DYRK / CLK protacs and uses thereof

Abstract

The present invention relates to bifunctional compounds, which find utility to degrade and (inhibit) one or more of the following kinases: DYRK1A, DYRK1B, DYRK2, DYRK3, DYRK4, CLK1, CLK2, CLK3, CLK4, CDK7, CDK8 / 19, PI3K, PDGFrA / B, mTOR, HIPKs, and / or CMGC kinases leading to inhibition of WNT signaling. In particular, the present invention is directed to compounds, which contain on one end an E3 ubiquitin ligase binding moiety which binds to an E3 ubiquitin ligase and on the other end a moiety which binds one or more of the following kinases: DYRK1A, DYRK1B, DYRK2, DYRK3, DYRK4, CLK1, CLK2, CLK3, CLK4, CDK7, CDK8 / 19, PI3K, PDGFrA / B, mTOR, HIPKs, and / or CMGC kinases leading to inhibition of WNT signaling, such that the one or more kinases is placed in proximity to the ubiquitin ligase to effect degradation (and inhibition) of the one or more kinases. The bifunctional compounds serve as therapeutics for the treatment of Alzheimer's disease, down syndrome, diabetes, an autoimmune disease, an inflammatory disorder (e.g., airway inflammation, osteoarthritis (e.g., knee related osteoarthritis)), cancer (e.g., glioblastoma, prostate cancer, metastatic breast cancer, metastatic lung cancer, multiple myeloma, secondary metastatic tumors of the brain, colorectal cancer, acute myeloid leukemia, myelodysplastic syndrome), a viral infection (e.g., SARS-CoV-2 infection (e.g., COVID-19)), and other diseases.

Description

[0001] DYRK / CLK PROTACS AND USES THEREOF

[0002] CROSS-REFERENCE TO RELATED APPLICATION

[0003] The present application claims priority to U.S. Provisional Application No. 63 / 543,343, filed October 10, 2023, which is incorporated herein by reference in its entirety.

[0004] FIELD OF THE INVENTION

[0005] The present invention relates to bifunctional compounds, which find utility to degrade and (inhibit) one or more of the following kinases: DYRK1A, DYRK1B, DYRK2, DYRK3, DYRK4, CLK1, CLK2, CLK3, CLK4, CDK7, CDK8 / 19, PI3K, PDGFrA / B, mTOR, homeodomain-interacting kinases (HIPKs), and / or CMGC kinases leading to inhibition of WNT signaling. In particular, the present invention is directed to compounds, which contain on one end an E3 ubiquitin ligase binding moiety which binds to an E3 ubiquitin ligase and on the other end a moiety which binds one or more of the following kinases: DYRK1 A, DYRK1B, DYRK2, DYRK3, DYRK4, CLK1, CLK2, CLK3, CLK4, CDK7, CDK8 / 19, PI3K, PDGFrA / B, mTOR, HIPKs, and / or CMGC kinases leading to inhibition of WNT signaling, such that the one or more kinases is placed in proximity to the ubiquitin ligase to effect degradation (and inhibition) of the one or more kinases. The bifunctional compounds serve as therapeutics for the treatment of Alzheimer’s disease, down syndrome, diabetes, an autoimmune disease, an inflammatory disorder (e.g., airway inflammation, osteoarthritis (e.g., knee related osteoarthritis), cancer (e.g., glioblastoma, prostate cancer, metastatic breast cancer, metastatic lung cancer, multiple myeloma, secondary metastatic tumors of the brain, colorectal cancer), a viral infection (e.g., SARS-CoV-2 infection (e.g., COVID-19)), diabetes, acute myeloid leukemia, myelodysplastic syndrome, and other diseases.

[0006] INTRODUCTION

[0007] It’s estimated that 2 million new cases of cancer will be shortly diagnosed per year with over 600,000 deaths in the US per annum. DYRK and CLK kinase inhibition affords opportunities across a spectrum of malignancies. Moreover, small molecule inhibition of DYRK and CLK kinase may play a role in mitigating disease progression of autoimmune disease and inflammatory disorders, exemplified by osteo-arthritis. DYRK1A has been revealed to play a key role in dementia and down syndrome pathogenesis. With > 40 million patients suffering, dementia is currently a leading unmet medical need and costly burden on public health. Seventy percent of these cases have been attributed to Alzheimer’s disease (AD), a neurodegenerative pathology whose most evident symptom is a progressive decline in cognitive functions. The underlying treatment of learning and / or memory disorders is a huge and significantly unmet medical need and also includes learning and memory repair after incidents of stroke or significant brain damage.

[0008] The present invention addresses these needs.

[0009] SUMMARY OF THE INVENTION

[0010] Dual-specificity tyrosine phosphorylation-regulated kinase 1 A (DYRK1 A) is a highly conserved protein kinase that is ubiquitously expressed in human tissues and acts as a master regulator of cell proliferation and differentiation (Soppa Becker 2015). Enhanced activity of DYRK1A results in altered brain development and cognitive deficits and is also associated with neurodegeneration in Alzheimer’s disease (Wegiel et al 2011, Lindberg & Meijer 2021, Deboever et al). Apart from its role neurological disorders, DYRK1 A dysregulation has been linked to tumor progression in certain types of cancer (Boni et al, Rammohan, Deboever). DYRK1A is also a target of interest in inflammatory diseases (Desmukh 2019, Kim 2022), diabetes (Kumar 2021) and viral infection (Kisaka et al 2020, Strine 2023). DYRK1B is a closely related protein kinase that shares 85% sequence identity with DYRK1A in the catalytic domain (Leder 1999, 2003). DYRK1B is expressed in a more limited spectrum of cell types than DYRK1A and is mainly recognized as a potential target in cancer therapy (Becker 2018).

[0011] The overactivity of DYRK1 A and DYRK1B in human diseases attracts much attention towards the development of DYRK1 inhibitors. Numerous pharmacological inhibitors of DYRK1A have been developed, and first have entered clinical trials (Lindberg et al 2023). The available DYRK1 inhibitors are classical type 1 kinase inhibitors that interfere with enzymatic activity by occupying the ATP binding site (Lindberg et al 2023). Owing to the structural similarity of the ATP binding site, they hardly discriminate DYRK1A and DYRK1B (Lindberg 2023).

[0012] Small-molecule-induced degradation of disease-causing proteins has emerged as an alternative approach of drug discovery. PROTACs (PROteolysis TArgeting Chimeras) are heterobifunctional molecules that recruit an E3 ubiquitin ligase in proximity to the target protein to promote its ubiquitination and subsequent proteasomal degradation. Due to the catalytic nature of this process, PROTACs can induce the degradation of super stoichiometric quantities of the target protein, while occupancy -based pharmacological effects require higher and sustained intracellular drug concentrations. Moreover, PROTACs eliminate not only the catalytic activity but also non-enzymatic functions of protein kinases, such as scaffolding. As a novel drug development strategy, PROTAC technology has received great attention from the pharmaceutical industry, where several companies have small-molecule degraders in preclinical and early clinical development (Nalawansha et al. 2020, Bekes et al 2022, He et al 2022).

[0013] Protein kinases have been among the early targets of small-molecule PROTAC development (Bondeson 2015). Since then, many PROTACs have been developed by combining classical kinase inhibitors with E3 ligase binding molecules (PROTAC-DB, ref. Donovan, Nemec et al 2022). Presently, protein kinases account for about half of the total PROTAC targets (He et al. 2022). However, no PROTACs have so far been reported that target DYRK1 A or DYRK1B. In fact, extensive efforts to map the degradable kinome identified DYRK1A as a kinase that was refractory to degradation by a panel of 91 chemically diverse, multitargeted degraders in seven different cell lines (Donovan et al. 2020).

[0014] While no PROTAC for DYRKs has been reported, targeted degradation of DYRK1 A has been achieved through a different mechanism. The ATP-competitive kinase inhibitor referred to as CaNDY (CDC37 association inhibitor for DYRK1 A) interferes with chaperone binding of DYRK1A, which results in the destabilization and degradation of DYRK1 A (Sonamoto et al 2015). Another kinase inhibitor with DYRK1 -degrading activity is the benzimidazole analog DYR219 (Branca et al. 2017, Velazquez et al 2019). DYR219 caused a significant reduction of DYRK1A protein levels by an unknown mechanism, an effect that likely contributes to its in vivo efficacy in a mouse model of Alzheimer’s disease (Velazquez 2019). However, long incubation times and high concentrations of CaNDY and DYR219 were required to induce the degradation of DYRK1 A, which limits their use as chemical probes.

[0015] Experiments conducted during the course of developing embodiments for the present invention resulted in the development of DYRK1 A / B degraders by combining a new ATP competitive DYRK1 inhibitor and a CRBN binding moiety. The lead compound, DYR684, induced the efficient degradation of DYRK1 A and DYRK1B in a proteasome dependent fashion. Furthermore, DYR684 reduced cellular DYRK1 activity more effectively than traditional kinase inhibitors and sensitized SH-SY5Y neuroblastoma cells to the cytotoxic effect of cisplatin. . Additional inhibiting warheads were combined with CRBN and shown to have inhibitory activity against one or more of the following kinases: DYRK1A, DYRK1B, DYRK2, DYRK3, DYRK4, CLK1, CLK2, CLK3, CLK4, CDK7, CDK8 / 19, PI3K, PDGFrA / B, mTOR, HIPKs, and / or CMGC kinases leading to inhibition of WNT signaling.

[0016] Accordingly, the present invention relates to bifunctional compounds, which find utility to degrade and (inhibit) one or more of the following kinases: DYRK1A, DYRK1B, DYRK2, DYRK3, DYRK4, CLK1, CLK2, CLK3, CLK4, CDK7, CDK8 / 19, PI3K, PDGFrA / B, mTOR, HIPKs, and / or CMGC kinases leading to inhibition of WNT signaling. In particular, the present invention is directed to compounds, which contain on one end an E3 ubiquitin ligase binding moiety which binds to an E3 ubiquitin ligase and on the other end a moiety which binds one or more of the following kinases: DYRK1A, DYRK1B, DYRK2, DYRK3, DYRK4, CLK1, CLK2, CLK3, CLK4, CDK7, CDK8 / 19, PI3K, PDGFrA / B, mTOR, HIPKs, and / or CMGC kinases leading to inhibition of WNT signaling, such that the one or more kinases is placed in proximity to the ubiquitin ligase to effect degradation (and inhibition) of the one or more kinases. The bifunctional compounds serve as therapeutics for the treatment of Alzheimer’s disease, down syndrome, diabetes, an autoimmune disease, an inflammatory disorder (e.g., airway inflammation, osteoarthritis (e.g., knee related osteoarthritis)), cancer (e.g., glioblastoma, prostate cancer, metastatic breast cancer, metastatic lung cancer, multiple myeloma, secondary metastatic tumors of the brain, colorectal cancer, acute myeloid leukemia, myelodysplastic syndrome), a viral infection (e.g., SARS-CoV-2 infection (e.g., COVID-19)), and other diseases.

[0017] The present inventors have identified kinase targets which are capable of being degraded by PROTACs in particular the targets DYRK1A, DYRK1B, DYRK2, DYRK3, DYRK4, CLK1, CLK2, CLK3, CLK4, CDK7, CDK8 / 19, PI3K, PDGFrA / B, mTOR, HIPKs, and / or CMGC kinases leading to inhibition of WNT signaling signaling.

[0018] Indeed, experiments conducted during the course of developing embodiments for the present invention designed, synthesized and biologically evaluated various PROTACs compounds capable of inhibiting one or more of DYRK1 A, DYRK1B, DYRK2, DYRK3, DYRK4, CLK1, CLK2, CLK3, CLK4, CDK7, CDK8 / 19, PI3K, PDGFrA / B, mTOR, HIPKs, and / or CMGC kinases leading to inhibition of WNT signaling.

[0019] The DYRK / CLK inhibitors and degraders described herein can also be considered as potential therapeutics for the treatment of developmental diseases such as Down syndrome, and neurodegenerative diseases such as Parkinson’s disease, and Huntington’s disease. Moreover, the DYRK / CLK PROTACs compounds of the present invention have been also implicated as potential therapeutics for the treatment of glioblastomas and further potential utility is highlighted in the oncology arena (see, e.g., lonescu et al., Mini-reviews in Medicinal Chemistry, 2012, 12, 1315-1329). Moreover, the DYRK / CLK PROTACs compounds of the present invention have been also implicated as potential therapeutics for the treatment of viral infections (e.g., SARS-CoV-2 infection (e.g., COVID-19), and other diseases).

[0020] These novel DYRK / CLK PROTAC compounds may also have utility as general cognitive enhancers, given the published findings that DYRK1A can phosphorylate sirtuin 1, a key regulator of learning and memory (see, e.g., Michan et al., J. Neurosci. 2010, 30(29), 9695-9707; Guo et al., J Biol. Chem. 2010, 285 (17), 13223-13232). Moreover, the effectiveness of small molecule inhibition of DYRK1A in mitigating both insoluble tau aggregates and amyloid plaques has been demonstrated (see, e.g., Branca et al., Aging Cell, 2017, 16(5), 1146-1154). The mechanistic rational for this was detailed previously (Smith et al., ACS Chem. Neuroscience, 2012, 3(11), 857-872). These novel DYRK / CLK Protac compounds may also have further utility as results identify DYRK1 A as a physiologically relevant regulator of Tregcell differentiation and suggest a broader role for other DYRK family members in immune homeostasis. As such, new roles may be found in autoimmune diseases such as inflammatory bowel disease and type 1 diabetes (see, e.g., Khor B, et al., eLife 2015;4:e05920).

[0021] These novel DYRK / CLK PROTACS may have utility for a variety of malignancies described herein underpinned by the roles that DYRKs and CLK play in mRNA splicing. This is exemplified for PROTACS derived from cirtuvivint currently in Phase lb clinical trials (Tam BY et al., The CLK inhibitor SM08502 induces anti-tumor activity and reduces Wnt pathway gene expression in gastrointestinal cancer models. Cancer Lett. 2020;473 : 186- 197.) Moreover, PROTACS employing a lorecivivint-like warhead would have utility for osteoporosis of the knee or shoulder (Haiyang K et. al. Effect of lorecivivint on osteoarthritis: A systematic review and meta-analysis. Heliyon. 2023; 9(8): el 8682. doi: 10.1016 / j.heliyon.2023.el 8682). Abemaciclib is a dual CDK4 / 6 inhibitor toward breast cancer and other malignancies but is also a pan-DYRK / CLK inhibitor. As such, abemaciclib PROTACS herein may have significant utility in cancer through degradation of CDKs and through degradation of the DYRK and CLK families (Johnston SRD et al., Abemaciclib Combined With Endocrine Therapy for the Adjuvant Treatment of HR+, HER2-, Node- Positive, High-Risk, Early Breast Cancer (monarchE). J Clin Oncol. 2020;38(34):3987- 3998.)

[0022] Accordingly, the present invention relates to bifunctional compounds, which find utility to degrade and (inhibit) one or more of the following kinases: DYRK1A, DYRK1B, DYRK2, DYRK3, DYRK4, CLK1, CLK2, CLK3, CLK4, CDK7, CDK8 / 19, PI3K, PDGFrA / B, mTOR, HIPKs, and / or CMGC kinases leading to inhibition of WNT signaling (and CDKs). In particular, the present invention is directed to compounds, which contain on one end an E3 ubiquitin ligase binding moiety which binds to an E3 ubiquitin ligase and on the other end a moiety which binds one or more of the following kinases: DYRK1A, DYRK1B, DYRK2, DYRK3, DYRK4, CLK1, CLK2, CLK3, CLK4, CDK7, CDK8 / 19, PI3K, PDGFrA / B, mTOR, HIPKs, and / or CMGC kinases leading to inhibition of WNT signaling, such that the one or more kinases is placed in proximity to the ubiquitin ligase to effect degradation (and inhibition) of the one or more kinases. The bifunctional compounds serve as therapeutics for the treatment of Alzheimer’s disease, down syndrome, diabetes, an autoimmune disease, an inflammatory disorder (e.g., airway inflammation, osteoarthritis (e.g., knee related osteoarthritis)), cancer (e.g., glioblastoma, prostate cancer, metastatic breast cancer, metastatic lung cancer, multiple myeloma, secondary metastatic tumors of the brain, colorectal cancer, acute myeloid leukemia, myelodysplastic syndrome), a viral infection (e.g., SARS-CoV-2 infection (e.g., COVID-19)), and other diseases.

[0023] The present disclosure describes compounds, including compositions comprising the same, which function to recruit endogenous proteins to an E3 ubiquitin ligase enzyme (e.g., Von Hippel-Lindau (VHL) E3 ubiquitin ligase -dioxopiperidin-3-yl)-5-fluoroisoindoline- 1,3-dione

[0024] and subsequent degradation, and methods of using the same. In particular, the present disclosure provides bifunctional or proteolysis targeting chimeric (PROTAC) compounds, which find utility as modulators of targeted ubiquitination and degradation of one or more of DYRK1 A, DYRK1B,

[0025] DYRK2, DYRK3, DYRK4, CLK1, CLK2, CLK3, CLK4, CDK7, CDK8 / 19, PI3K, PDGFrA / B, mTOR, HIPKs, and / or CMGC kinases leading to inhibition of WNT signaling.

[0026] In certain embodiments, the present invention provides compounds which function to recruit endogenous proteins (one or more of DYRK1A, DYRK1B, DYRK2, DYRK3, DYRK4, CLK1, CLK2, CLK3, CLK4, CDK7, CDK8 / 19, PI3K, PDGFrA / B, mTOR, HIPKs, and / or CMGC kinases leading to inhibition of WNT signaling) to E3 Ubiquitin Ligase for ubiquitination and degradation.

[0027] In certain embodiments, the compounds have the following general structure: Target Binding Moiety (TBM)— L— ULM , ULM— L— TBM, or TBM-ULM.

[0028] In some embodiments, the TBM is a small molecule inhibitor of one or more of DYRK1A, DYRK1B, DYRK2, DYRK3, DYRK4, CLK1, CLK2, CLK3, CLK4, CDK7, CDK8 / 19, PI3K, PDGFrA / B, mTOR, HIPKs, and / or CMGC kinases leading to inhibition of WNT signaling, ULM is an E3 ligase binding moiety (e.g., a VHL binding moiety, a cereblon binding moiety, a XIAP binding moiety, or a MDM2 binding moiety), and L is a bond or a linker moiety which links the TBM and ULM.

[0029] In certain embodiments, the small molecule inhibitor of one or more of DYRK1 A, DYRK1B, DYRK2, DYRK3, DYRK4, CLK1, CLK2, CLK3, CLK4, CDK7, CDK8 / 19, PI3K, PDGFrA / B, mTOR, HIPKs, and / or CMGC kinases leading to inhibition of WNT signaling is selected from 4-(3-(3-fluoro-4-(4-methylpiperazin-l-yl)phenyl)-2-methyl-3H-imidazo[4,5- b]pyridin-5-yl)pyridin-2-amine cyclopentyl-N,N-dimethyl-2-((5-(piperazin-l-yl)pyridin-2-yl)amino)-7H-pyrrolo[2,3- pyrazol-4-yl)isoquinolin-3-yl)-2-(4-methylpiperazin-l-yl)isonicotinamide

[0030] methyl- 1 H-pyrazol-4-y l)isoquinolin-3 -y 1) - 1 - methyl-lH-pyrazol-4-yl)isoquinolin-3-yl)piperidine-4-carboxamide ) - adamantan- 1 -yl) amino)-5 - (benzo [d] thiazol-

[0031] (benzo[d]thiazol-6-ylmethylene)-2-(((lr,3r)-3-ethoxyadamantan-l-yl)amino)-3,5-dihydro-4H- methy 1-N - (5 - (3 -(7 -phenyl- 1 H- imidazo[4,5-c]pyridin-2-yl)-lH-indazol-5-yl)pyridin-3-yl)butanamide -ethoxyphenyl)-lH-imidazo[4,5- c]pyridin-2-yl)-lH-indazol-5-yl)pyridin-3-yl)-3-methylbutanamide -(benzo[d]thiazol-6-ylmethylene)-2-((l -hydroxy-

[0032] 4-methylpentan-2-yl)amino)-3,5-dihydro-4H-imidazol-4-one methyl- lH-pyrazol-4-yl)isoquinolin-3-yl)-2-(4-methylpiperazin-l-yl)isonicotinamide -fhioro-4-(4-methylpiperazin-l- yl)phenyl)quinazolin-6-yl)pyridin-2-amine (

[0033] In certain embodiments, the small molecule inhibitor of one or more of DYRK1 A,

[0034] DYRK1B, DYRK2, DYRK3, DYRK4, CLK1, CLK2, CLK3, CLK4, CDK7, CDK8 / 19, PI3K, PDGFrA / B, mTOR, HIPKs, and / or CMGC kinases leading to inhibition of WNT signaling is

[0035]

[0036]

[0037] In some embodiments, the TBM covalently couples with the linker at any location within the TBM. In some embodiments related to the small molecule inhibitors of one or more of

[0038] DYRK1A, DYRK1B, DYRK2, DYRK3, DYRK4, CLK1, CLK2, CLK3, CLK4, CDK7,

[0039] CDK8 / 19, PI3K, PDGFrA / B, mTOR, HIPKs, and / or CMGC kinases leading to inhibition of

[0040] WNT signaling, the TBM covalently couples with the linker at the following location:,

[0041] In some embodiments related to the small molecule inhibitors of one or more of

[0042] DYRK1A, DYRK1B, DYRK2, DYRK3, DYRK4, CLK1, CLK2, CLK3, CLK4, CDK7,

[0043] CDK8 / 19, PI3K, PDGFrA / B, mTOR, HIPKs, and / or CMGC kinases leading to inhibition of

[0044] WNT signaling, the TBM covalently couples with the linker at the following “

[0045]

[0046]

[0047] In some embodiments, the ULM covalently couples with the linker at any location within the ULM. For example, in some embodiments, the ULM covalently couples with the linker at

[0048]

[0049] (2, 6-dioxopiperidin-3-yl)-5-fluoroisoindoline-l, 3-dione), oxoisoindolin-2-yl)piperidine-2, 6-dione).

[0050] As such, in certain embodiments, the description provides compounds having one of the following general structures: TBM — L — VHL binding moiety, TBM — L — Cereblon binding moiety, TBM — L — XIAP binding moiety, and TBM — L — MDM2 binding moiety.

[0051] In certain embodiments, the linker is a bond or a chemical linker that covalently couples the TBM and the ULM. In some embodiments, the linker is selected from:

[0052]

[0053] In some embodiments, TBM-L-ULM is represented by TBM- A-B -modified cereblon, wherein A is represented by ; wherein ; wherein n is 0, 1 or 2; wherein R is OH or H; wherein X is CH2 or CO; wherein each X1is either N or CH; wherein X2is CO, CH2, or NH; wherein X3is NH or 0. It will be understood that the general structures are exemplary, and the respective moieties can be arranged spatially in any desired order or configuration, e.g., ULM-L-TBM, TBM-L-ULM, TBM-ULM, ULM-TBM respectively.

[0054] In certain additional embodiments, the compounds comprise a plurality of E3 ligase binding moieties and / or a plurality of TBMs.

[0055] In certain embodiments, the description provides a bifunctional compound having a structure as described herein, a salt, a polymorph, and a prodrug thereof.

[0056] In certain embodiments, the bifunctional compound is selected from:,

[0057]

[0058]

[0059] In another aspect, the description provides compositions comprising compounds as described herein, and a pharmaceutically acceptable carrier. In certain embodiments, the compositions are therapeutic or pharmaceutical compositions comprising an effective amount of a compound as described herein and a pharmaceutically acceptable carrier. In certain embodiments, the therapeutic or pharmaceutical compositions comprise an additional biologically active agent, e.g., an agent effective for the treatment of a condition associated with aberrant expression of one or more of DYRK1 A, DYRK1B, DYRK2, DYRK3, DYRK4, CLK1 , CLK2, CLK3, CLK4, CDK7, CDK8 / 19, PI3K, PDGFrA / B, mTOR, HIPKs, and / or CMGC kinases leading to inhibition of WNT signaling (e.g., Alzheimer’s disease, down syndrome, diabetes, an autoimmune disease, an inflammatory disorder (e.g., airway inflammation, osteoarthritis (e.g., knee related osteoarthritis)), cancer (e.g., glioblastoma, prostate cancer, metastatic breast cancer, metastatic lung cancer, multiple myeloma, secondary metastatic tumors of the brain, colorectal cancer), a viral infection (e.g., SARS-CoV-2 infection (e.g., COVID- 19)), and other diseases).

[0060] In any of the aspects or embodiments described herein, the therapeutic compositions comprising compounds described herein can be in any suitable dosage form, e.g., solid, or liquid, and configured to be delivered by any suitable route, e.g., oral, parenteral, intravenous, intraperitoneal, subcutaneous, intramuscular, etc.

[0061] In another aspect, the disclosure provides methods of modulating protein ubiquitination and degradation in a subject, e.g., a cell, a tissue, mammal, or human patient, the method comprising administering an effective amount of a compound as described herein or a composition comprising an effective amount of the same to a subject, wherein the compound or composition comprising the same is effective in modulating protein ubiquitination and degradation of the protein in the subject. In certain embodiments, the protein is one or more of DYRK1A, DYRK1B, DYRK2, DYRK3, DYRK4, CLK1, CLK2, CLK3, CLK4, CDK7, CDK8 / 19, PI3K, PDGFrA / B, mTOR, HIPKs, and / or CMGC kinases leading to inhibition of WNT signaling.

[0062] In another aspect, the disclosure provides methods of modulating protein ubiquitination and degradation of one or more of DYRK1 A, DYRK1B, DYRK2, DYRK3, DYRK4, CLK1 , CLK2, CLK3, CLK4, CDK7, CDK8 / 19, PI3K, PDGFrA / B, mTOR, HIPKs, and / or CMGC kinases leading to inhibition of WNT signaling in a subject, e.g., a cell, a tissue, mammal, or human patient, the method comprising administering an effective amount of a compound as described herein or a composition comprising an effective amount of the same to a subject, wherein the compound or composition comprising the same is effective in modulating protein ubiquitination and degradation of one or more of DYRK1A, DYRK1B, DYRK2, DYRK3, DYRK4, CLK1, CLK2, CLK3, CLK4, CDK7, CDK8 / 19, PI3K, PDGFrA / B, mTOR, HIPKs, and / or CMGC kinases leading to inhibition of WNT signaling in the subject.

[0063] In another aspect, the disclosure provides methods of treating or ameliorating a symptom of a disease related to one or more of DYRK1 A, DYRK1B, DYRK2, DYRK3, DYRK4, CLK1, CLK2, CLK3, CLK4, CDK7, CDK8 / 19, PI3K, PDGFrA / B, mTOR, HIPKs, and / or CMGC kinases leading to inhibition of WNT signaling activity in a subject, e.g., a cell, a tissue, mammal, or human patient, the method comprising administering an effective amount of a compound as described herein or a composition comprising an effective amount of the same to a subject in need thereof, wherein the compound or composition comprising the same is effective in treating or ameliorating a symptom of a disease related to one or more of DYRK1A, DYRK1B, DYRK2, DYRK3, DYRK4, CLK1, CLK2, CLK3, CLK4, CDK7, CDK8 / 19, PI3K, PDGFrA / B, mTOR, HIPKs, and / or CMGC kinases leading to inhibition of WNT signaling activity in the subject. In a preferred embodiment, the subject is a human.

[0064] In another aspect, the disclosure provides methods for identifying the effects of the degradation of proteins of interest in a biological system (e.g., one or more of DYRK1 A, DYRK1B, DYRK2, DYRK3, DYRK4, CLK1, CLK2, CLK3, CLK4, CDK7, CDK8 / 19, PI3K, PDGFrA / B, mTOR, HIPKs, and / or CMGC kinases leading to inhibition of WNT signaling) using compounds according to the present invention.

[0065] In certain embodiments, the present invention provides methods for administering an effective amount of a compound as described herein or a composition comprising an effective amount of the same (alone or with additional agents (e.g., remdesivir)) to a subject (e.g., a human subject) (e.g., a human subject suffering from or at risk of suffering from viral infection) (a human subject suffering from or at risk of suffering from a condition related to a viral infection for purposes of treating, preventing and / or ameliorating the viral infection and / or symptoms of a viral infection (e.g., SARS-CoV-2 infection (e.g., COVID-19)). In such embodiments, administration results in suppression of pro-inflammatory cytokine activity (e.g., IL-6 activity) within the subject. In some embodiments, administration results in enhancement of NK cell activity within the subject. In some embodiments, administration results in enhancement of neutrophil activity within the subject. In some embodiments, administration results in inhibition of viral entry into the subject’s cells through inhibiting binding of the virus with heparin sulfate proteoglycan within such cells.

[0066] In such embodiments, the methods are not limited to treating, preventing and / or ameliorating a particular type or kind of viral infection or symptoms of a particular type or kind of viral infection. In some embodiments, the viral infection is a SARS-CoV-2 related viral infection (e.g., COVID-19). In some embodiments, the viral infection is any infection related to influenza, HIV, HIV-1, HIV-2, drug-resistant HIV, Junin virus, Chikungunya virus, Yellow Fever virus, Dengue virus, Pichinde virus, Lassa virus, adenovirus, Measles virus, Punta Toro virus, Respiratory Syncytial virus, Rift Valley virus, RHDV, SARS coronavirus, Tacaribe virus, and West Nile virus. In some embodiments, the viral infection is associated with any virals having Mproprotease activity and / or expression.

[0067] In some embodiments, the one or more symptoms related to viral infection includes, but is not limited to, fever, fatigue, dry cough, myalgias, dyspnea, acute respiratory distress syndrome, and pneumonia. In certain embodiments, the present invention provides methods of treating cancer in a subject in need thereof, the method comprising administering to the subject 1) a compound as described herein, and 2) at least one anticancer agent. In some embodiments, the cancer is one or more types of cancer selected from glioblastoma, prostate cancer, metastatic breast cancer, metastatic lung cancer, multiple myeloma, secondary metastatic tumors of the brain, colorectal cancer, acute myeloid leukemia, myelodysplastic syndrome. In some embodiments, the at least one anticancer agent comprises a chemotherapeutic agent, an immune checkpoint inhibitor, or radiation therapy. In some embodiments, the immune checkpoint inhibitor is one or more selected from: pembrolizumab, nivolumab, cemipilimab, atezolizumab, avelumab, durvalumab, and ipilimumab.

[0068] In another aspect, the description provides kits comprising compounds or compositions as described herein. The kit may be promoted, distributed, or sold as a unit for performing the methods of the present invention. In addition, the kits of the present invention may preferably contain instructions which describe a suitable use. Such kits can be conveniently used, e.g., in clinical settings, to treat patients.

[0069] Where applicable or not specifically disclaimed, any one of the embodiments described herein are contemplated to be able to combine with any other one or more embodiments, even though the embodiments are described under different aspects of the invention. As such, the preceding general areas of utility are given by way of example only and are not intended to be limiting on the scope of the present disclosure and appended claims. Additional objects and advantages associated with the compositions, methods, and processes of the present invention will be appreciated by one of ordinary skill in the art in light of the instant claims, description, and examples. For example, the various aspects and embodiments of the invention may be utilized in numerous combinations, all of which are expressly contemplated by the present description. These additional advantages objects and embodiments are expressly included within the scope of the present invention. The publications and other materials used herein to illuminate the background of the invention, and in particular cases, to provide additional details respecting the practice, are incorporated by reference.

[0070] DEFINITIONS

[0071] The following terms are used to describe the present invention. In instances where a term is not specifically defined herein, that term is given an art-recognized meaning by those of ordinary skill applying that term in context to its use in describing the present invention. The articles “a” and “an” as used herein and in the appended claims are used herein to refer to one or to more than one (i.e., to at least one) of the grammatical object of the article unless the context clearly indicates otherwise. By way of example, “an element” means one element or more than one element.

[0072] The phrase “and / or,” as used herein in the specification and in the claims, should be understood to mean “either or both” of the elements so conjoined, i.e., elements that are conjunctively present in some cases and disjunctively present in other cases. Multiple elements listed with “and / or” should be construed in the same fashion, i.e., “one or more” of the elements so conjoined. Other elements may optionally be present other than the elements specifically identified by the “and / or” clause, whether related or unrelated to those elements specifically identified. Thus, as a non-limiting example, a reference to “A and / or B”, when used in conjunction with open-ended language such as “comprising” can refer, in one embodiment, to A only (optionally including elements other than B); in another embodiment, to B only (optionally including elements other than A); in yet another embodiment, to both A and B (optionally including other elements); etc.

[0073] As used herein in the specification and in the claims, “or” should be understood to have the same meaning as “and / or” as defined above. For example, when separating items in a list, “or” or “and / or” shall be interpreted as being inclusive, i.e., the inclusion of at least one, but also including more than one, of a number or list of elements, and, optionally, additional unlisted items. Only terms clearly indicated to the contrary, such as “only one of’ or “exactly one of,” or, when used in the claims, “consisting of,” will refer to the inclusion of exactly one element of a number or list of elements. In general, the term “or” as used herein shall only be interpreted as indicating exclusive alternatives (i.e., “one or the other but not both”) when preceded by terms of exclusivity, such as “either,” “one of,” “only one of,” or “exactly one of.”

[0074] The term “about” and the like, as used herein, in association with numeric values or ranges, reflects the fact that there is a certain level of variation that is recognized and tolerated in the art due to practical and / or theoretical limitations. For example, minor variation is tolerated due to inherent variances in the manner in which certain devices operate and / or measurements are taken. In accordance with the above, the phrase “about” is normally used to encompass values within the standard deviation or standard error.

[0075] In the claims, as well as in the specification above, all transitional phrases such as “comprising,” “including,” “carrying,” “having,” “containing,” “involving,” “holding,” “composed of,” and the like are to be understood to be open-ended, i.e., to mean including but not limited to. Only the transitional phrases “consisting of’ and “consisting essentially of’ shall be closed or semi-closed transitional phrases, respectively, as set forth in the United States Patent Office Manual of Patent Examining Procedures, Section 2111.03.

[0076] As used herein in the specification and in the claims, the phrase “at least one,” in reference to a list of one or more elements, should be understood to mean at least one element selected from anyone or more of the elements in the list of elements, but not necessarily including at least one of each and every element specifically listed within the list of elements and not excluding any combinations of elements in the list of elements. This definition also allows that elements may optionally be present other than the elements specifically identified within the list of elements to which the phrase “at least one” refers, whether related or unrelated to those elements specifically identified. Thus, as a nonlimiting example, “at least one of A and B” (or, equivalently, “at least one of A or B,” or, equivalently “at least one of A and / or B”) can refer, in one embodiment, to at least one, optionally including more than one, A, with no B present (and optionally including elements other than B); in another embodiment, to at least one, optionally including more than one, B, with no A present (and optionally including elements other than A); in yet another embodiment, to at least one, optionally including more than one, A, and at least one, optionally including more than one, B (and optionally including other elements); etc.

[0077] It should also be understood that, in certain methods described herein that include more than one step or act, the order of the steps or acts of the method is not necessarily limited to the order in which the steps or acts of the method are recited unless the context indicates otherwise.

[0078] The terms “co-administration” and “co-administering” or “combination therapy” can refer to both concurrent administration (administration of two or more therapeutic agents at the same time) and time varied administration (administration of one or more therapeutic agents at a time different from that of the administration of an additional therapeutic agent or agents), as long as the therapeutic agents are present in the patient to some extent, preferably at effective amounts, at the same time. In certain preferred aspects, one or more of the present compounds described herein, are co-administered in combination with at least one additional bioactive agent, especially including an anticancer agent. In particularly preferred aspects, the co-administration of compounds results in synergistic activity and / or therapy, including anticancer activity.

[0079] The term “effective” can mean, but is in no way limited to, that amount / dose of the active pharmaceutical ingredient, which, when used in the context of its intended use, effectuates or is sufficient to prevent, inhibit the occurrence, ameliorate, delay or treat (alleviate a symptom to some extent, preferably all) the symptoms of a condition, disorder or disease state in a subject in need of such treatment or receiving such treatment. The term effective subsumes all other effective amount or effective concentration terms, e.g., “effective amount / dose,” “pharmaceutically effective amount / dose” or “therapeutically effective amount / dose,” which are otherwise described or used in the present application.

[0080] The effective amount depends on the type and severity of disease, the composition used, the route of administration, the type of mammal being treated, the physical characteristics of the specific mammal under consideration, concurrent medication, and other factors which those skilled in the medical arts will recognize. The exact amount can be ascertainable by one skilled in the art using known techniques (see, e.g., Lieberman, Pharmaceutical Dosage Forms (vols. 1- 3, 1992); Lloyd, The Art, Science and Technology of Pharmaceutical Compounding (1999); Pickar, Dosage Calculations (1999); and Remington: The Science and Practice of Pharmacy, 20th Edition, 2003, Gennaro, Ed., Lippincott, Williams & Wilkins).

[0081] The term “pharmacological composition,” “therapeutic composition,” “therapeutic formulation” or “pharmaceutically acceptable formulation” can mean, but is in no way limited to, a composition or formulation that allows for the effective distribution of an agent provided by the invention, which is in a form suitable for administration to the physical location most suitable for their desired activity, e.g., systemic administration.

[0082] The term “pharmaceutically acceptable” or “pharmacologically acceptable” can mean, but is in no way limited to, entities and compositions that do not produce an adverse, allergic or other untoward reaction when administered to an animal, or a human, as appropriate.

[0083] The term “pharmaceutically acceptable carrier” or “pharmacologically acceptable carrier” can mean, but is in no way limited to, any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like, compatible with pharmaceutical administration. Suitable carriers are described in the most recent edition of Remington's Pharmaceutical Sciences, a standard reference text in the field, which is incorporated herein by reference. Preferred examples of such carriers or diluents include, but are not limited to, water, saline, Ringer's solutions, dextrose solution, and 5% human serum albumin. Liposomes and non-aqueous vehicles such as fixed oils may also be used. The use of such media and agents for pharmaceutically active substances is well known in the art. Except insofar as any conventional media or agent is incompatible with the active compound, use thereof in the compositions is contemplated. Supplementary active compounds can also be incorporated into the compositions.

[0084] The term “systemic administration” refers to a route of administration that is, e.g., enteral or parenteral, and results in the systemic distribution of an agent leading to systemic absorption or accumulation of drugs in the blood stream followed by distribution throughout the entire body. Suitable forms, in part, depend upon the use or the route of entry, for example oral, transdermal, or by injection. Such forms should not prevent the composition or formulation from reaching a target cell (i.e., a cell to which the negatively charged polymer is desired to be delivered to). For example, pharmacological compositions injected into the blood stream should be soluble. Other factors are known in the art, and include considerations such as toxicity and forms which prevent the composition or formulation from exerting its effect. Administration routes which lead to systemic absorption include, without limitations: intravenous, subcutaneous, intraperitoneal, inhalation, oral, intrapulmonary and intramuscular. The rate of entry of a drug into the circulation has been shown to be a function of molecular weight or size. The use of a liposome or other drug carrier comprising the compounds of the instant invention can potentially localize the drug, for example, in certain tissue types, such as the tissues of the reticular endothelial system (RES). A liposome formulation which can facilitate the association of drug with the surface of cells, such as, lymphocytes and macrophages is also useful.

[0085] The term “local administration” refers to a route of administration in which the agent is delivered to a site that is apposite or proximal, e.g., within about 10 cm, to the site of the lesion or disease.

[0086] The term “compound”, as used herein, unless otherwise indicated, refers to any specific chemical compound disclosed herein and includes tautomers, regioisomers, geometric isomers, and where applicable, stereoisomers, including optical isomers (enantiomers) and other stereoisomers (diastereomers) thereof, as well as pharmaceutically acceptable salts and derivatives (including prodrug forms) thereof where applicable, in context. Within its use in context, the term compound generally refers to a single compound, but also may include other compounds such as stereoisomers, regioisomers and / or optical isomers (including racemic mixtures) as well as specific enantiomers or enantiomerically enriched mixtures of disclosed compounds. The term also refers, in context to prodrug forms of compounds which have been modified to facilitate the administration and delivery of compounds to a site of activity. It is noted that in describing the present compounds, numerous substituents and variables associated with same, among others, are described.

[0087] It is understood by those of ordinary skill that molecules which are described herein are stable compounds as generally described hereunder. When the bond “=” is shown, both a double bond and single bond are represented within the context of the compound shown.

[0088] As used herein, “derivatives” can mean compositions formed from the native compounds either directly, by modification, or by partial substitution. As used herein, “analogs” can mean compositions that have a structure similar to, but not identical to, the native compound. The term “Ubiquitin Ligase” refers to a family of proteins that facilitate the transfer of ubiquitin to a specific substrate protein, targeting the substrate protein for degradation. For example, Von HippeLLindau E3 Ubiquitin Ligase or VCB E3 Ubiquitin Ligase is protein that alone or in combination with an E2 ubiquitin-conjugating enzyme causes the attachment of ubiquitin to a lysine on a target protein, and subsequently targets the specific protein substrates for degradation by the proteasome. Thus, E3 ubiquitin ligase alone or in complex with an E2 ubiquitin conjugating enzyme is responsible for the transfer of ubiquitin to targeted proteins. In general, the ubiquitin ligase is involved in polyubiquitination such that a second ubiquitin is attached to the first; a third is attached to the second, and so forth. Polyubiquitination marks proteins for degradation by the proteasome. However, there are some ubiquitination events that are limited to mono-ubiquitination, in which only a single ubiquitin is added by the ubiquitin ligase to a substrate molecule. Mono-ubiquitinated proteins are not targeted to the proteasome for degradation, but may instead be altered in their cellular location or function, for example, via binding other proteins that have domains capable of binding ubiquitin. Further complicating matters, different lysines on ubiquitin can be targeted by an E3 to make chains. The most common lysine is Lys48 on the ubiquitin chain. This is the lysine used to make polyubiquitin, which is recognized by the proteasome.

[0089] The term “subject” is used throughout the specification to describe a cell, tissue, or animal, preferably a human or a domesticated animal, to whom treatment, including prophylactic treatment, with the compositions according to the present invention is provided. For treatment of those infections, conditions or disease states which are specific for a specific animal such as a human patient, the term patient refers to that specific animal, including a domesticated animal such as a dog or cat or a farm animal such as a horse, cow, sheep, etc. In general, in the present invention, the term patient refers to a human patient unless otherwise stated or implied from the context of the use of the term.

[0090] DETAILED DESCRIPTION OF THE INVENTION

[0091] The following is a detailed description provided to aid those skilled in the art in practicing the present invention. Those of ordinary skill in the art may make modifications and variations in the embodiments described herein without departing from the spirit or scope of the present disclosure. All publications, patent applications, patents, figures and other references mentioned herein are expressly incorporated by reference in their entirety.

[0092] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The terminology used in the description is for describing particular embodiments only and is not intended to be limiting of the invention.

[0093] Where a range of values is provided, it is understood that each intervening value, to the tenth of the unit of the lower limit unless the context clearly dictates otherwise (such as in the case of a group containing a number of carbon atoms in which case each carbon atom number falling within the range is provided), between the upper and lower limit of that range and any other stated or intervening value in that stated range is encompassed within the invention. The upper and lower limits of these smaller ranges may independently be included in the smaller ranges is also encompassed within the invention, subject to any specifically excluded limit in the stated range. Where the stated range includes one or both of the limits, ranges excluding either both of those included limits are also included in the invention.

[0094] Alternative to enzyme inhibition, small-molecule-induced protein degradation has emerged as a promising pharmacological modality for inactivating disease-relevant protein kinases. DYRK1 A and DYRK1B are closely related protein kinases that are involved in pathological processes such as neurodegeneration, cancer development and adaptive immune homeostasis.

[0095] Experiments conducted during the development of embodiments for the present invention resulted in the development of DYRK1 proteolysis targeting chimeras (PROTACs) that combine a new ATP-competitive DYRK1 inhibitor with ligands for the E3 ubiquitin ligase component Cereblon (CRBN) to induce ubiquitinylation and subsequent proteasomal degradation of DYRK1 A and DYRK1B. A lead compound (DYR684) promoted fast, efficient, potent and selective degradation of overexpressed and endogenous DYRK1A in cell-based assays (UPS- and unfoldase dependent manner). Although DYR684 was also active against DYRK1B, it was observed that an enzymatically inactive splicing variant of DYRK1B (p65) was resistant to degradation. Compared to competitive kinase inhibition, targeted degradation of DYRK1 by DYR684 provided improved suppression of downstream signaling. Moreover, DYR684 sensitized SH-SY5Y neuroblastoma cells to the cytotoxic effects of the anticancer drug cisplatin. Collectively, our results identify DYRK1 A and DYRK1B as viable targets for PROTAC-mediated degradation and qualify DYR684 as a suitable chemical probe for functional studies of the catalytically active DYRK1 A and DYRK1B variants. Additional inhibiting warheads were combined with CRBN and shown to have inhibitory activity against one or more of the following kinases: DYRK1A, DYRK1B, DYRK2, DYRK3, DYRK4, CLK1, CLK2, CLK3, CLK4, CDK7, CDK8 / 19, PI3K, PDGFrA / B, mTOR, HIPKs, and / or CMGC kinases leading to inhibition of WNT signaling. Accordingly, the present invention relates to bifunctional compounds, which find utility to degrade and (inhibit) one or more of the following kinases: DYRK1A, DYRK1B, DYRK2, DYRK3, DYRK4, CLK1, CLK2, CLK3, CLK4, CDK7, CDK8 / 19, PI3K, PDGFrA / B, mTOR, HIPKs, and / or CMGC kinases leading to inhibition of WNT signaling. In particular, the present invention is directed to compounds, which contain on one end an E3 ubiquitin ligase binding moiety which binds to an E3 ubiquitin ligase and on the other end a moiety which binds one or more of the following kinases: DYRK1A, DYRK1B, DYRK2, DYRK3, DYRK4, CLK1, CLK2, CLK3, CLK4, CDK7, CDK8 / 19, PI3K, PDGFrA / B, mTOR, HIPKs, and / or CMGC kinases leading to inhibition of WNT signaling, such that the one or more kinases is placed in proximity to the ubiquitin ligase to effect degradation (and inhibition) of the one or more kinases. The bifunctional compounds serve as therapeutics for the treatment of Alzheimer’s disease, down syndrome, diabetes, an autoimmune disease, an inflammatory disorder (e.g., airway inflammation, osteoarthritis (e.g., knee related osteoarthritis)), cancer (e.g., glioblastoma, prostate cancer, metastatic breast cancer, metastatic lung cancer, multiple myeloma, secondary metastatic tumors of the brain, colorectal cancer, acute myeloid leukemia, myelodysplastic syndrome), a viral infection (e.g., SARS-CoV-2 infection (e.g., COVID-19)), and other diseases.

[0096] In one aspect, the present invention provides compounds useful for regulating activity of one or more of the following kinases: DYRK1A, DYRK1B, DYRK2, DYRK3, DYRK4, CLK1, CLK2, CLK3, CLK4, CDK7, CDK8 / 19, PI3K, PDGFrA / B, mTOR, HIPKs, and / or CMGC kinases leading to inhibition of WNT signaling. The composition comprises a ubiquitin pathway protein binding moiety (preferably for an E3 ubiquitin ligase, alone or in complex with an E2 ubiquitin conjugating enzyme which is responsible for the transfer of ubiquitin to targeted proteins) according to a defined chemical structure and a protein targeting moiety which are linked or coupled together, preferably through a linker, wherein the ubiquitin pathway protein binding moiety recognizes an ubiquitin pathway protein and the targeting moiety recognizes one or more of the noted kinases.

[0097] In certain embodiments, the present invention provides compounds which function to recruit endogenous proteins (one or more of DYRK1A, DYRK1B, DYRK2, DYRK3, DYRK4, CLK1, CLK2, CLK3, CLK4, CDK7, CDK8 / 19, PI3K, PDGFrA / B, mTOR, HIPKs, and / or CMGC kinases leading to inhibition of WNT signaling) to E3 Ubiquitin Ligase for ubiquitination and degradation.

[0098] In certain embodiments, the compounds have the following general structure: Target Binding Moiety (TBM)— L— ULM , ULM— L— TBM, or TBM-ULM. In some embodiments, the TBM is a small molecule inhibitor of one or more of DYRK1A, DYRK1B, DYRK2, DYRK3, DYRK4, CLK1, CLK2, CLK3, CLK4, CDK7, CDK8 / 19, PI3K, PDGFrA / B, mTOR, HIPKs, and / or CMGC kinases leading to inhibition of WNT signaling, ULM is an E3 ligase binding moiety (e.g., a VHL binding moiety, a cereblon binding moiety, a XIAP binding moiety, or a MDM2 binding moiety), and L is a bond or a linker moiety which links the TBM and ULM.

[0099] In certain embodiments, the small molecule inhibitor of one or more of DYRK1 A,

[0100] DYRK1B, DYRK2, DYRK3, DYRK4, CLK1, CLK2, CLK3, CLK4, CDK7, CDK8 / 19, PI3K,

[0101] PDGFrA / B, mTOR, HIPKs, and / or CMGC kinases leading to inhibition of WNT signaling is selected from 4-(3-(3-fluoro-4-(4-methylpiperazin-l-yl)phenyl)-2-methyl-3H-imidazo[4,5- cyclopentyl-N,N-dimethyl-2-((5-(piperazin-l-yl)pyridin-2-yl)amino)-7H-pyrrolo[2,3- methyl- 1H- pyrazol-4-yl)isoquinolin-3-yl)-2-(4-methylpiperazin-l-yl)isonicotinamide methyl- 1 H-pyrazol-4-y l)isoquinolin-3 -y 1) - 1 - methyl- lH-pyrazol-4-yl)isoquinolin-3-yl)piperidine-4-carboxamide ) - adamantan- 1 -yl) amino)-5 - (benzo [d] thiazol- (benzo[d]thiazol-6-ylmethylene)-2-(((lr,3r)-3-ethoxyadamantan-l-yl)amino)-3,5-dihydro-4H- methy 1-N - (5 - (3 -(7 -phenyl- 1 H- imidazo[4,5-c]pyridin-2-yl)-lH-indazol-5-yl)pyridin-3-yl)butanamide -ethoxyphenyl)-lH-imidazo[4,5- c]pyridin-2-yl)-lH-indazol-5-yl)pyridin-3-yl)-3-methylbutanamide -(benzo[d]thiazol-6-ylmethylene)-2-((l-hydroxy-

[0102] 4-methylpentan-2-yl)amino)-3,5-dihydro-4H-imidazol-4-one methyl-lH-pyrazol-4-yl)isoquinolin-3-yl)-2-(4-methylpiperazin-l-yl)isonicotinamide -fluoro-4-(4-methylpiperazin-l- yl)phenyl)quinazolin-6-yl)pyridin-2-amine

[0103] In certain embodiments, the small molecule inhibitor of one or more of DYRK1 A,

[0104] DYRK1B, DYRK2, DYRK3, DYRK4, CLK1, CLK2, CLK3, CLK4, CDK7, CDK8 / 19, PI3K,

[0105] PDGFrA / B, mTOR, HIPKs, and / or CMGC kinases leading to inhibition of WNT signaling is selected from

[0106]

[0107] In some embodiments, the TBM covalently couples with the linker at any location within the TBM.

[0108] In some embodiments related to the small molecule inhibitors of one or more of

[0109] DYRK1A, DYRK1B, DYRK2, DYRK3, DYRK4, CLK1, CLK2, CLK3, CLK4, CDK7, CDK8 / 19, PI3K, PDGFrA / B, mTOR, HIPKs, and / or CMGC kinases leading to inhibition of WNT signaling, the TBM covalently couples with the linker at the following “ ” location:,

[0110]

[0111] In some embodiments related to the small molecule inhibitors of one or more of

[0112] DYRK1A, DYRK1B, DYRK2, DYRK3, DYRK4, CLK1, CLK2, CLK3, CLK4, CDK7, CDK8 / 19, PI3K, PDGFrA / B, mTOR, HIPKs, and / or CMGC kinases leading to inhibition of

[0113] WNT signaling, the TBM covalently couples with the linker at the following “

[0114]

[0115] In some embodiments, the ULM covalently couples with the linker at any location within the ULM. For example, in some embodiments, the ULM covalently couples with the linker at the following

[0116]

[0117] 0 (cereblon), 0 (2-(2,6-dioxopiperidin-3-yl)-5-

[0118] fluoroisoindoline-1, 3-dione), -oxoisoindolin-2-yl)piperidine-

[0119] 2, 6-dione).

[0120] As such, in certain embodiments, the description provides compounds having one of the following general structures: TBM — L — VHL binding moiety, TBM — L — Cereblon binding moiety, TBM — L — XIAP binding moiety, and TBM — L — MDM2 binding moiety. In certain embodiments, the linker is a bond or a chemical linker that covalently couples the TBM and the ULM.

[0121]

[0122] In some embodiments, TBM-L-ULM is represented by TBM- A-B -modified cereblon,

[0123] . Mn>

[0124] TBM- § - XvX wherein A is represented by or ; wherein wherein modified-

[0125] ; wherein n is 0, 1 or 2; wherein R is OH or H; wherein X is CH2 or CO; wherein each X1is either N or CH; wherein X2is CO, CH2, or NH; wherein X3is NH or 0.

[0126] It will be understood that the general structures are exemplary and the respective moieties can be arranged spatially in any desired order or configuration, e.g., ULM-L-TBM, TBM-L-ULM, TBM-ULM, ULM-TBM respectively.

[0127] In certain additional embodiments, the compounds comprise a plurality of E3 ligase binding moieties and / or a plurality of TBMs.

[0128] In certain embodiments, the description provides a bifunctional compound having a structure as described herein, a salt, a polymorph, and a prodrug thereof.

[0129]

[0130]

[0131] Without being bound by any particular theory, it is hypothesized that due at least in part to the proximity of the one or more kinases selected from DYRK1 A, DYRK1B, DYRK2, DYRK3, DYRK4, CLK1, CLK2, CLK3, CLK4, CDK7, CDK8 / 19, PI3K, PDGFrA / B, mTOR,

[0132] HIPKs, and / or CMGC kinases leading to inhibition of WNT signaling and the E3 ubiquitin ligase, the one or more kinases is ubiquitinated by the ubiquitin ligase and degraded.

[0133] The von Hippel-Lindau (VHL) tumor suppressor comprises the substrate recognition subunit / E3 ligase complex VCB, which includes elongins B and C, and a complex including Cullin-2 and Rbxl. The primary substrate of VHL is Hypoxia Inducible Factor la (HIF-la), a transcription factor that upregulates genes such as the pro- angiogenic growth factor VEGF and the red blood cell inducing cytokine erythropoietin in response to low oxygen levels. We generated the first small molecule ligands of Von Hippel Lindau (VHL) to the substrate recognition subunit of the E3 ligase, VCB, an important target in cancer, chronic anemia and ischemia, and obtained crystal structures confirming that the compound mimics the binding mode of the transcription factor HIF-la, the major substrate of VHL.

[0134] Inhibitors of Apoptosis Protein (IAPS) are guardian ubiquitin ligases that keep classic pro-apoptotic proteins in check, and regulates not only caspases and apoptosis, but also modulates inflammatory signaling and immunity, copper homeostasis, mitogenic kinase signaling, cell proliferation, as well as cell invasion and metastasis. IAPs act as a direct caspase inhibitor, and directly bind to the active site pocket of CASP3 and CASP7 and obstruct substrate entry. IAPS also inactivate CASP9 by keeping it in a monomeric, inactive state. IAP acts as an E3 ubiquitin-protein ligase regulating NF-kappa-B signaling and the target proteins for its E3 ubiquitin-protein ligase activity include: RIPK1, CASP3, CASP7, CASP8, CASP9, MAP3K2 / MEKK2, DIABLO / SMAC, AIFM1, CCS and BIRC5 / survivin. Ubiquitination of CCS leads to enhancement of its chaperone activity toward its physiologic target, SOD1, rather than proteasomal degradation. Ubiquitination of MAP3K2 / MEKK2 and AIFM1 does not lead to proteasomal degradation. IAP plays a role in copper homeostasis by ubiquitinating C0MMD1 and promoting its proteasomal degradation and can also function as E3 ubiquitin-protein ligase of the NEDD8 conjugation pathway, targeting effector caspases for neddylation and inactivation. IAP regulates the BMP signaling pathway and the SMAD and MAP3K7 / TAK1 dependent pathways leading to NF-kappa-B and JNK activation.

[0135] IAPs are an important regulator of innate immune signaling via regulation of Nodlike receptors (NLRs), and protects cells from spontaneous formation of the ripoptosome, a large multi-protein complex that has the capability to kill cancer cells in a caspase-dependent and caspase-independent manner. Suppresses ripoptosome formation by ubiquitinating RIPK1 and CASP8. Acts as a positive regulator of Wnt signaling and ubiquitinates TLE1, TLE2, TLE3, TLE4 and AES. Ubiquitination of TLE3 results in inhibition of its interaction with TCF7L2 / TCF4 thereby allowing efficient recruitment and binding of the transcriptional coactivator beta-catenin to TCF7L2 / TCF4 that is required to initiate a Wnt-specific transcriptional program. Inhibitors of the IAP, which are useful in making compounds as described herein, are known in the art.

[0136] Mouse double minute 2 homolog (MDM2) also known as E3 ubiquitin-protein ligase Mdm2 is a protein that in humans is encoded by the MDM2 gene. Mdm2 is an important negative regulator of the p53 tumor suppressor. Mdm2 protein functions both as an E3 ubiquitin ligase that recognizes the N-terminal trans-activation domain (TAD) of the p53 tumor suppressor and an inhibitor of p53 transcriptional activation. Inhibitors of the MDM2-p53 interaction, which are useful in making compounds as described herein, include the cis-imidazoline analog nutlin.

[0137] Cereblon is a protein that in humans is encoded by the CRBN gene. CRBN orthologs are highly conserved from plants to humans, which underscores its physiological importance. Cereblon forms an E3 ubiquitin ligase complex with damaged DNA binding protein 1 (DDB1), Cullin-4A (CUL4A), and regulator of cullins 1 (R0C1). This complex ubiquitinates a number of other proteins. Through a mechanism which has not been completely elucidated, cereblon ubiquitination of target proteins results in increased levels of fibroblast growth factor 8 (FGF8) and fibroblast growth factor 10 (FGF10). FGF8 in turn regulates a number of developmental processes, such as limb and auditory vesicle formation. The net result is that this ubiquitin ligase complex is important for limb outgrowth in embryos. In the absence of cereblon, DDB 1 forms a complex with DDB2 that functions as a DNA damage-binding protein.

[0138] Thalidomide, which has been approved for the treatment of a number of immunological indications, has also been approved for the treatment of certain neoplastic diseases, including multiple myeloma. In addition to multiple myeloma, thalidomide and several of its analogs are also currently under investigation for use in treating a variety of other types of cancer. While the precise mechanism of thalidomide's anti-tumor activity is still emerging, it is known to inhibit angiogenesis. Recent literature discussing the biology of the imides includes Lu et al Science 343, 305 (2014) and Krbnke et al Science 343, 301 (2014).

[0139] Significantly, thalidomide and its analogs (e.g., pomalidomide and lenalinomide), are known to bind cereblon. These agents bind to cereblon, altering the specificity of the complex to induce the ubiquitination and degradation of Ikaros (IKZF1) and Aiolos (IKZF3), transcription factors essential for multiple myeloma growth. Indeed, higher expression of cereblon has been linked to an increase in efficacy of imide drugs in the treatment of multiple myeloma. Therefore, thalidomide and its analogs are useful cereblon binding moieties for use in making compounds as described herein.

[0140] The present description includes, where applicable, the compositions comprising the pharmaceutically acceptable salts, in particular, acid or base addition salts of compounds of the present invention.

[0141] The term “pharmaceutically acceptable salt” is used throughout the specification to describe, where applicable, a salt form of one or more of the compounds described herein which are presented to increase the solubility of the compound in the gastic juices of the patient's gastrointestinal tract in order to promote dissolution and the bioavailability of the compounds. Pharmaceutically acceptable salts include those derived from pharmaceutically acceptable inorganic or organic bases and acids, where applicable. Suitable salts include those derived from alkali metals such as potassium and sodium, alkaline earth metals such as calcium, magnesium and ammonium salts, among numerous other acids and bases well known in the pharmaceutical art. Sodium and potassium salts are particularly preferred as neutralization salts of the phosphates according to the present invention.

[0142] The acids which are used to prepare the pharmaceutically acceptable acid addition salts of the aforementioned base compounds useful in this invention are those which form non-toxic acid addition salts, i.e., salts containing pharmacologically acceptable anions, such as the hydrochloride, hydrobromide, hydroiodide, nitrate, sulfate, bisulfate, phosphate, acid phosphate, acetate, lactate, citrate, acid citrate, tartrate, bitartrate, succinate, maleate, fumarate, gluconate, saccharate, benzoate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate and pamoate [i.e., l,T-methylene-bis-(2-hydroxy-3 naphthoate)]salts, among numerous others.

[0143] Pharmaceutically acceptable base addition salts may also be used to produce pharmaceutically acceptable salt forms of the compounds or derivatives according to the present invention. The chemical bases that may be used as reagents to prepare pharmaceutically acceptable base salts of the present compounds that are acidic in nature are those that form nontoxic base salts with such compounds. Such non-toxic base salts include, but are not limited to those derived from such pharmacologically acceptable cations such as alkali metal cations (eg., potassium and sodium) and alkaline earth metal cations (eg, calcium, zinc and magnesium), ammonium or water-soluble amine addition salts such as N-methylglucamine-(meglumine), and the lower alkanolammonium and other base salts of pharmaceutically acceptable organic amines, among others.

[0144] In another aspect, the description provides compositions comprising compounds as described herein, including salts thereof, and a pharmaceutically acceptable carrier. In certain embodiments, the compositions are therapeutic or pharmaceutical compositions comprising an effective amount of a compound as described herein and a pharmaceutically acceptable carrier.

[0145] The amount of compound in a pharmaceutical composition of the instant invention that may be combined with the carrier materials to produce a single dosage form will vary depending upon the host and disease treated, the particular mode of administration. Generally, an amount between 0.1 mg / kg and 1000 mg / kg body weight / day of active ingredients is administered dependent upon potency of the agent. Toxicity and therapeutic efficacy of such compounds can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, e.g., for determining the LD50 (the dose lethal to 50% of the population) and the ED50 (the dose therapeutically effective in 50% of the population). The dose ratio between toxic and therapeutic effects is the therapeutic index and it can be expressed as the ratio LD50 / ED50. Compounds that exhibit large therapeutic indices are preferred. While compounds that exhibit toxic side effects may be used, care should be taken to design a delivery system that targets such compounds to the site of affected tissue in order to minimize potential damage to uninfected cells and, thereby, reduce side effects. The data obtained from the cell culture assays and animal studies can be used in formulating a range of dosage for use in humans. The dosage of such compounds lies preferably within a range of circulating concentrations that include the ED50 with little or no toxicity. The dosage may vary within this range depending upon the dosage form employed and the route of administration utilized. For any compound used in the method of the invention, the therapeutically effective dose can be estimated initially from cell culture assays. A dose may be formulated in animal models to achieve a circulating plasma concentration range that includes the IC50 (i.e., the concentration of the test compound which achieves a half-maximal inhibition of symptoms) as determined in cell culture. Such information can be used to more accurately determine useful doses in humans. Levels in plasma may be measured, for example, by high performance liquid chromatography.

[0146] The compositions of the present invention may be formulated in a conventional manner using one or more pharmaceutically acceptable carriers and may also be administered in controlled-release formulations. Pharmaceutically acceptable carriers that may be used in these pharmaceutical compositions include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins, such as human serum albumin, buffer substances such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes, such as prolamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethylcellulose, poly acrylates, waxes, polyethylene-polyoxypropylene-block polymers, polyethylene glycol and wool fat.

[0147] The active compound is included in the pharmaceutically acceptable carrier or diluent in an amount sufficient to deliver to a patient a therapeutically effective amount for the desired indication, without causing serious toxic effects in the patient treated. A preferred dose of the active compound for all of the herein-mentioned conditions is in the range from about 10 ng / kg to 300 mg / kg, preferably 0.1 to 100 mg / kg per day, more generally 0.5 to about 25 mg per kilogram body weight of the recipient / patient per day. A typical topical dosage will range from 0.01-5% wt / wt in a suitable carrier. The compound is conveniently administered in any suitable unit dosage form, including but not limited to one containing less than Img, 1 mg to 3000 mg, preferably 5 to 500 mg of active ingredient per unit dosage form. An oral dosage of about 25-250 mg is often convenient.

[0148] The active ingredient is preferably administered to achieve peak plasma concentrations of the active compound of about 0.00001-30 mM, preferably about 0.1-30 pM. This may be achieved, for example, by the intravenous injection of a solution or formulation of the active ingredient, optionally in saline, or an aqueous medium or administered as a bolus of the active ingredient. Oral administration is also appropriate to generate effective plasma concentrations of active agent.

[0149] The concentration of active compound in the drug composition will depend on absorption, distribution, inactivation, and excretion rates of the drug as well as other factors known to those of skill in the art. It is to be noted that dosage values will also vary with the severity of the condition to be alleviated. It is to be further understood that for any particular subject, specific dosage regimens should be adjusted over time according to the individual need and the professional judgment of the person administering or supervising the administration of the compositions, and that the concentration ranges set forth herein are exemplary only and are not intended to limit the scope or practice of the claimed composition. The active ingredient may be administered at once, or may be divided into a number of smaller doses to be administered at varying intervals of time.

[0150] If administered intravenously, preferred carriers are physiological saline or phosphate buffered saline (PBS).

[0151] In one embodiment, the active compounds are prepared with carriers that will protect the compound against rapid elimination from the body, such as a controlled release formulation, including implants and microencapsulated delivery systems. Biodegradable, biocompatible polymers can be used, such as ethylene vinyl acetate, poly anhydrides, poly glycolic acid, collagen, poly orthoesters, and polylactic acid. Methods for preparation of such formulations will be apparent to those skilled in the art.

[0152] Liposomal suspensions may also be pharmaceutically acceptable carriers. These may be prepared according to methods known to those skilled in the art, for example, as described in U.S. Pat. No. 4,522,811 (which is incorporated herein by reference in its entirety). For example, liposome formulations may be prepared by dissolving appropriate lipid(s) (such as stearoyl phosphatidyl ethanolamine, stearoyl phosphatidyl choline, arachadoyl phosphatidyl choline, and cholesterol) in an inorganic solvent that is then evaporated, leaving behind a thin film of dried lipid on the surface of the container. An aqueous solution of the active compound are then introduced into the container. The container is then swirled by hand to free lipid material from the sides of the container and to disperse lipid aggregates, thereby forming the liposomal suspension.

[0153] In any of the aspects or embodiments described herein, the therapeutic compositions comprising compounds described herein can be in any suitable dosage form configured to be delivered by any suitable route. For example, the compounds can be administered by any appropriate route, for example, orally, parenterally, intravenously, intradermally, subcutaneously, or topically, including trans dermally, in liquid, cream, gel, or solid form, rectally, nasally, buccally, vaginally or via an implanted reservoir or by aerosol form.

[0154] The term “parenteral” as used herein includes subcutaneous, intravenous, intramuscular, intra-articular, intra- synovial, intrasternal, intrathecal, intrahepatic, intralesional and intracranial injection or infusion techniques. Preferably, the compositions are administered orally, intraperitoneally or intravenously.

[0155] The compounds as described herein may be administered in single or divided doses by the oral, parenteral or topical routes. Administration of the active compound may range from continuous (intravenous drip) to several oral administrations per day (for example, Q.I.D.) and may include oral, topical, parenteral, intramuscular, intravenous, sub-cutaneous, transdermal (which may include a penetration enhancement agent), buccal, sublingual and suppository administration, among other routes of administration. Enteric coated oral tablets may also be used to enhance bioavailability of the compounds from an oral route of administration. The most effective dosage form will depend upon the pharmacokinetics of the particular agent chosen as well as the severity of disease in the patient.

[0156] Administration of compounds as sprays, mists, or aerosols for intra-nasal, intra-tracheal or pulmonary administration may also be used. Compounds as described herein may be administered in immediate release, intermediate release or sustained or controlled release forms. Sustained or controlled release forms are preferably administered orally, but also in suppository and transdermal or other topical forms. Intramuscular injections in liposomal form may also be used to control or sustain the release of compound at an injection site.

[0157] Sterile injectable forms of the compositions as described herein may be aqueous or oleaginous suspension. These suspensions may be formulated according to techniques known in the art using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally- acceptable diluent or solvent, for example as a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that may be employed are water, Ringer's solution and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose, any bland fixed oil may be employed including synthetic mono- or di-glycerides. Fatty acids, such as oleic acid and its glyceride derivatives are useful in the preparation of injectables, as are natural pharmaceutically-acceptable oils, such as olive oil or castor oil, especially in their polyoxyethylated versions. These oil solutions or suspensions may also contain a long-chain alcohol diluent or dispersant, such as Ph. Hely or similar alcohol.

[0158] The pharmaceutical compositions as described herein may be orally administered in any orally acceptable dosage form including, but not limited to, capsules, tablets, aqueous suspensions or solutions. In the case of tablets for oral use, carriers which are commonly used include lactose and com starch. Lubricating agents, such as magnesium stearate, are also typically added. For oral administration in a capsule form, useful diluents include lactose and dried corn starch. When aqueous suspensions are required for oral use, the active ingredient is combined with emulsifying and suspending agents. If desired, certain sweetening, flavoring or coloring agents may also be added. Oral compositions will generally include an inert diluent or an edible carrier. They may be enclosed in gelatin capsules or compressed into tablets. For the purpose of oral therapeutic administration, the active compound or its prodrug derivative can be incorporated with excipients and used in the form of tablets, troches, or capsules. Pharmaceutically compatible binding agents, and / or adjuvant materials are included as part of the composition.

[0159] The tablets, pills, capsules, troches and the like can contain any of the following ingredients, or compounds of a similar nature: a binder such as microcrystalline cellulose, gum tragacanth or gelatin; an excipient such as starch or lactose, a dispersing agent such as alginic acid, Primogel, or corn starch; a lubricant such as magnesium stearate or Sterotes; a glidant such as colloidal silicon dioxide; a sweetening agent such as sucrose or saccharin; or a flavoring agent such as peppermint, methyl salicylate, or orange flavoring. When the dosage unit form is a capsule, it can contain, in addition to material of the above type, a liquid carrier such as a fatty oil. In addition, dosage unit forms can contain various other materials which modify the physical form of the dosage unit, for example, coatings of sugar, shellac, or enteric agents.

[0160] The active compound or pharmaceutically acceptable salt thereof can be administered as a component of an elixir, suspension, syrup, wafer, chewing gum or the like. A syrup may contain, in addition to the active compounds, sucrose as a sweetening agent and certain preservatives, dyes and colorings and flavors.

[0161] Alternatively, the pharmaceutical compositions as described herein may be administered in the form of suppositories for rectal administration. These can be prepared by mixing the agent with a suitable non-irritating excipient, which is solid at room temperature but liquid at rectal temperature and therefore will melt in the rectum to release the drug. Such materials include cocoa butter, beeswax and polyethylene glycols.

[0162] The pharmaceutical compositions of this invention may also be administered topically. Suitable topical formulations are readily prepared for each of these areas or organs. Topical application for the lower intestinal tract can be affected in a rectal suppository formulation (see above) or in a suitable enema formulation. Topically acceptable transdermal patches may also be used. For topical applications, the pharmaceutical compositions may be formulated in a suitable ointment containing the active component suspended or dissolved in one or more carriers. Carriers for topical administration of the compounds of this invention include, but are not limited to, mineral oil, liquid petrolatum, white petrolatum, propylene glycol, polyoxyethylene, polyoxypropylene compound, emulsifying wax and water. In certain preferred aspects of the invention, the compounds may be coated onto a stent which is to be surgically implanted into a patient in order to inhibit or reduce the likelihood of occlusion occurring in the stent in the patient.

[0163] Alternatively, the pharmaceutical compositions can be formulated in a suitable lotion or cream containing the active components suspended or dissolved in one or more pharmaceutically acceptable carriers. Suitable carriers include, but are not limited to, mineral oil, sorbitan monostearate, polysorbate 60, cetyl esters wax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol and water.

[0164] For ophthalmic use, the pharmaceutical compositions may be formulated as micronized suspensions in isotonic, pH adjusted sterile saline, or, preferably, as solutions in isotonic, pH adjusted sterile saline, with our without a preservative such as benzylalkonium chloride. Alternatively, for ophthalmic uses, the pharmaceutical compositions may be formulated in an ointment such as petrolatum.

[0165] The pharmaceutical compositions of this invention may also be administered by nasal aerosol or inhalation. Such compositions are prepared according to techniques well-known in the art of pharmaceutical formulation and may be prepared as solutions in saline, employing benzyl alcohol or other suitable preservatives, absorption promoters to enhance bioavailability, fluorocarbons, and / or other conventional solubilizing or dispersing agents.

[0166] Solutions or suspensions used for parenteral, intradermal, subcutaneous, or topical application can include the following components: a sterile diluent such as water for injection, saline solution, fixed oils, polyethylene glycols, glycerine, propylene glycol or other synthetic solvents; antibacterial agents such as benzyl alcohol or methyl parabens; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as ethylenediaminetetraacetic acid; buffers such as acetates, citrates or phosphates and agents for the adjustment of tonicity such as sodium chloride or dextrose. The parental preparation can be enclosed in ampoules, disposable syringes or multiple dose vials made of glass or plastic.

[0167] It should also be understood that a specific dosage and treatment regimen for any particular patient will depend upon a variety of factors, including the activity of the specific compound employed, the age, body weight, general health, sex, diet, time of administration, rate of excretion, drug combination, and the judgment of the treating physician and the severity of the particular disease or condition being treated.

[0168] A patient or subject in need of therapy using compounds as described herein can be treated by administering to the patient (subject) an effective amount of the compound including pharmaceutically acceptable salts, solvates or polymorphs, thereof optionally in a pharmaceutically acceptable carrier or diluent, either alone, or in combination with other known agents.

[0169] Disease states of conditions which may be treated using compounds or compositions according to the present description include, but not limited to, for example, Alzheimer’s disease and cancer. In certain embodiments, the therapeutic or pharmaceutical compositions comprise an effective amount of an additional biologically or bioactive active agent, e.g., an agent effective for the treatment of cancer, that is co-administered.

[0170] The term “coadministration” or “combination therapy” shall mean that at least two compounds or compositions are administered to the patient at the same time, such that effective amounts or concentrations of each of the two or more compounds may be found in the patient at a given point in time. Although compounds according to the present invention may be coadministered to a patient at the same time, the term embraces both administration of two or more agents at the same time or at different times, provided that effective concentrations of all coadministered compounds or compositions are found in the subject at a given time. In certain preferred aspects of the present invention, one or more of the present compounds described above, are co-administered in combination with at least one additional bioactive agent, especially including an anticancer agent. In particularly preferred aspects of the invention, the co-administration of compounds results in synergistic therapeutic, including anticancer therapy.

[0171] In another aspect, the description provides a composition comprising an effective amount of two or more of the PROTAC compounds as described herein, and a pharmaceutically acceptable carrier. In certain embodiments, the composition further comprises an effective or synergistic amount of another bioactive agent that is not a PROTAC compound. Pharmaceutical compositions comprising combinations of an effective amount of at least one bifunctional compound according to the present invention, and one or more of the compounds otherwise described herein, all in effective amounts, in combination with a pharmaceutically effective amount of a carrier, additive or excipient, represents a further aspect of the present invention.

[0172] The term “bioactive agent” is used to describe an agent, other than the PROTAC compounds described herein, which is used in combination with the present compounds as an agent with biological activity to assist in effecting an intended therapy, inhibition and / or prevention / prophylaxis for which the present compounds are used. Preferred bioactive agents for use herein include those agents which have pharmacological activity similar to that for which the present compounds are used or administered and include for example, anti-cancer agents.

[0173] The term “additional anti -cancer agent” is used to describe an anti-cancer agent, which may be combined with PROTAC compounds according to the present description to treat cancer. These agents include, for example, everolimus, trabectedin, abraxane, TLK 286, AV-299, DN- 101, pazopanib, GSK690693, RTA 744, ON O91O.Na, AZD 6244 (ARRY-142886), AMN-107, TKI-258, GSK461364, AZD 1152, enzastaurin, vandetanib, ARQ-197, MK-0457, MLN8054, PHA-739358, R-763, AT-9263, a FLT-3 inhibitor, an androgen receptor inhibitor, a VEGFR inhibitor, an EGFR TK inhibitor, an aurora kinase inhibitor, a PIK-1 modulator, a Bcl-2 inhibitor, an HD AC inhbitor, a c-MET inhibitor, a PARP inhibitor, a Cdk inhibitor, an EGFR TK inhibitor, an IGFR-TK inhibitor, an anti-HGF antibody, a PI3 kinase inhibitors, an AKT inhibitor, a JAK / STAT inhibitor, a checkpoint- 1 or 2 inhibitor, a focal adhesion kinase inhibitor, a Map kinase kinase (mek) inhibitor, a VEGF trap antibody, pemetrexed, erlotinib, dasatanib, nilotinib, decatanib, panitumumab, amrubicin, oregovomab, Lep-etu, nolatrexed, azd2171, batabulin, ofatumumab, zanolimumab, edotecarin, tetrandrine, rubitecan, tesmilifene, oblimersen, ticilimumab, ipilimumab, gossypol, Bio 111, 131-I-TM-601, ALT-110, BIO 140, CC 8490, cilengitide, gimatecan, IL13-PE38QQR, INO 1001, IPdRi KRX-0402, lucanthone, LY317615, neuradiab, vitespan, Rta 744, Sdx 102, talampanel, atrasentan, Xr 311, romidepsin, ADS-100380, sunitinib, 5-fluorouracil, vorinostat, etoposide, gemcitabine, doxorubicin, liposomal doxorubicin, 5 '-deoxy-5 -fluorouridine, vincristine, temozolomide, ZK-304709, seliciclib; PD0325901, AZD-6244, capecitabine, L-Glutamic acid, N-[4-[2-(2-amino-4,7- dihydro-4-oxo-lH-pyrrolo[2,3-d]pyrimidin-5-yl)ethyl]benzoyl]-, disodium salt, heptahydrate, camptothecin, PEG-labeled irinotecan, tamoxifen, toremifene citrate, anastrazole, exemestane, letrozole, DES(diethylstilbestrol), estradiol, estrogen, conjugated estrogen, bevacizumab, IMC- 1C11, CHIR-258); 3- [5-(methylsulfonylpiperadinemethyl)-indolylj -quinolone, vatalanib, AG- 013736, AVE-0005, the acetate salt of [D-Ser(Bu t)6,Azgly 10] (pyro-Glu-His-Trp-Ser-Tyr-D- Ser(Bu t)-Leu-Arg-Pro-Azgly-NH2 acetate [C59H84N18O14 — (C2H4O2)Xwhere x=l to 2.4], goserelin acetate, leuprolide acetate, triptorelin pamoate, medroxyprogesterone acetate, hydroxyprogesterone caproate, megestrol acetate, raloxifene, bicalutamide, flutamide, nilutamide, megestrol acetate, CP-724714; TAK-165, HKI-272, erlotinib, lapatanib, canertinib, ABX-EGF antibody, erbitux, EKB-569, PKI-166, GW-572016, lonafamib, BMS-214662, tipifamib; amifostine, NVP-LAQ824, suberoyl analide hydroxamic acid, valproic acid, trichostatin A, FK-228, SU11248, sorafenib, KRN951, aminoglutethimide, amsacrine, anagrelide, L-asparaginase, Bacillus Calmette-Guerin (BCG) vaccine, adriamycin, bleomycin, buserelin, busulfan, carboplatin, carmustine, chlorambucil, cisplatin, cladribine, clodronate, cyproterone, cytarabine, dacarbazine, dactinomycin, daunorubicin, diethylstilbestrol, epirubicin, fludarabine, fludrocortisone, fluoxymesterone, flutamide, gleevec, gemcitabine, hydroxyurea, idarubicin, ifosfamide, imatinib, leuprolide, levamisole, lomustine, mechlorethamine, melphalan, 6-mercaptopurine, mesna, methotrexate, mitomycin, mitotane, mitoxantrone, nilutamide, octreotide, oxaliplatin, pamidronate, pentostatin, plicamycin, porfimer, procarbazine, raltitrexed, rituximab, streptozocin, teniposide, testosterone, thalidomide, thioguanine, thiotepa, tretinoin, vindesine, 13-cis-retinoic acid, phenylalanine mustard, uracil mustard, estramustine, altretamine, floxuridine, 5-deooxyuridine, cytosine arabinoside, 6-mecaptopurine, deoxycoformycin, calcitriol, valrubicin, mithramycin, vinblastine, vinorelbine, topotecan, razoxin, marimastat, COL-3, neovastat, BMS-275291, squalamine, endostatin, SU5416, SU6668, EMD121974, interleukin- 12, IM862, angiostatin, vitaxin, droloxifene, idoxyfene, spironolactone, finasteride, cimitidine, trastuzumab, denileukin diftitox, gefitinib, bortezimib, paclitaxel, cremophor-free paclitaxel, docetaxel, epithilone B, BMS-247550, BMS-310705, droloxifene, 4- hydroxytamoxifen, pipendoxifene, ERA- 923, arzoxifene, fulvestrant, acolbifene, lasofoxifene, idoxifene, TSE-424, HMR-3339, ZK186619, topotecan, PTK787 / ZK 222584, VX-745, PD 184352, rapamycin, 40-O-(2-hydroxyethyl)-rapamycin, temsirolimus, AP-23573, RAD001, ABT-578, BC-210, LY294002, LY292223, LY292696, LY293684, LY293646, wortmannin, ZM336372, L-779,450, PEG-filgrastim, darbepoetin, erythropoietin, granulocyte colonystimulating factor, zolendronate, prednisone, cetuximab, granulocyte macrophage colonystimulating factor, histrelin, pegylated interferon alfa-2a, interferon alfa-2a, pegylated interferon alfa-2b, interferon alfa-2b, azacitidine, PEG-L-asparaginase, lenalidomide, gemtuzumab, hydrocortisone, interleukin-11, dexrazoxane, alemtuzumab, all-transretinoic acid, ketoconazole, interleukin-2, megestrol, immune globulin, nitrogen mustard, methylprednisolone, ibritgumomab tiuxetan, androgens, decitabine, hexamethylmelamine, bexarotene, tositumomab, arsenic trioxide, cortisone, editronate, mitotane, cyclosporine, liposomal daunorubicin, Edwina- asparaginase, strontium 89, casopitant, netupitant, an NK-1 receptor antagonist, palonosetron, aprepitant, diphenhydramine, hydroxyzine, metoclopramide, lorazepam, alprazolam, haloperidol, droperidol, dronabinol, dexamethasone, methylprednisolone, prochlorperazine, granisetron, ondansetron, dolasetron, tropisetron, pegfilgrastim, erythropoietin, epoetin alfa, darbepoetin alfa and mixtures thereof.

[0174] In another aspect, the disclosure provides methods of modulating protein ubiquitination and degradation in a subject, e.g., a cell, a tissue, mammal, or human patient, the method comprising administering an effective amount of a compound as described herein or a composition comprising an effective amount of the same to a subject, wherein the compound or composition comprising the same is effective in modulating protein ubiquitination and degradation of the protein in the subject. In certain embodiments, the protein is one or more kinases selected from: DYRK1A, DYRK1B, DYRK2, DYRK3, DYRK4, CLK1, CLK2, CLK3, CLK4, CDK7, CDK8 / 19, PI3K, PDGFrA / B, mTOR, HIPKs, and / or CMGC kinases leading to inhibition of WNT signaling.

[0175] In certain embodiments, the description provides a method for regulating protein activity of one or more kinases selected from DYRK1A, DYRK1B, DYRK2, DYRK3, DYRK4, CLK1, CLK2, CLK3, CLK4, CDK7, CDK8 / 19, PI3K, PDGFrA / B, mTOR, HIPKs, and / or CMGC kinases leading to inhibition of WNT signaling in a patient in need comprising administering to said patient an amount of a compound as described herein to a patient.

[0176] In still additional embodiments, the description provides a method of treating a disease state or condition in a patient wherein dysregulated protein activity is responsible for said disease state or condition, said method comprising administering to said patient an effective amount of a compound as described herein to said patient in order to regulate said protein activity in said patient. In certain embodiments, the protein is one or more kinases selected from: DYRK1A, DYRK1B, DYRK2, DYRK3, DYRK4, CLK1, CLK2, CLK3, CLK4, CDK7, CDK8 / 19, PI3K, PDGFrA / B, mTOR, HIPKs, and / or CMGC kinases leading to inhibition of WNT signaling.

[0177] The terms “treat”, “treating”, and “treatment”, etc., as used herein, refer to any action providing a benefit to a patient for which the present compounds may be administered, including the treatment of any disease state or condition which is modulated through the protein to which the present compounds bind. Disease states or conditions, including cancer, which may be treated using compounds according to the present invention are set forth hereinabove. In another aspect, the disclosure provides methods of modulating protein ubiquitination and degradation in a subject, e.g., a cell, a tissue, mammal, or human patient, the method comprising administering an effective amount of a compound as described herein or a composition comprising an effective amount of the same to a subject, wherein the compound or composition comprising the same is effective in modulating protein ubiquitination and degradation of the protein in the subject. In certain embodiments, the protein is one or more of DYRK1A, DYRK1B, DYRK2, DYRK3, DYRK4, CLK1, CLK2, CLK3, CLK4, CDK7, CDK8 / 19, PI3K, PDGFrA / B, mTOR, HIPKs, and / or CMGC kinases leading to inhibition of WNT signaling.

[0178] In another aspect, the disclosure provides methods of modulating protein ubiquitination and degradation of one or more of DYRK1 A, DYRK1B, DYRK2, DYRK3, DYRK4, CLK1 , CLK2, CLK3, CLK4, CDK7, CDK8 / 19, PI3K, PDGFrA / B, mTOR, HIPKs, and / or CMGC kinases leading to inhibition of WNT signaling in a subject, e.g., a cell, a tissue, mammal, or human patient, the method comprising administering an effective amount of a compound as described herein or a composition comprising an effective amount of the same to a subject, wherein the compound or composition comprising the same is effective in modulating protein ubiquitination and degradation of one or more of DYRK1A, DYRK1B, DYRK2, DYRK3, DYRK4, CLK1, CLK2, CLK3, CLK4, CDK7, CDK8 / 19, PI3K, PDGFrA / B, mTOR, HIPKs, and / or CMGC kinases leading to inhibition of WNT signaling in the subject.

[0179] In another aspect, the disclosure provides methods of treating or ameliorating a symptom of a disease related to one or more of DYRK1 A, DYRK1B, DYRK2, DYRK3, DYRK4, CLK1, CLK2, CLK3, CLK4, CDK7, CDK8 / 19, PI3K, PDGFrA / B, mTOR, HIPKs, and / or CMGC kinases leading to inhibition of WNT signaling activity in a subject, e.g., a cell, a tissue, mammal, or human patient, the method comprising administering an effective amount of a compound as described herein or a composition comprising an effective amount of the same to a subject in need thereof, wherein the compound or composition comprising the same is effective in treating or ameliorating a symptom of a disease related to one or more of DYRK1A, DYRK1B, DYRK2, DYRK3, DYRK4, CLK1, CLK2, CLK3, CLK4, CDK7, CDK8 / 19, PI3K, PDGFrA / B, mTOR, HIPKs, and / or CMGC kinases leading to inhibition of WNT signaling activity in the subject. In a preferred embodiment, the subject is a human.

[0180] In another aspect, the disclosure provides methods for identifying the effects of the degradation of proteins of interest in a biological system (e.g., one or more of DYRK1 A, DYRK1B, DYRK2, DYRK3, DYRK4, CLK1, CLK2, CLK3, CLK4, CDK7, CDK8 / 19, PI3K, PDGFrA / B, mTOR, HIPKs, and / or CMGC kinases leading to inhibition of WNT signaling) using compounds according to the present invention.

[0181] In certain embodiments, the present invention provides methods for administering an effective amount of a compound as described herein or a composition comprising an effective amount of the same (alone or with additional agents (e.g., remdesivir)) to a subject (e.g., a human subject) (e.g., a human subject suffering from or at risk of suffering from viral infection) (a human subject suffering from or at risk of suffering from a condition related to a viral infection for purposes of treating, preventing and / or ameliorating the viral infection and / or symptoms of a viral infection (e.g., SARS-CoV-2 infection (e.g., COVID-19)). In such embodiments, administration results in suppression of pro-inflammatory cytokine activity (e.g., IL-6 activity) within the subject. In some embodiments, administration results in enhancement of NK cell activity within the subject. In some embodiments, administration results in enhancement of neutrophil activity within the subject. In some embodiments, administration results in inhibition of viral entry into the subject’s cells through inhibiting binding of the virus with heparin sulfate proteoglycan within such cells.

[0182] In such embodiments, the methods are not limited to treating, preventing and / or ameliorating a particular type or kind of viral infection or symptoms of a particular type or kind of viral infection. In some embodiments, the viral infection is a SARS-CoV-2 related viral infection (e.g., COVID-19). In some embodiments, the viral infection is any infection related to influenza, HIV, HIV-1, HIV-2, drug-resistant HIV, Junin virus, Chikungunya virus, Yellow Fever virus, Dengue virus, Pichinde vims, Lassa vims, adenovims, Measles vims, Punta Toro vims, Respiratory Syncytial vims, Rift Valley vims, RHDV, SARS coronavims, Tacaribe vims, and West Nile vims. In some embodiments, the viral infection is associated with any virals having Mproprotease activity and / or expression.

[0183] In some embodiments, the one or more symptoms related to viral infection includes, but is not limited to, fever, fatigue, dry cough, myalgias, dyspnea, acute respiratory distress syndrome, and pneumonia.

[0184] In certain embodiments, the present invention provides methods of treating cancer in a subject in need thereof, the method comprising administering to the subject 1) a compound as described herein, and 2) at least one anticancer agent. In some embodiments, the cancer is one or more types of cancer selected from glioblastoma, prostate cancer, metastatic breast cancer, metastatic lung cancer, multiple myeloma, secondary metastatic tumors of the brain, colorectal cancer, acute myeloid leukemia, myelodysplastic syndrome. In some embodiments, the at least one anticancer agent comprises a chemotherapeutic agent, an immune checkpoint inhibitor, or radiation therapy. In some embodiments, the immune checkpoint inhibitor is one or more selected from: pembrolizumab, nivolumab, cemipilimab, atezolizumab, avelumab, durvalumab, and ipilimumab. In another aspect, the description provides kits comprising compounds or compositions as described herein. The kit may be promoted, distributed, or sold as a unit for performing the methods of the present invention. In addition, the kits of the present invention may preferably contain instructions which describe a suitable use. Such kits can be conveniently used, e.g., in clinical settings, to treat patients exhibiting symptoms of, e.g., Alzheimer’s disease, down syndrome, diabetes, an autoimmune disease, an inflammatory disorder (e.g., airway inflammation, osteoarthritis (e.g., knee related osteoarthritis)), cancer (e.g., glioblastoma, prostate cancer, metastatic breast cancer, metastatic lung cancer, multiple myeloma, secondary metastatic tumors of the brain, colorectal cancer), a viral infection (e.g., SARS-CoV-2 infection (e.g., COVID-19)), and other diseases. EXAMPLES The following examples are illustrative, but not limiting, of the compounds, compositions, and methods of the present invention. Other suitable modifications and adaptations of the variety of conditions and parameters normally encountered in clinical therapy and which are obvious to those skilled in the art are within the spirit and scope of the invention. The use of pronouns such as “I”, “we”, and “our”, for example, refer to the inventors. Example I. This example provides synthesis and characterization information for compounds described herein. 1-(4-(6-(4-(4-(4-(((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)methyl)-1H- pyrazol-1-yl)piperidine-1-carbonyl)piperidin-1-yl)pyridin-3-yl)but-3-yn-1-yl)-N-(6-(1-methyl- 1H-pyrazol-4-yl)isoquinolin-3-yl)piperidine-4-carboxamide (1) DMF (1.5 mL) and triethylamine (20 mg, 28 μL, 5 eq, 0.20 mmol) were added sequentially to a dry 5 mL microwave vial under argon atmosphere equipped with a magnetic stir bar containing 1-(but-3-yn-1-yl)-N-(6-(1-methyl-1H-pyrazol-4-yl)isoquinolin-3-yl)piperidine-4-carboxamide (2) (15 mg, 1 eq, 40 μmol), 2-(2,6-dioxopiperidin-3-yl)-4-(((1-(1-(1-(5-iodopyridin-2- yl)piperidine-4-carbonyl)piperidin-4-yl)-1H-pyrazol-4-yl)methyl)amino)isoindoline-1,3-dione (30 mg, 1 eq, 40 μmol), PdCl2(dppf) (3.8 mg, 0.13 eq, 5.2 μmol) and copper(I) iodide (1.9 mg, 0.25 eq, 10 μmol). This was stirred for 24 hours at room temperature, then concentrated under reduced pressure and purified by flash column chromatography using a DCM / MeOH gradient granting 1-(4-(6-(4-(4-(4-(((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4- yl)amino)methyl)-1H-pyrazol-1-yl)piperidine-1-carbonyl)piperidin-1-yl)pyridin-3-yl)but-3-yn- 1-yl)-N-(6-(1-methyl-1H-pyrazol-4-yl)isoquinolin-3-yl)piperidine-4-carboxamide (1) (13 mg, 13 μmol, 32 %).1H NMR (500 MHz, CDCl3) δ 10.15 (bs, 1H), 8.93 (s, 1H), 8.72 (s, 1H), 8.55 (s, 1H), 8.21 (s, 1H), 7.88 (s, 1H), 7.85 (d, 1H), 7.81 (s, 1H), 7.74 (s, 1H), 7.57 (d, 1H), 7.52-7.49 (m, 2H), 7.45 (dd, 1H), 7.40 (s, 1H), 7.14 (d, 1H), 6.94 (d, 1H), 6.56 (d, 1H), 6.42 (t, 1H), 4.94 (dd, 1H), 4.73 (d, 1H), 4.37-4.32 (m, 5H), 4.07 (d, 1H), 3.98 (s, 3H), 3.22 (t, 1H), 3.02 (d, 2H), 2.94-2.81 (m, 3H), 2.79-2.72 (m, 3H), 2.64 (d, 2H), 2.58 (d, 2H), 2.37 (t, 1H), 2.24 (d, 1H), 2.16-2.07 (m, 3H), 1.99-1.76 (m, 7H), 0.86 (m, 4H), 0.71 (t, 1H).13C NMR (125 MHz, CDCl3) δ 173.9, 173.1, 171.8, 169.7, 169.5, 167.7, 157.7, 151.1, 150.5146.8, 146.5, 140.2, 138.8, 138.4, 137.4, 136.4, 135.3, 132.7, 128.3, 127.9, 125.8, 125.2, 124.5, 122.6, 121.7, 118.1, 117.0, 112.2, 110.6, 109.0, 107.8, 106.3, 59.2, 57.6, 53.6, 52.9, 49.1, 44.9, 39.4, 38.8, 37.8, 31.7, 29.8, 29.5, 28.9, 28.2, 23.0, 22.8, 18.0, 14.3. LCMS [(M+2H+) / 2] = 505.8. Experiments were conducted with Compound 1 demonstrating: DYRK1A KD 1.3nM. HEK293-DYRK1A(HiBiT) > 50% @300nM degradation. 1-(but-3-yn-1-yl)-N-(6-( l)piperidine-4-carboxamide (2) 4-bromobut-1-yne (65 mg, 43 μL, 1.1 eq, 490 μmol) was added to a 5 mL microwave vial equipped with a magnetic stir bar containing 4-((6-(1-methyl-1H-pyrazol-4-yl)isoquinolin-3- yl)carbamoyl)piperidin-1-ium 2,2,2-trifluoroacetate (3) (200 mg, 1 eq, 445 μmol) and potassium carbonate (185 mg, 3 eq, 1.34 mmol) in DMF. The reaction mixture was stirred at room temperature for 8 hours where the reaction mixture was then cooled down to room temperature, solvent removed under reduced pressure, and saturated sodium bicarbonate (5 mL) added. The mixture was extracted with DCM (3 x 15 mL) and the organic layers were recombined, dried over sodium sulfate, and concentrated under reduced pressure. The crude residue was purified by automated flash column chromatography using a DCM / MeOH gradient to afford 1-(but-3-yn- 1-yl)-N-(6-(1-methyl-1H-pyrazol-4-yl)isoquinolin-3-yl)piperidine-4-carboxamide (2) (62 mg, 0.16 mmol, 36 %).1H NMR (500 MHz, CDCl3) δ 8.89 (s, 1H), 8.54 (s, 1H), 7.99 (s, 1H), 7.89 (s, 1H), 7.86 (d, 1H), 7.84 (s, 1H), 7.75 (s, 1H), 7.59 (dd, 1H), 3.99 (s, 3H), 3.03 (d, 2H), 2.63 (t, 2H), 2.41 (t, 2H), 2.32 (t, 1H), 2.14 (t, 2H), 2.03-1.88 (m, 5H). LCMS [M+H+] = 388.0. 4-((6-(1-methyl-1H -pyrazol-4-yl)isoquinolin-3-yl)carbamoyl)piperidin-1-ium 2,2,2- trifluoroacetate (3) To a solution of tert-butyl 4-((6-(1-methyl-1H-pyrazol-4-yl)isoquinolin-3- yl)carbamoyl)piperidine-1-carboxylate (4) (850 mg, 1 eq, 1.95 mmol) in DCM (15 mL) was added trifluoroacetic acid (4.45 g, 3 mL, 20 eq, 39 mmol). The mixture was stirred for 2 hours at room temperature. The volatiles were evaporated in vacuo, granting 4-((6-(1-methyl-1H- pyrazol-4-yl)isoquinolin-3-yl)carbamoyl)piperidin-1-ium 2,2,2-trifluoroacetate (3) (870 mg, 1.94 mmol, 99 %) which was carried over without further purification.1H NMR (500 MHz, d- DMSO ) δ 10.72 (s, 1H), 9.09 (s, 1H), 8.72 (s, 1H), 8.46 (s, 1H), 8.40 (s, 1H), 8.13 (s, 1H), 8.09 (s, 1H), 8.05 (d, 1H), 7.80 (d, 1H), 3.94 (s, 3H), 3.41 (d, 2H), 2.96 (q, 2H), 2.88 (t, 1H), 2.04 (d, 2H), 1.88 (q, 2H). LCMS [M+H]+= 335.0. O tert-butyl 4-((6-(1-methyl-1H-pyrazol-4-yl)isoquinolin-3-yl)carbamoyl)piperidine-1-carboxylate (4) To a 24 mL microwave vial was added potassium phosphate (1.47 g, 3 eq, 6.91 mmol), tert- butyl 4-((6-bromoquinolin-3-yl)carbamoyl)piperidine-1-carboxylate (5) (1.0 g, 1 eq, 2.3 mmol), 1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (527 mg, 1.1 eq, 2.533 mmol), and PdCl2(dppf) (168 mg, 0.1 eq, 230.2 μmol). The vial was capped, evacuated, and backfilled with nitrogen. Dry 1,4-dioxane (9.5 mL) and degassed water (2 mL) were injected, and the vial heated to 90 °C for 3 hours. The reaction mixture was then diluted with DCM, filtered through celite, and washed with water and brine. The organic layer was dried with MgSO4, filtered, and the solvents removed in vacuo. The residue was purified by automated column chromatography (0-5% Methanol in DCM) to grant the product tert-butyl 4-((6-(1- methyl-1H-pyrazol-4-yl)quinolin-3-yl)carbamoyl)piperidine-1-carboxylate (4) (855 mg, 1.96 mmol, 85 %), as a light brown solid.1H NMR (500 MHz, d-DMSO ) δ 8.89 (s, 1H), 8.52 (s, 1H), 8.03 (s, 1H), 7.89 (s, 1H), 7.86 (d, 1H), 7.84 (s, 1H), 7.76 (s, 1H), 7.60 (d, 1H), 4.20 (bs, 2H), 3.99 (s, 3H), 2.82 (t, 2H), 2.47 (tt, 1H), 1.95 (d, 2H), (1.79 (qd, 2H), 1.48 (s, 9H). LCMS [M+H]+= 435.9. tert-butyl 4-((6-br dine-1-carboxylate (5) To a 100 mL round bottom flask was added HATU (2.56 g, 1.5 eq, 6.72 mmol), N-[(tert- butoxy)carbonyl]piperidine-4-carboxylic acid (1.54 g, 1.5 eq, 6.72 mmol) and DMF (20 mL). The mixture was stirred at room temperature for 10 min, then DIPEA (1.74 g, 2.3 mL, 3 eq, 13.45 mmol) was added. The mixture was stirred for another 10 minutes, and then DMAP (109 mg, 0.2 eq, 896.6 μmol) and 6-bromoisoquinolin-3-amine (1 g, 1 eq, 4.48 mmol) were added and the mixture heated to 60 °C overnight. The solvents were then removed in vacuo. The reaction mixture was diluted with EtOAc (50 mL) and washed with water 2x (50 mL) then brine (50 mL). The organic layer was concentrated under reduced pressure, and the residue purified by column chromatography (EtOAc / Hexanes) to grant the product tert-butyl 4-((6- bromoisoquinolin-3-yl)carbamoyl)piperidine-1-carboxylate (5) (1.51 g, 3.47 mmol, 77 %).1H NMR (500 MHz, d-DMSO) δ 10.68 (s, 1H), 9.15 (s, 1H), 8.44 (s, 1H), 8.17 (s, 1H), 7.99 (d, 1H), 7.63 (d, 1H), 4.00 (d, 2H), 2.74 (t, 2H), 1.79 (t, 2H), 1.50 (qd, 2H), 1.41 (s, 9H). 2-(2,6-dioxopiperidin-3-yl)-4-(((1-(1-(1-(5-(3-(4-((6-((5-fluoro-4-(4-fluoro-1-isopropyl-2- methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)amino)pyridin-3-yl)methyl)piperazin-1- yl)prop-1-yn-1-yl)pyridin-2-yl)piperidine-4-carbonyl)piperidin-4-yl)-1H-pyrazol-4- yl)methyl)amino)isoindoline-1,3-dione (6) DMF (333 μL), triethylamine (17 mg, 23 μL, 5 eq, 166.5 μmol) were added sequentially to a dry 5 mL microwave vial equipped with a magnetic stir bar containing 2-(2,6-dioxopiperidin-3-yl)- 4-(((1-(1-(1-(5-iodopyridin-2-yl)piperidine-4-carbonyl)piperidin-4-yl)-1H-pyrazol-4- yl)methyl)amino)isoindoline-1,3-dione (25 mg, 1 eq, 33.3 μmol), 5-fluoro-4-(4-fluoro-1- isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)-N-(5-((4-(prop-2-yn-1-yl)piperazin-1- yl)methyl)pyridin-2-yl)pyrimidin-2-amine (7) (17.2 mg, 1 eq, 33.3 μmol), PdCl2(dppf) (3.17 mg, 0.13 eq, 4.33 μmol) and copper(I) iodide (1.2 mg, 0.2 eq, 6.66 μmol). The solution was stirred for 24 hours at room temperature where it was concentrated under reduced pressure and purified by flash column chromatography to afford 2-(2,6-dioxopiperidin-3-yl)-4-(((1-(1-(1-(5-(3-(4-((6- ((5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2- yl)amino)pyridin-3-yl)methyl)piperazin-1-yl)prop-1-yn-1-yl)pyridin-2-yl)piperidine-4- carbonyl)piperidin-4-yl)-1H-pyrazol-4-yl)methyl)amino)isoindoline-1,3-dione (6). LCMS [M+H]+1140. Experiments were conducted with Compound 6 demonstrating: DYRK1A KD 26 nM. CDK4 KD 940 nM. CDK6 KD 320 nM. HEK293-DYRK1A(HiBiT) >25% degradation @300nM. 5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)-N-(5-((4-(prop-2-yn-1- yl)piperazin-1-yl)methyl)pyridin-2-yl)pyrimidin-2-amine (7) 3-bromoprop-1-yne (99 mg, 74 μL, 80% Wt, 1.1 eq, 664 μmol) was added to a 24 mL microwave vial equipped with a magnetic stir bar containing 5-fluoro-4-(4-fluoro-1-isopropyl-2- methyl-1H-benzo[d]imidazol-6-yl)-N-(5-(piperazin-1-ylmethyl)pyridin-2-yl)pyrimidin-2-amine (8) (289 mg, 1 eq, 604 μmol) and potassium carbonate (167 mg, 2 eq, 1.21 mmol) in DMF (6 mL). The reaction mixture was cooled at 0 °C for 6 hours where the reaction mixture was washed with saturated sodium bicarbonate (5 mL). The mixture was extracted with DCM (3x) and the organic layers were recombined, dried over sodium sulfate, and concentrated under reduced pressure. The crude residue was purified by automated flash column chromatography using a DCM / MeOH gradient to afford 5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H- benzo[d]imidazol-6-yl)-N-(5-((4-(prop-2-yn-1-yl)piperazin-1-yl)methyl)pyridin-2-yl)pyrimidin- 2-amine (7) (103 mg, 199 μmol, 33 %). LCMS [M+H]+517.1H NMR (500 MHz, Chloroform-d) δ 9.40 (s, 1H), 8.46 (d, J= 3.7 Hz, 1H) , 8.37 (d, J= 8.5 Hz, 1H), 8.32 (d, J= 2.0 Hz, 1H), 8.13 (d, J= 1.3 Hz, 1H), 7.72 (dd, J= 11.5, 1.3 Hz, 1H), 7.63 (dd, J= 8.6, 2.3 Hz, 1H), 4.70 (p, J=7.0 Hz, 1H), 3.42 (s, 2H) , 3.24 (d,J=3.2 Hz, 2H), 2.83 (s, 1H), 2.62, (s,3H), (2.6-2.4,m, 8H) , 1.66 (d, J=7.0 Hz, 6H). F N H N N N NH 5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)-N-(5-(piperazin-1- ylmethyl)pyridin-2-yl)pyrimidin-2-amine (8) tert-butyl 4-((6-((5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6- yl)pyrimidin-2-yl)amino)pyridin-3-yl)methyl)piperazine-1-carboxylate (9) (183 mg, 1 eq, 316 μmol) was dissolved in DCM (3.2 mL), TFA (1.1 g, 731 μL, 30 eq, 9.49 mmol) was added dropwise with stirring and the reaction was carried out at 25 °C for 2 hours. After the reaction was complete, 50 mL of heptane was added and reaction mixture was concentrated in vacuo. Then the crude was dissolved in 50 mL of EtOAc and washed with 1 M NaOH (x3), collecting the organic layer which was washed with saturated sodium chloride, and dried over sodium sulfate to afford 5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)-N-(5- (piperazin-1-ylmethyl)pyridin-2-yl)pyrimidin-2-amine (8) (140 mg, 293 μmol, 93 %). LCMS [M+H]+479.1H NMR (500 MHz, Chloroform-d) δ 8.51 (s, 1H), 8.43 (d, J= 3.7 Hz, 1H) , 8.39 (d, J=8.7 Hz, 1H), 8.16, (d, J= 2.2 Hz, 1H), 8.14 (d, J= 1.4 Hz, 1H), 7.77 (d, J=11.4 Hz, 1H) , 7.67 (dd, J= 8.6, 2.3 Hz, 1H), 4.70 (p, J=7.0 Hz, 1H), 3.42 (s, 2H), 2.83 (t, 4H), 2.63 (s, 3H), 2.39 (s, 4H), 1.66 (d, J=6.9 Hz, 6H). F N O tert-butyl 4-((6-((5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6- yl)pyrimidin-2-yl)amino)pyridin-3-yl)methyl)piperazine-1-carboxylate (9) A 100 mL dry round bottom flask under atmosphere of argon containing tert-butyl 4-((6- aminopyridin-3-yl)methyl)piperazine-1-carboxylate (10) (453 mg, 1 eq, 1.55 mmol) in tert-butyl alcohol (7.8 mL) was added Pd2(dba)3 (142 mg, 0.1 eq, 155 μmol), xantphos (45 mg, 0.05 eq, 78 μmol), and 6-(2-chloro-5-fluoropyrimidin-4-yl)-4-fluoro-1-isopropyl-2-methyl-1H- benzo[d]imidazole (500 mg, 1 eq, 1.55 mmol) and Cs2CO3 (1.51 g, 3 eq, 4.65 mmol). The resulting solution was stirred at 110 °C for 16 hours. The reaction was then quenched by the addition of 25 mL of water and was extracted with DCM (3 x 50mL) and the organic layers were combined and dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude residue was purified by automated column chromatography using a CombiFlash to give tert-butyl 4-((6-((5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H- benzo[d]imidazol-6-yl)pyrimidin-2-yl)amino)pyridin-3-yl)methyl)piperazine-1-carboxylate (9) (183 mg, 316 μmol, 20 %). LCMS [M+H]+579.1H NMR (500 MHz, Chloroform-d) δ 8.51 (s, 1H), 8.43 (d, J= 3.7 Hz, 1H) , 8.39 (d, J=8.5 Hz, 1H), 8.25, (d, J= 2.2 Hz, 1H), 8.14 (d, J= 1.4 Hz, 1H), 7.77 (dd, J=11.4, 1.32 Hz, 1H) , 7.67 (dd, J= 8.6, 2.3 Hz, 1H), 4.70 (sep, J=6.9 Hz 1H), 3.46 (S, 2H) , 3.40 (s,4H), 2.67 (s, 3H), 2.38 (s, 4H) , 1.69 (d, J=6.8 Hz, 6H), 1.43 (s, 9H). tert-butyl 4-((6-am opy - -y e y p pe a ine-1-carboxylate (10) 6-aminonicotinaldehyde (1 g, 1 eq, 8.2 mmol) and tert-butyl piperazine-1-carboxylate (2.3 g, 1.5 eq, 12.3 mmol) were added into a 250 mL round bottom flask. To this, DCE (82 mL) was added and allowed to stir at 20 °C for 2 hours. Then sodium triacetoxyborohydride (2.8 g, 1.6 eq, 13 mmol) was added and stirred overnight at room temperature. After the reaction was complete, a saturated aqueous NaHCO3solution (30 mL) was added dropwise to the reaction flask to quench the reaction, extracted with dichloromethane (3 x 75 mL), and the organic phase was collected. It was dried with anhydrous sodium sulfate, concentrated, and purified by rapid preparative liquid chromatography (DCM:MeOH 10:1) to obtain tert-butyl 4-((6-aminopyridin-3- yl)methyl)piperazine-1-carboxylate (10) (2.3 g, 7.98 mmol, 97 %). LCMS [M+H]+293.1H NMR (500 MHz, Chloroform-d) δ 7.79 (d, J = 2.3 Hz, 1H), 7.25 (dd, J = 8.5, 2.3 Hz, 1H), 6.34 (d, J = 8.4 Hz, 1H), 4.82 (s, 2H), 3.22 (s, 2H), 2.66 (t, J = 5.1 Hz, 4H), 2.22 (t, J = 5.2 Hz, 4H), 1.32 (s, 9H). 4-(((1- ihydro-1H- imid idine-4- carbonyl)piperidin-4-yl)-1H-pyrazol-4-yl)methyl)amino)-2-(2,6-dioxopiperidin-3- yl)isoindoline-1,3-dione (11) To a 5 mL microwave vial equipped with a stir bar was added CuI (2.9 mg, 0.12 eq, 15μmol), PdCl2(dppf) (8 mg, 0.08 eq, 10μmol), (R,Z)-5-(benzo[d]thiazol-6-ylmethylene)-2-(4-methyl-1- (prop-2-yn-1-yloxy)pentan-2-yl)-3,5-dihydro-4H-imidazol-4-one (12) (47 mg, 1 eq, 127μmol), and 2-(2,6-dioxopiperidin-3-yl)-4-(((1-(1-(1-(5-iodopyridin-2-yl)piperidine-4- carbonyl)piperidin-4-yl)-1H-pyrazol-4-yl)methyl)amino)isoindoline-1,3-dione (96 mg, 1 eq, 127 μmol). The vial was capped and purged with argon (3x), then DMF (2.5 mL) and TEA (39 mg, 53 μL, 3 eq, 382 μmol) were added. The reaction was allowed to stir at room temperature for 24 hours, upon which the solvent was removed in vacuo. The residue was taken up in DCM (10 mL) and water (10 mL), the organic layer washed with brine (10 mL), dried over sodium sulfate, and purified via automated column chromatography (10% MeOH in DCM) to grant 4-(((1-(1-(1- (5-(3-(((R)-2-(((Z)-4-(benzo[d]thiazol-6-ylmethylene)-5-oxo-4,5-dihydro-1H-imidazol-2- yl)amino)-4-methylpentyl)oxy)prop-1-yn-1-yl)pyridin-2-yl)piperidine-4-carbonyl)piperidin-4- yl)-1H-pyrazol-4-yl)methyl)amino)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (11) (49 mg, 38 %), as a dark yellow powder.1H NMR (500 MHz, CDCl3) δ 8.96 (s, 1H), 8.91 (s, 1H), 8.20 (s, 1H), 8.06 (s, 1H), 7.54 – 7.39 (m, 3H), 7.13 (d, J = 7.2 Hz, 1H), 6.93 (d, J = 8.5 Hz, 1H), 6.67 (s, 1H), 6.43 (q, J = 12.7 Hz, 2H), 4.92 (dd, J = 12.3, 5.3 Hz, 1H), 4.72 (s, 1H), 4.42 (s, 1H), 4.36 (d, J = 5.5 Hz, 2H), 4.35 – 4.19 (m, 1H), 4.05 (s, 1H), 3.78 (t, J = 15.2 Hz, 2H), 3.22 (s, 1H), 2.96 – 2.67 (m, 7H), 2.24 (s, 1H), 2.19 – 2.03 (m, 2H), 1.91 (s, 2H), 1.85 – 1.66 (m, 3H), 1.63 (s, 12H), 1.26 (p, J = 3.9 Hz, 2H), 1.06 – 0.83 (m, 5H). LCMS [M+2H]2+503.4. Experiments were conducted with Compound 11 demonstrating: HEK293- DYRK1A(HiBiT) > 50% @100nM degradation. Degradation. DYRK1A KD > 1nM. (R,Z)-5-(benzo[d]thiazol-6-ylme hyl-1-(prop-2-yn-1-yloxy)pentan-2- yl)amino)-3, dazol-4-one (12) To a 5 mL microwave vial was added ammonium chloride (30 mg, 2 eq, 553 μmol), (R)-4- methyl-1-(prop-2-yn-1-yloxy)pentan-2-amine (13) (172 mg, 4 eq, 1.1 mmol), and (Z)-5- (benzo[d]thiazol-6-ylmethylene)-2-(ethylthio)-3,5-dihydro-4H-imidazol-4-one (80 mg, 1 eq, 276 μmol). The vial was capped and purged with argon, following which anisole (0.9 mL) was added and the reaction was heated to 125 °C overnight. After time elapsed the mixture was concentrated in vacuo directly onto silica gel and purified using automated column chromatography (alumina, 10% MeOH in DCM) to grant (R,Z)-5-(benzo[d]thiazol-6- ylmethylene)-2-((4-methyl-1-(prop-2-yn-1-yloxy)pentan-2-yl)amino)-3,5-dihydro-4H-imidazol- 4-one (12) (47 mg, 44 %), as a dark yellow powder.1H NMR (500 MHz, DMSO) δ 10.50 (s, 1H), 9.34 (s, 1H), 8.91 (s, 1H), 8.19 (d, J = 8.6 Hz, 1H), 8.02 (d, J = 8.6 Hz, 1H), 7.42 (s, 1H), 6.42 (s, 1H), 4.32 (s, 1H), 4.21 (s, 2H), 3.55 (d, J = 15.8 Hz, 2H), 3.45 (t, J = 2.4 Hz, 1H), 1.68 (s, 1H), 1.60 – 1.29 (m, 2H), 1.06 – 0.77 (m, 6H). LCMS [M+H]+383.1. (R)-4-methyl-1-( pentan-2-amine (13) To a 100 mL round bottom flask containing a solution of tert-butyl (R)-(4-methyl-1-(prop-2-yn- 1-yloxy)pentan-2-yl)carbamate (14) (771 mg, 1 eq, 3 mmol) in DCM (30 mL) was added trifluoroacetic acid (10.3 g, 6.98 mL, 30 eq, 91 mmol) dropwise. The reaction was allowed to stir for 2 hours, upon which the mixture was diluted with ethyl acetate and saturated sodium bicarbonate solution. The organic layer was separated, dried over sodium sulfate, and concentrated in vacuo to grant (R)-4-methyl-1-(prop-2-yn-1-yloxy)pentan-2-amine (13), a brown oil, in quantitative yield which was used without further purification.1H NMR (400 MHz, Chloroform-d) δ 4.21 (s, 2H), 3.74 (dd, 1H), 3.57 (t, 1H), 3.41 (m, 1H), 2.48 (t, 1H), 1.72 (m, 1H), 1.54 (m, 2H), 0.94 (dd, 6H). LCMS [M+H]+156.1. tert-butyl (R)-(4-meth pentan-2-yl)carbamate (14) To a 20 mL microwave vial was added 3-bromoprop-1-yne (793 mg, 590 μL, 1.5 eq, 6.67 mmol) to tert-butyl (R)-(1-hydroxy-4-methylpentan-2-yl)carbamate (15) (966 mg, 1 eq, 4.45 mmol) in THF (14.5 mL). The reaction was stirred for 10 minutes, following which TBAI (82 mg, 0.05 eq, 222 μmol), KI (111 mg, 0.15 eq, 667 μmol), and solid KOH (299 mg, 1.2 eq, 5.33 mmol) were sequentially added. The reaction mixture was allowed to stir overnight at room temperature. The mixture was then diluted with ethyl acetate (25 mL) and saturated sodium bicarbonate solution (25 mL). The organic layer was separated and then washed with water (25 mL) and brine (25 mL). The organic layer was dried over magnesium sulfate and purified via automated column chromatography (0-25% ethyl acetate in hexanes) to grant tert-butyl (R)-(4- methyl-1-(prop-2-yn-1-yloxy)pentan-2-yl)carbamate (14)(771 mg, 3 mmol, 68 %), as a clear oil.1H NMR (400 MHz, Chloroform-d) δ 4.62 (bs, 1H), 4.15 (t, 2H), 3.81 (bs, 1H), 3.51 (m, 2H), 2.43 (t, 1H), 1.65 (m, 1H), 1.44 (s, 9H), 1.37 (m, 2H), 0.92 (dd, 6H). LCMS [M-(t- butyl)+H]+200.1. tert-butyl (R)-(1-hydr ntan-2-yl)carbamate (15) To a 100 mL flask containing 2-amino-4-methylpentan-1-ol (2.00 g, 1 eq, 17.1 mmol) in THF (40 mL) was added boc anhydride (4.1 g, 4.3 mL, 1.1 eq, 19 mmol) dropwise. The reaction was allowed to stir overnight, following which the volatiles were removed in vacuo. The resulting oil was purified via automated column chromatography (0-40% EtOAc in Hexanes) to grant tert- butyl (R)-(1-hydroxy-4-methylpentan-2-yl)carbamate (15) (2.20 g, 10.1 mmol, 59 %), as a clear oil.1H NMR (400 MHz, Chloroform-d) δ 4.54 (bs, 1H), 3.78-3.47 (m, 3H), 2.41 (bs, 1H), 1.66 (m, 1H), 1.45 (s, 9H), 1.34-1.28 (m, 2H), 0.93 (dd, 6H). (Z)-5-(benzo[d]thiazol-6-ylmethy -3,5-dihydro-4H-imidazol-4-one (16) To a mixture of (Z)-5-(benzo[d]thiazol-6-ylmethylene)-2-thioxoimidazolidin-4-one (17) (1.37 g, 1 eq, 5.24 mmol), potassium carbonate (363 mg, 0.5 eq, 2.62 mmol) in MeCN (26 mL) was added iodoethane (985 mg, 510 μL, 1.2 eq, 6.29 mmol) at room temperature. After addition the reaction mixture was heated to 60 °C where it stirred for 19 hours. After time elapsed the reaction mixture was concentrated and recrystallized in DCM affording (Z)-5-(benzo[d]thiazol- 6-ylmethylene)-2-(ethylthio)-3,5-dihydro-4H-imidazol-4-one (16) (1.17 g, 4.04 mmol, 77%).1H NMR (500 MHz, DMSO-d6) δ 11.95 (s, 1H), 9.58 (s, 1H), 9.02 (s, 1H), 8.57 (d, J = 8.7 Hz, 1H), 8.24 (d, J = 8.8 Hz, 1H), 7.00 (s, 1H), 3.93 (s, 2H), 1.55 (d, J = 12.9 Hz, 3H). LCMS [M+H]+290. (Z)-5-(benzo[d]thiazol-6 y y oxoimidazolidin-4-one (17) To a mixture of 2-thioxoimidazolidin-4-one (581 mg, 1 eq, 5 mmol) and benzo[d]thiazole-6- carbaldehyde (816 mg, 1 eq, 5 mmol) in EtOH (16 mL) was added piperidine (426 mg, 494 μL, 1 eq, 5 mmol) and the mixture was stirred at 110 °C for 1.5 hour. After concentration, the residue was washed with DCM, collecting the filtrate to afford (Z)-5-(benzo[d]thiazol-6- ylmethylene)-2-thioxoimidazolidin-4-one (17) (1.19 g, 4.56 mmol, 91 %). LCMS [M+H]+262.9. 2 -1H- p -yl)- N-(6-(1-methyl-1H-pyrazol-4-yl)isoquinolin-3-yl)isonicotinamide (18) DMF (1.7 mL) and triethylamine (22 mg, 31 μL, 5 eq, 0.22 mmol) were added sequentially to a dry 5 mL microwave vial equipped with a magnetic stir bar containing N-(6-(1-methyl-1H- pyrazol-4-yl)isoquinolin-3-yl)-2-(4-(prop-2-yn-1-yl)piperazin-1-yl)isonicotinamide (19) (20 mg, 1 eq, 44 μmol), 2-(2,6-dioxopiperidin-3-yl)-4-(((1-(1-(1-(5-iodopyridin-2-yl)piperidine-4- carbonyl)piperidin-4-yl)-1H-pyrazol-4-yl)methyl)amino)isoindoline-1,3-dione (33 mg, 1 eq, 44 μmol), PdCl2(dppf) (4 mg, 0.13 eq, 5.8 μmol) and copper(I) iodide (2 mg, 0.25 eq, 11 μmol). The reaction mixture was stirred for 24 hours at room temperature. Upon completion, the reaction was extracted in EtOAc. The organics were combined and washed with water (x2) and brine (x3), dried over sodium sulfated, and filtered over a celite pad, and concentrated in vacuo. The crude product was dry-loaded onto silica gel and purified by flash column chromatography on a 12 g column (0-10% MeOH / DCM) to afford 2-(4-(3-(6-(4-(4-(4-(((2-(2,6-dioxopiperidin-3- yl)-1,3-dioxoisoindolin-4-yl)amino)methyl)-1H-pyrazol-1-yl)piperidine-1-carbonyl)piperidin-1- yl)pyridin-3-yl)prop-2-yn-1-yl)piperazin-1-yl)-N-(6-(1-methyl-1H-pyrazol-4-yl)isoquinolin-3- yl)isonicotinamide (18) (9.8 mg, 9.1 µmol, 21 %) as a white solid. LCMS [M / 2+H]+538.1H NMR (500 MHz, DMSO) δ 11.09 (s, 1H), 11.05 (s, 1H), 9.12 (s, 1H), 8.60 (s, 1H), 8.38 (d, J = 5.2 Hz, 1H), 8.26 (d, J = 5.1 Hz, 1H), 8.17 (d, J = 2.4 Hz, 1H), 8.14 (s, 1H), 8.11 (s, 1H), 8.07 (d, J = 8.6 Hz, 1H), 7.81 (dd, J = 8.5, 1.7 Hz, 1H), 7.79 (s, 1H), 7.59 – 7.55 (m, 1H), 7.53 (dd, J = 8.9, 2.4 Hz, 1H), 7.48 (s, 1H), 7.46 (s, 1H), 7.16 (dd, J = 6.9, 5.1 Hz, 2H), 7.04 (d, J = 7.0 Hz, 1H), 6.80 (d, J = 8.0 Hz, 2H), 5.04 (dd, J = 12.8, 5.4 Hz, 1H), 4.45 (d, J = 13.1 Hz, 1H), 4.36 (d, J = 5.7 Hz, 3H), 4.31 (d, J = 13.2 Hz, 2H), 4.11 (d, J = 13.6 Hz, 1H), 3.91 (s, 3H), 3.65 (s, 4H), 3.57 (s, 2H), 3.30 (s, 2H), 3.18 (t, J = 12.8 Hz, 1H), 2.98 – 2.86 (m, 4H), 2.65 (d, J = 4.7 Hz, 5H), 2.03 (s, 2H), 1.96 (s, 1H), 1.81 (d, J = 12.6 Hz, 1H), 1.66 (d, J = 12.6 Hz, 3H), 1.56 – 1.45 (m, 2H), 1.23 (s, 1H). Experiments were conducted with Compound 18 demonstrating: HEK293- DYRK1A(HiBiT) >50% degradation @300nM . HEK293-DYRK1B(HiBiT) > 50% degradation @100nM. DYRK1A KD 1.3nM. HCEC WNT Top Flash Reporter EC50 < 1µM. N-(6-(1-methyl-1 2-yn-1-yl)piperazin-1- 3-bromoprop-1-yne (25 mg, 19 μL, 80% Wt, 1 eq, 0.17 mmol) was added to a 5 mL microwave vial equipped with a magnetic stir bar containing N-(6-(1-methyl-1H-pyrazol-4-yl)isoquinolin-3- yl)-2-(piperazin-1-yl)isonicotinamide, Trifluoroacetic acid (20) (90 mg, 1 eq, 0.17 mmol) and potassium carbonate (59 mg, 2.5 eq, 0.43 mmol) in DMF (1.7 mL). The reaction mixture was heated at 60 °C for 16 hours. Upon completion the reaction mixture was then cooled down to room temperature and washed with saturated sodium bicarbonate (5 mL). The mixture was extracted with DCM (3 x 15 mL) and the organic layers were recombined, dried over sodium sulfate, and concentrated under reduced pressure. The crude residue was purified by automated flash column chromatography using a DCM / MeOH gradient to afford N-(6-(1-methyl-1H- pyrazol-4-yl)isoquinolin-3-yl)-2-(4-(prop-2-yn-1-yl)piperazin-1-yl)isonicotinamide (19) (41 mg, 91 µmol, 53 %) as a white solid. LCMS [M+H]+452.1H NMR (500 MHz, DMSO) δ 11.05 (s, 1H), 9.13 (s, 1H), 8.59 (s, 1H), 8.38 (s, 1H), 8.26 (d, J = 5.1 Hz, 1H), 8.14 (d, J = 1.5 Hz, 1H), 8.11 (s, 1H), 8.07 (d, J = 8.5 Hz, 1H), 7.81 (dd, J = 8.5, 1.7 Hz, 1H), 7.47 (s, 1H), 7.16 (dd, J = 5.2, 1.3 Hz, 1H), 3.91 (s, 3H), 3.64 (t, J = 5.0 Hz, 4H), 3.35 (d, J = 2.5 Hz, 2H), 3.18 (t, J = 2.4 Hz, 1H), 2.57 (t, J = 5.0 Hz, 4H).13C NMR (126 MHz, DMSO) δ 164.86, 159.27, 150.74, 148.24, 147.31, 142.88, 137.81, 136.91, 135.06, 129.05, 128.20, 124.73, 124.35, 121.23, 120.56, 110.88, 108.07, 105.24, 79.20, 75.90, 50.92, 46.09, 44.53, 40.11, 40.02, 39.95, 39.85, 39.78, 39.69, 39.61, 39.52, 39.44, 39.35, 39.19, 39.02, 38.81. N-(6-(1-methyl-1H-pyr n-1-yl)isonicotinamide (20) A mixture of DCM:TFA (4:1, 7 mL) was added to the crude mixture of tert-butyl 4-(4-((6-(1- methyl-1H-pyrazol-4-yl)isoquinolin-3-yl)carbamoyl)pyridin-2-yl)piperazine-1-carboxylate (21) (93 mg, 1 eq, 180 μmol) and stirred at room temperature for 2 hours. After completion, excess TFA was evaporated off in vacuo and afforded N-(6-(1-methyl-1H-pyrazol-4-yl)isoquinolin-3- yl)-2-(piperazin-1-yl)isonicotinamide, Trifluoroacetic acid (74 mg, 140 μmol, 78 %) as a TFA salt. This compound was used without any further purification. LCMS [M+H]+414.1H NMR (500 MHz, DMSO) δ 11.08 (s, 1H), 9.34 (s, 1H), 9.12 (d, J = 1.0 Hz, 1H), 8.60 (d, J = 1.0 Hz, 1H), 8.37 (s, 1H), 8.31 (dd, J = 5.1, 0.7 Hz, 1H), 8.15 – 8.06 (m, 2H), 8.05 (s, 1H), 7.80 (dd, J = 8.5, 1.7 Hz, 1H), 7.56 (t, J = 1.1 Hz, 1H), 7.26 (dd, J = 5.1, 1.3 Hz, 1H), 3.91 (s, 3H), 3.86 (t, J = 5.3 Hz, 4H), 3.25 (s, 4H). tert-butyl 4-(4-((6-( 1-methyl-1H-pyrazol-4-yl)isoquinolin-3-yl)carbamoyl)pyridin-2- yl)piperazine-1-carboxylate (21) Argon was purged for 15 minutes through a stirring solution of tert-butyl 4-(4-((6- bromoisoquinolin-3-yl)carbamoyl)pyridin-2-yl)piperazine-1-carboxylate (22) (216 mg, 1 eq, 422 μmol), 1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (105 mg, 1.2 eq, 506 μmol), and potassium phosphate (179 mg, 2 eq, 843 μmol) in Dioxane:H2O (2.11 mL, 9:1). Pd(dppf)Cl2(28 mg, 0.08 eq, 34 μmol) was added to it and further purging was done for 10 minutes. The reaction mixture was allowed to stir at 100 °C for 18 hours. Upon completion, the reaction mixture was poured over water and product was extracted with ethyl acetate, washed with brine, dried over sodium sulfate, filtered, and concentrated in vacuo. The crude residue was purified by automated flash column chromatography using a DCM / MeOH gradient (0-2%) to afford tert-butyl 4-(4-((6-(1-methyl-1H-pyrazol-4-yl)isoquinolin-3-yl)carbamoyl)pyridin-2- yl)piperazine-1-carboxylate (21) (93 mg, 180 µmol, 43%). LCMS [M+H]+514. This compound was carried forward without any further characterization. tert-butyl 4-(4-((6-bromoiso yl)piperazine-1-carboxylate (22) A solution of 2-(4-(tert-butoxycarbonyl)piperazin-1-yl)isonicotinic acid (23) (63 mg, 1 eq, 0.20 mmol), 2-(3H-[1,2,3]triazolo[4,5-b]pyridin-3-yl)-1,1,3,3-tetramethylisouronium hexafluorophosphate(V) (93 mg, 1.2 eq, 0.24 mmol) and N-ethyl-N-isopropylpropan-2-amine (28 mg, 39 μL, 1.06 eq, 0.22 mmol) were stirred in DCM (1.2 mL, 0.2 M) at room temperature for 60 minutes before addition of the N-ethyl-N-isopropylpropan-2-amine (28 mg, 39 μL, 1.06 eq, 0.22 mmol). The reaction was stirred overnight at 45 °C. The next day the reaction was quenched with water and extracted with DCM. The combined organics were dried with sodium sulfate, filtered, and concentrated. The resulting residue was dry-loaded onto silica and purified via flash column chromatography (0-30% Hex:EA) to afford tert-butyl 4-(4-((6- bromoisoquinolin-3-yl)carbamoyl)pyridin-2-yl)piperazine-1-carboxylate (22) (67 mg, 131 µmol, 64 %). LCMS [M+H]+513.1H NMR (500 MHz, CDCl3) δ 9.24 (s, 1H), 8.99 (d, J = 0.9 Hz, 1H), 8.71 (s, 1H), 8.42 – 8.31 (m, 2H), 8.06 (d, J = 1.8 Hz, 1H), 7.80 (d, J = 8.7 Hz, 1H), 7.67 – 7.59 (m, 1H), 7.10 (dd, J = 5.2, 1.2 Hz, 1H), 3.57 (d, J = 5.5 Hz, 8H), 1.49 (d, J = 1.1 Hz, 9H). OOHC N Boc 2-(4-(tert-butoxycar n-1-yl)isonicotinic acid (23) To a 5 mL microwave vial with tert-butyl 4-(4-cyanopyridin-2-yl)piperazine-1-carboxylate (24) (1.69 g, 1 eq, 5.86 mmol) was added ethanol (14 mL). The reaction was allowed to stir for 5 minutes before sodium hydroxide (586 mg, 12.2 mL, 1.2 molar, 2.5 eq, 14.7 mmol) was added. This reaction mixture was refluxed overnight. Upon completion the mixture was cooled to room temperature and the EtOH was removed under vacuum. The remaining solution was cooled in an ice bath, treated with water, and adjusted PH to ~3 with citric acid. The mixture was extracted with EtOAc, washed with brine, dried over sodium sulfate, filtered, and concentrated. The crude mixture was dry-loaded onto silica and purified via flash chromatography (DCM:MeOH 0-5%) to give 2-(4-(tert-butoxycarbonyl)piperazin-1-yl)isonicotinic acid (23) (1.43 g, 4.65 mmol, 79 %). LCMS [M+H]+308.1H NMR (500 MHz, CDCl3) δ 8.33 (d, J = 5.1 Hz, 1H), 7.27 (d, J = 1.9 Hz, 1H), 7.20 (d, J = 5.1 Hz, 1H), 3.67 – 3.51 (m, 8H), 1.49 (s, 9H). CN N Boctert-butyl 4-(4-cyano iperazine-1-carboxylate (24) To a 100 mL round bottom flask, tert-butyl piperazine-1-carboxylate (2 g, 1 eq, 8 mmol), potassium carbonate (3 g, 3 eq, 0.02 mol), and 2-fluoroisonicotinonitrile (1 g, 1 eq, 8 mmol) were added and dissolved in DMF (30 mL). The reaction mixture was stirred for 3 hours at 110 °C. Upon completion, the reaction mixture was cooled and extracted in EtOAc, washed with water (x2) and brine (x3), dried over sodium sulfate, and concentrated in vacuo. The crude mixture was dry-loaded onto silica and purified via flash column chromatography (EtOAc:Hex 0-10%) to afford tert-butyl 4-(4-cyanopyridin-2-yl)piperazine-1-carboxylate (24) (1.79 g, 6.21 mmol, 80 %). LCMS [M+H]+289. This compound was used without further characterization. NH2N N 4-(((1-(1 erazin-1- yl)prop-1 -yn-1-yl)pyridin-2-yl)piperidine-4-carbonyl)piperidin-4-yl)-1H-pyrazol-4- yl)methyl)amino)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (25) DMF (2.0 mL) and triethylamine (27 mg, 37 μL, 5 eq, 0.26 mmol) were added sequentially to a dry 5 mL microwave vial equipped with a magnetic stir bar containing 4-(4-(3-fluoro-4-(4- (prop-2-yn-1-yl)piperazin-1-yl)phenyl)quinazolin-6-yl)pyridin-2-amine (26) (23 mg, 1 eq, 52 μmol), 2-(2,6-dioxopiperidin-3-yl)-4-(((1-(1-(1-(5-iodopyridin-2-yl)piperidine-4- carbonyl)piperidin-4-yl)-1H-pyrazol-4-yl)methyl)amino)isoindoline-1,3-dione (39 mg, 1 eq, 52 μmol), PdCl2(dppf) (5.0 mg, 0.13 eq, 6.8 μmol) and copper(I) iodide (2.5 mg, 0.25 eq, 13 μmol). This was stirred for 24 hours at room temperature where it was concentrated under reduced pressure and purified by flash column chromatography to afford 4-(((1-(1-(1-(5-(3-(4-(4-(6-(2- aminopyridin-4-yl)quinazolin-4-yl)-2-fluorophenyl)piperazin-1-yl)prop-1-yn-1-yl)pyridin-2- yl)piperidine-4-carbonyl)piperidin-4-yl)-1H-pyrazol-4-yl)methyl)amino)-2-(2,6-dioxopiperidin- 3-yl)isoindoline-1,3-dione (25) (20 mg, 19 μmol, 36 %). LCMS [M+H]+1061.1H NMR (400 MHz, DMSO-D6) δ 9.29 (s, 1H), 8.30 – 8.22 (m, 2H), 8.15 (d, J = 1.8 Hz, 2H), 7.99 (d, J = 5.3 Hz, 1H), 7.76 (d, J = 2.6 Hz, 1H), 7.69 (d, J = 2.0 Hz, 1H), 7.64 (d, J = 7.7 Hz, 2H), 7.54 – 7.49 (m, 2H), 7.43 (s, 1H), 7.23 (t, J = 8.7 Hz, 1H), 7.12 (d, J = 8.6 Hz, 1H), 7.00 (d, J = 7.1 Hz, 1H), 6.83 (dd, J = 5.4, 1.7 Hz, 1H), 6.78 (d, J = 8.5 Hz, 2H), 6.71 (d, J = 1.6 Hz, 1H), 6.06 (s, 2H), 5.01 – 4.94 (m, 1H), 4.32 (t, J = 5.9 Hz, 4H), 4.08 (d, J = 13.3 Hz, 2H), 3.55 (m, 3H), 3.21 (q, J = 4.7 Hz, 6H), 2.96 – 2.88 (m, 3H), 2.69 (t, J = 4.7 Hz, 4H), 2.51 (d, J = 3.8 Hz, 2H), 1.98 (dd, J = 19.2, 11.0 Hz, 4H), 1.63 (d, J = 11.9 Hz, 4H), 1.47 (t, J = 14.3 Hz, 2H). Experiments were conducted with Compound 25 demonstrating: HEK293- DYRK1A(HiBiT) > 25% @300 nM degradation. HEK293-DYRK1B(HiBiT) > 25% @100 nM. Degradation. DYRK1A KD 19 nM. N N F 4-(4-(3-fluoro-4-(4-(prop-2-yn-1-yl l)quinazolin-6-yl)pyridin-2-amine (26) 3-bromoprop-1-yne (22 mg, 17 μL, 80% Wt, 1 eq, 0.15 mmol) was added to a 5 mL microwave vial equipped with a magnetic stir bar containing 4-(4-(3-fluoro-4-(piperazin-1- yl)phenyl)quinazolin-6-yl)pyridin-2-amine (27) (60 mg, 1 eq, 0.15 mmol) and potassium carbonate (41 mg, 2 eq, 0.30 mmol) in DMF. The reaction mixture was heated at 60 °C for 16 hours where the reaction mixture was then cooled down to room temperature and washed with saturated sodium bicarbonate (5 mL). The mixture was extracted with DCM (3 x 15 mL) and the organic layers were recombined, dried over sodium sulfate, and concentrated under reduced pressure. The crude residue was purified by automated flash column chromatography using a DCM / MeOH gradient to afford 4-(4-(3-fluoro-4-(4-(prop-2-yn-1-yl)piperazin-1- yl)phenyl)quinazolin-6-yl)pyridin-2-amine (26) (23 mg, 52 μmol, 35 %). LCMS [M+H]+439.1H NMR (400 MHz, CDCl3) δ 9.35 (s, 1H), 8.35 (d, J = 2.2 Hz, 1H), 8.18 (d, J = 8.7 Hz, 1H), 8.15 – 8.08 (m, 2H), 7.63 – 7.53 (m, 2H), 7.13 (t, J = 8.4 Hz, 1H), 6.92 (dd, J = 5.5, 1.6 Hz, 1H), 6.77 (d, J = 1.6 Hz, 1H), 4.93 (s, 2H), 3.40 (d, J = 2.5 Hz, 2H), 3.32 (t, J = 4.9 Hz, 4H), 2.81 (t, J = 4.9 Hz, 4H), 2.31 (q, J = 2.4 Hz, 1H).13C NMR (101 MHz, CDCl3) δ 158.69, 155.16, 152.01, 151.45, 149.61, 147.75, 138.05, 132.58, 130.40, 129.97, 126.82, 125.11, 123.04, 120.90, 118.92, 118.20, 117.98, 112.76, 107.03, 78.50, 73.71, 51.88, 50.02, 47.01. N F 4-(4-(3-fluoro-4-(piperazi lin-6-yl)pyridin-2-amine (27) A 5 mL microwave vial was loaded with tert-butyl 4-(4-(6-bromoquinazolin-4-yl)-2- fluorophenyl)piperazine-1-carboxylate (28) (110 mg, 1 eq, 226 μmol), tert-butyl (4-(4,4,5,5- tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-yl)carbamate (80 mg, 1.1 eq, 248 μmol), PdCl2(dppf) (18 mg, 0.1 eq, 23 μmol), and tripotassium phosphate (72 mg, 1.5 eq, 339 μmol), the vial capped then degassed for 10 minutes then injected with solvents degassed 1,4- Dioxane (0.75 mL):Water (0.13 mL), (6:1 v / v), degassed further for 10 minutes and heated to 110 °C for 3 hours in an oil bath. Upon completion, the solution was diluted with DCM (5 mL). TFA (3.67 g, 2.48 mL, 66 eq, 32.2 mmol) was added to the solution and allowed to stir at 25 °C for 2 hours. Upon completion the solution was basified with 1 M NaOH, and extracted with DCM (3 x 20 mL). The organic layer was removed under vacuum and purified via column chromatography (DCM: MeOH 0-5%) to afford 4-(4-(3-fluoro-4-(piperazin-1- yl)phenyl)quinazolin-6-yl)pyridin-2-amine (27) (136 mg, 340 μmol, 70%) as a yellow solid. LCMS [M+H]+400.1H NMR (400 MHz, DMSO-D6) δ 9.27 (s, 1H), 8.29 (d, J = 2.0 Hz, 1H), 8.26 – 8.21 (m, 2H), 7.98 (d, J = 5.4 Hz, 1H), 7.66 – 7.60 (m, 2H), 7.18 (t, J = 8.8 Hz, 1H), 6.81 (dd, J = 5.3, 1.8 Hz, 1H), 6.71 (d, J = 1.7 Hz, 1H), 6.02 (s, 1H), 3.07 (t, J = 4.9 Hz, 4H), 2.84 (q, J = 7.0 Hz, 4H), 1.03 (s, 1H).13C NMR (101 MHz, CDCl3) δ 167.13, 159.24, 156.49, 154.87, 154.32, 151.19, 149.06, 148.71, 138.37, 132.74, 129.65, 126.87, 124.97, 123.38, 122.99, 118.84, 117.93, 112.59, 106.76, 50.40, 45.96. N F tert-butyl 4-(4-(6-(2-aminopyr 4-yl)-2-fluorophenyl)piperazine-1- carboxylate (28) In a vial 6-bromo-4-chloroquinazoline (346 mg, 1 eq, 1.42 mmol), tert-butyl 4-(2-fluoro-4- (4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)piperazine-1-carboxylate (29) (578 mg, 1 eq, 1.42 mmol), PdCl2(dppf) (52 mg, 0.05 eq, 71 μmol), tripotassium phosphate (906 mg, 3 eq, 4.27 mmol) and purged for 10 minutes. Dioxane (12 mL) and water (2.0 mL) (6:1) were added to the vial and purged for an additional 10 minutes. The reaction was placed in the oil bath at 90 °C for 16 hours. Upon completion, the reaction was cooled and diluted with EtOAc. The organic layer was washed with brine (20 mL) and water (20 mL). The recovered organic layer was filtered through celite and dried with sodium sulfate. The solvent was removed under vacuum and then separated through column chromatography (0-10% DCM / MeOH) to afford tert-butyl 4-(4-(6-bromoquinazolin-4-yl)-2-fluorophenyl)piperazine-1-carboxylate (28) (502 mg, 1.03 mmol, 73 %) as a yellow solid. LCMS [M+H]+487.1H NMR (500 MHz, CDCl3) δ 9.32 (s, 1H), 8.30 (dd, J = 1.7, 0.9 Hz, 1H), 7.99 – 7.95 (m, 2H), 7.56 – 7.49 (m, 2H), 7.09 (t, J = 8.6 Hz, 1H), 3.63 (t, J = 5.0 Hz, 4H), 3.17 (t, J = 5.0 Hz, 4H), 1.48 (s, 9H).13C NMR (126 MHz, CDCl3) δ 165.73, 156.23, 154.69, 154.26, 149.81, 142.04, 137.31, 130.72, 130.39, 128.92, 126.71, 123.89, 121.71, 118.96, 117.98, 80.06, 50.15, 50.12, 28.45. F tert-butyl 4-(2-fluoro-4-(4,4,5,5-tetra methyl-1,3,2-dioxaborolan-2-yl)phenyl)piperazine-1- carboxylate (29) A microwave vial containing Bis(pinacolato)diborane (203 mg, 1.2 eq, 798 μmol), PdCl2(dppf) (26 mg, 0.05 eq, 33 μmol), and potassium acetate (196 mg, 3 eq, 2.00 mmol) was capped, purged with argon, then tert-butyl 4-(4-bromo-2-fluorophenyl)piperazine-1-carboxylate (30) (239 mg, 1 eq, 665 μmol) was added in DMF (1.6 mL) . The mixture was purged with argon and heated to 80 °C for 5 hours in an oil bath. The reaction was cooled, diluted with EtOAc, and poured into a separatory funnel containing EtOAc and saturated ammonium chloride solution. The mixture was shaken and filtered through celite. After multiple extractions with EtOAc (500 mL EtOAc), the organics were washed with saturated ammonium chloride solution (40 mL x 2) and brine (40 mL) to produce tert-butyl 4-(2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2- yl)phenyl)piperazine-1-carboxylate (29) (240 mg, 591 μmol, 89 %) as a white solid. LCMS [M+H]+406.1H NMR (500 MHz, CDCl3) δ 7.31 – 7.24 (m, 1H), 7.23 (dd, J = 13.3, 1.5 Hz, 1H), 6.67 (d, J = 8.2 Hz, 1H), 3.37 (t, J = 5.0 Hz, 4H), 2.86 (t, J = 5.0 Hz, 4H), 1.26 (s, 9H), 1.11 (s, 12H).13C NMR (126 MHz, CDCl3) δ 155.94, 154.68, 153.98, 142.36, 131.37, 121.98, 118.17, 83.81, 79.85, 50.15, 28.42, 24.83. Br F tert-butyl 4-(4-bromo-2-f iperazine-1-carboxylate (30) In a 100 mL round bottom flask purged and maintained with an inert atmosphere of argon, was added 4-bromo-2-fluoro-1-iodobenzene (1.68 g, 1 eq, 5.57 mmol) in toluene (32 mL). Then Pd2(dba)3(153 mg, 0.03 eq, 167 μmol), cesium carbonate (4.54 g, 2.5 eq, 13.9 mmol), 4,5- Bis(diphenylphosphino)-9,9-dimethyl xanthene (322 mg, 0.1 eq, 557 μmol), and tert-butyl piperazine-1-carboxylate (1.04 g, 1 eq, 5.57 mmol) were added. The resulting solution was stirred at 100 °C for 24 hours. The reaction was then quenched by adding 25 mL of water and extracted with 25 mL of ethyl acetate. The organic layers were combined and dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude residue was purified by automated column chromatography using a CombiFlash (0-15% EtOAc / Hex) to give tert-butyl 4-(4-bromo-2-fluorophenyl)piperazine-1-carboxylate (30) (634 mg, 1.76 mmol, 32 %) as a light-yellow solid. LCMS [M+H]+359.1H NMR (500 MHz, DMSO) δ 7.47 – 7.43 (m, 1H), 7.31 (ddd, J = 8.6, 2.3, 1.0 Hz, 1H), 7.00 (t, J = 9.0 Hz, 1H), 3.46 (t, J = 5.0 Hz, 4H), 2.96 – 2.91 (m, 4H), 1.42 (s, 9H). INCORPORATION BY REFERENCE The entire disclosure of each of the patent documents and scientific articles referred to herein is incorporated by reference for all purposes. EQUIVALENTS The invention may be embodied in other specific forms without departing from the spirit or essential characteristics thereof. The foregoing embodiments are therefore to be considered in all respects illustrative rather than limiting the invention described herein. Scope of the invention is thus indicated by the appended claims rather than by the foregoing description, and all changes that come within the meaning and range of equivalency of the claims are intended to be embraced therein.

Claims

What Is Claimed Is:

1. A bifunctional compound comprising the chemical structure: TBM-L-ULM or TBM-ULM, wherein: TBM is a binding moiety for one or more of DYRK1A, DYRK1B, DYRK2, DYRK3, DYRK4, CLK1, CLK2, CLK3, CLK4, CDK7, CDK8 / 19, PI3K, PDGFrA / B, mTOR, HIPKs, and / or CMGC kinases leading to inhibition of WNT signaling; L is a bond or a chemical linker that covalently compounds the TBM and the ULM; and ULM is an E3 ubiquitin ligase binding moiety.

2. The bifunctional compound of claim 1, wherein TBM is a small molecule inhibitor of one or more of DYRK1A, DYRK1B, DYRK2, DYRK3, DYRK4, CLK1, CLK2, CLK3, CLK4, CDK7, CDK8 / 19, PI3K, PDGFrA / B, mTOR, HIPKs, and / or CMGC kinases leading to inhibition of WNT signaling.

3. The bifunctional compound of claim 2, wherein the small molecule inhibitor of one or more of DYRK1A, DYRK1B, DYRK2, DYRK3, DYRK4, CLK1, CLK2, CLK3, CLK4, CDK7, CDK8 / 19, PI3K, PDGFrA / B, mTOR, HIPKs, and / or CMGC kinases leading to inhibition of WNT signaling is selected from ,;alently coupleswith the linker.

4. The bifunctional compound of claim 2, wherein the small molecule inhibitor of one or more of DYRK1A, DYRK1B, DYRK2, DYRK3, DYRK4, CLK1, CLK2, CLK3, CLK4, CDK7, CDK8 / 19, PI3K, PDGFrA / B, mTOR, HIPKs, and / or CMGC kinases leading to inhibition ofWNT signaling is selected from: ,,wherei ” indicates the location where the small molecule covalently coupleswith the linker5. The bifunctional compound of claim 1, wherein the ULM covalently couples with the linker at any location within the ULM.

6. The bifunctional compound of claim 1, wherein the ULM is selected from:),F nd hthe7. The bifunctional compound of claim 1, wherein the linker is selected from:the linker is selected from:and8. The bifunctional compound of claim 1, wherein TBM-L-ULM is represented by TBM-A-B-modified-cereblon; wherein A is represente ;;wherein R is OH or H; wherein X is CH2 or CO; wherein each X1is either N or CH; wherein X2is CO, CH2, or NH; wherein X3is NH or O.

9. The bifunctional compound of claim 1, wherein the bifunctional compound is selected N (CH2)n,and, n Example I.

11. A pharmaceutical composition comprising a compound of Claim 1.

12. A method of treating, ameliorating, or preventing a disorder related to one or more of DYRK1A activity, DYRK1B activity, DYRK2 activity, DYRK3 activity, DYRK4 activity, CLK1 activity, CLK2 activity, CLK3 activity, and CLK4 activity in a patient comprising administering to said patient a therapeutically effective amount of the pharmaceutical composition of Claim 11.

13. The method of Claim 12, wherein said disorder is selected from Alzheimer’s disease, Down syndrome, Huntington’s disease, Parkinson’s disease, an autoimmune disease, an inflammatory disorder (e.g., airway inflammation, osteoarthritis (e.g., knee related osteoarthritis)), diabetes, or cancer (e.g., glioblastoma, prostate cancer, metastatic breast cancer, metastatic lung cancer, multiple myeloma, secondary metastatic tumors of the brain, colorectal cancer, acute myeloid leukemia, myelodysplastic syndrome), a viral infection (e.g., SARS-CoV- 2 infection (e.g., COVID-19)), and other diseases).

14. The method of Claim 12, wherein said patient is a human patient.

15. The method of Claim 12, further comprising administering to said patient one or more agents for treating Alzheimer’s disease, Down syndrome, Huntington’s disease, Parkinson’s disease, autoimmune disease, an inflammatory disorder (e.g., airway inflammation, osteoarthritis (e.g., knee related osteoarthritis)), diabetes, or cancer (e.g., glioblastoma, prostate cancer, metastatic breast cancer, metastatic lung cancer, multiple myeloma, secondary metastatic tumors of the brain, colorectal cancer), a viral infection (e.g., SARS-CoV-2 infection (e.g., COVID- 19)), and other diseases).

16. A kit comprising a compound of Claim 1 and instructions for administering said compound to a patient having a disorder related to one or more of DYRK1A activity, DYRK1B activity, DYRK2 activity, DYRK3 activity, DYRK4 activity, CLK1 activity, CLK2 activity, CLK3 activity, and CLK4 activity.

17. The kit of Claim 16, wherein the disorder is Alzheimer’s disease, Down syndrome, Huntington’s disease, Parkinson’s disease, autoimmune disease, inflammatory disorder (e.g., airway inflammation, osteoarthritis (e.g., knee related osteoarthritis)), diabetes, or cancer (e.g., glioblastoma, prostate cancer, metastatic breast cancer, metastatic lung cancer, multiple myeloma, secondary metastatic tumors of the brain, colorectal cancer, acute myeloid leukemia, myelodysplastic syndrome), a viral infection (e.g., SARS-CoV-2 infection (e.g., COVID-19)), and other diseases).

18. The kit of Claim 16, further comprising one or more agents for treating Alzheimer’s disease, Down syndrome, Huntington’s disease, Parkinson’s disease, autoimmune disease, aninflammatory disorder (e.g., airway inflammation, osteoarthritis (e.g., knee related osteoarthritis)), diabetes, or cancer (e.g., glioblastoma, prostate cancer, metastatic breast cancer, metastatic lung cancer, multiple myeloma, secondary metastatic tumors of the brain, colorectal cancer), a viral infection (e.g., SARS-CoV-2 infection (e.g., COVID-19)), and other diseases).

19. A method for inhibiting one or more of DYRK1A activity, DYRK1B activity, DYRK2 activity, DYRK3 activity, DYRK4 activity, CLK1 activity, CLK2 activity, CLK3 activity, and CLK4 activity in a subject, comprising administering to said patient a therapeutically effective amount of the pharmaceutical composition of Claim 11.

20. The method of Claim 19, wherein administration of the compound results in inhibition of one or more of DYRK1A related PI3K / Akt signaling; DYRK1A related tau phosphorylation; DYRK1A related NFAT phosphorylation; DYRK1A related ASK1 / JNK1 pathway activation; DYRK1A related p53 phosphorylation; DYRK1A related Amph 1 phosphorylation; DYRK1A related Dynamin 1 phosphorylation; DYRK1A related Synaptojanin phosphorylation; DYRK1A related presenilin 1 (the catalytic sub-unit of γ-secretase) activity; DYRK1A related amyloid precursor protein phosphorylation; DYRK1A related SIRT1 activation; DYRK2 related heat shock factor 1 and 26S proteasome activities; DYRK3 related mTOR activity; DYRK3 phosphorylation (e.g., PRAS40); DYRK1B activity; CMGC / CLK kinase activity; CLK1 activity; CLK2 activity; CLK3 activity; CLK4 activity; and WNT signaling.

21. The method of Claim 19, wherein the subject is human subject suffering from or at risk for developing a disorder related to DYRK1A, DYRK1B, DYRK2, DYRK3, and / or CLK1, CLK2, CLK3 and CLK4 activity.

22. The method of Claim 19, wherein the disorder is Alzheimer’s disease, Down syndrome, Huntington’s disease, Parkinson’s disease, autoimmune disease, an inflammatory disorder (e.g., airway inflammation, osteoarthritis (e.g., knee related osteoarthritis)), diabetes, or cancer (e.g., glioblastoma, prostate cancer, metastatic breast cancer, metastatic lung cancer, multiple myeloma, secondary metastatic tumors of the brain, colorectal cancer), a viral infection (e.g., SARS-CoV-2 infection (e.g., COVID-19)), and other diseases).

23. A method of treating cancer in a subject in need thereof, the method comprising administering to the subject 1) a therapeutically effective amount of the pharmaceutical composition of Claim 11, and 2) at least one anticancer agent.

24. The method of claim 23, wherein the cancer is one or more types of cancer selected from glioblastoma, prostate cancer, metastatic breast cancer, metastatic lung cancer, multiple myeloma, secondary metastatic tumors of the brain, colorectal cancer, acute myeloid leukemia, myelodysplastic syndrome.

25. The method of claim 23, wherein the at least one anticancer agent comprises a chemotherapeutic agent, an immune checkpoint inhibitor, or radiation therapy.

26. The method of claim 25, wherein the immune checkpoint inhibitor is one or more selected from: pembrolizumab, nivolumab, cemipilimab, atezolizumab, avelumab, durvalumab, and ipilimumab.

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