was purified by column chromatography on silica gel (eluted with PE:EA = 100:0 to 25: 1) to give tert-butyl N-(l-(3-nitrophenyl)piperidin- 4-yl)carbamate (6.0 g, 40.8% yield, 87.87% purity) as a yellow solid. LCMS (ESI ⁺ ) m/z 322.1 [M+H] ⁺ . ‘H NMR (400 MHz, DMSO-d ₆ ) 57.62 (t, J = 2.2 Hz, 1H), 7.53 (dd, J = 7.8, 1.0 Hz, 1H), 7.44 (t, J = 8.1 Hz, 1H), 7.40 (d, J = 1.5 Hz, 1H), 6.87 (br d, J = 7.8 Hz, 1H), 3.77 (br d, J = 13.2 Hz, 2H), 3.45 (br dd, J = 8.4, 1.9 Hz, 1H), 2.92 - 2.84 (m, 2H), 1.81 (br d, J = 11.0 Hz, 2H), 1.50 - 1.42 (m, 2H), 1.39 (s, 9H).

[0237] Step 2: To a solution of tert-butyl N-[l-(3-nitrophenyl)-4- piperidyl]carbamate (6.0 g, 16.41 mmol) in THF (30 mL) was added Pd/C (17.4 g, 16.41 mmol, 30% purity) at 25 °C. The reaction was stirred at 25 °C for 12 h. TLC showed the starting material consumed and a new spot was detected. The mixture was filtered and the filter cake was washed with THF (50 mL). The collected filtrate was concentrated to give tert-butyl N-[l-(3-aminophenyl)-4-piperidyl]carbamate (4.0 g, 41.84% yield) as a yellow solid. ¹ H NMR (400 MHz, DMSO-d ₆ ) 8 = 6.90 - 6.78 (m, 1H). 6.73 (br d, J = 7.6 Hz, 1H), 6.17 - 6.07 (m, 1H), 5.99 (dd, 7 = 7.7, 1.2 Hz, 1H), 4.81 (s, 1H), 3.60 (t, 7 = 6.4 Hz, 1H), 3.51 (br d, 7 = 12.7 Hz, 1H), 3.40 - 3.17 (m, 2H), 2.86 (br d, 7 = 12.3 Hz, 1H), 2.67 - 2.58 (m, 1H), 2.39 (dt, 7 = 12.0, 2.0 Hz. 1H), 1.80 - 1.70 (m, 2H). 1.61 (br d. 7 = 11.0 Hz, 1H). 1.38 (d, 7 = 5.9 Hz, 9H), 1.26 - 1.12 (m, 1H).

[0238] Step 3: To a mixture of tert-butyl (l-(3-aminophenyl)piperidin-4- yl)carbamate (4.0 g, 6.86 mmol) in dioxane (40 mL) were added 3-bromopiperidine-2, 6-dione (3.9 g, 20.59 mmol), DIEA (1.7 g, 13.73 mmol, 2.4 mL) and Nal (205.7 mg, 1.37 mmol) at 25 °C. The mixture was stirred at 60 °C for 12 h. LCMS showed the reaction was completed and the desired compound was detected. The mixture was diluted with water (30 mL) and extracted with DCM (3 x 30 mL). The organic combined organic layers were washed with brine (3 x 30 mL) and dried over Na^SCL. After filtering, the filtrate was concentrated under reduced pressure to give tert-butyl (l-(3-((2,6-dioxopiperidin-3-yl)amino)phenyl)piperidin-4- yl)carbamate (1.5 g, 3.11 mmol, 45.3% yield) as a yellow solid. LCMS (EST ⁺ ) m/z 403.2 [M+H] ⁺ .

[0239] Step 4: To a solution of tert-butyl (l-(3-((2,6-dioxopiperidin-3- yl)amino)phenyl)piperidin-4-yl) carbamate (1.5 g, 3.11 mmol) in DCM (8 mL) was added TFA (0.8 mL) at 25 °C. The reaction was stirred at 25 °C for 12 h. LCMS showed the starting material was consumed and the main product with desired MS was detected. The mixture was concentrated under reduced pressure to give 3-((3-(4-aminopiperidin-l- yl)phenyl)amino)piperidine-2, 6-dione (Intermediate 33) (0.4 g, 19.5% yield, 43.96% purity) as a yellow solid. LCMS (ESI ⁺ ) m/z 303.2 [M+H] ⁺ .

Intermediate 34 2-allyl-l-[(7R)-7-ethyl-7-hydroxy-5,6-dihydrocyclopenta[b] pyridin-2-yl]-6-[4-(4-oxo-l- piperidyl)anilino]pyrazolo[3,4-d]pyrimidin-3-one (FIG. 34)

[0240] Step 1: A solution of tert-butyl (4-(4-oxopiperidin-l-yl)phenyl)carbamate (1.00 g, 3.44 mmol) in DCM (10 mL) and TFA (1 mL) at 25 °C. The reaction was stirred at 25 °C for 12 h. LCMS showed the reaction was completed and the desired product was detected. The mixture was concentrated under reduced pressure to give l-(4- aminophenyl)piperidin-4- one (0.50 g, 76.31% yield) as a yellow solid. LCMS (ESI ⁺ ) m/z 191.2 [M+H] ⁺ .

[0241] Step 2: To a solution of l-(4-aminophenyl)piperidin-4-one (0.50 g, 2.63 mmol) in toluene (3 mL) were added DIEA (622.15 mg, 4.81 mmol) and 2-allyl-l-[(7R)-7- ethyl-7 -hydroxy-5, 6-dihydrocyclopenta[b]pyridin-2-yl]-6-methylsulfonyl-pyrazol o[3, 4- d]pyrimidin-3-one (Huang et al., J. Med. Chem. (2021) 64: 13004-13024, 1.00 g, 2.41 mmol) at 25 °C. The reaction was stirred at 25 °C for 12 h. LCMS showed the reaction was completed and the desired product was detected. The mixture was diluted with water (10 mL) and extracted with EA (3 x 10 mL). The organic combined organic layers were washed with brine (3 x 10 mL) and dried over Na SCL. The crude product was purified by column chromatography on silica gel (eluted with PE:EA = 100:0 to 20: 1) to give 2-allyl-l-[(7R)-7- ethyl-7 -hydroxy-5, 6-dihydrocyclopenta[b]pyridin-2-yl] -6- [4-(4-oxo- 1 - piperidyl)anilino]pyrazolo[3,4-d]pyrimidin-3-one (0.30 g, 23.71% yield) as a yellow solid.

LCMS (ESI ⁺ ) m/z 526.2 [M+H] ⁺ .

Intermediate 35

3-[4-(2-aminoethylamino)phenyl]piperidine- 2,6-dione (FIG. 35)

[0242] Step 1: To a solution of 3-(4-aminophenyl)piperidine-2, 6-dione (0.60 g, 1.72 mmol) and tert-butyl N-(2-bromoethyl)carbamate (384.8 mg, 1.72 mmol) in DMF (12.0 mL) was added DIEA (443.9 mg, 3.43 mmol) at 20 °C. The mixture was stirred at 80 °C for 12 h under N2. LCMS showed the starting material was consumed completely. The mixture was concentrated in vacuum to give a residue. The residue was purified by prep-HPLC (NH4HCO3 condition) to afford tert-butyl N-[2-[4-(2,6-dioxo-3- piperidyl)anilino]ethyl] carbamate (60.0 mg, 9.38% yield) as a white solid. LCMS (ESI ⁺ ) m/z 348.2 [M+H] ⁺ . 'H NMR (400 MHz, DMSO-d ₆ ) 5 10.72 (s. 1H). 6.98 - 6.78 (m, 3H), 6.52 (d. J = 8.5 Hz. 2H), 5.54 (br t, J - 5.2 Hz, 1H), 3.64 (dd, J - 10.5, 4.9 Hz, 1H), 3.05 (td, J - 11.0, 5.5 Hz, 4H), 2.66 - 2.55 (m, 1H). 2.47 - 2.40 (m, 1H), 2.14 - 2.05 (m, 1H), 2.05 - 1.94 (m, 1H), 1.38 (s, 9H).

[0243] Step 2: A mixture of tert-butyl N-[2-[4-(2,6-dioxo-3- piperidyl)anilino]ethyl]carbamate (60.0 mg, 161.15 pmol, 93.31% purity) in DCM (2.0 mL) and TFA (0.2 mL) was stirred at 20 °C for 2 h. TLC showed the starting material was consumed completely. The mixture was concentrated in vacuum to give 3-[4-(2- aminoethylamino)phenyl]piperidine-2,6-dione (TFA salt, 42.0 mg, 70.61% yield) as a yellow oil. Intermediate 36

2-allyl-6-[4-(4-amino-l-pipcridyl)anilino]-l-[(7R)-7-cthy l-7-hydroxy-5,6- dihydrocy clopenta[b]pyridin-2-yl] pyrazolo[3 ,4-d]pyrimidin-3 -one (FIG. 36)

[0244] Step 1: To a solution of N-[l-(4-aminophenyl)-4-piperidyl]-2,2,2- trifluoro-acetamide (829.76 mg, 2.89 mmol) and 2-allyl-l-[(7R)-7-ethyl-7-hydroxy-5,6- dihydrocyclopcnta[b]pyridin-2-yl]-6-mcthylsulfonyl-pyrazolo[ 3,4-d]pyrimidin-3-onc (1.00 g, 2.41 mmol) in DMF (10 mL) was added DIEA (622.16 mg, 4.81 mmol). The reaction was stirred at 20 °C for 12 h. LCMS showed the reaction was completed. The mixture was diluted with water (20 mL) and extracted with EA (3 x 30 mL). The organic layer was dried over Na SCM and filtered. The filtrate was concentrated under reduced pressure to give N-[l-[4- [[2-allyL 1 - [(7R)-7 -ethyl-7 -hydroxy-5, 6-dihydrocyclopenta[b]pyridin-2-yl] -3- oxopyrazolo[3,4-d]pyrimidin-6-yl]amino]phenyl]-4-piperidyl]- 2,2,2-trifluoro-acetamide (1.10 g, 73.4% yield) as a yellow solid. LCMS (ESI ⁺ ) m/z 623.2 [M+H] ⁺ . ^} H NMR (400 MHz, DMSO-d ₆ ) 5 10.12 (br s, 1H), 9.35 (br d, 7 = 7.8 Hz, 1H), 8.82 (s, 1H), 7.93 (br d, J = 7.3 Hz, 1H), 7.76 - 7.64 (m, 1H). 7.58 (br s. 2H), 6.94 (br d, J = 9.0 Hz, 2H), 5.71 - 5.61 (m, 1H), 5.04 (s, 1H), 4.99 (dd, J = 10.3, 1.0 Hz, 1H), 4.92 - 4.82 (m, 1H), 4.77 (br d, J = 14.4 Hz, 1H), 4.57 (br dd, J = 15.4, 5.6 Hz, 1H), 3.69 (br d, J = 12.5 Hz, 2H), 3.04 - 2.91 (m, 1H), 2.83 - 2.65 (m, 3H), 2.26 - 2.16 (m. 1H). 2.07 - 2.00 (m, 2H), 1.94 - 1.83 (m, 2H), 1.78 - 1.61 (m, 3H), 0.87 (t, J = 7.4 Hz, 3H).

[0245] Step 2: To a solution of N-[l-[4-[[2-allyl-l-[(7R)-7-ethyl-7-hydroxy-5,6- dihydrocyclopenta[b]pyridin-2-yl] -3 -oxo-pyrazolo [3 ,4-d]pyrimidin-6-y 1] am i no] phcny 1] -4- piperidyl]-2,2,2-trifluoro-acetamide (1.00 g, 1.61 mmol) in MeOH (50 mL) was added K2CO3 (1.78 g. 12.85 mmol). The reaction was stirred at 20 °C for 12 h. LCMS showed the reaction was completed. The mixture was concentrated under reduced pressure to give 2-allyl-6-[4- (4-amino-l-pipcridyl)anilino]-l-[(7R)-7-cthyl-7-hydroxy-5,6- dihydrocyclopcnta[b]pyridin- 2-yl]pyrazolo[3,4-d]pyrimidin-3-one (Intermediate 36) (0.96 g, 90.8% yield) as a white solid. LCMS (ESI ⁺ ) mJz 527.3 [M+H] ⁺ . ’ H NMR (400 MHz, DMS0-d ₆ ) 5 10.22 - 9.95 (m, 1H), 8.87 - 8.75 (m, 1H), 7.93 (br d, J = 7.3 Hz, 1H), 7.69 (d, J = 8.1 Hz, 1H), 7.64 - 7.44 (m, 2H), 6.91 (br d, J= 8.9 Hz, 2H), 5.77 - 5.56 (m. 1H), 5.14 - 5.03 (m, 1H). 5.00 (br d. J= 10.1 Hz, 1H), 4.86 (br d, J = 17.2 Hz, 1H), 4.75 (br d, J = 12.8 Hz, 1H), 4.57 (br dd, J = 15.6, 5.6 Hz, 1H), 3.62 - 3.44 (m, 2H), 3.30 (br s, 2H), 3.09 - 2.90 (m, 2H), 2.83 - 2.71 (m, 2H), 2.71 - 2.60 (m, 2H), 2.26 - 2.12 (m. 1H). 2.07 - 1.99 (m, 1H), 1.95 - 1.83 (m, 1H). 1.83 - 1.63 (m. 3H), 1.40 - 1.28 (m, 1H), 0.87 (br t, J - 7.4 Hz, 3H).

Intermediate 37

4-(3-(4-oxopiperidin-l-yl)phenyl)piperidine-2, 6-dione (FIG. 37)

[0246] Step 1: To a solution of 4-(3-bromophenyl)piperidine-2, 6-dione (1 g, 3.73 mmol) and l,4-dioxa-8-azaspiro[4.5]decane (194.96 mg, 1.36 mmol) in dioxane (20 mL) were added CS2CO3 (1.33 g, 4.08 mmol) and l,3-bis[2,6-bis(l-propylbutyl)phenyl]-4,5-dichloro- imidazol-2-ylidene]-dichloro-(3-chloropyridin- 1-ium- l-yl)palladium (92.79 mg, 136.16 pmol) 25 °C under N2. The reaction was stirred at 70 °C under N2 for 12 h. LCMS showed the reaction was completed and the desired product was detected. The reaction was diluted with water (100 mL) and extracted with EA (100 mL). The organic layer was dried over Na2SC>4. After filtering, the filtrate was concentrated under reduced pressure to give the crude product. The crude product was purified by column chromatography on silica gel (eluted with PE:EA = 99: 1 to 10:90) to give 4-(3-(l,4-dioxa-8-azaspiro[4.5]decan-8-yl)phenyl)piperidine- 2, 6-dione (0.16 g, 484.29 pmol, 35.57% yield) as a yellow solid. LCMS (ESI ⁺ ) m/z 331.2 [M+H] ⁺ .

[0247] Step 2: A solution of 4-(3-(l,4-dioxa-8-azaspiro[4.5]decan-8- yl)phenyl)piperidine-2, 6-dione (0.15 g, 454.03 pmol) in HCOOH (1.5 mL) was stirred at 90 °C for 12 h. LCMS showed the reaction was completed and the desired product was detected. The reaction was cooled to 25 °C and quenched with saturated NaHCCL solution. The resulting mixture was extracted with EA (100 mL). The organic layer was dried over Na2SO4. After filtered, the filtrate was concentrated under reduced pressure to give 4-(3-(4- oxopiperidin-l -yl)phenyl)piperidine-2, 6-dione (Intermediate 37) (0.11 g, 384.18 pmol, 84.62% yield), which was used to next step without further purification. LCMS (ESI ⁺ ) m/z 287.1 [M+H] ⁺ . ^! H NMR (400 MHz, DMSO-d ₆ ) 5 10.83 (br s, 1H). 7.19 (t, J = 7.9 Hz. 1H), 6.98 (s, 1H), 6.89 (dd, 7 = 2.0, 8.1 Hz, 1H), 6.70 (d, 7 = 7.7 Hz, 1H), 3.60 (t, 7 = 5.9 Hz, 4H), 3.40 - 3.34 (m, 1H), 2.82 (dd, 7 = 11.4, 16.8 Hz, 2H), 2.66 (d, 7 = 4.3 Hz, 1H), 2.61 (d, 7 = 4.4 Hz. 1H), 2.40 (t. 7 = 5.9 Hz, 4H).

Intermediate 38

4-[4-[4-[[2-allyl-l-[(7R)-7-ethyl-7-hydroxy-5,6-dihydrocy clopenta[b]pyridin-2-yl]-3-oxo- pyrazolo [3 ,4-d]pyrimidin-6-yl] amino]phenyl]piperazin- 1 -yl]butanal (FIG. 38)

[0248] To a solution of 2-allyl-l-[(7R)-7-cthyl-7-hydroxy-5,6- dihydrocyclopenta[b]pyridin-2-yl]-6-(4-piperazin-l-ylanilino )pyrazolo[3,4-d]pyrimidin-3- one (Intermediate 8) (0.1 g, 195.08 umol) in THF (5 mL) were added butanedial (503.83 mg, 5.85 mmol) and NaBH(OAc)3 (82.69 mg, 390.16 umol,) at 20 °C for 2 h under N2. LCMS showed the starling material was consumed completely. The mixture was poured into H2O (5 mL). The mixture was extracted with EA (3 x 5 mL). The organic phase was washed with brine (10 mL), dried over anhydrous NarSCL and concentrated in vacuum to give a residue,, which was purified by prep-TLC to afford 4-[4-[4-[[2-allyl-l-[(7R)-7-ethyl-7-hydroxy-5,6- dihydrocyclopenta[b]pyridin-2-yl]-3-oxo-pyrazolo[3,4-d]pyrim idin-6- yl]amino]phenyl]piperazin-l-yl]butanal (Intermediate 38) (0.082 g, 101.60 umol, 52.08% yield, 72.20% purity) as a yellow solid. LCMS (ESI ⁺ ) m/z 583.4 [M+H] ⁺ . Intermediate 40

3-[4-[4- [[4-(4- aminophcnyl)pipcrazin-l-yl]mcthyl]-4-hydroxy-l- piperidyl]anilino]piperidine-2, 6-dione (FIG. 39)

[0249] Step 1: To a mixture of tert-butyl 4-oxopiperidine- 1 -carboxylate (20.00 g, 100.38 mmol) in ACN (200 mL) were added trimethylsulfoxonium iodide (26.51 g, 120.45 mmol) and KOH (8.45 g, 150.57 mmol) at 20 °C, and the mixture was stirred at 20 °C for 12 h. TLC (PE:EA = 1: 1) showed the starting material was consumed completely. The mixture was poured into H ₂ O (150 mL). The mixture was extracted with EA (3 x 150 mL). The organic phase was washed with brine (150 mL), dried over anhydrous NaiSOr and concentrated in vacuum to give a residue. The residue was used for nest step reaction directly without purification. l-(4-nitrophcnyl)pipcrazinc (20.00 g, 46.71% yield) was obtained as a yellow oil. ¹ H NMR (400 MHz, DMSO-d ₆ ) ¹ H NMR (400 MHz, DMSO-d ₆ ) 53.66 (br d, J = 12.5 Hz, 2H), 2.99 (br s, 2H), 2.89 - 2.54 (m, 2H), 1.55 - 1.42 (m, 4H), 1.38 (s, 9H).

[0250] Step 2: To a solution of tert-butyl N-(l-oxaspiro[2.5]octan-6-yl)carbamate (10.00 g, 23.44 mmol, 50.00% purity) and l-(4-nitrophenyl)piperazine (5.83 g, 28.13 mmol) in MeOH (100 mL) was added DIEA (9.09 g, 70.33 mmol, 12.25 mL). The reaction was stirred at 60 °C for 12 h. LCMS showed the reaction was completed and the desired product was detected. The mixture was concentrated under reduced pressure at 40 °C to give the crude product. The crude product was purified by column chromatography on silica gel (eluted with PE:EA =4 0:60) to give the pure tert-butyl 4-hydroxy-4-((4-(4-nitrophenyl)piperazin-l- yl)methyl)piperidine- 1 -carboxylate (8.00 g, 81.15% yield) was obtained as yellow oil. LCMS (ESI ⁺ ) m/z 421.3 [M+H] ⁺ . ’H NMR (400 MHz, DMSO-d ₆ ) 8 8.04 (d, J = 9.4 Hz, 2H), 7.00 (d, J = 9.4 Hz, 2H), 4.27 (s. 1H), 3.60 (br d, J = 12.8 Hz, 2H), 3.47 - 3.38 (m. 4H). 3.10 (br s, 2H), 2.67 - 2.60 (m, 4H), 2.30 (s, 2H), 1.48 - 1.42 (m, 4H), 1.38 (s, 9H).

[0251] Step 3: To a solution of tert-butyl 4-hydroxy-4-[[4-(4- nitrophenyl)piperazin-l-yl]methyl]piperidine -1 -carboxylate (8.00 g, 19.02 mmol, 100% purity) in THF (160 mL) was added Pd/C (10%, 2.02 g) under N2 atmosphere. The suspension was degassed and purged with H2 (3x). The mixture was stirred under H2 (15 psi) at 20 °C for 12 h. LCMS showed the reaction was completed and the desired product was detected. The mixture was concentrated under reduced pressure at 40 °C to give tert-butyl 4-[[4-(4- aminophenyl)piperazin-l-yl]methyl]-4-hydroxy -piperidine- 1 -carboxylate (7.00 g, 88.02% yield) as a white solid. LCMS (ESI ⁺ ) m/z 391.1 [M+H] ⁺ .

[0252] Step 4: To a solution of tert-butyl 4-[[4-(4-aminophenyl)piperazin-l- yl]methyl]-4-hydroxy-piperidine -1 -carboxylate (7.00 g, 16.75 mmol, 93.42% purity) in DCM (70 mL) were added TFAA (3.87 g, 18.42 mmol, 2.56 mL) and TEA (3.39 g, 33.49 mmol, 4.66 mL). The reaction was stirred at 20 °C for 12 h. LCMS showed the reaction was completed and the desired product was detected. The mixture was concentrated under reduced pressure at 40 °C to give a product, which was used in the next step without further purification. Tert-butyl 4-hydroxy-4-[[4-[4-[(2,2,2-trifluoroacetyl)amino]phenyl]pipe razin- l-yl]methyl] piperidine- 1 -carboxylate (6.00 g, 67.77% yield, 92.02% purity) was obtained as a yellow oil. LCMS (ESI ⁺ ) m/z 487.3 [M+H] ⁺ . ’H NMR (400 MHz. CDCI3) 5 7.92 (br s, 1H), 7.52 - 7.40 (m, 2H), 6.92 (d, J = 8.9 Hz, 2H), 4.03 - 3.83 (m, 2H), 3.37 - 3.06 (m, 6H), 2.90 (s, 3H), 2.74 - 2.68 (m, 1H), 2.61 - 2.28 (m, 2H), 1.62 (br s, 3H), 1.47 (s, 9H).

[0253] Step 5: A mixture of tert-butyl 4-hydroxy-4-[[4-[4-[(2,2,2-trifluoroacetyl) amino]phenyl] piperazin- l-yl]methyl]piperidine-l -carboxylate (6.00 g, 11.35 mmol, 92.02% purity) in TFA (60 mL) and DCM (6 mL) was stirred at 20 °C for 12 h. LCMS showed the reaction was completed and the desired product was detected. The mixture was concentrated under reduced pressure at 40 °C to give a product, which was used in the next step without further purification. 2,2,2-trifluoro-N-(4-(4-((4-hydroxypiperidin-4-yl)methyl)pip erazin-l- yl) phenyl)acetamide (4.00 g, 85.65% yield) was obtained as a black solid. LCMS (ESI ⁺ ) m/z 387.2 [M+H] ⁺ .

[0254] Step 6: To a solution of 2,2,2-trifluoro-N-[4-[4-[(4-hydroxy-4- piperidyl)methyl]piperazin-l-yl] phenyl] acetamide (3.85 g, 9.36 mmol, TFA salt) and 1- fluoro-4-nitro-benzene (1.10 g, 7.80 mmol, 827.07 L) in DMF (20 mL) was added DIEA (5.04 g, 38.98 mmol, 6.79 mL). The reaction was stirred at 80 °C for 12 h. LCMS showed the reaction was completed and the desired product was detected. The mixture was poured into water (20 mL) and extracted with EA (3 x 20 mL). The combined organic layer was washed with brine (20 mL), dried over sodium sulfate, filtered and concentrated under reduced pressure to give 2,2,2-trifluoro-N-[4-[4-[[4-hydroxy-l-(4-nitrophenyl)-4- pipcridyl]mcthyl]pipcraz in- l-yl]phcnyl] acetamide (2.00 g, 46.05% yield, 91.09% purity) as a white solid. LCMS (ESI ⁺ ) m/z 508.3 [M+H] ⁺ . ’ H NMR (400 MHz, DMS0-d ₆ ) 5 11.02 (s, 1H), 8.02 (d, J = 9.5 Hz, 2H), 7.48 (d, J = 8.9 Hz, 2H), 7.01 (d, J = 9.5 Hz, 2H), 6.93 (d, J = 9.0 Hz, 2H), 4.40 (s, 1H), 3.76 (br d, J = 13.2 Hz, 2H), 3.09 (br d, J = 4.2 Hz, 5H), 2.66 (br s, 4H), 2.34 (s, 2H), 1.61 (br d, J = 3.9 Hz. 4H).

[0255] Step 7: To a solution of 2,2,2-trifluoro-N-[4-[4-[[4-hydroxy-l-(4- nitrophenyl)-4-piperidyl]methyl] piperazin- l-yl]phenyl] acetamide (2.00 g, 3.59 mmol) in MeOH (20 mL) was added K2CO3 (992.24 mg, 7.18 mmol). The mixture was stirred at 20 °C for 2 h. LCMS showed the reaction was completed and the desired product was detected. The mixture was concentrated under reduced pressure at 40 °C to give 4-[[4-(4- aminophenyl)piperazin-l-yl] methyl] -l-(4-nitrophenyl)piperidin-4-ol (1.50 g, 36.12% yield) as a white solid. LCMS (ESI ⁺ ) m/z 412.3 [M+H] ⁺ .

[0256] Step 8: To a solution of 4-[[4-(4-aminophenyl)piperazin-l-yl]methyl]-l- (4-nitrophenyl)piperidin-4-ol (1.50 g, 1.30 mmol, 35.57% purity) in DCM (15 mL) were added BOC2O (311.27 mg, 1.43 mmol) and TEA (262.41 mg, 2.59 mmol). The mixture was stirred at 20 °C for 12 h. The mixture was concentrated under reduced pressure at 40 °C to give tert-butyl N-[4-[4-[[4-hydroxy- l-(4-nitrophenyl)-4-piperidyl]methyl]piperazin- 1- yl]phenyl] carbamate (0.80 g, 96.13% yield) as a yellow oil. LCMS (ESI ⁺ ) m/z 512.4 [M+H] ⁺ .

[0257] Step 9: To a solution of tert-butyl N-[4-[4-[[4-hydroxy-l-(4-nitrophenyl)- 4-piperidyl]methyl]piperazin-l-yl]phenyl]carbamate (0.80 g, 1.25 mmol) in THF (10 mL) was added Pd/C (10%, 182.38 mg) under N2 atmosphere. The suspension was degassed and purged with H2 (3x). The mixture was stirred under H2 (15 psi) at 20 °C for 2 h. LCMS showed the reaction was completed and the desired product was detected. The mixture was filtered over celite. The filtrate was concentrated under reduced pressure to give tert-butyl N- [4-[4-[[l-(4-aminophenyl)-4-hydroxy-4-piperidyl]methyl]piper azin-l-yl]phenyl] carbamate (0.60 g, 88.62% yield) as a yellow oil. The product was used to the next step without further purification. LCMS (ESI ⁺ ) m/z 482.4 [M+H] ⁺ .

[0258] Step 10: To a solution of tert-butylN-[4-[4-[[l-(4-aminophenyl)-4- hydroxy-4-piperidyl]methyl] piperazin- l-yl]phenyl]carbamate (0.60 g, 1.10 mmol) in dioxane (8 mL) and 3-bromopiperidine-2, 6-dione (212.10 mg, 1.10 mmol) were added DIEA (428.30 mg, 3. 1 mmol) and Nal (16.56 mg, 110.46 pmol,). The mixture was stirred at 100 °C for 12 h. LCMS showed the reaction was completed and the desired product was detected. The mixture was poured into water (10 mL) and extracted with EA (3 x 10 mL). The combined organic layer was washed with brine (10 mL), dried over sodium sulfate, filtered and concentrated under reduced pressure to give the crude product. The crude product was purified by column chromatography on silica gel (eluted with PE:EA = 1: 1) to give tert- butylN-[4-[4-[[l-[4-[(2,6-dioxo-3-piperidyl)amino]phenyl]-4- hydroxy-4- piperidyl]methyl]piperazin-l-yl]phenyl]carbamate (0.60 g, 86.75% yield) as a white solid. LCMS (ESI ⁺ ) m/z 593.4 [M+H] ⁺ .

[0259] Step 11: A solution of tert-butyl N-[4-[4-[[l-[4-[(2,6-dioxo-3- piperidyl)amino]phenyl]-4-hydroxy -4-piperidyl]methyl]piperazin-l-yl]phenyl]carbamate (0.3 g, 479.11 pmol, 94.66% purity) in TFA (0.3 mL) and DCM (3 mL) was stirred at 20 °C for 12 h. The mixture was concentrated under reduced pressure at 40 °C to give 3 -[4- [4- [[4- (4- aminophenyl)piperazin- l-yl]methyl]-4-hydroxy- 1 -piperidyl] anilino]piperidine-2, 6-dione (0.15 g, 33.17% yield) as a yellow oil. LCMS (ESI ⁺ ) m/z 493.2 [M+H] ⁺ . ¹ H NMR (400 MHz, DMSO-d ₆ ) 8 10.75 (br s, 1H), 9.01 (br s, 1H), 7.27 (br d, J = 8.2 Hz, 2H), 6.85 - 6.72 (m, 4H), 6.59 (br d, J = 8.7 Hz, 2H), 5.33 (d, J = 7.2 Hz, 1H), 4.23 - 4.13 (m, 1H), 4.07 (s, 1H), 3.09 - 2.98 (m, 6H), 2.95 - 2.83 (m, 2H), 2.76 - 2.64 (m, 5H), 2.60 - 2.53 (m, 2H), 2.32 (s, 2H), 2.10 (dt, J = 8.7, 4.2 Hz, 1H), 1.91 - 1.75 (m, 1H), 1.73 - 1.61 (m, 2H). 1.56 (br d, J = 12.3 Hz, 2H).

Intermediate 43

2-[4-[[2-allyl-l-[(7R)-7-ethyl-7-hydroxy-5,6-dihydrocyclo penta[b]pyridin-2-yl]-3-oxo- pyrazolo[3,4-d]pyrimidin-6-yl]amino]phenyl]ethyl methanesulfonate (FIG. 40)

[0260] Step 1: To a mixture of 2-(4-aminophenyl)ethanol (120.0 mg, 874.77 pmol) and (R)-2-allyl-l-(7-ethyl-7-hydroxy-6,7-dihydro-5H-cyclopenta[b ]pyridin-2-yl)-6- (methylsulfonyl)-l ,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (436.1 mg, 1.05 mmol) in DMF (3 mL) was added DIEA (226.1 mg, 1.75 mmol) in one portion at 20 °C under N2. The mixture was stirred at 20 °C for 12 h. LCMS showed the reaction was completed and the desired product was detected. The mixture was concentrated under reduced pressure at 40 °C to give the crude product. The crude product was purified by column chromatography on silica gel (eluted with PE:EA = 100: 1 to 0: 1) to give 2-allyl-l-[(7R)-7-ethyl-7-hydroxy-5,6- [b]pyridin-2-yl]-6-[4-(2-hydroxyethyl)anilino]pyrazolo[3,4-d ]pyrimidin-3-one (330.0 mg, 75.93%). LCMS (ESI ⁺ ) ni/z 473.2 [M+H] ⁺ .

[0261] Step 2: To a solution of product obtained from above Step 1 (330.0 mg, 664.21 pmol) in DCM (6 mL) were added TEA (134.4 mg, 1.33 mmol) and MsCl (83.7 mg, 730.63 pmol) in one portion at 25 °C under N2. The mixture was stirred at 25 °C for 1 h. TLC showed the starting material consumed and a new spot was detected. The reaction was purified by prep-TLC (PE:EA = 1:1, Rf=0.5) to give 2-[4-[[2-allyl-l-[(7R)-7-ethyl-7- hydroxy- 5,6-dihydrocyclopenta[b]pyridin-2-yl]-3-oxo-pyrazolo[3,4-d]p yrimidin-6- yl]amino]phenyl]ethyl methanesulfonate (Intermediate 43) (160.0 mg, 43%).

Intermediate 44

3- [4-(l,4-diazepan-l-yl)anilino]piperidine-2, 6-dione (FIG. 41)

[0262] Step 1: To a solution of tert-butyl 4-(4-aminophenyl)-l,4-diazepane-l- carboxylate (1.5 g, 5.04 mmol,) and 3-bromopiperidine-2, 6-dione (1.0 g, 5.54 mmol) in DMF (40 mL) was added NaHCCh (846.9 mg, 10.08 mmol) at 20 °C. The mixture was stirred at 60 °C for 12 h. LCMS showed the reaction was completed and the desired product was detected. The mixture was concentrated in vacuum to give tert-butyl 4-[4-[(2,6-dioxo-3- piperidyl)amino]phenyl]-l,4-diazepane-l -carboxylate (1.0 g, 42%), which was used to next step without further purification. LC/MS (ESI) m/z 403.3 [M+H] ⁺ .

[0263] Step 2: To a solution of product obtained from above Step 1 (0.60 g, 1.29 mmol) in DCM (20 mL) and TFA (2 mL) was stirred at 25 °C for 2 h. LCMS showed the reaction was completed and the desired product was detected. The mixture was concentrated under reduced pressure at 40 °C to give the crude product. The crude product was purified by prcp-HPLC to give 3-[4-(l,4-diazcpan-l-yl)anilino]pipcridinc-2,6-dionc (150.0 mg, 36%). LC/MS (ESI) m/z 303.3 [M+H] ⁺ .

Intermediate 47 and Intermediate 48

(R)-2-(4-((2-allyl-l-((R)-7-ethyl-7-hydroxy-6,7-dihydro-5 H-cyclopenta[b]pyridin-2-yl)-3- oxo-2, 3-dihydro-lH-pyrazolo[3,4-d]pyrimidin-6-yl)amino)phenyl)-2-h ydroxyethyl methane sulfonate and (S)-2-(4-((2-allyl-l-((R)-7-ethyl-7-hydroxy-6,7-dihydro-5H- cyclopenta[b]pyridin-2-yl)-3-oxo-2,3-dihydro-lH-pyrazolo[3,4 -d]pyrimidin-6- yl)amino)phenyl)-2-hydroxyethyl methane sulfonate (FIG. 42)

[0264] Step 1: 2-allyl-6-[4-(l,2-dihydroxyethyl)anilino]-l-[(7R)-7-ethyl-7- hydroxy-5,6-dihydrocyclopenta[b]pyridin-2-yl]pyrazolo[3,4-d] pyrimidin-3-one (31.35% yield) was synthesized from l-(4-aminophenyl)ethane-l,2-diol and (R)-2-allyl-l-(7-ethyl-7- hydroxy-6,7-dihydro-5H-cyclopenta[b]pyridin-2-yl)-6-(methyls ulfonyl)-l,2-dihydro-3H- pyrazolo[3,4-d]pyrimidin-3-one using 2-propanol as solvent. LC/MS (ESI) m/z 489.3 [M+HJ ⁺ .

[0265] Step-2: [2-[4-[[2-allyl-l-[(7R)-7-ethyl-7-hydroxy-5,6- dihydrocyclopenta[b]pyridin-2-yl]-3-oxo-pyrazolo[3,4-d]pyrim idin-6-yl]amino]phenyl]-2- hydroxy-ethyl] methanesulfonate (26% yield) was synthesized from the product obtained in above Step 1 by following a similar procedure as described in Step 2 for the synthesis of Intermediate 43. The product obtained was further subjected to SFC purification to afford [(2R)-2-[4-[[2-allyl-l-[(7R)-7-ethyl-7-hydroxy-5,6-dihydrocy clopenta[b]pyridin-2-yl]-3- oxo-pyrazolo[3,4-d]pyrimidin-6-yl]amino]phenyl]-2-hydroxy-et hyl] methanesulfonate (Intermediate 47) (39% yield) and [(2S)-2-[4-[[2-allyl-l-[(7R)-7-ethyl-7-hydroxy-5,6- dihydrocyclopenta[b]pyridin-2-yl]-3- oxo-pyrazolo[3,4-d]pyrimidin-6-yl]amino]phenyl]-2 - hydroxy-ethyl] methanesulfonate (Intermediate 48) (42% yield). Intermediate 50 3-[4-[4-[[l-(4-aminophenyl)-4- piperidyl]amino]-l-piperidyl]anilino]piperidine-2, 6-dione (FIG. 43)

[0266] Step 1: tert-butyl N-[4-[4-[[l-[4-[(2,6-dioxo-3-piperidyl)amino]phenyl]- 4-piperidyl]amino]-l-piperidyl]phenyl]carbamate (0.3 g, 27.85%) was synthesized from 3- ((4-(4-aminopiperidin-l-yl)phenyl)amino)piperidine-2, 6-dione and tert-butyl N-[4-(4-oxo-l- piperidyl)phenyl] carbamate . LC/MS (LCMS) m/z 577.4 [M+H] ⁺ .

[0267] Step 2: 3-[4-[4-[[l-(4-aminophenyl)-4- piperidyl]amino]-l- piperidyl]anilino]piperidine-2, 6-dione (53.26% yield) was synthesized from the product obtained in above Step 1 by following a similar procedure as described in Step 2 for the synthesis of Intermediate 44. LC/MS (ESI) m/z 477.3 [M+H] ⁺ .

Intermediate 51 (2-(3-(2,6-dioxopiperidin-3-yl)benzyl)-2-azaspiro[3.3]heptan -6-yl)methyl methanesulfonate (FIG. 44)

[0268] Step 1: To a mixture of 2,6-dibenzyloxy-3-(4,4,5,5-tetramethyl-l,3,2- dioxaborolan-2-yl)pyridine (7.29 g, 8.73 mmol) and 2-(3-bromophenyl)-l,3-dioxolane (2.00 g, 8.73 mmol) in Dioxane (90 mL) and H2O (9 mL) were added Pd(dppf)C12 (638.8 mg, 873.09 umol) and K2CO3 (2.41 g, 17.46 mmol) at 25 °C under N2. The mixture was stirred at 100 °C for 12 h. LCMS showed the reaction was completed and the desired product was detected. The mixture was extracted with EA (3 x 100 mL). The combined organic was washed with brine (100 mL), dried over Na2SO4, filtered and concentrated under reduced pressure. The crude product was purified by silica gel chromatography (eluted with PE: EA = 100: 1 to 0: 1 , Rf = 0.4) to give 2,6-dibenzyloxy-3-[3-(l ,3-dioxolan-2-yl)phenyl] pyridine (2.00 g 50.43%). LC/MS (ESI) m/z 440.3 [M+H] ⁺ .

[0269] Step 2: To a solution of 2,6-dibenzyloxy-3-[3-(l,3-dioxolan-2- yl)phenyl]pyridine (2.00 g, 4.40 mmol, 96.72% purity) in THF (40 mL) were added Pd/C (0.47 g, 0.44 mmol, 10% purity) and TEA (445.5 mg, 4.40 mmol) at dropwise atmosphere under N2. The reaction was stirred at 25 °C for 12 h under H2 (15 psi). LCMS showed the reaction was completed and the desired product was detected. The mixture was filtered and the filter cake was washed with THF (50 mL), the collected filtrate was concentrated to give 3-[3-(l,3-dioxolan-2-yl)phenyl] piperidine-2, 6-dione (1.29 g, 55.20%). LC/MS (ESI) m/z 262.23 [M+H] ⁺ .

[0270] Step 3: To a solution of 3-[3-(l,3-dioxolan-2-yl)phenyl]piperidine-2,6- dione (1.29 g, 4.94 mmol) in THF (20 mL) was added aqueous HC1 (39.52 mmol 10.05 mL, 2 M) in one portion at 25 °C under N2. The mixture was stirred at 55 °C for 3 h. TLC showed the reaction was completed and new spot was detected. The mixture was concentrated under reduced pressure to give the crude product. The crude product was purified by column chromatography on silica gel (eluted with PE: EA = 100: 1 to 0:1, Rf = 0.4) to afford 3-(2,6- dioxo-3-piperidyl)benzaldehyde (521 mg, 86.86%). ^! H NMR (400 MHz, DMSO-d ₆ ) 8 10.86 (s, 1H), 7.40 - 7.22 (m, 5H), 5.71 (s, 1H), 4.09 - 4.01 (m, 2H), 4.00 - 3.93 (m, 2H), 2.73 - 2.63 (m, 1H), 2.51 - 2.46 (m, 1H), 2.21 (dq. J = 12.3, 4.5, Hz 1H), 2.10 - 1.98 (m, 1H).

[0271] Step 4: 3-[3-[[6-(hydroxymethyl)-2-azaspiro[3.3]heptan-2-yl]methyl] phenyl] piperidine-2, 6-dione (19.53% yield) was synthesized from the product obtained in above Step 3 and 2-azaspiro[3.3]heptan-6-ylmethanol. LC/MS (ESI) m/z 329.2 [M+H] ⁺ .

[0272] Step 5: [2-[[3-(2,6-dioxo-3-piperidyl)phenyl]methyl]-2- azaspiro[3.3]heptan-6-yl]methyl methanesulfonate (Intermediate 51) (47.19% yield) as a white solid was synthesized from the product obtained in above Step-4 by following a similar procedure as described in Step 2 for the synthesis of Intermediate 43. LC/MS (ESI) m/z 407.2 [M+H] ⁺ . Intermediate 52

3-(4-(2-(4-(4-aminophcnyl)pipcrazin-l-yl)cthyl)phcnyl)pip cridinc-2,6-dionc (FIG. 45)

[0273] Step 1: To a solution of 2-[4-(2,6-dibenzyloxy-3-pyridyl)phenyl]ethanol (WO2023/098656 Al ) (0.7 g, 1.07 mmol) in DCM (40.0 mL) was added DMP (1.44 g, 3.4 mmol, 1.05 mL) at 0 °C. The reaction was stirred at 25 °C for 2 h. TLC (PE: EA = 1:1, Rf =0.5) showed the starting material consumed and a new spot was detected. The reaction was quenched with HoO (50.0 mL) at 25 °C and extracted with DCM (3 x 50.0 mL). The organic layer was dried over NaiSCk. After filtration, the filtrate was concentrated under reduced pressure to give the crude product, which was purified by column chromatography on silica gel (eluted with PE: EA = 100: 1 to 1: 1) to give 2-[4-(2,6-dibenzyloxy-3- pyridyl)phenyl] acetaldehyde (0.6 g, 86.14%). LC/MS (ESI) m/z 410.2 [M+H] ⁺ .

[0274] Step 2: l-[2-[4-(2,6-dibenzyloxy-3-pyridyl) phenyl]ethyl]-4-(4- nitrophenyl)piperazine (45.44% yield) was synthesized from product obtained from above Step 1 and l-(4-nitrophenyl)piperazine.

[0275] Step 3: 3-[4-[2-[4-(4-aminophenyl)piperazin-l- yl]ethyl]phenyl]piperidine-2, 6-dione (76.52% yield) was synthesized from product obtained from above Step 2 and by following a similar procedure as described in Step 2 for the synthesis of Intermediate 46.

Intermediate 53 l-((l-(4-(2,6-dioxopipcridin-3-yl)phcnyl)pipcridin-4-yl)mcth yl)azctidin-3-yl methanesulfonate (FIG. 46)

[0276] Step l-[4-(2,6-dibenzyloxy-3-pyridyl)phenyl]piperidine-4- carbaldehyde (31.38% yield) was synthesized by following a similar procedure as described in Step 1 for the synthesis of Intermediate 52. LC/MS (ESI) m/z 479.23 [M+H] ⁺ .

[0277] Step 2: l-[[l-[4-(2,6- dibenzyloxy-3-pyridyl)phenyl]-4- piperidyl] methyl] azetidin-3-ol (53 % yield) was synthesized from the product obtained in above Step 1. LC/MS (ESI) m/z 536.3 [M+H] ⁺ .

[0278] Step 3: 3-[4-[4-(hydroxymethyl)-l-piperidyl]phenyl]piperidine-2, 6-dione (95% yield) was synthesized from the product obtained in above Step 2. LC/MS (ESI) m/z 358.2 [M+H] ⁺ .

[0279] Step 4: [l-[[l-[4-(2,6-dioxo-3-piperidyl)phenyl]-4- piperidyl] methyl] azetidin-3-yl] methane sulfonate (41% yield) was synthesized from the product obtained in above Step 3. LC/MS (ESI) m/z 436.2 [M+H] ⁺ .

In termediate 54

4-(2,6-dioxopiperidin-3-yl)benzaldehyde (FIG. 47)

[0280] Step 1: To a solution of methyl 2-[4-[[tertbutyl(dimethyl)silyl]oxymethyl]phenyl]acetate (2.1 g, 7.13 mmol) in DMF (80 mL) were added prop-2-enamide (1.01 g, 14.26 mmol) and tBuOK (1.60 g. 14.26 mmol) at 0 °C. The reaction was stirred at 0 °C for 1 h. TLC (PE:EA = 1: 1, RIM). ) showed the reaction was completed and the desired product was detected. The reaction was quenched with saturated NH4CI solution and extracted with EA (50 mL). The organic layer was washed with brine (30 mL), dried over NaiSCU and filtered. The filtrate was concentrated to give the crude product, which was purified by flash column on silica gel to give 3-[4-[[tert- butyl(dimethyl)silyl]oxymethyl]phenyl]piperidine-2, 6-dione (1.6 g, 67%). 'H NMR (400 MHz, CDCI3) 5 7.93 (br s, 1H), 7.23 (d, J = 8.1 Hz, 2H), 7.07 (d, J = 8.1 Hz, 2H), 4.63 (s, 2H), 3.69 (dd, J = 5.3, 9.2 Hz. 1H). 2.68 - 2.46 (m, 2H), 2.24 - 2.10 (m, 2H), 0.84 (s. 9H). 0.00 (s, 6H).

[0281] Step 2: To a solution of 3-[4-[[tert- butyl(dimethyl)silyl]oxymethyl]phenyl]piperidine-2, 6-dione (1.6 g, 4.80 mmol) in THF (20 mL) was added aqueous HC1 solution (2 N, 2 mL) at 25 °C. The reaction was stirred at 25°C for 1 h. TLC (PE:EA = 1: 1, Rf=0.2) showed the reaction was completed. The reaction was concentrated under reduced pressure to give 3-(4-(hydroxymethyl)phenyl)piperidine-2,6- dione (0.8 g, 76%), which was used to next step without further purification.

[0282] Step 3: To a solution of 3-[4-(hydroxymethyl)phenyl]piperidine-2, 6-dione (0.42 g, 1.92 mmol) in DCM (30 mL) was added DMP (1.22 g, 2.87 mmol) at 0 °C. The reaction was stirred at 0 °C for 1 h. TLC showed the reaction was completed. The reaction was quenched with saturated NaHCCh solution and extracted with DCM (3 x 15 mL). The organic layer was dried over Na SCL and filtered. The filtrate was concentrated to give the crude product, which was purified by column chromatography on silica gel (eluted with PE:EA = 50: 1 to 5: 1) to give 4-(2,6-dioxo-3-piperidyl)benzaldehyde (Intermediate 54) (0.3 g, 72%).

Intermediate 55

(2-(4-(2,6-dioxopiperidin-3-yl)benzyl)-2-azaspiro[3.3]hep tan-6-yl)methyl methanesulfonate

(FIG. 48)

[0283] Step 1: To a solution of 2-azaspiro[3.3]heptan-6-ylmethanol (0.3 g, 1.89 mmol) in THF (10 mL) were added 4-(2,6-dioxo-3-piperidyl)benzaldehyde (Intermediate 54, 409.9 mg, 1.89 mmol) and NaBH(OAc)s (799.8 mg, 3.77 mmol) and TBSC1 (284.4 mg, 1.89 mmol, 231.2 pL) at 25 °C. The reaction was stirred at 25 °C for 12 h. LCMS showed the reaction was completed and the desired compound was detected. The mixture was diluted with water (10 mL) and extracted with DCM (3 x 10 mL). The combined organic layers were washed with brine (3 x 10 mL) and dried over NaoSCU- After filtration, the filtrate was concentrated under reduced pressure to give 3-[4-[[6-(hydroxymethyl)-2- azaspiro[3.3]heptan-2-yl] methyl]phenyl]piperidine-2, 6-dione (0.1 g, 12.5%). LC/MS (ESI) m/z 329.2 [M+H] ⁺ .

[0284] Step 2: [2-[[4-(2,6-dioxo-3-piperidyl)phenyl] methyl]-2- azaspiro[3.3]heptan-6-yl]methyl methanesulfonate (Intermediate 55) (61% yield) was synthesized from the product obtained in above Step 2 by following a similar procedure as described in Step 2 for the synthesis of Intermediate 43. LC/MS (ESI) m/z 407.2 [M+H] ⁺ .

Intermediate 56

3-((4-(2,6-dioxopiperidin-3-yl)phenyl)amino)propyl methanesulfonate (FIG. 49)

[0285] Step 1: To a mixture of 3-(4-aminophenyl)piperidine-2, 6-dione (0.70 g, 2.00 mmol, 58.45% purity) and 3-bromopropan-l-ol (278.5 mg, 2.00 mmol) in DMF (14 mL) was added DIEA (517.8 mg, 4.01 mmol) in one portion at 25 °C under N2. The mixture was stirred at 80 °C for 12 h. LCMS showed the reaction was completed and the desired product was detected. The mixture was concentrated under reduced pressure at 60 °C to give the crude product. The crude product was purified by prep-TLC (PE: EA = 0: 1, Rf = 0.4) to give 3-[4- (3-hydroxypropylamino)phenyl]piperidine-2, 6-dione (0.15 g, 15.49%).

[0286] Step 2: 3 -[4-(2,6-dioxo-3-piperidyl)anilino] propyl methanesulfonate (47% yield) was synthesized from the product obtained in above Step 1 by following a similar procedure as described in Step 2 for the synthesis of Intermediate 43. LC/MS (ESI) m/z 436.2 [M+H] ⁺ . Intermediate 57

4-(2,6-dioxopipcridin-3-yl)phcncthyl mcthancsulfonatc (FIG. 50)

[0287] 4-(2,6-dioxopiperidin-3-yl)phenethyl methanesulfonate (Intermediate 57) (57% yield) was synthesized from 3- [4-(2-hydroxyethyl)phenyl]piperidine-2, 6-dione by following a similar procedure as described in Step 2 for the synthesis of Intermediate 43. LC/MS (ESI) m/z 329.1 [M+H] ⁺ .

Intermediate 58

2-((4-(2,6-dioxopiperidin-3-yl)phenyl)amino)ethyl methanesulfonate (FIG. 51)

[0288] 2-[4-(2,6-dioxo-3-piperidyl)anilino]ethyl methanesulfonate (22.27%) was synthesized from 3-[4-(2-hydroxyethylamino)phenyl]piperidine-2, 6-dione.

Intermediate 60

3-[4-(2,6-dioxo-3-piperidyl)phenyl]propanal (FIG. 52)

[0289] Step 1: To a mixture of 3-(4-bromophenyl)propan-l-ol (0.5 g, 2.32 mmol) in Dioxane:H2O = 10: 1 (11 mL) were added 2,6-bis(benzyloxy)-3-(4,4,5,5-tetramethyl-l,3,2- dioxaborolan-2-yl)-pyridine (1.16 g, 2.79 mmol), Pd(dppf)Ch (85.05 mg, 116.23 pmol) and K2CO3 (642.56 mg, 4.65 mmol,) at 20 °C. The mixture was stirred at 100 °C for 12 h under N2. LCMS showed the starting material consumed and the desired product was detected. The mixture was poured into H2O (10 mL). The mixture was extracted with EA (3 x 10 mL). The organic phase was washed with brine (5 mL), dried over anhydrous NaiSCL and concentrated in vacuum to give reduced pressure to give a residue. The crude product was purified by column chromatography on silica gel (eluted with PE: EA = 100: 1 to 10: 1) to give 3-[4-(2,6- dibenzyloxy-3-pyridyl)phenyl]propan-l-ol (0.5 g, 37.13%) as a white solid. LC/MS (ESI) m/z 426.2 [M+H] ⁺ .

[0290] Step 2: 3-[4-(3-hydroxypropyl)phenyl]piperidine-2, 6-dione (38% yield) was synthesized from the product obtained in above Step 1 by following a similar procedure as described in Step 2 for the synthesis of Intermediate 46.

[0291] Step 3: 3-[4-(2,6-dioxo-3-piperidyl)phenyl]propanal (Intermediate 60) (50% yield) was synthesized from the product obtained in above Step 2 by following a similar procedure as described in Step 1 for the synthesis of Intermediate 52.

Example 1: 3-((4-(4-(4-((2-allyl-l-((R)-7-ethyl-7-hydroxy-6,7-dihydro-5 H- cyclopenta[b]pyridin-2-yl)-3-oxo-2,3-dihydro-lH-pyrazolo[3,4 -d]pyrimidin-6- yl)amino)benzyl)piperazin-l-yl)phenyl) amino)piperidine-2, 6-dione (Compound 1)

[0292] To a mixture of Intermediate 1 (80.0 mg, 170 pmol, 84% purity) in i- PrOH (1 mL) were added a solution of 2-allyl-l-[(7R)-7-ethyl-7-hydroxy-5,6- dihydrocyclopenta[£]pyridin-2-yl]-6-methylsulfonyl-pyrazolo [3,4-< ]pyrimidin-3-one (Huang et al., J. Med. Chem. (2021) 64: 13004-13024, 70.8 mg, 170 pmol) in z’-PrOH (0.5 mL) at 20 °C. Then the mixture was stirred at 80 °C for 12 h under N2. LCMS showed the reaction was completed and the desired product was detected. The mixture was purified by prep- HPLC (NH4HCO3 condition) to afford Compound 1, 3-[4-[4-[[4-[[2-allyl-l-[(7R)-7-ethyl- 7-hydroxy-5,6-dihydrocyclopenta[/?]pyridin-2-yl]-3-oxo-pyraz olo[3,4-<7]pyrimidin-6- yl]amino]phenyl]methyl]piperazin-l-yl]anilino]piperidine-2, 6-dione (10.3 mg, 8% yield, 97.17% purity). LC/MS (ESI) m/z 729.4 [M+H] ⁺ , RT: 2.696 min. ’ H NMR (400 MHz, DMSO-d ₆ ) 8 10.74 (br s, 1H, NH), 10.25 (br s, 1H, NH), 8.88 (s, 1H), 7.92 (d, J = 8.1 Hz, 1H), 7.70 (d, J = 8.1 Hz, 1H), 7.70 (d, J = 8.4 Hz, 1H), 7.27 (d, J = 8.4 Hz, 2H), 6.74 (d, J = 8.9 Hz, 2H), 6.60 (d, J = 8.9 Hz, 2H), 5.68 (tdd, J = 16.8, 10.6, 5.8 Hz, 1H), 5.37 (d, J = 7.3 Hz, 1H), 5.05 (s, 1H, OH), 5.00 (dd, J = 10.3, 1.0 Hz, 1H), 4.86 (dd, J = 17.1, 1.0 Hz, 1H), 4.76 (br dd, J = 16.1, 4.9 Hz, 1H), 4.56 (br dd, J = 16.0, 6.1 Hz, 1H), 4.18 (ddd, 7 = 11.3, 6.9, 4.8 Hz, 1H), 3.47 (s, 2H), 3.04 - 2.87 (m, 5H), 2.85 - 2.65 (m, 2H), 2.62 - 2.53 (m, 1H), 2.49 - 2.44 (m, 4H), 2.20 (ddd, J = 13.5, 8.3, 5.8 Hz, 1H), 2.10 (ddd, J = 13.3. 8.7, 4.4 Hz. 1H). 2.01 (ddd, 7 - 13.4, 8.4, 5.3 Hz, 1H), 1.94 - 1.77 (m, 2H), 1.76 - 1.64 (m, 1H), 0.87 (t. 7 - 7.4 Hz, 3H).

Example 2: 3-((4-(4-(4-((2-allyl-l-((R)-7-ethyl-7-hydroxy-6,7-dihydro-5 H- cyclopenta[b]pyridin-2-yl)-3-oxo-2,3-dihydro-lH-pyrazolo[3,4 -dJpyrimidin-6- yl)amino)phenethyl)piperazin-l-yl) phenyl)amino)piperidine-2, 6-dione (Compound 2)

[0293] Compound 2 was prepared by following the similar procedure described in Example 1 using Intermediate 2 (0.120 g, 294 pmol ) in place of Intermediate 1, afforded the desired titled product Example 2, 3-((4-(4-(4-((2-allyl-l-((R)-7-ethyl-7-hydroxy-6,7- dihydro-5/7-cyclopenta[Z?]pyridin-2-yl)-3-oxo-2,3-dihydro-l/ 7-pyrazolo[3,4-<7]pyrimidin-6- yl)amino)phenethyl)piperazin- l-yl)phenyl)amino)piperidine-2, 6-dione after purification (13.8 mg. 18.6 pmol, 6.3% yield. 100% purity). 'l l NMR (400 MHz, DMSO-d ₆ ) 5 10.75 (s, 1H. NH). 10.21 (br s. 1H, NH), 8.87 (s. 1H). 7.91 (d, J = 8.1 Hz, 1H), 7.70 (d. J = 8.1 Hz. 1H), 7.65 (br d, J - 8.4 Hz, 2H), 7.20 (d, J = 8.4 Hz, 2H), 6.76 (d, J = 8.9 Hz, 2H), 6.61 (d, J = 8.9 Hz, 2H), 5.74 - 5.61 (m. 1H), 5.37 (br d, J = 6.0 Hz, 1H, NH), 5.05 (br s, 1H, OH), 5.00 (dd, J = 9.2, 1.0 Hz, 1H), 4.86 (dd, J = 17.1, 1.0 Hz, 1H), 4.75 (br dd, J = 15.7, 3.8 Hz, 1H), 4.56 (br dd, J = 16.0, 6.1 Hz, 1H), 4.24 - 4.13 (m, 1H), 3.01 - 2.90 (m, 5H), 2.80 - 2.68 (m, 4H), 2.60 - 2.48 (m. 7H), 2.26 - 2.15 (m, 1H), 2.15 - 2.06 (m, 1H). 2.01 (ddd. J = 13.4, 8.4, 5.3 Hz, 1H), 1.95 - 1.77 (m, 2H), 1.76 - 1.64 (m, 1H), 0.87 (t, J = 7.4 Hz, 2H). Example 3: 3-((4-(4-(4-((2-allyl-l-((R)-7-cthyl-7-hydroxy-6,7-dihydro-5 H- cyclopenta[b]pyridin-2-yl)-3-oxo-2,3-dihydro-lH-pyrazolo[3,4 -d]pyrimidin-6- yl)amino)phenyl)piperazin-l-yl)phenyl) amino )piperidine-2,6-dione (Compound 3)

[0294] Compound 3 was prepared by following the similar procedure described in Example 1 using Intermediate 3 (70.0 mg, 175 pmol 94.91% purity) in place of Intermediate 1. The mixture was filtered and the filtrate was concentrated to give a residue, which was purified by prep-HPLC (NH4HCO3 condition) to give Compound 3, 3-((4-(4-(4- ((2-allyl-l-((R)-7-ethyl-7-hydroxy-6,7-dihydro-5H-cyclopenta [/?]pyridin-2-yl)-3-oxo-2,3- dihydro-lH-pyrazolo[3,4-<7]pyrimidin-6-yl)amino)phenyl)pi perazin-l- yl)phenyl)amino)piperidine-2, 6-dione (12.0 mg, 9.4% yield, 98.42% purity). LC/MS (ESI ) m/z 713.4 [M-H]-, RT: 2.681 min. 'H NMR (400 MHz, DMSO-d ₆ ) 8 10.51 - 10.38 (m, 1H, NH), 9.74 (s, 1H, NH), 8.78 (s, 1H), 7.87 (d, J = 8.2 Hz, 1H), 7.69 (d, J = 8.2 Hz, 1H), 7.58 (d, J = 8.9 Hz, 2H), 6.96 (d, J = 8.9 Hz, 2H), 6.84 (d, J = 8.8 Hz, 2H), 6.69 (d, J = 8.8 Hz, 2H), 5.78 - 5.63 (m, 1H), 5.17 (d, J = 7.0 Hz, 1H). 5.02 (d, J = 10.4 Hz, 1H), 4.94 (d. J = 17.6 Hz, 1H), 4.76 - 4.68 (m, 1H), 4.71 (s, 1H, OH), 4.62 - 4.54 (m, 1H), 4.17 (ddd, J = 10.7, 6.3, 4.9 Hz, 1H), 3.30 - 3.21 (m, 4H), 3.16 - 3.08 (m, 4H), 3.08 - 3.04 (m, 1H), 3.00 - 2.95 (m, 1H), 2.85 - 2.76 (m. 1H), 2.73 - 2.63 (m, 1H). 2.28 - 2.13 (m, 2H), 2.12 - 2.02 (m, 1H). 1.98 - 1.82 (m, 2H), 1.81 - 1.69 (m, 1H), 0.90 (t, J = 7.5 Hz, 3H). Example 4: 3-((4-(4-(3-(4-((2-allyl-l-((R)-7-ethyl-7-hydroxy-6,7-dihydr o-5H-cyclopenta[b] pyridin-2-yl)-3-oxo-2,3-dihydro-lH-pyrazolo[3,4-d]pyrimidin- 6yl)amino)phenyl)propyl) piperazin- l-yl)phenyl)amino)piperidine-2, 6-dione (Compound 4)

[0295] Compound 4 was prepared by following the similar procedure described in Example 1 using Intermediate 4 (80.0 mg, 193 pmol) in place of Intermediate 1. The mixture was filtered and the filtrate was concentrated to give a residue, which was purified by prep-HPLC (FA condition) to give Compound 4, 3-((4-(4-(3-(4-((2-allyl-l-((R)-7-ethyl-7- hydroxy-6,7-dihydro-5/7-cyclopenta[Z?]pyridin-2-yl)-3-oxo-2, 3-dihydro- 1 H-pyrazolo[3,4- d]pyrimidin-6-yl)amino)phenyl)propyl)piperazin-l-yl)phenyl)a mino)piperidine-2, 6-dione, (18.4 mg, 24.1 pmol, 7.8% yield, 99% purity). LC/MS (ESI ⁺ ) mJz 757.4 [M+H] ⁺ . RT: 2.155 min. ’ H NMR (400 MHz, DMSO-d ₆ ) 8 10.75 (br s, 1H, NH), 10.21 (br s, 1H, NH), 8.87 (s, 1H), 7.92 (d, J = 8.1 Hz, 1H), 7.70 (d, J = 8.1 Hz, 1H), 7.65 (br d. J = 8.1 Hz, 2H), 7.17 (d, J = 8.1 Hz, 2H), 6.75 (d, J = 8.8 Hz, 2H), 6.61 (d, J = 8.8 Hz, 2H), 5.73 - 5.61 (m, 1H), 5.37 (d, J = 7.2 Hz, 1H), 5.05 (br s, 1H), 5.00 (d, J = 9.7 Hz, 1H), 4.86 (br d, J = 17.4 Hz, 1H), 4.81 - 4.70 (m, 1H). 4.56 (brdd, J = 16.0, 6.0 Hz. 1H), 4.23 - 4.13 (m, 1H). 3.02 - 2.96 (m, 1H). 2.94 (br s, 4H), 2.83 - 2.65 (m, 2H), 2.60 - 2.54 (m, 3H), 2.49 - 2.45 (m, 4H), 2.31 (br t, J = 7.0 Hz, 2H), 2.20 (ddd, J = 13.4, 8.1, 5.6 Hz, 1H), 2.15 - 2.06 (m, 1H), 2.01 (ddd, J = 13.4, 8.4, 5.3 Hz, 1H), 1.95 - 1.80 (m. 2H). 1.80 - 1.65 (m, 3H), 0.87 (t, J = 7.4 Hz. 3H).

Example 5: 3-((3-(4-(4-((2-allyl-l-((R)-7-ethyl-7-hydroxy-6,7-dihydro-5 H- cyclopenta[b]pyridin-2-yl)-3-oxo-2,3-dihydro-lH-pyrazolo[3,4 -d]pyrimidin-6- yl)amino)phenyl)piperazin-l-yl)phenyl) amino)piperidine-2,6-dione (Compound 5) [0296] Compound 5 was prepared by following the similar procedure described in Example 1 using Intermediate 6 (0.250 g, 333 pmol, 50.47% purity) in place of Intermediate 1. The mixture was filtered and the filtrate was concentrated to give a residue, which was purified by prep-HPLC (NH4HCO3 condition) to give Compound 5, 3-((3-(4-(4- ((2-allyl-l-((R)-7-ethyl-7-hydroxy-6,7-dihydro-5H-cyclopenta [/?]pyridin-2-yl)-3-oxo-2,3- dihydro-lH-pyrazolo[3,4-<7]pyrimidin-6-yl)amino)phenyl)pi perazin-l- yl)phenyl)amino)piperidine-2, 6-dione, (24.4 mg, 10% yield, 100% purity). LC/MS (ESI ⁺ ) m/z 715.3 [M+H] ⁺ , RT: 2.179 min. ‘H NMR (400 MHz. DMSO-d ₆ ) 5 10.77 (br s, 1H, NH), 10.32 - 10.03 (m, 1H, NH), 8.83 (s, 1H), 7.94 (br d, J = 5.6 Hz, 1H), 7.70 (d, J = 8.1 Hz, 1H), 7.67 - 7.54 (m, 2H), 6.99 (d, 7 = 8.1 Hz, 1H), 6.95 (dd, J = 8.0, 8.0 Hz, 1H), 6.34 (s, 1H), 6.26 (dd. J = 8.0, 1.6 Hz, 1H). 6.18 (dd, J = 8.0, 1.6 Hz, 1H), 5.73 - 5.61 (m, 2H), 5.05 (s, 1H), 5.00 (dd, J = 10.2, 1.2 Hz, 1H), 4.86 (dd, J = 17.1, 1.2 Hz, 1H), 4.82 - 4.70 (m, 1H), 4.62 - 4.51 (m, 1H), 4.38 - 4.29 (m, 1H), 3.23 (s, 8H), 3.03 - 2.93 (m, 1H), 2.83 - 2.70 (m, 2H), 2.59 (td, J = 17.6. 4.2 Hz, 1H), 2.27 - 2.16 (m. 1H), 2.11 (td, J = 8.6, 4.1 Hz, 1H), 2.02 (ddd, J = 13.5, 8.5, 5.3 Hz, 1H), 1.95 - 1.80 (m, 2H), 1.71 (qd, J = 13.8, 7.1 Hz, 1H), 0.87 (t, J = 7 A Hz, 3H).

Example 6: 3-((4-(4-((4-(4-((2-allyl- 1 -((R)-7 -ethyl-7 -hydroxy-6,7 -dihydro-5H- cyclopenta[b]pyridin-2-yl)-3-oxo-2,3-dihydro-lH-pyrazolo[3,4 -d]pyrimidin-6- yl)amino)phenyl)piperazin-l-yl)methyl)piperidin-l-yl)phenyl) amino)piperidine-2, 6-dione

(Compound

[0297] To a mixture of Intermediate 7 (0.34 g, 648 pmol, 60.12% purity) in DCM (2 mL) were added Intermediate 8 (332 mg, 648 pmol,) and DIEA (168 mg, 13.0 pmol,) at 25 °C. The mixture was stirred at 25 °C for 0.5 h under N2. AcOH (77.9 mg, 13.0 pmol) and NaBH(OAc)a (275 mg, 1.30 mmol) was added to the mixture at 0 °C. The mixture was stirred at 25 °C for 12 h under N2. LCMS showed the reaction was completed and the desired product was detected. The mixture was filtered and the filter was concentrated to give the residue. The residue was purified by prep-HPLC (NH4HCO3 Condition) to give Compound 6, 3-((4- (4-((4-(4-((2-allyl-l-((/?)-7-cthyl-7-hydroxy-6,7-dihydro-5H -cyclopcnta[h]pyridin-2-yl)-3- oxo-2, 3-dihydro-lH-pyrazolo[3,4-7]pyrimidin-6-yl)amino)phenyl)pipe razin-l- yl)methyl)piperidin-l-yl)phenyl)amino) piperidine-2, 6-dione, (43.2 mg, 8.2% yield, 100% purity). LC/MS (ESI ⁺ ) m/z 812.4 [M+H] ⁺ , RT: 1.936 min. NMR (400 MHz, DMSO-d ₆ ) 5 10.75 (s, 1H. NH). 10.19 - 10.03 (m, 1H. NH). 8.82 (s, 1H). 7.93 (br d. J = 7.6 Hz, 1H). 7.69 (d, 7 = 8.1 Hz, 1H), 7.58 (br s, 2H), 6.92 (br d, 7 = 8.9 Hz, 2H), 6.76 (d, J= 8.9 Hz, 2H), 6.60 (d, J = 8.8 Hz, 2H), 5.73 - 5.60 (m, 1H), 5.35 (d, J = 7.3 Hz, 1H. NH), 5.05 (s, 1H), 5.00 (dd, J = 10.2, 1.0 Hz, 1H), 4.85 (br d, J = 17.4 Hz, 1H), 4.80 - 4.70 (m, 1H), 4.57 (br dd, J = 15.7, 5.5 Hz, 1H), 4.22 - 4.13 (m, 1H), 3.42 - 3.32 (m, 3H), 3.31 (s, 4H), 3.10 (br s, 4H), 3.02 - 2.92 (m, 1H), 2.82 - 2.68 (m, 2H). 2.62 - 2.53 (m, 2H), 2.48 - 2.45 (m, 2H), 2.26 - 2.15 (m, 3H), 2.10 (dt, J = 8.9, 4.9 Hz, 1H), 2.02 (ddd, J = 13.4, 8.5, 5.3 Hz, 1H), 1.88 (br dd, J = 12.7, 5.5 Hz, 1H), 1.86 - 1.75 (m, 3H), 1.70 (br dd, J = 13.6, 7.3 Hz, 1H), 1.61 (br d, J = 3.9 Hz, 1H), 0.87 (t, 7 = 7.4 Hz, 3H).

Example 7 : 3-(4-(4-(( 1 - (4-((2- ally 1- 1 -((R)-7 -ethyl-7 -hydroxy-6,7 -dihydro-5H-cyclopenta[b] pyridin-2-yl)-3-oxo-2,3-dihydro-lH-pyrazolo[3,4-d]pyrimidin- 6-yl)amino)phenyl)piperidin- 4-yl) amino)piperidin-l-yl)phenyl)piperidine-2,6-dione (Compound 7)

[0298] Compound 7 was prepared by following the similar procedure described in Example 1 using Intermediate 10 (0.150 g, 0.325 mmol) in place of Intermediate 1. The mixture was stirred at rt for 48 h. The crude was diluted with saturated sodium bicarbonate solution (50 mL) and extracted with 10% MeOH in DCM (2 x 50 mL). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to get the crude product. The crude product was purified by silica gel flash chromatography using 0 to 10% MeOH in DCM as eluent to get enriched product (250 mg) which was further purified by prcp-HPLC (Column/dimcnsions: X-SELECT C18 (19*250*5um) Mobile phase A: 0.1 % formic acid in in water; Mobile phase B: Acetonitrile; Gradient (Time/%B): 0.01/10, 1/10, 10/30, 15/30, 15.01/100, 18/100, 18.01/10, 20/10. Flow rate: 16 mL/min.; Solubility: Acetonitrile + THF + water) to obtain Compound 7, 3-(4-(4-((l-(4-((2-allyl-l-((7?)-7-ethyl- 7 -hydroxy-6, 7-dihydro-5/7-cyclopenta[i] pyridin-2-yl)-3-oxo-2,3-dihydro-l/7-pyrazolo[3,4- ^]pyrimidin-6-yl)amino)phenyl)piperidin-4-yl)amino)piperidin -l-yl)phenyl)piperidine-2,6- dione (57 mg, 22%). LC/MS (ESI ⁺ ) /z 797.59 [M+H] ⁺ . ¹ H NMR (400 MHz, DMSO-d ₆ ) 5 10.77 (s, 1H, NH), 10.11 (br s, 1H, NH), 8.81 (s, 1H), 7.93 (d, J = 7.2 Hz, 1H), 7.69 (d, J =

8.4 Hz, 1H), 7.57 (br s, 2H), 7.04 (d, J = 8.8 Hz, 2H), 6.93 (d, J = 8.4 Hz. 2H), 6.90 (d, J =

8.8 Hz, 2H), 5.72 - 5.60 (m, 1H), 5.05 (br s, 1H, OH), 5.00 (d, J = 9.6 Hz, 1H), 4.86 (d, J =

17.2 Hz, 1H), 4.81 - 4.70 (m, 1H), 4.62 - 4.50 (m, 1H), 3.75 - 3.57 (m, 5H), 3.03 - 2.85 (m,

3H), 2.85 - 2.57 (m, 7H), 2.50 - 2.45 (m, 1H). 2.25 - 1.83 (m, 8H), 1.77 - 1.65 (m, 1H). 1.48 - 1.33 (m, 4H), 0.87 (t, J = 7.2 Hz, 3H). Formate ion peak observed in ¹ H NMR.

Example 8: 3-(4-(4-(( 1 - (4-((2- ally 1- 1 -((R)-7-ethyl-7 -hydroxy-6,7 -dihydro-5H-cyclopenta[b] pyridin-2-yl)-3-oxo-2,3-dihydro-lH-pyrazolo[3,4-d]pyrimidin- 6-yl)amino)phenyl)piperidin- 4-yl) methyl)piperazin-l-yl)phenyl)piperidine-2, 6-dione (Compound 8)

[0299] Compound 8 was prepared by following the similar procedure described in Example 1 using Intermediate 12 (0.100 g, 0.217 mmol) in place of Intermediate 1. The mixture was stirred at rt for 48 h. The crude was diluted with saturated sodium bicarbonate solution (50 mL) and extracted with 10% MeOH in DCM (2 x 50 mL). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to get the crude product. The crude product was purified by silica gel flash chromatography using 0 to 10% MeOH in DCM as eluent to get enriched product (250 mg) which was further purified by prep-HPLC (Column/dimensions: X-SELECT C18 (19*250*5um) Mobile phase A: 5mM ammonium acetate in water. Mobile phase B: Acetonitrile; Gradient (Time/%B): 0.01/25, 1/25, 15/60, 17.5/60, 17.6/100, 20/100, 20.1/25, 23/25. Flow rate: 18 mL/min.; Solubility: THF + ACN + water) to obtain Compound 8, 3-(4-(4-((l-(4-((2-allyl-l-((7?)-7- ethyl-7-hydroxy-6,7-dihydro-5/7-cyclopenta[/?]pyridin-2-yl)- 3-oxo-2,3-dihydro-l/7- pyrazolo[3,4-<7]pyrimidin-6yl)amino)phcnyl)pipcridin-4-yl )mcthyl)pipcrazin-l- yl)phenyl)piperidine-2, 6-dione (9 mg. 5%). LC/MS (ESI ⁺ ) m z 797.50 [M+H] ⁺ . ’H NMR (400 MHz, DMSO-d ₆ ) 8 10.77 (s, 1H, NH), 10.11 (br s, 1H, NH), 8.81 (s, 1H), 7.93 (d, J = 7.2 Hz, 1H), 7.70 (d, J = 8.4 Hz, 1H), 7.56 (br s, 2H), 7.06 (d, J = 8.8 Hz, 2H), 6.92 (d, J = 8.8 Hz. 2H), 6.89 (d, J = 8.4 Hz. 2H), 5.73 - 5.60 (m. 1H). 5.05 (s, 1H. OH). 5.00 (d, J = 9.2 Hz, 1H), 4.87 (d, J = 17.2 Hz, 1H), 4.80 - 4.70 (m, 1H), 4.60 - 4.50 (m, 1H), 3.758 - 3.69 (m, 1H), 3.64 (br d, J = 12.0 Hz, 2H), 3.17(br s, 4H), 3.01 - 2.92 (m, 1H), 2.85 - 2.72 (m, 1H), 2.69 - 2.55 (m, 7H), 2.50 - 2.45 (m, 1H), 2.25 - 2.04 (m, 4H), 2.04 - 1.95 (m, 2H), 1.94 - 1.83 (m, 1H), 1.83 (br d, J = 12.0 Hz, 2H), 1.76 - 1.64 (m, 2H), 1.32 - 1.17 (m, 2H), 0.87 (t, J = 7.2 Hz, 3H).

Example 9: 3-(4-(4-(((l-(4-((2-allyl-l-((R)-7-ethyl-7-hydroxy-6,7-dihyd ro-5H- cyclopenta[b]pyridin-2-yl)-3-oxo-2,3-dihydro-lH-pyrazolo[3,4 -d]pyrimidin-6- yl)amino)phenyl)piperidin-4-yl) methyl)amino)piperidin-l-yl)phenyl)piperidine-2, 6-dione

(Compound 9)

[0300] Compound 9 was prepared by following the similar procedure described in Example 1 using Intermediate 14 (0.190 g, 0.399 mmol) in place of Intermediate 1. The mixture was stirred at rt for 48 h. The crude was diluted with saturated sodium bicarbonate solution (50 mL) and extracted with 10% MeOH in DCM (2 x 50 mL). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to get the crude product. The crude product was purified by silica gel flash chromatography using 0 to 10% MeOH in DCM as eluent to get enriched product (250 mg) which was further purified by prep-HPLC (Column/dimensions: X-SELECT C18 (19*250*5um); Mobile phase A: 10 mM ammonium acetate in water; Mobile phase B: acetonitrile; Gradient (Time/%B): 0.01/25, 2/25, 10/45, 15/45, 15.1/100, 20.5/100, 20.6/45, 22.5/45. Flow rate: 18 mL/min.; Solubility: ACN + water + THF) to obtain Compound 9, 3-(4-(4-((( 1 -(4-((2-allyl- l-((R)-7-ethyl-7-hydroxy-6,7-dihydro-5H-cyclopenta[/?]pyridi n-2-yl)-3-oxo-2,3-dihydro- lH-pyrazolo[3,4-<7]pyrimidin-6-yl)amino)phcnyl)pipcridin- 4-yl)mcthyl)amino)pipcridin-l- yl)phenyl)piperidine-2, 6-dione (35 mg, 11%). LC/MS (ESI ⁺ ) m/z 811.62 [M+H] ⁺ . NMR (400 MHz, DMSO-d ₆ ) 6 10.77 (s, 1H, NH), 10.11 (br s, 1H, NH), 8.81 (s, 1H), 7.92 (d, J = 7.2 Hz, 1H), 7.70 (d, J = 8.4 Hz, 1H), 7.56 (br s, 2H), 7.03 (d, J = 8.4 Hz, 2H), 9.91 (d, J = 9.0 Hz. 1H), 6.88 (d, J = 9.0 Hz. 1H), 5.72 - 5.60 (m. 1H). 5.05 (s, 1H. OH). 5.00 (d, J = 9.2 Hz, 1H), 4.87 (d, J = 17.2 Hz, 1H), 4.80 - 4.70 (m, 1H), 4.60 - 4.50 (m, 1H), 3.75 - 3.67 (m, 1H), 3.65 - 3.55 (m, 4H), 3.02 - 2.92 (m, 1H). 2.85 - 2.55 (m, 7H), 2.52 - 2.40 (m, 1H), 2.25 - 2.15 (m, 1H), 2.15 - 2.06 (m, 1H), 2.05 - 1.95 (m, 2H), 1.93 - 1.75 (m, 7H), 1.73 - 1.65 (m, 1H), 1.55 - 1.42 (m, 1H), 1.40 - 1.18 (m, 4H), 0.87 (t, J = 7.2 Hz, 3H).

Example 10: 3-(4-(2-((( 1 -(4-((2-allyl- 1 -((R)-7 -ethyl-7 -hydroxy-6,7 -dihydro-5H- cyclopenta[b]pyridin-2-yl)-3-oxo-2,3-dihydro-lH-pyrazolo[3,4 -d]pyrimidin-6- yl)amino)phenyl)piperidin-4-yl) methyl)amino)ethoxy)phenyl)piperidine-2, 6-dione

(Compound 10)

[0301] Compound 10 was prepared by following the similar procedure described in Example 1 using Intermediate 16 (0.210 g, 0.481 mmol) in place of Intermediate 1. The mixture was stirred at rt for 48 h. The crude was diluted with saturated sodium bicarbonate solution (50 mL) and extracted with 10% MeOH in DCM (2 x 50 mL). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to get the crude product. The crude product was purified by silica gel flash chromatography using 0 to 10% MeOH in DCM as eluent to get enriched product (250 mg) which was further purified by RP-HPLC (Column/dimensions: X-SELECT C18 (19*250*5um); Mobile phase A: 5mM ammonium acetate in water; Mobile phase B: acetonitrile; Gradient (Time/%B): 0.01/20, 1/20, 10/45, 14/45, 14.1/100, 17/100, 17.1/20, 20/20; Flow rate: 17 mL/min.; Solubility: ACN + THF + water) to obtain Compound 10, 3-(4-(2-(((l-(4-((2-allyl- l-((R)-7- ethyl-7-hydroxy-6,7-dihydro-5Z/-cyclopenta[/?]pyridin-2-yl)- 3-oxo-2,3-dihydro-l/7- pyrazolo[3,4-ri]pyrimidin-6-yl)amino)phenyl)piperidin-4- yl)mcthyl)amino)cthoxy)phcnyl)pipcridinc-2,6-dionc (23 mg, 6%). LC/MS (ESI ⁺ ) m/z 772.58 10.77 (s. 1H, NH), 10.11 (br s, 1H, NH), 8.81 (s, 1H), 7.93 (d, J = 72 Hz, 1H), 7.69 (d, J = 8.4 Hz, 1H), 7.56 (br s, 2H), 7.13 (d, J = 8.8 Hz, 2H), 6.93-6.89 (m, 4H), 5.72 - 5.60 (m, 1H), 5.01 (s, 1H, OH), 5.00 (d, J = 9.2 Hz, 1H), 4.87 (d. J = 17.2 Hz, 1H). 4.80 - 4.70 (m. 1H), 4.60 - 4.50 (m, 1H), 4.01(t, J = 5.6 Hz. 2H), 3.78 (dd, J - 11.4, 5.0 Hz, 1H), 3.63 (br d, J - 12.0 Hz, 2H), 3.03 - 2.92 (m, 1H), 2.88 (br s, 2H), 2.85 - 2.70 (m, 1H), 2.70 - 2.52 (m, 3H). 2.48 - 2.42 (m, 1H), 2.25 - 2.10 (m, 2H), 2.07 - 1.95 (m, 2H), 1.94 - 1.75 (m, 5H), 1.75 - 1.65 (m, 1H), 1.60 - 1.47 (m, 1H), 1.30 - 1.18 (m, 2H), 0.87 (t, J= 7.2 Hz, 3H).

Example 11 : 3 -(4- (( 1 -(( 1 -(4-((2-allyl- 1 -((R)-7 -ethyl-7 -hydroxy-6,7 -dihydro-5H- cyclopenta[b]pyridin-2-yl)-3-oxo-2,3-dihydro-lH-pyrazolo[3,4 -d]pyrimidin-6- yl)amino)phenyl)piperidin-4-yl) methyl)piperidin-4-yl)oxy)phenyl)piperidine-2, 6-dione

(Compound 11)

[0302] Compound 11 was prepared by following the similar procedure described in Example 1 using Intermediate 17 (0.210 g, 0.481 mmol) in place of Intermediate 1. The mixture was stirred at rt for 48 h. The crude was diluted with saturated sodium bicarbonate solution (50 mL) and extracted with 10% MeOH in DCM (2 x 50 mL). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to get the crude product. The crude product was purified by silica gel flash chromatography using 0 to 10% MeOH in DCM as eluent to get enriched product (250 mg) which was further purified by RP-HPLC (Column/dimensions: X-SELECT C18 (19*250*5um); Mobile phase A: lOmM ammonium acetate in water; Mobile phase B: 100% Acetonitrile; Gradient (Time/%B): 0.01/20, 1/30, 15/60, 15.1/100. 17/100, 17.1/30, 20/30; Flow rate: 17 mL/min.; Solubility: water + ACN + THF) to obtain Compound 11, 3-(4-(( 1 -((1 -(4-((2-allyl- 1-((7?)-7- ethyl-7-hydroxy-6,7-dihydro-5/7-cyclopenta[/?]pyridin-2-yl)- 3-oxo-2,3-dihydro-l/7- pyrazolo[3,4-<7]pyrimidin-6-yl)amino)phcnyl)pipcridin-4-y l)mcthyl)pipcridin-4- yl)oxy)phenyl)piperidine-2,6-dione (16 mg, 6%). LC/MS (ESI ⁺ ) m/z 812.60 [M+H] ⁺ . ’H NMR (400 MHz, DMSO-d ₆ ) 5 10.77 (s, 1H, NH), 10.11 (br s, 1H, NH), 8.81 (s, 1H), 7.92 (d, 7 = 7.2 Hz, 1H), 7.69 (d. J = 8.4 Hz, 1H), 7.56 (br s, 2H), 7.11 (d, J = 8.8 Hz, 2H), 6.93 - 6.86 (m, 4H), 5.72 - 5.60 (m. 1H), 5.05 (s. 1H, OH), 5.00 (d. J = 9.2 Hz, 1H), 4.87 (d. J = 17.2 Hz. 1H), 4.80 - 4.70 (m, 1H), 4.60 - 4.50 (m, 1H), 4.40 - 4.30 (m, 1H), 3.77 (dd, J - 11.4, 5.0 Hz, 1H), 3.63 (br d, J = 12.0 Hz, 2H), 3.03 - 2.90 (m, 1H), 2.83 - 2.73 (m, 1H). 2.73 - 2.55 (m, 5H), 2.48 - 2.42 (m, 1H), 2.25 - 2.10 (m, 5H), 2.08 - 1.85 (m, 4H), 1.82 - 1.75 (m, 5H), 1.76

- 1.66 (m, 1H), 1.68 - 1.56 (m, 2H), 1.30 - 1.14 (m, 2H), 0.87 (t, 7 = 7.2 Hz, 3H).

Example 12: 4-((4-(4-((2-allyl-l-((R)-7-ethyl-7-hydroxy-6,7-dihydro-5H- cyclopenta[b]pyridin-2-yl)-3-oxo-2,3-dihydro-lH-pyrazolo[3,4 -d]pyrimidin-6- yl)amino)phenyl)piperazin-l-yl)methyl) -[l,3'-bipiperidine]-2',6'-dione (Compound 12)

[0303] Compound 12 was prepared by following the similar- procedure described in Example 1 using Intermediate 19 (0.120 g, 209 pmol, 67.05% purity) in place of Intermediate 1 . The mixture was stirred at 80 °C for 12 h under N2. LCMS showed the reaction was completed and the desired product was detected. The mixture was filtered and the filter was concentrated to give the residue which was purified by RP-HPLC (FA condition) to obtain Compound 12, 4-((4-(4-((2-allyl-l-((R)-7-ethyl-7-hydroxy-6,7-dihydro-577- cyclopenta[Z?]pyridin-2-yl)-3-oxo-2,3-dihydro-l/7-pyrazolo[3 .4-<7]pyrimidin-6- yl)amino)phenyl)piperazin-l-yl)methyl)-[l,3'-bipiperidine]-2 ',6'-dione, (24.1 mg, 16% yield, 100.00% purity). LC/MS (ESI ⁺ ) ni/z 721.4 [M+H] ⁺ , RT: 1.892 min. NMR (400 MHz, DMSO-de) 5 10.56 (s, 1H, NH), 10.11 (br s, 1H, NH), 8.81 (s, 1H), 7.92 (br d, J = 7.5 Hz, 1H), 7.69 (d, J = 8.1 Hz, 1H), 7.57 (br d, J = 1.3 Hz, 2H), 6.91 (br d, J = 9.0 Hz, 2H), 5.66 (tdd, J = 17.0, 10.4, 6.0 Hz. 1H), 5.05 (s, 1H, OH), 4.99 (dd, J = 10.4, 1 .1 Hz, 1H). 4.85 (br dd, 7 = 17.0, 1.1 Hz, 1H), 4.80 - 4.68 (m, 1H), 4.56 (br dd, 7 = 15.8, 5.3 Hz, 1H), 3.37 (dd, 7 = 10.9, 4.3 Hz, 1H), 3.08 (br s, 4H), 3.02 - 2.91 (m, 1H), 2.83 - 2.70 (m, 3H), 2.63 - 2.52 (m, 3H), 2.48 (br s, 4H). 2.46 - 2.35 (m, 1H), 2.25 - 2.13 (m. 1H), 2.17 (d, 7 = 7.2 Hz, 2H), 2.08 - 1.95 (m, 2H), 1.94 - 1.80 (m, 2H), 1.76 - 1.63 (m, 3H), 1.58 - 1.45 (m, 1H), 1.15 - 1.01 (m, 2H), 0.87 (t, 7 = 7.4 Hz, 3H).

Example 13: 3-((3-(4-((4-(4-((2-allyl-l-((R)-7-ethyl-7-hydroxy-6,7-dihyd ro-5H- cyclopenta[b]pyridin-2-yl)-3-oxo-2,3-dihydro-lH-pyrazolo[3.4 -d]pyrimidin-6- yl)amino)phenyl)piperazin-l-yl)methyl)piperidin-l-yl)phenyl) amino)piperidine-2, 6-dione

(Compound

[0304] Compound 13 was prepared by following the similar procedure described in Example 6 using Intermediate 22 (150 mg, 228 pmol, 48.01% purity) in place of Intermediate 7 to react with Intermediate 8. The mixture was stirred at rt for 2 h. LCMS showed the starting material was consumed and main product with desired MS was detected. The mixture was concentrated under reduced pressure to give a residue. The residue was purified by prep-HPLC (FA condition) to give Compound 13, 3-((3-(4-((4-(4-((2-allyl- 1- ((R)-7-ethyl-7-hydroxy-6.7-dihydro-5//-cyclopenta[Z?]pyridin -2-yl)-3-oxo-2,3-dihydro-l/7- pyrazolo[3 ,4-<7]pyrimidin-6-yl)amino)phenyl)piperazin- 1 -yl)methyl)piperidin- 1 - yl)phenyl)amino)piperidine-2, 6-dione (11.5 mg, 6% yield, 100% purity) was obtained. LC/MS (ESI ⁺ ) m/z 812.4 [M+H] ⁺ , RT: 1.982 min. ’H NMR (400 MHz, DMSO-d ₆ ) 8 10.75 (br s, 1H, NH), 10.12 (br s, 1H, NH), 8.82 (s, 1H), 7.93 (br d, 7= 7.4 Hz, 1H), 7.69 (d, 7= 8.1 Hz, 1H), 7.58 (br s, 2H), 6.92 (br d, 7 = 8.8 Hz, 2H), 6.90 (t, 7 = 8.1 Hz, 1H), 6.27 (s, 1H), 6.19 (dd, 7 = 8.1, 1.2 Hz, 1H), 6.11 (dd. 7 = 8.1, 1.2 Hz. 1H), 5.72 - 5.61 (m, 1H). 5.59 (d, 7 = 7.5 Hz, 1H, NH), 5.05 (s, 1H, OH), 4.99 (dd, 7 = 10.4, 0.8 Hz, 1H), 4.85 (br d, 7 = 17.6 Hz, 1H), 4.81 - 4.68 (m, 1H), 4.62 - 4.52 (m, 1H), 4.34 - 4.26 (m, 1H). 3.60 (br d, 7 = 11.4 Hz, 2H), 3.10 (br s, 4H), 3.02 - 2.93 (m, 1H), 2.82 - 2.69 (m, 2H), 2.65 - 2.55 (m, 3H), 2.50 (br s, 4H), 2.25 - 2.16 (m, 3H), 2.14 - 2.07 (m, 1H), 2.06 - 1.97 (m, 1H), 1.95 - 1.82 (m, 2H), 1.78 (br d, 7= 12.5 Hz, 2H), 1.74 - 1.63 (m, 2H), 1.27 - 1.14 (m, 2H), 0.87 (t, 7 = 7.4 Hz, 3H).

Example 14: 3-(4-(6-(( 1 -(4-((2-allyl- 1 -((R)-7 -ethyl-7 -hydroxy-6,7 -dihydro-5H- cyclopenta[b]pyridin-2-yl)-3-oxo-2,3-dihydro-lH-pyrazolo[3,4 -d]pyrimidin-6- yl)amino)phenyl)piperidin-4-yl) methyl)-2,6-diazaspiro[3.3]heptan-2-yl)phenyl)piperidine-

2,6-dione (Compound 14)

[0305] Compound 14 was prepared by following the similar’ procedure described in Example 1 using Intermediate 24 (100 mg, 126 pmol, 59.48% purity) in place of Intermediate 1, afforded the desired titled product Compound 14, 3-(4-(6-((l-(4-((2-allyl- l-((7?)-7-ethyl-7-hydroxy-6,7-dihydro-5//-cyclopenta[/?Jpyri din-2-yl)-3-oxo-2,3-dihydro- lH-pyrazolo[3,4-7]pyrimidin-6-yl)amino)phenyl)piperidin-4-yl )methyl)-2,6- diazaspiro[3.3]heptan-2-yl)phenyl)piperidine-2,6-dione, (23 mg, 22% yield, 97.53% purity). LCMS (ESI ⁺ ) m/z 809.3 [M+H] ⁺ , RT: 1.982 min. ‘H NMR (400 MHz, DMSO-d ₆ ) 8 10.74 (s, 1H, NH), 10.10 (br s, 1H, NH), 8.81 (s, 1H), 7.92 (br d, J = 7.5 Hz, 1H), 7.69 (br d, J = 8.1 Hz, 1H), 7.56 (br s, 2H), 7.00 (br d, J = 8.1 Hz, 2H), 6.90 (br d, J = 8.6 Hz. 2H), 6.38 (br d, J = 8.3 Hz, 2H), 5.71 - 5.61 (m, 1H), 5.05 (s, 1H, OH), 5.00 (d, J = 10.4Hz, 1H), 4.85 (d, J = 17.2 Hz, 1H), 4.81 - 4.70 (m, 1H), 4.63 - 4.50 (m, 1H), 3.84 (br s, 4H), 3.69 (br dd, J = 10.6, 4.8 Hz, 1H), 3.60 (br d, J = 10.9 Hz, 1H), 3.27 (br s, 4H). 3.01 - 2.92 (m, 1H), 2.84 - 2.73 (m, 1H), 2.65 - 2.53 (m, 3H), 2.48 - 2.40 (m, 1H), 2.27 (br d, J = 6.3 Hz, 2H), 2.26 - 2.16 (m, 1H), 2.13 - 2.06 (m, 1H), 2.06 - 1.95 (m. 2H), 1.93 - 1.84 (m, 1H), 1.80 - 1.65 (m, 3H), 1.43 - 1.32 (m, 1H), 1.30 - 1.13 (m, 2H), 0.87 (br t, J = 7.3 Hz, 3H). Example 15: 3-(4-(4-((l-(4-((2-allyl-l-((R)-7-ethyl-7-hydroxy-6,7-dihydr o-5H- cyclopenta[b]pyridin-2-yl)-3-oxo-2,3-dihydro-lH-pyrazolo[3,4 -d]pyrimidin-6- yl)amino)phenyl)piperidin-4-yl) amino)butoxy)phenyl)piperidine-2, 6-dione (Compound 16)

[0306] Compound 15 was prepared by following the similar procedure described in Example 1 using Intermediate 26 (388 mg, 0.860 mmol) in place of Intermediate 1. The mixture was stirred at rt for 48 h. The crude was diluted with saturated sodium bicarbonate solution (50 mL) and extracted with 10% MeOH in DCM (2 x 50 mL). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to get the crude product. The crude product was purified by silica gel flash chromatography using 0 to 10% MeOH in DCM as eluent to get enriched product (250 mg) which was further purified by RP-HPLC (Column/dimensions: XB RIDGE C8 (19*150*5um); Mobile phase A: lOmM ammonium acetate in water; Mobile phase B: acetonitrile; Gradient (Time/%B): 0.1/10, 1/10, 10/40, 13/40, 13.1/100, 17/100, 17.1/10, 19/10; Flow rate: 18 mL/min.; Solubility: Acetonitrile + THF + water) to obtain Compound 15, 3-(4-(4-((l-(4-((2-allyl- 1- ((7?)-7-ethyl-7-hydroxy-6,7-dihydro-5H-cyclopenta[/?]pyridin -2-yl)-3-oxo-2,3-dihydro-lH- pyrazolo[3,4-ri]pyrimidin-6-yl)amino)phenyl)piperidin-4- yl)amino)butoxy)phenyl)piperidine-2, 6-dione (111 mg, 18%). LCMS (ESI ⁺ ) m/z 786.64 [M+H] ⁺ . ’ H NMR (400 MHz, DMSO-d ₆ ) 8 10.77 (br s, 1H, NH), 10.11 (br s, 1H, NH), 8.81 (s, 1H), 7.93 (d, J = 7.2 Hz, 1H), 7.69 (d, J = 8.0 Hz, 1H), 7.55 (br s, 2H), 7.11 (d, J = 8.4 Hz, 2H), 6.91 (d, J = 9.2 Hz, 2H), 6.88 (d. J = 8.4 Hz, 2H), 5.73 - 5.60 (m, 1H), 5.01 (br s, 1H, OH), 4.99 (d, J = 9.6 Hz, 1H), 4.85 (d, J = 17.2 Hz, 1H), 4.82 - 4.70 (m, 1H), 4.62 - 4.50 (m, 1H), 3.96 (t, J= 6.4 Hz, 2H), 3.77 (dd. J= 11.4, 5.0 Hz, 1H), 3.62 - 3.52 (m, 2H), 3.03 - 2.92 (m, 2H), 2.83 - 2.72 (m, 1H), 2.72 - 2.63 (m, 3H), 2.61 (t, J = 6.8 Hz, 2H), 2.52 - 2.43 (m, 1H), 2.25 - 2.07 (m, 2H), 2.06 - 1.95 (m, 2H), 1.95 - 1.83 (m, 3H), 1.80 - 1.65 (m, 3H), 1.61

- 1.50 (m, 2H), 1.40 - 1.30 (m, 2H), 0.87 (t, J = 7.4 Hz, 3H).

Example 16: 3-(4-(4-((4-(4-((2-allyl-l-((R)-7-ethyl-7-hydroxy-6,7-dihydr o-5H- cyclopenta[b]pyridin-2-yl)-3-oxo-2,3-dihydro-lH-pyrazolo[3,4 -d]pyrimidin-6- yl)amino)phenyl)piperazin-l-yl)methyl)piperidin-l-yl)phenyl) piperidine-2, 6-dione

(Compound 16)

[0307] Compound 16 was prepared by following the similar procedure described in Example 6 using l-(4-(2,6-dioxopiperidin-3-yl)phenyl)piperidine-4-carbaldehy de

(Reference: WO 2022/012622, P150, 58.6 mg, 0.195 mmol) in place of Intermediate 7 to react with Intermediate 8. The mixture was stirred at rt for 16 h. The reaction was quenched with water (35 mL) and extracted with dichloromethane (2 x 50 mL). The organic layer was washed with water (20 mL) and saturated sodium bicarbonate solution (20 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by prep-HPLC (Column/dimensions: x-BRIDGE C8; Mobile phase A: 5mM ammonium acetate; Mobile phase B: Acetonitrile; Gradient (Time/%B): 0.01/20, 1/20, 10/45, 15/45, 15.1/100, 19/100, 19.1/20, 22/20; solubility: Acetonitrile + THF + water; Flow rate: 18 mL/minutes) to get Compound 16, 3-(4-(4-((4-(4-((2-allyl-l-((R)-7-ethyl-7-hydroxy-6,7- dihydro-5/7-cyclopenta[Z?]pyridin-2-yl)-3-oxo-2,3-dihydro-l/ /-pyrazolo[3,4-i/]pyrimidin-6- yl)amino)phenyl)piperazin-l-yl)methyl)piperidin-l-yl)phenyl) piperidine-2, 6-dione (33 mg, 21%). LCMS (ESI ⁺ ) m/z 797.73 [M+H] ⁺ . 'H NMR (400 MHz, DMSO-d ₆ ) 8 10.77 (br s, 1H, NH), 10.11 (br s, 1H, NH), 8.82 (s, 1H), 7.93 (d. J = 6.8 Hz, 1H), 7.70 (d, J = 8.0 Hz. 1H), 7.56 (br s, 2H), 7.03 (d, J = 8.4 Hz, 2H), 6.92 (d, J = 8.8 Hz, 2H), 6.89 (d, J = 8.8 Hz, 2H), 5.75 - 5.60 (m, 1H), 5.05 (br s, 1H, OH), 5.00 (d, J = 10.4 Hz, 1H), 4.86 (d, J = 17.2 Hz, 1H), 4.82 - 4.70 (m, 1H), 4.62 - 4.50 (m, 1H), 3.72 (dd, J = 11.0, 5.0 Hz, 1H), 3.67 (br d, J = 12.4 Hz, 2H), 3.11 (br s, 4H), 3.05 - 2.93 (m, 1H), 2.85 - 2.73 (m, 1H), 2.72 - 2.57 (m, 4H), 2.53 -

-I l l- 2.43 (m, 3H), 2.38 - 2.07 (m, 3H), 2.06 - 1.95 (m, 3H), 1.93 - 1.83 (m, 1H), 1.81 (br d, 7 = 11.6 Hz, 2H), 1.78 - 1.65 (m, 3H), 1.30 - 1.15 (m, 2H), 0.87 (t, 7 = 7.2 Hz, 3H).

Example 17: 3-[4-[4-[4-[[2-allyl-l-[(7R)-7-ethyl-7-hydroxy-5,6- dihydrocyclopenta[b]pyridin-2-yl]-3-oxo-pyrazolo[3,4-d]pyrim idin-6- yl]amino]phenyl]piperazin-l-yl]phenyl]piperidine-2, 6-dione (Compound 17)

[0308] To a solution of 2-allyl-l-[(7R)-7-ethyl-7-hydroxy-5,6- dihydrocyclopenta[b] pyridin-2-yl]-6-methylsulfonyl-pyrazolo[3,4-d]pyrimidin-3-on e (186.6 mg, 449.24 pmol) in iPrOH (2 mL) was added 3-[4-[4-(4-aminophenyl)piperazin-l- yl]phenyl]piperidine-2, 6-dione (Intermediate 27) (100.0 mg, 224.62 pmol) at 25 °C. The reaction was stirred at 80 °C for 12 h. LCMS showed the reaction was completed and the desired product was detected. The mixture was filtered, and the filter was concentrated to give a residue. The residue was purified by prep-HPLC (NH4HCO3 Condition) to afford 3- [4- [4- [4- [ [2 - ally 1- 1 - [(7R)-7 -cthyl-7-hy droxy-5 ,6-dihydrocyclopcnta[b]pyridin-2-y 1] -3-oxo- pyrazolo[3,4-d]pyrimidin-6-yl]amino]phenyl]piperazin-l-yl]ph enyl]piperidine-2, 6-dione (Compound 17) (30.0 mg, 19.09% yield, 100.00% purity) as a yellow solid. LCMS (ESI) m/z 700.3 [M+H], RT: 2.266 min. ’H NMR (400 MHz, DMSO-d ₆ ) 5 10.78 (s, 1H), 10.25 - 10.05 (m, 1H). 8.83 (s, 1H). 8.03 - 7.85 (m, 1H). 7.70 (d, 7 = 8.1 Hz. 1H), 7.66 - 7.49 (m. 2H). 7.09 (d, J = 8.6 Hz, 2H), 6.99 (br t, J = 9.3 Hz, 4H), 5.67 (tdd, J = 16.9, 10.6, 5.9 Hz, 1H), 5.09 - 4.95 (m, 2H), 4.85 (br d. J = 17.1 Hz, 1H). 4.81 - 4.69 (m, 1H), 4.57 (br dd, J = 15.4, 4.9 Hz, 1H), 3.75 (dd, J = 11.1, 4.9 Hz, 1H), 3.29 - 3.23 (m, 8H), 3.04 - 2.91 (m, 1H), 2.83 - 2.72 (m, 1H), 2.68 - 2.60 (m, 1H), 2.47 - 2.41 (m, 1H), 2.27 - 2.09 (m, 2H), 2.08 - 1.96 (m, 2H), 1.89 (qd, J = 13.8, 7.0 Hz. 1H), 1.78 - 1.64 (m, 1H), 0.87 (t, J = 7.4 Hz, 3H). Example 18: 3-[4-[4-[2-[4-[[2-allyl-l-[(7R)-7-ethyl-7-hydroxy-5,6- dihydrocyclopenta[b]pyridin-2-yl]-3-oxo-pyrazolo[3,4-d]pyrim idin-6- yl]amino]phenyl]ethyl]piperazin- l-yl]phenyl]piperidine-2, 6-dione (Compound 18)

[0309] To a solution of 2-allyl-l-[(7R)-7-ethyl-7-hydroxy-5,6- dihydrocyclopenta[b]pyridin-2-yl]-6-methylsulfonyl-pyrazolo[ 3,4-d]pyrimidin-3-one (200.5 mg, 482.51 pmol) in iPrOH (2 mL) was added 3-[4-[4-[2-(4-aminophenyl)ethyl]piperazin-l- yl]phenyl] piperidine-2, 6-dione (Intermediate 28) (0.1 g, 241.25 pmol) at 20 °C. The reaction was stirred at 80 °C for 12 h. LCMS showed the reaction was completed and the desired product was detected. The mixture was filtered, and the filter was concentrated to give the residue. The residue was purified by prep-HPLC (NH4HCO3 Condition) to afford 3- [4- [4-[2-[4- [ [2- ally 1- 1 - [(7R)-7 -ethyl-7 -hydroxy-5 ,6-dihydrocyclopenta[b]pyridin-2-yl] -3- oxo-pyrazolo[3,4-d]pyrimidin-6-yl]amino]phenyl]ethyl]piperaz in-l-yl]phenyl]piperidine- 2, 6-dione (Compound 18) (22.0 mg, 12.5% yield, 100% purity) as a yellow solid. LCMS (ESI ⁺ ) m/z 728.3 [M+H] ⁺ . ^X H NMR (400 MHz, DMSO-d ₆ ) 8 10.77 (s, 1H), 10.29 - 10.14 (m, 1H), 8.87 (s, 1H), 7.91 (d, J= 8.1 Hz, 1H), 7.74 - 7.58 (m, 3H), 7.21 (d, J= 8.5 Hz, 2H), 7.05 (d, J = 8.8 Hz, 2H), 6.90 (d, J = 8.8 Hz. 2H), 5.67 (tdd, J = 16.8, 10.6. 5.9 Hz, 1H), 5.05 (s, 1H), 5.00 (dd, J = 10.3, 1.3 Hz, 1H), 4.86 (dd, 7= 17.0, 1.3 Hz, 1H), 4.75 (br dd, J= 15.6, 4.4 Hz, 1H), 4.56 (br dd, 7 = 16.1, 6.1 Hz, 1H), 3.73 (dd, 7 = 10.9, 4.9 Hz, 1H), 3.18 - 3.07 (m, 4H), 2.99 - 2.89 (m. 1H), 2.81 - 2.70 (m, 3H). 2.68 - 2.61 (m, 1H), 2.61 - 2.53 (m, 6H). 2.48 - 2.41 (m, 1H), 2.24 - 2.06 (m, 2H), 2.05 - 1.95 (m, 2H), 1.94 - 1.83 (m, 1H), 1.76 - 1.64 (m, 1H), 0.86 (t, 7 = 7.4 Hz, 3H). Example 19: 3-[[4-[4-[4-[[2-allyl-l-[(7R)-7-ethyl-7-hydroxy-5,6- dihydrocyclopenta[b]pyridin-2-yl]-3-oxo-pyrazolo[3,4-d]pyrim idin-6- yl]amino]phenyl]piperazin-l-yl]cyclohexyl]amino]piperidine-2 , 6-dione (Compound 19)

[0310] To a solution of 2-allyl-l-[(7R)-7-ethyl-7-hydroxy-5,6- dihydrocyclopenta[b]pyridin-2-yl] -6-methylsulfonyl-pyrazolo[3,4-d]pyrimidin-3-one (151.3 mg, 364.20 pmol) in iPrOH (2 mL) was added 3-[[4-[4-(4-aminophenyl)piperazin-l- yl]cyclohexyl] amino] piperidine-2, 6-dione (Intermediate 29) (100.0 mg, 182.10 pmol, 70.20% purity) at 25 °C. The reaction was stirred at 80 °C for 12 h. LCMS showed the reaction was completed and the desired product was detected. The mixture was filtered, and the filter was concentrated to give the residue. The residue was purified by prep-HPLC (NH4HCO3 Condition) to afford 3-[[4-[4-[4-[[2-allyl-l-[(7R)-7-ethyl-7-hydroxy-5,6- dihydrocyclopenta[b]pyridin-2-yl]-3-oxo-pyrazolo[3,4-d]pyrim idin-6- yl]amino]phenyl]piperazin-l-yl]cyclohexyl]amino]piperidine-2 , 6-dione (Compound 19) (15.0 mg, 11.43% yield, 100% purity) as a yellow solid. LCMS (ES1 ⁺ ) m/z 721.4 [M+H] ⁺ . 'H NMR (400 MHz, DMSO-d ₆ ) 5 10.70 (br s, 1H), 10.21 - 10.01 (m, 1H), 8.82 (s, 1H), 7.92 (br d, J = 7.4 Hz, 1H), 7.69 (d, J = 8.3 Hz, 1H), 7.57 (br s, 2H). 6.91 (br d, J = 8.9 Hz, 2H), 5.67 (tdd, J = 16.8, 10.5, 5.9 Hz, 1H), 5.04 (s, 1H), 4.99 (dd, J = 10.2, 0.9 Hz, 1H), 4.85 (br d, J= 17.9 Hz, 1H), 4.81 - 4.68 (m, 1H), 4.56 (br dd, J = 15.8, 5.8 Hz, 1H), 3.42 (dd, J = 10.4, 4.8 Hz, 1H), 3.09 (br s, 4H), 3.02 - 2.91 (m, 1H), 2.85 - 2.72 (m, 2H). 2.65 - 2.57 (m, 4H), 2.57 - 2.52 (m, 3H), 2.25 - 2.15 (m, 2H), 2.02 (ddd, J= 12.9, 7.6, 5.2 Hz, 2H), 1.94 - 1.84 (m, 1H), 1.76 - 1.58 (m, 6H), 1.48 (br d, J = 7.4 Hz, 4H), 0.87 (t. J = 7.4 Hz, 3H). Example 20: (R)-4-(4-(4-(((l-(4-((2-allyl-l-(7-ethyl-7-hydroxy-6,7-dihyd ro-5H- cyclopenta[b]pyridin-2-yl)-3-oxo-2,3-dihydro-lH-pyrazolo[3,4 -d]pyrimidin-6- yl)amino)phenyl)piperidin-4-yl)methyl)amino)piperidin-l-yl)p henyl)piperidine-2, 6-dione

(Compound 20)

[0311] To a solution of 4-(4-(4-oxopiperidin-l-yl)phenyl)piperidine-2, 6-dione (Intermediate 30) (0.05 g, 174.63 pmol) and (R)-2-allyl-6-((4-(4-(aminomethyl)piperidin-l- yl)phenyl)amino)-l-(7-ethyl-7-hydroxy-6,7-dihydro-5H-cyclope nta[b]pyridin-2-yl)-l,2- dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (Intermediate 31) (94.41 mg, 174.63 pmol) in DCM (1 mL) was added AcOH (1.05 mg, 17.46 pmol, 1 pL) to adjust pH=5-6. Then NaBH(OAc)3 (74.02 mg, 349.25 pmol) was added at 25 °C after 2 h. The reaction was stirred at 20 °C for 12 h. LCMS showed the reaction was completed and the desired product was detected. The mixture was concentrated under reduced pressure at 40 °C to give the crude product. The crude product was purified by prep-HPLC (NH4HCO3 Condition) to give the pure product (R)-4-(4-(4-(((l-(4-((2-allyl-l-(7-ethyl-7-hydroxy-6,7-dihyd ro-5H- cyclopenta[b]pyridin-2-yl)-3-oxo-2,3-dihydro-lH-pyrazolo[3,4 -d]pyrimidin-6- yl)amino)phenyl)piperidin-4-yl)methyl)amino)piperidin-l-yl)p henyl)piperidine-2,6-dione (Compound 20) (0.012 g, 14.48 pmol, 8.29% yield, 97.88% purity) was obtained as a white solid. LCMS (ESI ⁺ ) ni/z 833.5 [M+Na] ⁺ . ’H NMR (400 MHz. DMSO-d ₆ ) 5 10.79 (s, 1H), 10.17 - 10.03 (m, 1H), 8.81 (s, 1H), 7.97 - 7.86 (m, 1H), 7.69 (d, J = 8.1 Hz, 1H), 7.56 (br d, J = 3.5 Hz, 2H), 7.11 (d, J = 8.8 Hz, 2H), 6.97 - 6.83 (m, 4H), 5.67 (tdd, J = 16.8, 10.5, 6.0 Hz, 1H), 5.04 (s. 1H), 5.02 - 4.97 (m, 1H), 4.85 (br d, J = 17.4 Hz, 1H), 4.81 - 4.69 (m, 1H), 4.61 - 4.51 (m, 1H), 3.71 - 3.52 (m, 4H), 3.01 - 2.93 (m, 1H), 2.79 - 2.65 (m, 5H), 2.64 - 2.55 (m, 4H), 2.47 - 2.46 (m, 1H), 2.25 - 2.11 (m, 2H), 2.06 - 1.97 (m, 1H), 1.93 - 1.78 (m, 5H), 1.70 (br dd, J = 13.6, 7.4, Hz, 1H), 1.57 - 1.41 (m, 2H), 1.38 - 1.19 (m, 5H), 0.87 (t, J = 7.4 Hz, 3H).

Example 21 : 3-((3-(4-(( 1 -(4-((2-allyl- 1 -((R)-7 -ethyl-7 -hydroxy-6,7 -dihydro-5H- cyclopenta[b]pyridin-2-yl)-3-oxo-2,3-dihydro-lH-pyrazolo[3,4 -d]pyrimidin-6- yl)amino)phenyl)piperidin-4-yl)amino)piperidin-l-yl)phenyl)a mino)piperidine-2, 6-dione (Compound 21)

[0312] To a solution of 3-((3-(4-aminopiperidin-l-yl)phenyl)amino)piperidine- 2, 6-dione (Intermediate 33) (0.2 g, 290.77 pmol) in MeOH (0.5 rnL) were added (R)-2-allyl- l-(7-ethyl-7-hydroxy-6,7-dihydro-5H-cyclopenta[b]pyridin-2-y l)-6-((4-(4-oxopiperidin-l- yl)phenyl)amino)-l,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-o ne (Intermediate 34) (50.9 mg, 96.92 pmol) and DIEA (125.2 mg, 969.23 pmol, 168.8 pL) AcOH (11.6 mg, 193.85 pmol, 11.10 pL) NaBHsCN (9.1 mg, 145.38 pmol) at 25 °C. The reaction was stirred at 25 °C for 12 h. LCMS showed the reaction was completed and the desired product was detected. The mixture was filtered, and the filter was concentrated to give the residue. The residue was purified by prep-HPLC (NH4HCO3 Condition) to afford 3-((3-(4-((l-(4-((2-allyl-l-((R)-7- ethyl-7-hydroxy-6,7-dihydro-5H-cyclopenta[b]pyridin-2-yl)-3- oxo-2,3-dihydro-lH- pyrazolo[3,4-d]pyrimidin-6-yl)amino)phenyl)piperidin-4-yl)am ino)piperidin-l- yl)phenyl)amino)piperidine-2, 6-dione (Compound 21) (9.0 mg, 10.8% yield, 95.18% purity) as a yellow solid. LCMS (ESI ⁺ ) mJz 812.4 [M+H] ⁺ . NMR (400 MHz, DMSO-d ₆ ) 6 10.75 (br s, 1H), 10.18 - 10.00 (m, 1H), 8.81 (s, 1H), 7.93 (br d, J = 7.5 Hz, 1H), 7.69 (d, J = 8.1 Hz, 1H), 7.56 (br s, 2H), 6.97 - 6.90 (m, 2H), 6.90 - 6.85 (m, 1H), 6.27 (s, 1H), 6.19 (br d, J = 8.1 Hz, 1H), 6.11 (br d, J = 7.9 Hz, 1H), 5.73 - 5.62 (m. 1H), 5.62 - 5.54 (m, 1H), 5.05 (s, 1H), 5.00 (br d, J = 10.3 Hz, 1H), 4.85 (br d, J = 17.1 Hz, 1H), 4.81 - 4.67 (m, 1H), 4.64 - 4.49 (m, 1H), 4.37 - 4.23 (m, 1H), 3.68 - 3.48 (m, 4H). 3.01 - 2.91 (m, 1H), 2.85 - 2.62 (m, 8H), 2.56 (br s, 2H), 2.26 - 2.15 (m, 1H), 2.14 - 2.06 (m. 1H), 2.01 (ddd, J = 13.3, 8.4. 5.2 Hz, 1H). 1.96 - 1.76 (m, 6H), 1.70 (br dd, J = 13.6, 7.4 Hz, 1H), 1.40 - 1.22 (m, 4H), 0.87 (t, J = 7.4 Hz,

3H).

Example 22: 3 - [4- [2- [ [ 1 - [4- [ [2 - ally 1- 1 - [(7R)-7 -ethyl-7-hydroxy-5 ,6- dihydrocyclopentas [b ] pyridin-2-yl] -3 -oxo-pyrazolo [3 ,4-d] pyrimidin-6-yl] amino] phenyl] -4- piperidyl] amino]ethylamino]phenyl]piperidine-2, 6-dione (Compound 22)

[0313] To a mixture of 3-[4-(2-aminoethylamino)phenyl]piperidine-2, 6-dione (Intermediate 35) (42.0 mg, 77.88 pmol, 67% purity, TFA salt) and 2-allyl-l-[(7R)-7-ethyl- 7-hydroxy-5.6-dihydrocyclopenta[b]pyridin-2-yl]-6-[4-(4-oxo- l- piperidyl)anilino]pyrazolo[3,4-d]pyrimidin-3-one (Intermediate 34) (40.0 mg, 76.10 pmol) in DCM (1.0 mL) was added NaBH(OAc)s (32.3 mg, 152.21 pmol) at 20 °C under N2. The mixture was stirred at 20 °C for 12 h. LCMS showed the reaction was completed and the desired product was detected. The mixture was concentrated under reduced pressure at 40 °C to give the crude product. The crude product was purified by prep-HPLC (NH4HCO3 Condition) to give 3-[4-[2-[[l-[4-[[2-allyl-l-[(7R)-7-ethyl-7-hydroxy-5,6- dihydrocyclopentas[b]pyridin-2-yl]-3-oxo-pyrazolo[3,4-d]pyri midin-6-yl]amino]phenyl]-4- piperidyl]amino]ethylamino]phenyl]piperidine-2, 6-dione (Compound 22) (18.0 mg, 31.25% yield) as a white solid. LCMS (ESI ⁺ ) m/z 379.3 [M/2+H] ⁺ . ‘H NMR (400 MHz, DMSO-d ₆ ) 5 10.73 (s, 1H), 10.22 - 9.94 (m, 1H), 8.81 (s, 1H), 7.99 - 7.86 (m, 1H), 7.69 (d, J = 8.1 Hz,

1H), 7.63 - 7.47 (m, 2H), 7.01 - 6.82 (m, 4H), 6.54 (d, J = 8.6 Hz, 2H), 5.72 - 5.61 (m, 1H), 5.51 - 5.46 (m, 1H), 5.07 - 4.96 (m, 2H), 4.90 - 4.70 (m, 2H), 4.62 - 4.51 (m, 1H), 3.67 - 3.56 (m, 4H), 3.09 (br d, J = 5.6 Hz, 2H), 3.01 - 2.93 (m, 1H), 2.77 (br t, J = 5.8 Hz, 3H). 2.73 - 2.65 (m, 4H), 2.44 - 2.39 (m, 1H), 2.24 - 2.16 (m, 1H), 2.11 - 2.04 (m, 1H). 2.00 (ddd. J = 13.1, 8.3, 4.3 Hz, 2H), 1.94 - 1.86 (m, 3H), 1.75 - 1.67 (m, 1H), 1.42 - 1.32 (m, 2H), 0.87 (t, 7 = 7.3 Hz, 3H).

Example 23: (R)-4-(3-(4-((l-(4-((2-allyl-l-(7-ethyl-7-hydroxy-6,7-dihydr o-5H- cyclopenta[b]pyridin-2-yl)-3-oxo-2,3-dihydro-lH-pyrazolo[3,4 -d]pyrimidin-6- yl)amino)phenyl)piperidin-4-yl)amino)piperidin-l-yl)phenyl)p iperidine-2, 6-dione

(Compound 23)

[0314] To a solution of 4-(3-(4-oxopiperidin-l-yl)phenyl)piperidine-2, 6-dione (Intermediate 37) (32.62 mg, 113.93 pmol) and (R)-2-allyl-6-((4-(4-aminopiperidin-l- yl)phenyl)amino)-l-(7-ethyl-7-hydroxy-6,7-dihydro-5H-cyclope nta[b]pyridin-2-yl)-l,2- dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (Intermediate 36) (0.05 g, 94.94 pmol) in DCM (1 mL) and MeOH (0.2 mL) was added AcOH (570.15 pg, 9.49 pmol) to adjust pH=5-6. Then NaBH(OAc)3 (40.24 mg, 189.89 pmol) was added at 25°C after 2 h. The reaction was stirred at 20 °C for 12 h. LCMS showed the reaction was completed and the desired product was detected. The mixture was concentrated under reduced pressure at 40 °C to give the crude product. The crude product was purified by prep-HPLC (NH3.H2O Condition) to give the pure product. (R)-4-(3-(4-((l-(4-((2-allyl-l-(7-ethyl-7-hydroxy-6,7-dihydr o-5H- cyclopenta[b]pyridin-2-yl)-3-oxo-2,3-dihydro-lH-pyrazolo[3,4 -d]pyrimidin-6- yl)amino)phenyl)piperidin-4-yl)amino)piperidin-l-yl)phenyl)p iperidine-2, 6-dione (Compound 23) (0.019 g, 36.39 pmol, 38.33% yield, 99.72% purity) was obtained as a white solid. LCMS (ESI ⁺ ) m/z 797.5 [M+H] ⁺ . ’H NMR (400 MHz, DMSO-d ₆ ) 5 10.82 (br s, 1H), 10.11 (br d, J = 1.6 Hz. 1H). 8.81 (s, 1H), 7.93 (br d, J = 6.8 Hz, 1H), 7.69 (d. 7 = 8.1 Hz, 1H), 7.56 (br s, 2H), 7.14 (t, J = 7.9 Hz, 1H), 6.95 - 6.85 (m, 3H), 6.80 (br d, J = 8.3 Hz, 1H), 6.66 (d, 7 = 7.4 Hz, 1H), 5.73 - 5.61 (m, 1H), 5.05 (br s, 1H), 4.99 (br d, 7 = 10.1 Hz, 1H), 4.85 (br d, 7 = 17.1 Hz, 1H), 4.76 (br d, 7 = 11.0 Hz, 1H). 4.61 - 4.52 (m, 1H), 3.69 - 3.55 (m, 4H), 3.02 - 2.91 (m, 1H), 2.86 - 2.57 (m, 12H), 2.24 - 2.15 (m, 1H), 2.O8 - 1.97 (m, 1H), 1.95

- 1.82 (m, 5H), 1.70 (br dd, 7=13.7, 7.3 Hz, 1H), 1.43 - 1.20 (m, 5H), 0.87 (t, J = 7.4 Hz, 3H).

Example 24: (R)-3-(4-(4-(((l-(4-((2-allyl-l-((R)-7-ethyl-7-hydroxy-6,7-d ihydro-5H- cyclopenta[b]pyridin-2-yl)-3-oxo-2,3-dihydro-lH-pyrazolo[3,4 -d]pyrimidin-6- yl)amino)phenyl)piperidin-4-yl)methyl)amino)piperidin-l-yl)p henyl)piperidine-2, 6-dione

(Compound 24A) and (S)-3-(4-(4-(((l-(4-((2-allyl-l-((R)-7-ethyl-7-hydroxy-6,7-d ihydro-

5H-cyclopenta[b]pyridin-2-yl)-3-oxo-2,3-dihydro-lH-pyrazo lo[3,4-d]pyrimidin-6- yl)amino)phenyl)piperidin-4-yl)methyl)amino)piperidin-l-yl)p henyl)piperidine-2, 6-dione

(Compound 24B)

(Compound 24B)

[0315] 3-[4-[4-[[l-[4-[[2-allyl-l-[(7R)-7-ethyl-7-hydroxy-5,6- dihydrocyclopenta[b]pyridin-2-yl] -3 -oxo-pyrazolo [3 ,4-d]pyrimidin-6-y 1] aminojpheny 1] -4- piperidyl]methylamino]-l-piperidyl]phenyl]piperidine-2, 6-dione (Compound 9) (100 mg) was purified by chiral SFC purification (Column/dimensions : CHIRALPAK-AS-H (30*250)mm,5p; % CCh: 50%; % Co solvent: 50% (0.2% 7N Methanolic ammonia in ACN : MeOH) (1 :1 ), Total flow: 110 mL/min.; Back Pressure: 100 bar; UV: 307 nm; Solubility: McOH; No. of injections: 3; Total purification time: 01:30 Hrs; Run time per injection: 24:50 minutes; Instrument details: Make/Model: SFC- 150-11. Nitrogen gas was purged into the collecting fraction to remove dissolved ammonia) to afford Peak 1: 48.0 mg (Compound 24A) as a pale yellow solid and Peak 2: 45.0 mg (Compound 24B) as a pale yellow solid. Note: Stereochemistry was assigned arbitrarily.

[0316] Peak 1: (R)-3-(4-(4-(((l-(4-((2-allyl-l-((R)-7-ethyl-7-hydroxy-6,7- dihydro-5H-cyclopenta[b]pyridin-2-yl)-3-oxo-2,3-dihydro-lH-p yrazolo[3,4-d]pyrimidin-6- yl)amino)phenyl)piperidin-4-yl)methyl)amino)piperidin-l-yl)p henyl)piperidine-2, 6-dione (Compound 24A). LCMS (ESI ) m/z 809.56 10.76 (s, 1H), 10.11 (br s, 1H), 8.81 (s, 1H), 7.93 (d, J= 7.2 Hz, 1H), 7.70 (d, J= 8.4 Hz. 1H), 7.56 (br s, 2H), 7.03 (d, J = 8.4 Hz, 2H), 6.93-6.87 (m, 4H), 5.72-5.60 (m, 1H), 5.05 (s, 1H), 5.00 (d. J = 9.2 Hz, 1H), 4.87 (d, J = 17.2 Hz, 1H), 4.80-4.70 (m, 1H), 4.60-4.50 (m, 1H), 3.75-3.68 (m, 1H), 3.66-3.55 (m, 4H). 3.01-2.95 (m. 1H). 2.82-2.52 (m. 8H), 2.50-2.40 (m, 2H), 2.21-1.98 (m, 4H), 1.95-1.80 (m, 5H), 1.73-1.68 (m, 1H), 1.60-1.48 (m, 1H), 1.40-1.18 (m, 5H), 0.87 (t, 7 = 7.2 Hz. 3H).

[0317] Peak 2: (S)-3-(4-(4-(((l-(4-((2-allyl-l-((R)-7-ethyl-7-hydroxy-6,7- dihydro-5H-cyclopenta[b]pyridin-2-yl)-3-oxo-2,3-dihydro-lH-p yrazolo[3,4-d]pyrimidin-6- yl)amino)phenyl)piperidin-4-yl)methyl)amino)piperidin-l-yl)p henyl)piperidine-2, 6-dione (Compound 24B). LCMS (ESI ) m/z 809.56 [M-H]’. ‘H NMR (400 MHz, DMSO-^): 8 10.76 (s, 1H), 10.11 (br s, 1H), 8.81 (s, 1H), 7.93 (d, 7= 7.6 Hz, 1H), 7.70 (d, 7= 8.4 Hz, 1H), 7.56 (br s, 2H), 7.03 (d, 7 = 8.4 Hz, 2H), 6.93-6.87 (m. 4H), 5.72-5.60 (m, 1H), 5.05 (s. 1H), 5.00 (d, 7 = 9.2 Hz, 1H), 4.87 (d, 7 = 17.2 Hz, 1H), 4.80-4.70 (m, 1H), 4.60-4.50 (m, 1H), 3.75-3.68 (m, 1H), 3.66-3.55 (m, 4H), 3.01-2.95 (m, 1H), 2.82-2.52 (m, 8H), 2.50-2.40 (m, 2H), 2.21-1.98 (m, 4H). 1.95-1.80 (m. 5H), 1.73-1.68 (m, 1H), 1.60-1.48 (m, 1H), 1.40-1.18 (m, 5H), 0.87 (t, 7= 7.2 Hz, 3H). Example 25: (R)-4-(3-(4-(((l-(4-((2-allyl-l-(7-ethyl-7-hydroxy-6,7-dihyd ro-5H- cyclopenta[b]pyridin-2-yl)-3-oxo-2,3-dihydro-lH-pyrazolo[3,4 -d]pyrimidin-6- yl)amino)phenyl)piperidin-4-yl)methyl)amino)piperidin-l-yl)p henyl)piperidine-2, 6-dione

(Compound 25)

[0318] To a solution of 4-(3-(4-oxopiperidin-l-yl)phenyl)piperidine-2, 6-dione (Intermediate 37) (0.05 g, 174.63 pmol) and (R)-2-allyl-6-((4-(4-(aminomethyl)piperidin-l- yl)phenyl)amino)-l-(7-ethyl-7-hydroxy-6,7-dihydro-5H-cyclope nta[b]pyridin-2-yl)-l,2- dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (Intermediate 31) (47.21 mg, 87.31 pmol) in DCM (1 mL) was added AcOH (524.33 pg, 8.73 pmol) to adjust pH=5-6. Then NaBH(OAc)3 (37.01 mg, 174.63 pmol) was added at 20 °C after 2 h. The reaction was stirred at 20 °C for 12 h. LCMS showed the reaction was completed and the desired product was detected. The mixture was concentrated under reduced pressure at 40 °C to give the crude product. The crude product was purified by prep-HPLC (NH3H2O Condition) to give the pure product. (R)- 4-(3-(4-(((l-(4-((2-allyl-l-(7-ethyl-7-hydroxy-6,7-dihydro-5 H-cyclopenta[b]pyridin-2-yl)-3- oxo-2,3-dihydro-lH-pyrazolo[3,4-d]pyrimidin-6-yl)amino)pheny l)piperidin-4- yl)methyl)amino)piperidin-l-yl)phenyl)piperidine-2, 6-dione (Compound 25) (0.01 g, 11.81 pmol, 13.53% yield, 94.14% purity) was obtained as a white solid. LCMS (ESI ⁺ ) m/z 811.5 [M+H] ⁺ . ’H NMR (400 MHz. DMSO-d ₆ ) 8 10.81 (br s, 1H). 10.10 (br d. J = 4.1 Hz. 1H), 8.81 (s, 1H), 7.92 (br d, 7= 7.1 Hz, 1H), 7.69 (d, J = 8.1 Hz, 1H), 7.56 (br s, 2H), 7.14 (t, J = 7.9 Hz, 1H), 6.94 - 6.86 (m, 3H), 6.80 (dd, 7 =8.3, 1.8 Hz, 1H), 6.66 (d, 7= 7.6 Hz, 1H), 5.72 - 5.61 (m, 1H), 5.04 (s, 1H), 4.99 (d, 7 = 9.4 Hz, 1H), 4.85 (br d, 7 = 17.0 Hz, 1H), 4.80 - 4.70 (m, 1H), 4.60 - 4.51 (m, 1H), 3.63 (br d, 7 = 12.1 Hz, 4H), 3.02 - 2.92 (m, 1H), 2.84 - 2.68 (m, 5H), 2.64 (br d, 7 = 4.3 Hz. 2H), 2.60 (br d. 7 = 4.1 Hz, 2H), 2.55 (br d, 7 = 9.9 Hz, 2H), 2.48 (br s, 1H), 2.20 (ddd, 7=13.6, 8.3, 5.8 Hz, 1H), 2.01 (ddd, 7=13.4, 8.3, 5.3 Hz, 1H), 1.92 - 1.88 (m, 3H), 1.87 - 1.78 (m, 3H), 1.70 (br dd, J =13.6, 7.4 Hz, 1H), 1.52 - 1.44 (m, 1H), 1.38 - 1.21 (m, 4H), 0.87 (t, J = 7.4 Hz, 3H).

Example 26: (R)-4-(4-(4-((l-(4-((2-allyl-l-(7-ethyl-7-hydroxy-6,7-dihydr o-5H- cyclopenta[b]pyridin-2-yl)-3-oxo-2,3-dihydro-lH-pyrazolo[3,4 -d]pyrimidin-6- yl)amino)phenyl)piperidin-4-yl)amino)piperidin-l-yl)phenyl)p iperidine-2, 6-dione

(Compound 26)

[0319] To a solution of 4-(4-(4-oxopiperidin-l-yl)phenyl)piperidine-2, 6-dione (Intermediate 30) (0.05 g, 174.63 pmol) and (R)-2-allyl-6-((4-(4-aminopiperidin-l- yl)phenyl)amino)-l-(7-ethyl-7-hydroxy-6,7-dihydro-5H-cyclope nta[b]pyridin-2-yl)-l,2- dihydro-3H-pyrazolo[3.4-d]pyrimidin-3-one (Intermediate 36) (72.85 mg, 145.52 pmol) in DCM (1 mL) and MeOH (0.2 mL) was added AcOH (873.89 pg, 14.55 pmol) to adjust pH=5- 6. Then NaBH(OAc)3 (61.68 mg, 291.04 pmol) was added at 25 °C after 2 h and the reaction was stirred at 20 °C for 12 h. LCMS showed the reaction was completed and the desired product was detected. The mixture was concentrated under reduced pressure at 40 °C to give the crude product. The crude product was purified by prep-HPLC (FA Condition) to give the pure product. (R)-4-(4-(4-(( 1 -(4 -((2- ally 1- 1 -(7 -ethyl-7-hydroxy-6,7 -dihydro-5 H- cyclopenta[b]pyridin-2-yl)-3-oxo-2,3-dihydro-lH-pyrazolo[3,4 -d]pyrimidin-6- yl)amino)phenyl)piperidin-4-yl)amino)piperidin-l-yl)phenyl)p iperidine-2, 6-dione (Compound 26) (0.0275 g, 33.62 pmol, 23.10% yield, 97.42% purity) was obtained as a yellow solid. LCMS (ESI ⁺ ) m/z 797.5 [M+H] ⁺ . ¹ H NMR (400 MHz, DMSO-d ₆ ) 8 10.79 (s, 1H), 10.27 - 9.92 (m, 1H), 8.81 (s, 1H), 7.92 (br d, J = 6.6 Hz, 1H). 7.69 (d, J = 8.1 Hz. 1H), 7.56 (br s, 2H), 7.12 (d, J = 8.6 Hz, 2H), 6.95 - 6.87 (m, 4H), 5.72 - 5.61 (m, 1H), 5.05 (br s, 1H), 4.99 (br d, J = 10.4 Hz, 1H), 4.85 (br d, J = 17.1 Hz, 1H), 4.81 - 4.70 (m, 1H), 4.61 - 4.51 (m, 1H), 3.67 - 3.58 (m. 4H). 3.01 - 2.93 (m, 1H), 2.86 - 2.66 (m. 10H), 2.61 (br dd, J = 4.4. 16.6 Hz, 3H), 2.20 (ddd, J =13.5, 8.2, 5.8 Hz, 1H), 2.01 (ddd, J =13.3, 8.3, 5.2 Hz, 1H), 1.94

- 1.84 (m, 5H), 1.70 (br dd, J =13.7, 7.3 Hz, 1H), 1.43 - 1.30 (m, 4H), 0.87 (t, J = 7.4 Hz, 3H).

Example 27: 3 - [4- [4- [4- [4- [ [2 - ally 1- 1 - [ (7 S )-7 -ethy 1-7 -hydroxy-5 ,6- dihydrocyclopenta[b]pyridin-2-yl]-3-oxo-pyrazolo[3,4-d]pyrim idin-6- yl]amino]phenyl]piperazin-l-yl]butylamino]phenyl]piperidine- 2, 6-dione (Compound 27)

[0320] To a solution of 4-[4-[4-[[2-allyl-l-[(7R)-7-ethyl-7-hydroxy-5,6- dihydrocyclopenta[b]pyridin-2-yl]-3-oxo-pyrazolo[3,4-d]pyrim idin-6- yl]amino]phenyl]piperazin-l-yl]butanal (Intermediate 38) (0.082 g, 101.60 pmol, 72.20% purity) in THF (2 mL) were added 3-(4-aminophenyl)piperidine-2, 6-dione (commercially available) (41.50 mg, 203.21 pmol), NaBH(OAc)s (43.07 mg, 203.21 pmol,) at 20 °C for 2 h under N2. LCMS showed the starting material was consumed completely. The mixture was concentrated in vacuum to give a residue, which was purified by prep-HPLC to afford 3-[4- [4-[4-[4-[[2-allyl-l-[(7S)-7-ethyl-7-hydroxy-5,6-dihydrocycl openta[b]pyridin-2-yl]-3-oxo- pyrazolo[3,4-d]pyrimidin-6-yl]amino]phenyl]piperazin-l-yl]bu tylamino]phenyl]piperidine- 2, 6-dione (Compound 27) (0.021 g, 25.92% yield, 96.67% purity) as a yellow solid. LCMS (ESI ⁺ ) m/z 771.4 [M+H] ⁺ . NMR (400 MHz, DMSO-d ₆ ) 8 10.72 (br s, 1H), 10.12 (br s, 1H), 8.82 (s, 1H), 7.97 - 7.83 (m. 1H), 7.69 (br d, J = 8.1 Hz. 1H). 7.57 (br s, 2H), 6.96 - 6.85 (m, 4H), 6.52 (br d, J = 8.1 Hz, 2H), 5.76 - 5.59 (m, 1H), 5.52 (br t, J = 4.8 Hz, 1H), 5.05 (s, 1H), 4.99 (br d, J = 10.1 Hz, 1H), 4.85 (br d, J = 17.1 Hz, 1H), 4.80 - 4.70 (m, 1H). 4.62 - 4.51 (m, 1H), 3.63 (br dd, J = 10.3, 4.9 Hz, 1H), 3.09 (br s, 4H), 3.00 (m, 4H), 2.83 - 2.72 (m, 1H), 2.69 - 2.56 (m, 2H), 2.45 (br s, 1H), 2.41 (br s, 1H), 2.35 (br s, 2H), 2.25 - 2.15 (m, 1H), 2.13 - 1.95 (m, 4H), 1.93 - 1.83 (m, 1H), 1.70 (qd. J = 13.7, 6.9 Hz, 1H), 1.57 (br s, 4H). 0.87 (br t, J = 7.3 Hz, 3H). Example 28: 3-((4-(4-((4-(4-((2-allyl-l-((R)-7-ethyl-7-hydroxy-6,7-dihyd ro-5H- cyclopenta[b]pyridin-2-yl)-3-oxo-2,3-dihydro-lH-pyrazolo[3,4 -d]pyrimidin-6- yl)amino)phenyl)piperazin-l-yl)methyl)-4-hydroxypiperidin-l- yl)phenyl)amino (piperidine-

2,6-dione (Compound 28)

[0321] To a solution of tert-butyl 4-[[4-(4-aminophenyl)piperazin-l-yl]methyl]-4- hydroxy-piperi dine- 1 -carboxylate (Intermediate 40) (0.15 g, 200.46 pmol, 52.19% purity) and 2-allyl-l-[(7R)-7-ethyl-7-hydroxy-5,6-dihydrocyclopenta[b]py ridin-2-yl]-6- methylsulfonyl-pyrazolo[3,4-d]pyrimidin- 3-one (166.57 mg, 400.93 pmol) in DMF (1.5 mL) was added DIEA (51.82 mg, 400.93 pmol, 69.83 pL) at 20 °C. The mixture was stirred at 20 °C for 12 h. LCMS showed the reaction was completed and the desired product was detected. The mixture was filtered and the filter was concentrated to give the residue. The residue was purified by prep-HPLC (FA Condition) to afford 3-((4-(4-((4-(4-((2-allyl-l-((R)-7-ethyl-7- hydroxy-6,7-dihydro-5H-cyclopenta[b]pyridin-2-yl)-3-oxo-2,3- dihydro-lH-pyrazolo[3,4- d]pyrimidin-6-yl)amino)phcnyl)pipcrazin- 1 -y l)mcthyl)-4-hydroxypipcridin- 1 - yl)phenyl)amino)piperidine-2, 6-dione (Compound 28) (40.0 mg, 22.95% yield, 95.23% purity) as a white solid. LCMS (ESI ⁺ ) m/z 414.8 [M/2+H] ⁺ . ¹ H NMR (400 MHz, DMSO-d ₆ ) 5 10.76 (s, 1H), 10.23 - 10.00 (m, 1H). 8.82 (s, 1H), 7.98 - 7.89 (m, 1H), 7.69 (d, 7 = 8.2 Hz, 1H), 7.65 - 7.47 (m, 2H), 6.91 (br d, J = 8.9 Hz, 2H), 6.76 (d, J = 8.8 Hz, 2H), 6.60 (d, J = 8.9 Hz, 2H), 5.67 (tdd. 7 - 5.9, 10.6, 16.8 Hz, 1H), 5.34 (d, 7= 7.1 Hz, 1H), 5.06 (s, 1H), 4.99 (d, 7 = 10.1 Hz, 1H), 4.85 (br d, 7 = 17.5 Hz, 1H), 4.81 - 4.67 (m, 1H), 4.57 (br dd, 7 = 4.1, 14.9 Hz, 1H), 4.18 (ddd, 7 = 4.8, 6.8, 11.2 Hz, 1H), 4.09 (s, 1H), 3.18 - 2.84 (m, 9H), 2.83 - 2.63 (m, 6H), 2.57 (td, 7 = 4.2, 17.6 Hz, 1H), 2.34 (s, 2H), 2.20 (ddd, 7 = 5.7, 8.1, 13.4 Hz, 1H), 2.15 - 2.06 (m, 1H), 2.01 (ddd, 7 = 5.4. 8.4, 13.4 Hz, 1H), 1.96 - 1.77 (m, 2H), 1.76 - 1.64 (m, 3H), 1.61 - 1.52 (m, 2H), 0.87 (t, 7 = 7.5 Hz, 3H). Example 29: 3-((4-(4-(4-((2-allyl-l-((R)-7-ethyl-7-hydroxy-6,7-dihydro-5 H- cyclopenta[b]pyridin-2-yl)-3-oxo-2,3-dihydro-lH-pyrazolo[3,4 -d]pyrimidin-6- yl)amino)phenethyl)-l,4-diazepan-l-yl)phenyl)amino)piperidin e-2, 6-dione (Compound 29)

[0322] To a solution of 2-[4-[[2-allyl-l-[(7R)-7-ethyl-7-hydroxy-5,6- dihydrocyclopenta[b]pyridin-2-yl]-3-oxo-pyrazolo[3,4-d]pyrim idin-6-yl]amino]phenyl]ethyl methanesulfonate (Intermediate 43) (160.0 mg, 290.58 pmol) and 3-[4-(l,4-diazepan-l- yl)anilino]piperidine-2, 6-dione (Intermediate 44) (141.5 mg, 435.87 pmol,) in dioxane (5 mb) was added DIEA (75.1 mg, 581.15 pmol) and Nal (4.4 mg, 29.06 pmol) in one portion at 25 °C under N2. The mixture was stirred at 100 °C for 12 h. LCMS showed the reaction was completed and the desired product was detected. The mixture was concentrated under reduced pressure at 40 °C to give the crude product. The crude product was purified by prep- HPLC (NH4HCO3 condition) to afford Compound 29 (22.0 mg, 8.45%). LCMS (ESI ⁺ ) m/z 757.3 [M+H] ⁺ . NMR (400 MHz, DMSO-d ₆ ) 5 10.75 (s, 1H), 10.43 - 9.95 (m. 1H). 8.87 (s, 1H), 7.92 (d, J = 8.1 Hz, 1H), 7.76 - 7.55 (m, 3H), 7.17 (br d, J = 8.4 Hz, 2H), 6.65 - 6.58 (m, 2H), 6.58 - 6.51 (m, 2H), 5.75 - 5.60 (m, 1H), 5.13 - 5.03 (m, 2H), 5.00 (d, J = 10.1 Hz, 1H), 4.86 (brd, J = 17.5 Hz, 1H). 4.81 - 4.70 (m, 1H), 4.57 (br dd. J = 15.8. 6.3 Hz, 1H).4.16 - 4.07 (m, 1H), 3.40 (br d, J - 4.0 Hz, 4H), 3.03 - 2.92 (m, 1H), 2.83 - 2.73 (m, 3H), 2.72 - 2.64 (m, 5H), 2.63 - 2.55 (m, 3H), 2.26 - 2.17 (m, 1H), 2.16 - 2.08 (m, 1H), 2.02 (ddd, J = 13.4, 8.3, 5.4 Hz, 1H), 1.95 - 1.77 (m, 4H), 1.71 (qd, J = 13.9, 7.1 Hz, 1H), 0.87 (t, J = 7.3 Hz, 3H). Example 30: 3-((4-(4-((R)-2-(4-((2-allyl-l-((R)-7-ethyl-7-hydroxy-6,7-di hydro-5H- cyclopenta[b]pyridin-2-yl)-3-oxo-2,3-dihydro-lH-pyrazolo[3,4 -d]pyrimidin-6- yl)amino)phenyl)-2-hydroxyethyl)piperazin-l-yl)phenyl)amino) piperidine-2, 6-dione

(Compound 30)

[0323] To a solution of 3 -(4-piperazin-l-ylanilino)piperidine-2, 6-dione (WO 2021/083949) (138.03 mg, 478.69 pmol) in THF (1 mL) were added (R)-2-(4-((2-allyl-l-((R)- 7-ethyl-7-hydroxy-6,7-dihydro-5H-cyclopenta[b]pyridin-2-yl)- 3-oxo-2,3-dihydro-lH- pyrazolo[3,4-d]pyrimidin-6-yl)amino)phenyl)-2-hydroxyethyl methanesulfonate

(Intermediate 47) (138.01 mg, 119.67 pmol) and TEA (36.33 mg, 359.02 pmol, 49.97 uL) at 20 °C. Then the mixture was stirred at 65 °C for 24 h under N2. LCMS showed the reaction was completed and the desired product was detected. The mixture was concentrated in vacuum to give a residue. The residue was purified hy prep-HPLC (FA Condition) to afford Compound 30 (9.1 mg, 9.69%). LCMS (ESI ⁺ ) m/z 759.3 [M+H] ⁺ . DMSO-d ₆ ) 5 10.75 (s. 1H), 10.24 (br s, 1H), 8.88 (s, 1H), 7.91 (d. J = 8.1 Hz, 1H), 7.70 (dd, J = 8.3, 3.6 Hz, 3H), 7.31 (d, J = 8.5 Hz, 2H), 6.75 (d, J = 8.9 Hz, 2H), 6.61 (d, J = 8.9 Hz, 2H), 5.78 - 5.55 (m, 1H), 5.37 (br d, 7 = 7.1 Hz. 1H), 5.13 - 4.93 (m, 3H), 4.86 (dd, 7 = 17.1, 1.1 Hz, 1H), 4.71 (br dd, 7 = 7.8, 4.7 Hz, 2H), 4.58 (br d, 7 = 6.1 Hz, 1H), 4.25 - 4.11 (m, 1H), 3.03 - 2.82 (m, 5H), 2.81 - 2.67 (m, 2H), 2.62 (br s, 4H), 2.59 - 2.51 (m, 2H), 2.43 (br dd, 7 = 12.6, 4.5 Hz, 1H), 2.25 - 2.15 (m, 1H). 2.14 - 2.06 (m, 1H), 2.01 (br d, 7 = 5.1 Hz. 1H), 1.95 - 1.77 (m, 2H), 1.76 - 1.64 (m, 1H), 0.87 (t, 7 = 7.4 Hz, 3H). Example 31: 3-((4-(4-((S)-2-(4-((2-allyl-l-((R)-7-ethyl-7-hydroxy-6,7-di hydro-5H- cyclopenta[b]pyridin-2-yl)-3-oxo-2,3-dihydro-lH-pyrazolo[3,4 -d]pyrimidin-6- yl)amino)phenyl)-2-hydroxyethyl)piperazin-l-yl)phenyl)amino) piperidine-2, 6-dione

(Compound 31)

[0324] Compound 31 (5% yield) was synthesized from 3-(4-piperazin-l- ylanilino)piperidine-2, 6-dione and Intermediate 48 by following a similar procedure as described for the synthesis of Compound 30. LCMS (ESI ⁺ ) m/z 759.3 [M+H] ⁺ . ^! H NMR (400 MHz, DMSO-d ₆ ) 8 10.76 (br s. 1H), 10.25 (br s, 1H), 8.88 (s, 1H). 7.91 (br d, J = 8.1 Hz, 1H), 7.69 (br dd, 7 = 8.1, 3.3 Hz, 3H), 7.31 (br d, J = 8.5 Hz, 2H), 6.75 (br d, J = 8.7 Hz, 2H), 6.60 (br d, J = 8.7 Hz, 2H), 5.75 - 5.60 (m, 1H), 5.38 (br d, J = 7.3 Hz, 1H), 5.08 (s, 1H), 5.00 (br d, J = 10.1 Hz, 2H), 4.85 (br d, J = 17.2 Hz, 1H), 4.80 - 4.67 (m. 2H). 4.56 (br dd, J = 16.2, 5.9 Hz, 1H), 4.23 - 4.13 (m, 1H), 2.93 (br s, 5H), 2.81 - 2.68 (m, 3H), 2.66 - 2.57 (m, 4H), 2.57 - 2.53 (m, 1H), 2.43 (br dd, 7 = 12.6, 4.4 Hz, 1H), 2.24 - 2.15 (m, 1H), 2.14 - 2.06 (m, 1H), 2.00 (ddd, 7 =13.2, 8.2, 5.4 Hz, 1H). 1.94 - 1.77 (m, 2H), 1.75 - 1.64 (m. 1H). 0.86 (br t, 7 = 7.3 Hz, 3H).

Example 32: 3-((4-(4-((l-(4-((2-allyl-l-((R)-7-ethyl-7-hydroxy-6,7-dihyd ro-5H- cyclopenta[b]pyridin-2-yl)-3-oxo-2,3-dihydro-lH-pyrazolo[3,4 -d]pyrimidin-6- yl)amino)phenyl)piperidin-4-yl)amino)piperidin-l -yl)phenyl)amino)piperidine-2, 6-dione

(Compound 32) [0325] Compound 32 (30% yield) was synthesized from Intermediate 50 and (R)-2-allyl-l-(7-cthyl-7-hydroxy-6,7-dihydro-5H-cyclopcnta[b ]pyridin-2-yl)-6- (methylsulfonyl)-l,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-o ne using 2-propanol as solvent and in the presence of DIEA and at 20 °C for 12 h. LCMS (ESI ⁺ ) m/z 812.4 [M+H] ⁺ . 'H NMR (400 MHz, DMSO-d ₆ ) 8 10.75 (br s, 1H), 10.21 - 9.98 (m, 1H), 8.81 (s, 1H), 7.93 (br d, J = 7.4 Hz, 1H), 7.69 (d. J = 8.1 Hz, 1H), 7.56 (br s, 2H). 6.92 (br d, J = 9.0 Hz. 2H). 6.75 (d, J = 8.9 Hz, 2H), 6.60 (d, J = 8.9 Hz, 2H), 5.67 (tdd, J = 16.8, 10.5, 6.0 Hz, 1H), 5.35 (d, J = 7.3 Hz, 1H), 5.04 (s, 1H), 5.02 - 4.96 (m, 1H), 4.85 (br d, J = 17.0 Hz, 1H), 4.80 - 4.68 (m, 1H), 4.56 (br dd, J = 15.7, 5.4 Hz, 1H), 4.18 (ddd, J = 11.3, 6.9, 4.9 Hz, 1H), 3.59 (br d, J = 12.3 Hz, 2H), 3.36 (br s, 3H), 3.04 - 2.91 (m, 1H), 2.83 - 2.62 (m, 6H), 2.61 - 2.51 (m, 3H), 2.26 - 2.16 (m, 1H). 2.14 - 2.06 (m, 1H), 2.01 (ddd, J = 13.4, 8.4, 5.3 Hz, 1H). 1.94 - 1.78 (m, 6H), 1.70 (dd, J = 13.6, 7.3 Hz, 1H), 1.42 - 1.28 (m, 4H), 0.87 (t, J = 7.4 Hz, 3H)

Example 33: 3-(3-((6-((4-(4-((2-allyl-l-((R)-7-ethyl-7-hydroxy-6,7-dihyd ro-5H- cyclopenta[b]pyridin-2-yl)-3-oxo-2,3-dihydro-lH-pyrazolo[3,4 -d]pyrimidin-6- yl)amino)phenyl)piperazin-l-yl)methyl)-2-azaspiro[3.3]heptan -2- yl)methyl)phenyl)piperidine-2, 6-dione (Compound 34)

[0326] Compound 40 (10% yield) was synthesized from Intermediate 8 and Intermediate 51 by following a similar procedure as described in Example 29 for the synthesis of Compound 29. LCMS (ESI ⁺ ) mJz 823.4 [M+H] ⁺ . ¹ H NMR (400 MHz, DMSO- d ₆ ) 8 10.85 (s, 1H), 10.33 - 10.01 (m, 1H), 8.84 (s. 1H), 8.36 (s, 2H). 7.90 (br d, J = 8.1 Hz, 1H), 7.73 - 7.61 (m, 3H), 7.31 - 7.21 (m, 2H), 7.17 (s, 1H), 7.11 - 6.99 (m, 3H), 5.67 (tdd, J = 16.8, 10.7, 5.9 Hz, 1H), 5.00 (d, J = 9.8 Hz, 1H), 4.86 (br d, J = 17.3 Hz, 1H), 4.80 - 4.70 (m, 1H), 4.57 (br dd, J = 15.8, 5.1 Hz, 1H), 3.93 (br s, 2H), 3.88 - 3.77 (m, 7H), 3.67 (s, 2H), 3.55 - 3.50 (m, 4H), 3.01 - 2.91 (m, 1H), 2.83 - 2.73 (m, 1H), 2.65 (ddd, J = 17.1, 11.5, 5.3 Hz, 1H), 2.55 - 2.51 (m, 1H), 2.47 (s, 2H), 2.25 - 2.11 (m, 2H), 2.08 - 2.00 (m, 2H), 1.99 - 1.93 (m, 2H), 1.93 - 1.84 (m, 1H), 1.80 - 1.66 (m, 3H), 0.87 (t, J = 7.4 Hz, 3H). Example 34: 3-(4-(2-(4-(4-((2-allyl-l-((R)-7-cthyl-7-hydroxy-6,7-dihydro -5H- cyclopenta[b]pyridin-2-yl)-3-oxo-2,3-dihydro-lH-pyrazolo[3,4 -d]pyrimidin-6- yl)amino)phenyl)piperazin-l-yl)ethyl)phenyl)piperidine-2, 6-dione (Compound 34)

[0327] Compound 34 (21% yield) was synthesized from Intermediate 52 and (R)-2-allyl-l-(7-ethyl-7-hydroxy-6,7-dihydro-5H-cyclopenta[b ]pyridin-2-yl)-6- (methylsulfonyl)-l,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-o ne in the presence of i-PrOH and at 80 °C for 12 h under N ₂ . LCMS (ESI ⁺ ) m/z 728.4 [M+H] ⁺ . ’ H NMR (400 MHz, DMSO-d ₆ ) 5 10.81 (s. 1H), 10.18 - 10.06 (m, 1H), 8.82 (s, 1H). 7.93 (br d, J = 7.1 Hz, 1H), 7.69 (d, ./ = 8.1 Hz, 1H), 7.58 (br s, 2H), 7.24 - 7.17 (m, 2H), 7.17 - 7.09 (m, 2H), 6.93 (br d, J = 8.9 Hz, 2H), 5.67 (tdd, J = 16.8, 10.6, 5.9 Hz, 1H), 5.05 (s, 1H), 5.02 - 4.97 (m, 1H), 4.85 (br d, J = 17.1 Hz. 1H), 4.81 - 4.67 (m, 1H), 4.63 - 4.51 (m. 1H), 3.81 (dd, J = 11.3, 4.9 Hz, 1H), 3.11 (br s, 4H), 3.02 - 2.92 (m, 1H), 2.83 - 2.73 (m, 3H), 2.71 - 2.64 (m, 1H), 2.64 - 2.55 (m, 6H), 2.48 - 2.44 (m, 1H), 2.25 - 2.12 (m, 2H), 2.07 - 1.97 (m, 2H), 1.94 - 1.83 (m, 1H), 1.76 - 1.65 (m, 1H), 0.87 (t. J = 7.4 Hz, 3H).

Example 35: 3-(4-(4-((4-((2-allyl-l-((R)-7-ethyl-7-hydroxy-6,7-dihydro-5 H- cyclopenta[b]pyridin-2-yl)-3-oxo-2,3-dihydro-lH-pyrazolo[3,4 -d]pyrimidin-6- yl)amino)phenethyl)amino)piperidin-l-yl)phenyl)piperidine-2, 6-dione (Compound 35) [0328] Compound 35 (19% yield) was synthesized by following a similar procedure as described in Example 29 for the synthesis of Compound 29 from 3-[4-(4- amino-l-piperidyl)phenyl]piperidine-2, 6-dione (Intermediate 13) and 2-[4-[[2-allyl- 1- [(7R)-7-ethyl-7-hydroxy-5,6-dihydrocyclopenta[b]pyridin-2-yl ]-3-oxo-pyrazolo[3,4- d]pyrimidin-6-yl]amino]phenyl]ethyl methanesulfonate (Intermediate 43) using DMF as solvent. LCMS (ESI ⁺ ) m/z 742.3 [M+H] ⁺ . 'H NMR (400 MHz, DMSO-d ₆ ) 8 10.76 (br s. 1H), 10.20 (br d, 7=2.50 Hz, 1H), 8.87 (s, 1H), 7.91 (d, 7=8.13 Hz, 1H), 7.62-7.72 (m, 3H), 7.18 (br d, 7=8.25 Hz. 2H), 7.02 (d, 7=8.50 Hz. 2H), 6.87 (d, 7=8.63 Hz, 2H), 5.59-5.74 (m, 1H), 5.06 (s, 1H), 5.00 (br d, 7=10.26 Hz, 1H), 4.86 (br d, 7=17.13 Hz, 1H), 4.75 (br dd, 7=15.13, 3.50 Hz, 1H), 4.56 (br dd, 7=15.95, 6.19 Hz, 1H), 3.71 (br dd, 7=10.82, 4.94 Hz, 1H), 3.57 (br s. 2H). 2.92-3.01 (m, 1H), 2.75-2.82 (m, 3H), 2.73 (br d, 7=5.88 Hz, 1H), 2.63- 2.71 (m, 4H), 2.56-2.63 (m, 2H), 2.41-2.47 (m, 1H), 2.14-2.24 (m, 1H), 2.08-2.14 (m, 1H), 1.97-2.07 (m, 2H), 1.84-1.92 (m, 3H), 1.70 (dq, 7=13.91, 7.12 Hz, 1H), 1.25-1.37 (m, 2H), 0.87 (t, 7=7.32 Hz, 3H).

Example 36: 3-(4-(4-((3-(4-(4-((2-allyl-l-((R)-7-ethyl-7-hydroxy-6,7-dih ydro-5H- cyclopenta[b]pyridin-2-yl)-3-oxo-2,3-dihydro-lH-pyrazolo[3,4 -d]pyrimidin-6- yl)amino)phenyl)piperazin- 1 -yl)azetidin- 1 -yl)methyl)piperidin- 1 -yl)phenyl)piperidine-2,6- dione (Compound 36)

[0329] Compound 36 (9% yield) was synthesized from Intermediate 53 and

Intermediate 8 by following a similar procedure as described in Example 33 for the synthesis of Compound 33. LCMS (ESI ⁺ ) m/z. 852.5 [M+H] ⁺ . ¹ H NMR (400 MHz, DMSO-d ₆ ) 8 10.75 (s. 1H), 10.17 - 10.01 (m, 1H), 8.81 (s, 1H), 7.93 (br d, J = 7.9 Hz, 1H), 7.69 (d, 7 = 8.1 Hz,

1H), 7.58 (br d, 7 = 4.4 Hz. 2H), 7.03 (d, 7 = 8.6 Hz, 2H), 6.90 (br dd, 7 = 13.8, 8.9 Hz. 4H),

5.74 - 5.57 (m, 1H), 5.07 - 4.94 (m, 2H), 4.85 (br d, J = 17.4 Hz, 1H), 4.80 - 4.68 (m, 1H),

4.63 - 4.50 (m, 1H), 3.77 - 3.61 (m, 3H), 3.54 (br dd. J = 17.9, 10.4 Hz, 2H), 3.02 - 2.92 (m, 1H), 2.87 - 2.71 (m, 5H), 2.71 - 2.57 (m, 4H), 2.49 - 2.41 (m, 2H), 2.36 - 2.25 (m, 2H), 2.25 - 2.12 (m, 5H), 2.09 (br dd, J= 11.6, 3.4 Hz, 1H), 2.01 (ddd, J = 13.3, 8.2, 5.4 Hz, 2H), 1.89 (qd. J = 13.9, 7.1 Hz, 1H), 1.84 - 1.59 (m, 5H), 1.28 - 1.14 (m. 2H). 0.87 (t, J = 7.4 Hz, 3H).

Example 37: 3-(4-((6-((4-(4-((2-allyl-l-((R)-7-ethyl-7-hydroxy-6,7-dihyd ro-5H- cyclopenta[b]pyridin-2-yl)-3-oxo-2,3-dihydro-lH-pyrazolo[3,4 -d]pyrimidin-6- yl)amino)phenyl)piperazin-l-yl)methyl)-2-azaspiro[3.3]heptan -2- yl)methyl)phenyl)piperidine-2, 6-dione (Compound 37)

[0330] Compound 37 (4% yield) was synthesized from Intermediate 55 and Intermediate 8 by following a similar procedure as described in Example 33 for the synthesis of Compound 33. LCMS (ESI ⁺ ) m/z 823.4 [M+H] ⁺ . NMR (400 MHz. DMSO-d ₆ ) 8 11.05

- 10.53 (m, 1H), 10.31 - 10.06 (m, 1H), 8.84 (d, J = 1.3 Hz, 1H), 7.95 - 7.87 (m, 1H), 7.75 - 7.59 (m, 3H), 7.29 (d, J = 8.0 Hz, 1H), 7.20 (br d, J = 8.6 Hz, 1H), 7.18 - 7.08 (m, 2H), 7.02 (br d, J = 8.8 Hz, 2H). 5.75 - 5.59 (m, 1H), 5.00 (br d, J = 10.4 Hz. 1H), 4.86 (br d. J = 16.8 Hz, 1H), 4.80 - 4.66 (m, 1H), 4.62 - 4.50 (m, 1H), 4.48 - 4.28 (m, 1H), 3.92 (br s, 1H), 3.88 - 3.74 (m, 6H), 3.68 (br s. 1H), 3.66 (s, 2H), 3.48 (br d, J = 4.0 Hz, 3H), 3.42 (br s, 2H), 3.21 - 3.12 (m, 2H), 3.02 - 2.91 (m. 1H). 2.84 - 2.73 (m, 1H), 2.65 - 2.58 (m, 1H). 2.43 - 2.38 (m. 1H), 2.25 - 2.11 (m, 2H), 2.08 - 1.94 (m, 3H), 1.93 - 1.80 (m, 2H), 1.79 - 1.64 (m, 2H), 1.60

- 1.45 (m, 1H). 0.87 (t, J = 7.4 Hz, 3H).

Example 38: 3-(4-((3-(4-(4-((2-allyl-l-((R)-7-ethyl-7-hydroxy-6,7-dihydr o-5H- cyclopenta[b]pyridin-2-yl)-3-oxo-2,3-dihydro-lH-pyrazolo[3,4 -d]pyrimidin-6- yl)amino)phenyl)piperazin- 1 -yl)propyl)amino)phenyl)piperidine-2, 6-dione (Compound 38)

[0331] Compound 38 (12% yield) was synthesized from Intermediate 56 and 2- allyl- 1 - [(7R)-7 -ethyl-7 -hydroxy-5, 6-dihydrocyclopenta[b]pyridin-2-yl] -6-(4-piperazin- 1 - ylanilino)pyrazolo[3,4-d]pyrimidin-3-one by following a similar procedure as described in Example 33 for the synthesis of Compound 33. LCMS (ESI ⁺ ) m/z 757.4 [M+H] ⁺ . ¹ H NMR (400 MHz, DMSO-d ₆ ) 5 10.73 (s, 1H), 10.24 - 9.99 (m, 1H), 8.83 (s, 1H), 7.94 (br d, J - 5.7 Hz, 1H), 7.70 (d, J = 8.2 Hz, 1H), 7.59 (br d, J = 4.6 Hz, 2H), 7.00 - 6.83 (m, 4H), 6.53 (d, J = 8.5 Hz, 2H), 5.77 - 5.53 (m, 2H), 5.08 - 4.96 (m, 2H), 4.91 - 4.82 (m, 1H), 4.81 - 4.70 (m, 1H), 4.63 - 4.53 (m, 1H), 3.64 (dd, J = 10.5, 5.1 Hz, 1H), 3.12 (br s, 4H), 3.06 (q, J = 6.2 Hz, 2H), 3.02 - 2.93 (m, 1H), 2.83 - 2.74 (m, 1H), 2.66 - 2.58 (m. 1H), 2.54 (br s, 4H), 2.48 - 2.41 (m, 3H), 2.25 - 2.16 (m, 1H), 2.12 - 1.96 (m, 3H), 1.95 - 1.85 (m, 1H), 1.73 (qd, J = 14.0, 6.9 Hz, 3H), 0.88 (t, J = 7.4 Hz, 3H).

Example 39: 3-(4-((( 1 -(4-((2-allyl- 1 -((R)-7 -ethyl-7 -hydroxy-6,7 -dihydro-5H- cyclopenta[b]pyridin-2-yl)-3-oxo-2,3-dihydro-lH-pyrazolo[3,4 -d]pyrimidin-6- yl)amino)phenyl)piperidin-4-yl)amino)methyl)phenyl)piperidin e-2, 6-dione (Compound 39) [0332] To a mixture of Intermediate 36 (0.08 g, 105.26 pmol) in MeOH (2 mL) were added Intermediate 54 (27.44 mg, 126.31 pmol), AcOH (6.32 mg, 105.26 pmol, 6.03 pL) and NaBHsCN (7.94 mg, 126.31 pmol), and the mixture was stirred at 20 °C for 1 h under N2. LCMS showed the starting material was consumed completely and desired product was formed. The mixture was purified by prep-HPLC (FA condition) purification to afford Compound 39 (19.0 mg. 24.76%). LCMS (ESI ⁺ ) m/z 728.4 [M+H] ⁺ . ’H NMR (400 MHz. DMSO-d ₆ ) 8 10.85 (s, 1H), 10.21 - 10.05 (m, 1H), 8.82 (s, 1H), 8.23 (s, 1H), 8.03 - 7.82 (m, 1H), 7.70 (d, 7= 8.1 Hz, 1H), 7.57 (br d. J = 1.7 Hz, 2H), 7.35 (d, J = 7.9 Hz, 2H), 7.19 (d, J = 8.0 Hz, 2H), 6.93 (br d, J = 8.9 Hz, 2H), 5.74 - 5.61 (m, 1H), 5.20 - 5.02 (m, 1H), 5.00 (br d, J= 9.8 Hz, 1H), 4.86 (br d, J= 17.2 Hz, 1H), 4.81 - 4.69 (m, 1H), 4.64 - 4.49 (m, 1H), 3.86 (br d, J = 5.0 Hz, 1H), 3.83 (s, 2H), 3.62 (br d, J = 11.9 Hz, 2H), 3.03 - 2.92 (m, 1H). 2.84 - 2.74 (m, 1H), 2.73 - 2.58 (m, 4H), 2.49 - 2.44 (m, 1H), 2.27 - 2.11 (m, 2H), 2.08 - 1.82 (m, 5H), 1.71 (qd, J = 13.8, 7.0 Hz, 1H), 1.46 (q, J = 10.1 Hz, 2H), 0.88 (t, 7 = 7.3 Hz, 3H).

Example 40: 3-(4-(2-((l-(4-((2-allyl-l-((R)-7-ethyl-7-hydroxy-6,7-dihydr o-5H- cyclopenta[b]pyridin-2-yl)-3-oxo-2,3-dihydro-lH-pyrazolo[3,4 -d]pyrimidin-6- yl)amino)phenyl)piperidin-4-yl)amino)ethyl)phenyl)piperidine -2, 6-dione (Compound 40)

[0333] Compound 40 (15% yield) was synthesized from Intermediate 57 and 2- allyl-6- [4-(4-amino- 1 -piperidyl)anilino]- 1 - [(7S)-7 -ethyl-7 -hydroxy-5, 6- dihydrocyclopenta[b]pyridin-2-yl]pyrazolo[3.4-d]pyrimidin-3- one (Intermediate 36) by following a similar procedure as described in Example 29 for the synthesis of Compound 29. LCMS (ESI ⁺ ) m/z 742.3 [M+H] ⁺ . ’H NMR (400 MHz, DMSO-d ₆ ) 8 10.82 (br s, 1H), 10.23 - 10.02 (m, 1H). 8.81 (s, 1H), 7.93 (br d, J = 6.7 Hz, 1H), 7.69 (d. J = 8.1 Hz, 1H). 7.56 (br s, 2H), 7.29 - 7.08 (m, 4H), 6.92 (br d, J = 8.9 Hz, 2H), 5.66 (tdd, J = 16.8, 10.5, 5.9 Hz, 1H), 5.17 - 5.02 (m, 1H), 4.99 (dd, 7 = 10.2, 0.9 Hz, 1H), 4.85 (br d, 7 = 17.9 Hz, 1H), 4.81 - 4.66 (m, 1H), 4.56 (br dd, J = 15.5, 5.3 Hz, 1H), 3.81 (dd, J = 11.4, 4.9 Hz, 1H), 3.59 (br d, 7 = 12.2 Hz, 2H), 3.04 - 2.91 (m, 1H), 2.88 - 2.75 (m, 3H), 2.75 - 2.57 (m, 6H), 2.49 - 2.43 (m, 2H), 2.26 - 2.10 (m, 2H), 2.07 - 1.97 (m, 2H), 1.95 - 1.83 (m, 3H), 1.76 - 1.64 (m, 1H), 1.38 (q, J = 10.0 Hz, 2H), 0.87 (t, J = 7.4 Hz. 3H).

Example 41: 3-(4-((2-(4-(4-((2-allyl-l-((R)-7-ethyl-7-hydroxy-6,7-dihydr o-5H- cyclopenta[b]pyridin-2-yl)-3-oxo-2,3-dihydro-lH-pyrazolo[3,4 -d]pyrimidin-6- yl)amino)phenyl)piperazin-l-yl)ethyl)amino)phenyl)piperidine -2, 6-dione (Compound 41)

[0334] Compound 41 (25% yield) was synthesized by following a similar procedure as described for the synthesis of Compound 38 using 2-((4-(2,6-dioxopiperidin-3- yl)phenyl)amino)ethyl methanesulfonate (Intermediate 58) in place of 3-((4-(2,6- dioxopiperidin-3-yl)phenyl)amino)propyl methane sulfonate. LCMS (ESI ⁺ ) m/z 743.3 [M+H] ⁺ . ’ H NMR (400 MHz, DMSO-d ₆ ) 8 10.73 (br s. 1H), 10.12 (br s, 1H), 8.82 (s, 1H), 7.92 (br d, J = 5.1 Hz, 1H), 7.69 (br d, J = 8.1 Hz, 1H), 7.58 (br s, 2H), 6.92 (br d, J = 7.4 Hz, 4H), 6.56 (br d, J = 7.9 Hz, 2H), 5.75 - 5.59 (m, 1H), 5.37 (br s, 1H), 5.05 (s, 1H), 4.99 (br d, J = 10.3 Hz, 1H), 4.85 (br d, J = 17.1 Hz, 1H), 4.80 - 4.69 (m. 1H), 4.62 - 4.51 (m, 1H). 3.64 (br dd, J = 9.9, 4.4 Hz, 1H), 3.12 (br s, 6H), 3.04 - 2.89 (m, 2H), 2.83 - 2.72 (m, 1H), 2.58 (br s, 7H), 2.47 - 2.39 (m, 1H), 2.19 (br d, J = 5.6 Hz, 1H), 2.14 - 1.97 (m, 3H). 1.94 - 1.85 (m, 1H), 1.70 (br dd, J = 13.6,7.1 Hz, 1H), 0.87 (br t, 7 = 6.9 Hz, 3H).

Example 42: 3-(4-(3-(4-(4-((2-allyl-l-((R)-7-ethyl-7-hydroxy-6,7-dihydro -5H- cyclopenta[b]pyridin-2-yl)-3-oxo-2,3-dihydro-lH-pyrazolo[3,4 -d]pyrimidin-6- yl)amino)phenyl)piperazin- 1 -yl)propyl)phenyl)piperidine-2, 6-dione (Compound 42)

[0335] Compound 42 (51 % yield) was synthesized from Intermediate 60 and 2- allyl- 1 - [(7R)-7-ethyl-7-hydroxy-5,6-dihydrocyclopenta[b]pyridin-2-yl ] -6-(4-piperazin- 1 - ylanilino)pyrazolo[3,4-d]pyrimidin-3-one (Intermediate 8) by following a similar procedure as described in Example 32 for the synthesis of Compound 32. LCMS (ESI ⁺ ) m/z 742.3 [M+H] ⁺ . ’ H NMR (400 MHz, DMSO-d ₆ ) 5 10.81 (s. 1H), 10.22 - 9.99 (m, 1H), 8.82 (s. 1H), 7.93 (bi d, 7 - 7.1 Hz, 1H), 7.69 (d, 7- 8.1 Hz, 1H), 7.58 (br s, 2H), 7.21 - 7.16 (m, 2H), 7.15

- 7.10 (m, 2H), 6.92 (br d. J = 8.9 Hz, 2H), 5.74 - 5.59 (m, 1H), 5.04 (s, 1H), 4.99 (d, J = 9.9 Hz, 1H), 4.85 (br d, J = 17.0 Hz, 1H), 4.81 - 4.68 (m, 1H), 4.56 (br dd, J = 15.4, 5.4 Hz, 1H), 3.81 (dd, 7= 11.2, 4.9 Hz, 1H), 3.10 (br s, 4H), 3.03 - 2.90 (m, 1H), 2.84 - 2.73 (m, 1H), 2.71

- 2.56 (m, 3H), 2.55 - 2.51 (m. 4H), 2.47 - 2.43 (m, 1H), 2.35 (br t, 7 = 7.1 Hz, 2H). 2.25 - 2.11 (m, 2H), 2.09 - 1.97 (m, 2H), 1.95 - 1.83 (m, 1H), 1.83 - 1.65 (m, 3H), 0.87 (t, 7 = 7.4 Hz, 3H).

Example A

MOLT-4 and A-427 Cell Proliferation Assay

[0336] Cell proliferation was measured using the CellTiter-Glo® Luminescent Cell Viability Assay. The assay involved the addition of a single reagent (CellTiter-Glo® Reagent) directly to cells cultured in serum- supplemented medium. MOLT-4 cells (ATCC, CRL-1582) and A427 cells (ATCC, HTB-53) were cultured according to ATCC recommendations and were seeded at 6,000 cells for MOLT-4 and 3,000 cells for A427 per well.

[0337] Each compound evaluated was prepared as a DMSO stock solution (10 mM). Compounds were tested in duplicate on each plate, with a 10-point serial dilution curve (1 :3 dilution). The highest compound concentration was 10 pM (final), with a 0.1 % final DMSO concentration. Plates were then incubated at 37 °C, 5% CO2 for 72 h, and then were equilibrated at rt for approximately 30 mins. An equi-volume amount of CellTiter-Glo® Reagent (100 pL) was added to each well. Plates were mixed for 2 mins on an orbital shaker to induce cell lysis and then incubated at rt for 10 mins to stabilize the luminescent signal. Luminescence was recorded using a Spectramax i3x (Molecular Devices) plate reader according to CellTiter-Glo protocol. IC50 determination was performed using a non-linear regression, variable slope (four parameter), inhibitor v. response equation (Prism 9.0). IC50 values are provided in Table 1. For comparison, Table 1 also includes IC50 value for compound ZN-c3.

Table 1

For MOLT-4 and A427 CTG IC50: A = a single IC50 < 100 nM; B = a single IC50 >100 nM and < 200nM; C = a single IC50 >200 nM; ND = not determined

Example B

Protein degradation assay in MOLT-4 cells

[0338] 1 million MOLT-4 cells (ATCC, CRL-1582) were incubated with vehicle

(DMSO) or 10 pM, 1 pM, 0.1 pM or 0.01 pM concentrations of the indicated compounds for 5 h. After treatment, the cells were harvested in RIPA lysis buffer supplemented with 1% Phosphatase Inhibitor and Protease Inhibitor Cocktail and protein concentration was determined by BCA assay. An equal amount of protein (4.5 pg/lane) from each cell extract was loaded onto 12-230 kDa Separation 25 Capillary Cartridges from ProteinSimple and probed with WEE1 (1:200 dilution, final concentration of 1 pg/mL) and -Actin (1:500 dilution, final concentration of 0.6 pg/mL) antibodies on a ProteinSimple Jess system according to ProtcinSimplc/SimplcWcstcrn protocols. The area under the curve (AUC) from the resulting eletropherograms for each protein peak at the concentrations tested was used to calculate percent remaining for WEE1 using the equation percent remaining = lOO*(WEEl/0- Actin)/(WEElDMSo/p-ActinDMSo). For illustrative purposes, electopherograms were exported as virtual blots using Compass for SW v 6.0 (ProteinSimple). The WEE-1 antibody (sc-5285) was purchased from Santa Cruz Biotechnology and P-actin (MAB8929) was purchased from R&D Systems. Percent remaining of WEE1 after 5 hour treatment in MOLT-4 cells at the tested concentrations are provided in Table 2.

[0339] The ability for a heterobifunctional degrader to degrade a protein of interest (POI) is dependent upon the efficient formation of a ternary complex between a POI, the compound and the E3 ligase in a 1: 1: 1 ratio. At high compound concentrations, a phenomenon known as the “hook effect” occurs where POI and E3 binding sites are saturated, leading to POI and E3 proteins being bound by distinct degrader compounds, forming separate binary complexes. In this case, a 1:1:1 ternary complex cannot be formed efficiently between the POI and E3, and loss of degradation is observed. A “hook effect” was observed at higher concentrations with several compounds, however these results indicated that many of the exemplified compounds are potent degraders of WEE 1.

Table 2

For MOLT-4 cells WEE1 degradation assay: A = <30%; B = 30-60%; C = >60%; ND = not determined.

Example C

HiBiT Assay to assess IKZF1 and GSPT1 neosubstrate degradation

[0340] Glutarimide-containing heterobifunctional degraders are known to engage additional proteins due to neomorphic interactions with the E3 ligase. Neosubstrate degradation is monitored, as degradation of these additional targets may increase efficacy or contribute to undesired off-target effects. The assessment of two well-documented neosubstrates, IKZF1 and GSPT1 , is described using the HiBiT Assay.

[0341] 5xl0 ⁴ JURKAT-IKZFl-HiBiT (Promega CS3023169) or 5xl0 ⁴ HEK-293-

GSPTl-HiBiT (Promega CS3O231O5) were seeded in a volume of 90 pL into wells of white, flat opaque bottom 96-well plates and incubated for 1 hour (IKFZ1) or overnight (GSPT1) at 37 °C and 5% CO2. RPMI-1640 + 10% FBS + 1% Pen/Strep + 200pg/mL Hygromycin (IKZF1) or DMEM + 10% FBS + 1% Pen/Strep + 200pg/mL Hygromycin (GSPT1) constituted complete medium. One hour before compound treatment, 90 pL of a 1:50 dilution of lOOx Vivazine substrate (Promega N2581) in media was added to each well. Cells were incubated for 1 hour at 37 °C and 5% CO2, and baseline luciferase levels were recorded using a SpectraMax M5e multi-mode microplate reader. Cells were then treated with compounds in a 10-point half-log serial dilution format starting with a high dose of 10 pM using Pico8, with 0.1% DMSO normalization across all doses. 0.1% DMSO was included as a control. Total volume per well was 180 uL. Cells were incubated for 16 hours (IKZF1) or 24 hours (GSPT1) at 37 °C and 5% CO2. After the indicated incubation time, luciferase levels were read using SpectraMax M5e multi-mode microplate reader following the Cell-Titer Glo-luminescence protocol after each period of incubation. Data were corrected for background signal and initial seeding levels, and remaining IKZF1 or GSPT1 protein was expressed as % relative to DMSO. DC50 and D _max were determined using GraphPad Prizm Software (v9.1).

[0342] Table 3 summarizes DC50 and Dmax for both IKZF 1 ( 16 h) and GSPT1 (24 h) protein degradation.

Table 3

For GSPT1 and IKZF1 HiBiT protein degradation assay DC50: A = <100 nM; B = 100-500 nM; C = >500 nM; D _max : A = >70%; B = 40-70%; C = <40%.

[0343] Furthermore, although the foregoing has been described in some detail by way of illustrations and examples for purposes of clarity and understanding, it will be understood by those of skill in the art that numerous and various modifications can be made without departing from the spirit of the present disclosure. Therefore, it should be clearly understood that the forms disclosed herein are illustrative only and are not intended to limit the scope of the present disclosure, but rather to also cover all modification and alternatives coming with the true scope and spirit of the disclosure.