Inhibitors of HIF-alpha in tumor cells and method of use thereof

Abstract

This invention provides a compound having the following structure: ( I ) wherein the compound is used for anti-cancer treatments. The invention also provides an assay for selecting one or more compounds that inhibit interaction between ID2 protein and the tumor suppressor complex CRLVHL.

Description

Docket: 92169-A-PCT / GJG / YX / RBINHIBITORS OF HIF-ALPHA IN TUMOR CELLS AND METHOD OF USE THEREOF

[0001] This application claims the benefit of U. S. Provisional Application No. 63 / 720,555, filed November 14, 2024, the contents of which is hereby incorporated by reference.

[0002] Throughout this application, various publications are referenced, including referenced in parenthesis. The disclosures of all publications mentioned in this application in their entireties are hereby incorporated by reference into this application in order to provide additional description of the art to which this invention pertains and of the features in the art which can be employed with this invention.STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT

[0003] This invention was made with government support under CA239721 awarded by the National Institutes of Health. The government has certain rights in the invention.BACKGROUND OF THE INVENTION

[0004] Hypoxia-inducible factor- 1 alpha (“HIF-a”) transcription factors generally play the role of master regulators of cancer stem cells, and tumor angiogenesis, and also result in poor clinical outcome of most human neoplasms. HIF-a transcription factors have been demonstrated to drive tumor growth and angiogenesis in several cancers including gliomas (Soeda, 2009). One way in which these proteins are negatively regulated is by E3 ligase mediated degradation. The HIF-a E3 ligase consists of the protein Cul2 presenting the Elongin-B, Elongin-C and VHL complex (VBC), which is the HIF-a recognition motif (Maxwell, 2001). It has recently been disclosed that the ID2 protein binds to VHL and displaces it from the E3 ligase complex, preventing HIF-a degradation in glioma cells (Lee, 2016). The four ID proteins are transcriptional regulators of stem cells which are coopted in certain cancers to drive stem cell like behaviors and promote angiogenesis (Lasorella, 2014; Perk, 2005). The ID2 protein itself is associated with glioma sternness, tumor angiogenesis and is part of the genetic signature of terminal high grade gliomas (Lasorella, 2005; Niola, 2013; Vandeputte, 2002). It is anticipated that compounds able to block the interaction of ID2 and VHL have potential as anti-cancer therapeutics. Such compounds would restore the VHL E3 ligase function and inhibit tumor growth and angiogenesis.14935-6729-8938v.lTherefore, compounds that emerge as agonists of HIF-α promise to have therapeutic value as highly effective anti-tumor agents.24935-6729-8938v. 1BRIEF SUMMARY OF THE INVENTION

[0005] The present invention provides a compound having the following structure:whereinA is a ring;X is NH, O, or S;Ri is an aryl or heteroaryl;R2 and R3 are each independently aryl, heteroaryl, cycloalkyl or heterocycloalkyl;R4andR? are each independently alkyl, alkenyl, alkynyl, -CN; -OH, -O-alkyl, -NH2, -NH- alkyl, -NH-(CO)-alkyl, or -NH-(SO2)-alkyl;Rs is H, alkyl, alkenyl, alkynyl, -OH, -O-alkyl, -NH2, -NH-alkyl, -NH-(CO)-alkyl, or -NH- (SO2)-alkyl;R7 is aryl or heteroaryl;Rs is cycloalkyl, heterocycloalkyl, alkenyl, alkynyl, aryl, or heteroaryl;R9 and Rio are each independently H, alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, or heteroaryl;Y is CR17 or N; andR11, R12, R13, R14, RIS, Rie, and R17 are each independently H, alkyl, alkenyl, alkynyl, aryl, heterocycloalkyl, alkyl-heterocycloalkyl, heteroaryl, -(CO2)-alkyl, -(CO2)-aryl -NH2, -NH- alkyl, -NH-alkenyl, -NH-alkynyl, -NH-aryl, -NH-heteroaryl, -OH, -OAc, -O-C(O)alkyl,, halo, -CF3, -OCFs-O-alkyl, -O-alkenyl, -O-aryl, -S-(alkyl), -S-(alkenyl), -S-(alkynyl), - S(O)-(alkyl), -SO2-(alkyl), -SO2-(aryl), -SO2-(heteroaryl), alkyl-OH, alkyl-O-alkyl, alkyl- NH2, alkyl-NH-alkyl, or alkyl-N(alkyl)2.34935-6729-8938v. 1

[0006] The present invention provides an assay for selecting one or more compounds that inhibit interaction between ID2 protein and tumor suppressor complex CRLVHLfrom a pool of compounds; wherein the assay comprises donor beads and acceptor beads.

[0007] The present invention provides a method of inhibiting interaction between ID2 protein and tumor suppressor complex CRLVHLcomprising contacting the ID2 protein or the suppressor complex CRLVHIwith the compound having the following structure:whereinA is a ring;X is NH, O, or S;Ri is an aryl or heteroaryl;R2 and R3 are each independently aryl, heteroaryl, cycloalkyl or heterocycloalkyl;R4andR? are each independently alkyl, alkenyl, alkynyl, -CN; -OH, -O-alkyl, -NH2, -NH- alkyl, -NH-(CO)-alkyl, or -NH-(SO2)-alkyl;Re is H, alkyl, alkenyl, alkynyl, -OH, -O-alkyl, -NH2, -NH-alkyl, -NH-(CO)-alkyl, or -NH- (SO2)-alkyl;R7 is aryl or heteroaryl;Rs is cycloalkyl, heterocycloalkyl, alkenyl, alkynyl, aryl, or heteroaryl;R9 and Rio are each independently H, alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, or heteroaryl;Y is CR17 or N; andR11, R12, Ri3, Ri4, R15, Rie, and R17 are each independently H, alkyl, alkenyl, alkynyl, aryl, heterocycloalkyl, alkyl-heterocycloalkyl, heteroaryl, -(CO2)-alkyl, -(CO2)-aryl -NH2, -NH- alkyl, -NH-alkenyl, -NH-alkynyl, -NH-aryl, -NH-heteroaryl, -OH, -OAc, -O-C(O)alkyl,,44935-6729-8938v. 1halo, -CF3, -OCFs-O-alkyl, -O-alkenyl, -O-aryl, -S-(alkyl), -S-(alkenyl), -S-(alkynyl), - S(O)-(alkyl), -SO2-(alkyl), -SO2-(aryl), -SO2-(heteroaryl), alkyl-OH, alkyl-O-alkyl, alkyl- NH2, alkyl-NH-alkyl, or alkyl-N(alkyl)2.

[0008] The present invention provides a method of inhibiting proliferation of tumor or cancer cells comprising contacting ID2 protein or suppressor complex CRLVHLwith the compound having the following structure:whereinA is a ring;X is NH, O, or S;Ri is an aryl or heteroaryl;R2 and R3 are each independently aryl, heteroaryl, cycloalkyl or heterocycloalkyl;R4andRs are each independently alkyl, alkenyl, alkynyl, -CN; -OH, -O-alkyl, -NH2, -NH- alkyl, -NH-(CO)-alkyl, or -NH-(SO2)-alkyl;Re is H, alkyl, alkenyl, alkynyl, -OH, -O-alkyl, -NH2, -NH-alkyl, -NH-(CO)-alkyl, or -NH- (SO2)-alkyl;R7 is aryl or heteroaryl;Rs is cycloalkyl, heterocycloalkyl, alkenyl, alkynyl, aryl, or heteroaryl;R9 and Rio are each independently H, alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, or heteroaryl;Y is CR17 or N; andR11, R12, Ri3, Ri4, R15, Rie, and R17 are each independently H, alkyl, alkenyl, alkynyl, aryl, heterocycloalkyl, alkyl-heterocycloalkyl, heteroaryl, -(CO2)-alkyl, -(CO2)-aryl -NH2, -NH-54935-6729-8938v. 1alkyl, -NH-alkenyl, -NH-alkynyl, -NH-aryl, -NH-heteroaryl, -OH, -OAc, -O-C(O)alkyl,, halo, -CF3, -OCFs-O-alkyl, -O-alkenyl, -O-aryl, -S-(alkyl), -S-(alkenyl), -S-(alkynyl), - S(O)-(alkyl), -SO2-(alkyl), -SO2-(aryl), -SO2-(heteroaryl), alkyl-OH, alkyl-O-alkyl, alkyl- NH2, alkyl-NH-alkyl, or alkyl-N(alkyl)2.

[0009] The present invention provides a method of suppressing activity of tumor or cancer cells comprising contacting ID2 protein or suppressor complex CRLVHTwith the compound having the following structure:whereinA is a ring;X is NH, O, or S;Ri is an aryl or heteroaryl;R2 and R3 are each independently aryl, heteroaryl, cycloalkyl or heterocycloalkyl;R4andRs are each independently alkyl, alkenyl, alkynyl, -CN; -OH, -O-alkyl, -NH2, -NH- alkyl, -NH-(CO)-alkyl, or -NH-(SO2)-alkyl;Re is H, alkyl, alkenyl, alkynyl, -OH, -O-alkyl, -NH2, -NH-alkyl, -NH-(CO)-alkyl, or -NH- (SO2)-alkyl;R7 is aryl or heteroaryl;Rs is cycloalkyl, heterocycloalkyl, alkenyl, alkynyl, aryl, or heteroaryl;R9 and Rio are each independently H, alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, or heteroaryl;Y is CR17 or N; and64935-6729-8938v. 1Rn, R12, R13, R14, R15, Ri6, and R17 are each independently H, alkyl, alkenyl, alkynyl, aryl, heterocycloalkyl, alkyl-heterocycloalkyl, heteroaryl, -(CC>2)-alkyl, -(CO2)-aryl -NH2, -NH- alkyl, -NH-alkenyl, -NH-alkynyl, -NH-aryl, -NH-heteroaryl, -OH, -OAc, -O-C(O)alkyl,, halo, -CF3, -OCFs-O-alkyl, -O-alkenyl, -O-aryl, -S-(alkyl), -S-(alkenyl), -S-(alkynyl), - S(O)-(alkyl), -SO2-(alkyl), -SO2-(aryl), -SO2-(heteroaryl), alkyl-OH, alkyl-O-alkyl, alkyl- NH2, alkyl-NH-alkyl, or alkyl-N(alkyl)2.

[0010] The present invention provides a method of treating tumor or cancer in a subject comprising administering the compound having the following structure:whereinA is a ring;X is NH, O, or S;Ri is an aryl or heteroaryl;R2 and R3 are each independently aryl, heteroaryl, cycloalkyl or heterocycloalkyl;R4andRs are each independently alkyl, alkenyl, alkynyl, -CN; -OH, -O-alkyl, -NH2, -NH- alkyl, -NH-(CO)-alkyl, or -NH-(SO2)-alkyl;Re is H, alkyl, alkenyl, alkynyl, -OH, -O-alkyl, -NH2, -NH-alkyl, -NH-(CO)-alkyl, or -NH- (SO2)-alkyl;R7 is aryl or heteroaryl;Rs is cycloalkyl, heterocycloalkyl, alkenyl, alkynyl, aryl, or heteroaryl;R9 and Rio are each independently H, alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, or heteroaryl;Y is CR17 or N; and74935-6729-8938v. 1Rn, R12, R13, R14, R15, Ri6, and R17 are each independently H, alkyl, alkenyl, alkynyl, aryl, heterocycloalkyl, alkyl-heterocycloalkyl, heteroaryl, -(CC>2)-alkyl, -(CO2)-aryl -NH2, -NH- alkyl, -NH-alkenyl, -NH-alkynyl, -NH-aryl, -NH-heteroaryl, -OH, -OAc, -O-C(O)alkyl,, halo, -CF3, -OCFs-O-alkyl, -O-alkenyl, -O-aryl, -S-(alkyl), -S-(alkenyl), -S-(alkynyl), - S(O)-(alkyl), -SO2-(alkyl), -SO2-(aryl), -SO2-(heteroaryl), alkyl-OH, alkyl-O-alkyl, alkyl- NH2, alkyl-NH-alkyl, or alkyl-N(alkyl)2.

[0011] The present invention provides a method of selecting one or more compounds that inhibit interaction between ID2 protein and tumor suppressor complex CRIHLfrom a pool of compounds using an assay; wherein the assay comprises donor beads and acceptor beads, wherein the method comprises the following steps:a) adding, in any order:i) glutathione S-transferase tagged von Hippel-Lindau tumor suppressor protein and Elongin C protein complex (GST-VHL / EloC);ii) one or more compounds from the pool of compounds;iii) FLAG tags;iv) the acceptor beads; andv) the donor beads;b) exciting a luminescent signal at a wavelength between 670-690 nm, preferably at 680 nm; andc) obtaining an emission signal at a wavelength between 610-625 nm, preferably at 617 nm.84935-6729-8938v. 1BRIEF DESCRIPTION OF THE DRAWINGS

[0012] Figure 1. A representation of the AlphaScreen® binding assay used to identify and quantify inhibitor hit compounds, on a molecular scale. As shown, VHL and ID2 proteins are immobilized to donor and acceptor beads, respectively. Excitation of the donor beads by light of 680 nm wavelength is followed by energy transfer to the acceptor beads which in turn emit light at 520-620 nm wavelengths. Such emission is detectable and the intensity is quantifiable. While ID2 interacts with VHL, the donor and acceptor beads are within proximity to permit efficient energy transfer between the two. When the ID2-VHL interaction does not exist, either by inhibition or not colliding in solution, the resulting energy transfer upon excitation, and ensuing emission, does not occur. Inhibition of the ID2-VHL interaction by a compound is quantified by the relative intensity of the emission detected. Hit compounds were identified implementing a high-throughput screen using this assay.

[0013] Figure 2. SPR traces of 4 compounds demonstrating 1:1 binding mode. Each line represents a different concentration used to calculate the KD of each compound to the VHL complex, listed below each trace. Top panels: D411 series compounds. Bottom panels: left panelcompound 3112-5273; right panel- 3112-5273 neighbor compound (Tanimoto distance 0.0369).

[0014] Figure 3. Pharmacophore model of D411 molecule series indicating the location and type of productive intermolecular interactions.

[0015] Figure 4. SDS-PAGE gel scan of pull-down experiments demonstrating the inhibition of the ID2-VHL interaction in the presence or absence (DMSO) of hit compounds confirmed by SPR binding experiments, including D411 molecules. The highlighted compounds are distinguished by their specific type of interactions with the VHL complex. ID2 co-precipitated with VHL was detected by Western blot.

[0016] Figure 5. In silico model of 3112-5273 docking to ID2-Thr-27 binding to VCB complex identifying the likely binding site of molecules blocking the ID2-VHL interaction.94935-6729-8938v. 1DETAILED DESCRIPTION OF THE INVENTION

[0017] The present invention provides a compound having the following structure:whereinA is a ring;X is NH, O, or S;Ri is an aryl or heteroaryl;R2 and R3 are each independently aryl, heteroaryl, cycloalkyl or heterocycloalkyl;R4andR? are each independently alkyl, alkenyl, alkynyl, -CN; -OH, -O-alkyl, -NH2, -NH- alkyl, -NH-(CO)-alkyl, or -NH-(SO2)-alkyl;Rs is H, alkyl, alkenyl, alkynyl, -OH, -O-alkyl, -NH2, -NH-alkyl, -NH-(CO)-alkyl, or -NH- (SO2)-alkyl;R7 is aryl or heteroaryl;Rs is cycloalkyl, heterocycloalkyl, alkenyl, alkynyl, aryl, or heteroaryl;R9 and Rio are each independently H, alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, or heteroaryl;Y is CR17 or N; andR11, R12, R13, R14, RIS, Rie, and R17 are each independently H, alkyl, alkenyl, alkynyl, aryl, heterocycloalkyl, alkyl-heterocycloalkyl, heteroaryl, -(CO2)-alkyl, -(CO2)-aryl -NH2, -NH- alkyl, -NH-alkenyl, -NH-alkynyl, -NH-aryl, -NH-heteroaryl, -OH, -OAc, -O-C(O)alkyl,, halo, -CF3, -OCFs-O-alkyl, -O-alkenyl, -O-aryl, -S-(alkyl), -S-(alkenyl), -S-(alkynyl), - S(O)-(alkyl), -SO2-(alkyl), -SO2-(aryl), -SO2-(heteroaryl), alkyl-OH, alkyl-O-alkyl, alkyl- NH2, alkyl-NH-alkyl, or alkyl-N(alkyl)2.

[0018] In some embodiments, A is a carbocycle.

[0019] In some embodiments, A is cycloalkyl.104935-6729-8938v. 1

[0020] In some embodiments, A is C5-12 cycloalkyl.

[0021] In some embodiments, A is C5-10 cycloalkyl.

[0022] In some embodiments, A is cycloheptane or cyclopentane.

[0023] In some embodiments, A is cycloheptane.

[0024] In some embodiments, A is cyclopentane.

[0025] In some embodiments, the compound has the following structure:o

[0026] In some embodiments, Ri is heteroaryl.

[0027] In some embodiments, Ri is a 5-membered heterocycle.

[0028] In some embodiments, Ri is furan, oxazole, 1,3,4-oxadiazole, thiophene, or thiazole.

[0029] In some embodiments, Ri is furan or oxazole.

[0030] In some embodiments, Ri is furan.

[0031] In some embodiments, Ri is

[0032] In some embodiments, R2 is cycloalkyl or heterocycloalkyl.

[0033] In some embodiments, R2 is heterocycloalkyl.

[0034] In some embodiments, R2 is a 6-membered heterocycle.

[0035] In some embodiments, R2 is a substituted or unsubstituted piperidine, piperazine, dioxane, or morpholine.

[0036] In some embodiments, R2 is a substituted or unsubstituted piperazine or piperidine.

[0037] In some embodiments, R2 has the following structure:114935-6729-8938v. 1R19ZAR20wherein Z is CHR22 or NR22; andR18, R19, R20, R21, and R22 are each, independently H, halo, -CN, -CF3, -OCF3, -alkyl, - alkenyl, -alkynyl, -aryl, -heteroaryl, -NH2, -NH-alkyl, -NH-alkenyl, -NH-alkynyl, -NH- aryl, -NH-heteroaryl, -OH, -OAc, -O-C(O)alkyl, -O-alkyl, -O-alkenyl, -S-(alkyl), -S- (alkenyl), -S-(alkynyl), -S(O)-(alkyl), -SO2-(alkyl), -SO2-(aryl), -SO2-(heteroaryl), alkyl- OH, alkyl-O-alkyl, alkyl-NH2, alkyl-NH-alkyl, or alkyl-N(alkyl)2.

[0038] In some embodiments, Z is NR22.

[0039] In some embodiments, R22 is H, halo, -CN, -CF3, -OCF3, -alkyl, -alkenyl, -alkynyl, -aryl, -heteroaryl, -NH2, -NH-alkyl, -NH-alkenyl, -NH-alkynyl, -NH-aryl, -NH-heteroaryl, -OH, -OAc, -O-C(O)alkyl, -O-alkyl, -O-alkenyl, -S-(alkyl), -S-(alkenyl), -S-(alkynyl), -S(O)-(alkyl), -SO2-(alkyl), -SO2-(aryl), -SO2-(heteroaryl), alkyl-OH, alkyl-O-alkyl, alkyl-NH2, alkyl-NH-alkyl, or alkyl-N(alkyl)2.

[0040] In some embodiments, R22 is H, halo, -CN, -CF3, -OCF3, or -alkyl.

[0041] In some embodiments, R22 is alkyl.

[0042] In some embodiments, R22 is C1-6 alkyl.

[0043] In some embodiments, R22 is methyl.

[0044] In some embodiments, one of R18, R19, R20, and R21 is H.

[0045] In some embodiments, two of R18, R19, R20, and R21 are H.

[0046] In some embodiments, three of R18, R19, R20, and R21 are H.

[0047] In some embodiments, R18, R19, R20, and R21 are each H.

[0048] In some embodiments, R3 is aryl.

[0049] In some embodiments, R3 is a 5-8-membered aromatic ring.

[0050] In some embodiments, R3 is a 6-membered aromatic ring.124935-6729-8938v. 1

[0051] In some embodiments, R3 is a substituted or unsubstituted benzene.

[0052] In some embodiments, R3 has the following structure:wherein R23, R24, R25, R26, R27 are each, independently H, halo, -CN, -CF3, -OCF3, -alkyl, -alkenyl, -alkynyl, -aryl, -heteroaryl, -NH2, -NH-alkyl, -NH-alkenyl, -NH-alkynyl, -NH- aryl, -NH-heteroaryl, -OH, -OAc, -O-C(O)alkyl, -O-alkyl, -O-alkenyl, -S-(alkyl), -S- (alkenyl), -S-(alkynyl), -S(O)-(alkyl), -SO2-(alkyl), -SO2-(aryl), -SO2-(heteroaryl), alkyl- OH, alkyl-O-alkyl, alkyl-NH2, alkyl-NH-alkyl, or alkyl-N(alkyl)2.

[0053] In some embodiments, R25 is H, halo, -CN, -CF3, -OCF3, -alkyl, -alkenyl, -alkynyl, -aryl,-heteroaryl, -NH2, -NH-alkyl, -NH-alkenyl, -NH-alkynyl, -NH-aryl, -NH-heteroaryl, -OH, -OAc, -O-C(O)alkyl, -O-alkyl, -O-alkyl-haloalkyl, -O-alkenyl, -S-(alkyl), -S-(alkenyl), - S-(alkynyl), -S(O)-(alkyl), -SO2-(alkyl), -SO2-(aryl), -SO2-(heteroaryl), alkyl-OH, alkyl-O-alkyl, alkyl-NH2, alkyl-NH-alkyl, or alkyl-N(alkyl)2.

[0054] In some embodiments, R25 is H, halo, -CN, -CF3, -OCF3, -alkyl or -O-alkyl-halo.

[0055] In some embodiments, R25 is alkyl or -O-alkyl-halo.

[0056] In some embodiments, R25 is C1-6 alkyl or -O-C1-6 alkyl-halo.

[0057] In some embodiments, R25 is methyl or -O-CHF2.

[0058] In some embodiments, one of R23, R24, R26, and R27 is H.

[0059] In some embodiments, two of R23, R24, R26, and R27 are H.

[0060] In some embodiments, three of R23, R24, R26, and R27 are H.

[0061] In some embodiments, R23, R24, R26, and R27 are H.

[0062] In some embodiments, R24 is H, halo, -CN, -CF3, -OCF3, -alkyl, -alkenyl, -alkynyl, -aryl,134935-6729-8938v. 1-heteroaryl, -NH2, -NH-alkyl, -NH-alkenyl, -NH-alkynyl, -NH-aryl, -NH-heteroaryl, -OH, -OAc, -O-C(O)alkyl, -O-alkyl, -O-alkyl-haloalkyl, -O-alkenyl, -S-(alkyl), -S-(alkenyl), - S-(alkynyl), -S(O)-(alkyl), -SO2-(alkyl), -SO2-(aryl), -SO2-(heteroaryl), alkyl-OH, alkyl-O-alkyl, alkyl-NH2, alkyl-NH-alkyl, or alkyl-N(alkyl)2.

[0063] In some embodiments, one of R23, R25, R26, and R27 is H.

[0064] In some embodiments, two of R23, R25, R26, and R27 are H.

[0065] In some embodiments, three of R23, R25, R26, and R27 are H.

[0066] In some embodiments, R23, R25, R26, and R27 are H.

[0067] In some embodiments, R4 is alkyl, alkenyl, or alkynyl.

[0068] In some embodiments, R4 is alkyl.

[0069] In some embodiments, R4 is Ci-6 alkyl.

[0070] In some embodiments, R4 is methyl.

[0071] In some embodiments, the compound has the following structure:O X-R7

[0072] In some embodiments, R5 is alkynyl or -CN.

[0073] In some embodiments, R5 is -CN.

[0074] In some embodiments, Rs is H, halo, -CN, -CF3, -OCF3, -alkyl, -alkenyl, -alkynyl, -aryl,-heteroaryl, -NH2, -NH-alkyl, -NH-alkenyl, -NH-alkynyl, -NH-aryl, -NH-heteroaryl, -OH, -OAc, -O-C(O)alkyl, -O-alkyl, -O-alkyl-haloalkyl, -O-alkenyl, -S-(alkyl), -S-(alkenyl), - S-(alkynyl), -S(O)-(alkyl), -SO2-(alkyl), -SO2-(aryl), -SO2-(heteroaryl), alkyl-OH, alkyl- O-alkyl, alkyl-NH2, alkyl-NH-alkyl, or alkyl-N(alkyl)2.

[0075] In some embodiments, Rs is H or alkyl.

[0076] In some embodiments, Rs is H or methyl.144935-6729-8938v. 1

[0077] In some embodiments, R7 is aryl.

[0078] In some embodiments, R7 is C6-C8 aryl.

[0079] In some embodiments, R7 is Cg aryl.

[0080] In some embodiments, R7 is a substituted or unsubstituted benzene.

[0081] In some embodiments, R7 has the following structure:wherein R28, R29, R30, R31, R32 are each, independently H, halo, -CN, -CF3, -OCF3, -alkyl, -alkenyl, -alkynyl, -aryl, -heteroaryl, -NH2, -NH-alkyl, -NH-alkenyl, -NH-alkynyl, -NH- aryl, -NH-heteroaryl, -OH, -OAc, -O-C(O)alkyl, -O-alkyl, -O-alkenyl, -S-(alkyl), -S- (alkenyl), -S-(alkynyl), -S(O)-(alkyl), -SO2-(alkyl), -SO2-(aryl), -SO2-(heteroaryl), alkyl- OH, alkyl-O-alkyl, alkyl-NH2, alkyl-NH-alkyl, or alkyl-N(alkyl)2.

[0082] In some embodiments, R30 is H, halo, -CN, -CF3, -OCF3, -alkyl, -alkenyl, -alkynyl, -aryl,-heteroaryl, -NH2, -NH-alkyl, -NH-alkenyl, -NH-alkynyl, -NH-aryl, -NH-heteroaryl, -OH, -OAc, -O-C(O)alkyl, -O-alkyl, -O-alkyl-haloalkyl, -O-alkenyl, -S-(alkyl), -S-(alkenyl), - S-(alkynyl), -S(O)-(alkyl), -SO2-(alkyl), -SO2-(aryl), -SO2-(heteroaryl), alkyl-OH, alkyl- O-alkyl, alkyl-NH2, alkyl-NH-alkyl, or alkyl-N(alkyl)2.

[0083] In some embodiments, R30 is H, halo, -CN, -CF3, -OCF3, -alkyl or -O-alkyl-halo.

[0084] In some embodiments, R30 is alkyl.

[0085] In some embodiments, R30 is C1-6 alkyl.

[0086] In some embodiments, R30 is propyl.

[0087] In some embodiments, one of R28, R29, R31, and R32 is H.

[0088] In some embodiments, two of R28, R29, R31, and R32 are H.

[0089] In some embodiments, three of R28, R29, R31, and R32 are H.154935-6729-8938v. 1

[0090] In some embodiments, R28, R29, R31, and R32 are H.

[0091] In some embodiments, X is O.

[0092] The current invention provides a compound having the following structure:whereinR6 is H, alkyl, -O-alkyl, -NH-alkyl, -NH-(CO)-alkyl, or -NH-(SO2)-alkyl; andR25 is -alkyl, -NH-alkyl, -O-alkyl, -O-alkyl-haloalkyl, -S(O)-(alkyl), -SO2-(alkyl), alkyl- OH, -alkyl-O-alkyl, alkyl-NH2, alkyl-NH-alkyl, or alkyl-N(alkyl)2.

[0093] In some embodiments, the compound has the following structure:

[0094] In some embodiments, the compound has the following structure:164935-6729-8938v. 1

[0095] In some embodiments, the compound has the following structure:N O

[0096] In some embodiments, Rs has the following structure:iwwwherein R33, R34, R35, R36, R37 are each, independently H, halo, -CN, -CF3, -OCF3, -alkyl, -alkenyl, -alkynyl, -aryl, -heteroaryl, -NH2, -NH-alkyl, -NH-alkenyl, -NH-alkynyl, -NH- aryl, -NH-heteroaryl, -OH, -OAc, -O-C(O)alkyl, -O-alkyl, -O-alkenyl, -S-(alkyl), -S- (alkenyl), -S-(alkynyl), -S(O)-(alkyl), -SO2-(alkyl), -SO2-(aryl), -SO2-(heteroaryl), alkyl- OH, alkyl-O-alkyl, alkyl-NH2, alkyl-NH-alkyl, or alkyl-N(alkyl)2.

[0097] In some embodiments, R34 is H, halo, -CN, -CF3, -OCF3, -alkyl, -alkenyl, -alkynyl, -aryl,-heteroaryl, -NH2, -NH-alkyl, -NH-alkenyl, -NH-alkynyl, -NH-aryl, -NH-heteroaryl, -OH, -OAc, -O-C(O)alkyl, -O-alkyl, -O-alkyl-haloalkyl, -O-alkenyl, -S-(alkyl), -S-(alkenyl), - S-(alkynyl), -S(O)-(alkyl), -SO2-(alkyl), -SO2-(aryl), -SO2-(heteroaryl), alkyl-OH, alkyl- O-alkyl, alkyl-NH2, alkyl-NH-alkyl, or alkyl-N(alkyl)2; and R33, R35, R36, and R37 are H.

[0098] In some embodiments, R34 is H, halo, -CN, -CF3, -OCF3, -alkyl or -O-alkyl-halo.

[0099] In some embodiments, R34 is alkyl or halo.

[0100] In some embodiments, R34 is C1-6 alkyl or halo.174935-6729-8938v. 1

[0101] In some embodiments, R34 is methyl or F.

[0102] In some embodiments, R9 is H or alkyl.

[0103] In some embodiments, R9 is H or C1-6 alkyl.

[0104] In some embodiments, R9 is H or ethyl.

[0105] In some embodiments, Rio has the following structure:R38 R42R41R40wherein n is 0, 1, 2, or 3;R38, R39, R40, R41, R42 are each, independently H, halo, -CN, -CF3, -OCF3, -alkyl, -alkenyl, -alkynyl, -aryl, -heteroaryl, -NH2, -NH-alkyl, -NH-alkenyl, -NH-alkynyl, -NH-aryl, -NH- heteroaryl, -OH, -OAc, -O-C(O)alkyl, -O-alkyl, -O-alkenyl, -S-(alkyl), -S-(alkenyl), -S- (alkynyl), -S(O)-(alkyl), -SO2-(alkyl), -SO2-(aryl), -SO2-(heteroaryl), alkyl-OH, alkyl-O- alkyl, alkyl-NH2, alkyl-NH-alkyl, or alkyl-N(alkyl)2.

[0106] In some embodiments, n is 0 or 1.

[0107] In some embodiments, n is 0.

[0108] In some embodiments, n is 1.

[0109] In some embodiments, R38 and R41 are each independently H, halo, -CN, -CF3, -OCF3, -alkyl, -alkenyl, -alkynyl, -aryl, -heteroaryl, -NH2, -NH-alkyl, -NH-alkenyl, -NH-alkynyl, -NH-aryl, -NH-heteroaryl, -OH, -OAc, -O-C(O)alkyl, -O-alkyl, -O-alkyl-haloalkyl, -O-alkenyl, -S-(alkyl), -S-(alkenyl), -S-(alkynyl), -S(O)-(alkyl), -SO2-(alkyl), -SO2-(aryl), -SO2-(heteroaryl), alkyl-OH, alkyl-O-alkyl, alkyl-NH2, alkyl-NH-alkyl, or alkyl-N(alkyl)2; and R39, R40, and R42 are H.

[0110] In some embodiments, R38 and R41 are each independently H, halo, -alkyl or -O-alkyl.

[0111] In some embodiments, R38 is -O-alkyl and R41 is -alkyl.

[0112] In some embodiments, R38 is -O-C1-6 alkyl and R41 is -C1-6 alkyl.184935-6729-8938v. 1

[0113] In some embodiments, R38 is -O-methyl and R41 is methyl.

[0114] In some embodiments, R38 and R41 are each independently halo.

[0115] In some embodiments, R38 and R41 are F.

[0116] In some embodiments, R38, R39, R40, R41 and R42 are H.

[0117] In some embodiments, the compound has the following structure:

[0118] In some embodiments, the compound has the following structure:

[0119] In some embodiments, Ru is H, alkenyl, aryl, substituted styrenyl or heterocycloalkylaryl.

[0120] In some embodiments, RH is H, styrenyl-O-alkyl, or N-containing heterocycloalkylaryl.

[0121] In some embodiments, Ru is H, styrene-O-Ci-6 alkyl, or N-containing heterocycloalkyl-aryl, wherein the heterocylcoalkyl is piperidine.

[0122] In some embodiments, Ru is H, dimethoxy styrene or N-phenyl-piperidine.

[0123] In some embodiments, R12 is H or -(CO2)-alkyl.

[0124] In some embodiments, R12 is H or -(CO2)-Ci-6 alkyl.

[0125] In some embodiments, R12 is H or -(CO2)-ethyl.

[0126] In some embodiments, R13 is H or alkyl.

[0127] In some embodiments, R13 is H or alkyl-O-alkyl.194935-6729-8938v. 1

[0128] In some embodiments, R13 is H or C1-6 alkyl-O-Ci-6 alkyl.

[0129] In some embodiments, R13 is H or -CH2-O-isopropyl.

[0130] In some embodiments, R14 is H or alkyl.

[0131] In some embodiments, R14 is H or C1-6 alkyl.

[0132] In some embodiments, R14 is H or methyl.

[0133] In some embodiments, R15 is H, alkyl, alkyl-heterocycloalkyl-aryl, or -CF3.

[0134] In some embodiments, R15 is H, C1-6 alkyl, C1-6 alkyl-N-containing heterocycloalkylbenzene, or -CF3.[01351 In some embodiments, R15 is H, methyl, -CH2-piperidine- / M-trifluoromethylbenzene, or -CF3.

[0136] In some embodiments, Ri6 is H or -OH.

[0137] In some embodiments, R17 is H or -NH-aryl.

[0138] In some embodiments, R17 is H or -NH-benzene.

[0139] In some embodiments, R17 is H or -NH-p-aniline.

[0140] In some embodiments, R17 is H or -NH- / ?-N, N-dimethylaniline.

[0141] In some embodiments, the compound has the following structure:

[0142] The present invention provides a compound having the following structure:204935-6729-8938v. 1whereinA is a ring;X is CH2, N, or S;Ri is an aryl or heteroaryl;R2and R3 are each independently aryl, heteroaryl, cycloalkyl or heterocycloalkyl;R4 is alkenyl, alkynyl, -CN; -OH, -O-alkyl, -NH2, -NH-alkyl, -NH-(CO)-alkyl, or -NH- (SO2)-alkyl;Rs is alkyl, alkenyl, alkynyl, -CN; -OH, -O-alkyl, -NH2, -NH-alkyl, -NH-(CO)-alkyl, or - NH-(SO2)-alkyl;Re is H, alkyl, alkenyl, alkynyl, -OH, -O-alkyl, -NH2, -NH-alkyl, -NH-(CO)-alkyl, or -NH- (SO2)-alkyl;R7 is aryl or heteroaryl;Rs is cycloalkyl, heterocycloalkyl, alkenyl, alkynyl, aryl, or heteroaryl;R9 and Rio are each independently alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, or heteroaryl;Y is CR17; andR11, R12, R13, R14, R15, and Rie are each independently H, alkyl, alkenyl, alkynyl, aryl, heterocycloalkyl, alkyl-heterocycloalkyl, heteroaryl, -(CO2)-alkyl, -(CO2)-aryl -NH2, -NH- alkyl, -NH-alkenyl, -NH-alkynyl, -NH-aryl, -NH-heteroaryl, -OH, -OAc, -O-C(O)alkyl,, halo, -CF3, -OCFs-O-alkyl, -O-alkenyl, -O-aryl, -S-(alkyl), -S-(alkenyl), -S-(alkynyl), - S(O)-(alkyl), -SO2-(alkyl), -SO2-(aryl), -SO2-(heteroaryl), alkyl-OH, alkyl-O-alkyl, alkyl- NH2, alkyl -NH-alkyl, or alkyl-N(alkyl)2;R17 is alkyl, alkenyl, alkynyl, aryl, heterocycloalkyl, alkyl-heterocycloalkyl, heteroaryl, - (CO2)-alkyl, -(CO2)-aryl -NH2, -NH-alkyl, -NH-alkenyl, -NH-alkynyl, -NH-heteroaryl, -214935-6729-8938v. 1OH, -OAc, -O-C(O)alkyl,, halo, -CF3, -OCF3-O-alkyl, -O-alkenyl, -O-aryl, -S-(alkyl), -S- (alkenyl), -S-(alkynyl), -S(O)-(alkyl), -SO2-(alkyl), -SO2-(aryl), -SO2-(heteroaryl), alkyl- OH, alkyl-O-alkyl, alkyl-NH2, alkyl-NH-alkyl, or alkyl-N(alkyl)2.

[0143] The present invention provides a compound having the following structure:whereinA is a ring;X is NH, O, or S;Ri is an aryl or heteroaryl;R2 and R3 are each independently aryl, heteroaryl, cycloalkyl or heterocycloalkyl;R4andRs are each independently alkyl, alkenyl, alkynyl, -CN; -OH, -O-alkyl, -NH2, -NH- alkyl, -NH-(CO)-alkyl, or -NH-(SO2)-alkyl;Re is H, alkyl, alkenyl, alkynyl, -OH, -O-alkyl, -NH2, -NH-alkyl, -NH-(CO)-alkyl, or -NH- (SO2)-alkyl;R7 is aryl or heteroaryl;Rs is cycloalkyl, heterocycloalkyl, alkenyl, alkynyl, aryl, or heteroaryl;R9 and Rio are each independently H, alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, or heteroaryl;Y is CR17 or N; andRH, R12, Ri3, Ri4, R15, Ri6, and R17 are each independently H, alkyl, alkenyl, alkynyl, aryl, heterocycloalkyl, alkyl-heterocycloalkyl, heteroaryl, -(CO2)-alkyl, -(CO2)-aryl -NH2, -NH- alkyl, -NH-alkenyl, -NH-alkynyl, -NH-aryl, -NH-heteroaryl, -OH, -OAc, -O-C(O)alkyl,, halo, -CF3, -OCF3-O-alkyl, -O-alkenyl, -O-aryl, -S-(alkyl), -S-(alkenyl), -S-(alkynyl), -224935-6729-8938v. 1S(O)-(alkyl), -S02-(alkyl), -SC>2-(aryl), -SO2-(heteroaryl), alkyl-OH, alkyl-O-alkyl, alkyl- NH2, alkyl-NH-alkyl, or alkyl-N(alkyl)2;wherein the compound is other than

[0144] In some embodiments, the compound has the following structure:234935-6729-8938v. 14935-6729-8938v. 1

[0145] The present invention provides a pharmaceutical composition comprising the compounds of the present invention and a pharmaceutically acceptable carrier.

[0146] The present invention provides a method of inhibiting interaction between ID2 protein and tumor suppressor complex CRLVHLcomprising contacting the ID2 protein or the suppressor complex CRLVHLwith the compound having the following structure:whereinA is a ring;X is NH, O, or S;Ri is an aryl or heteroaryl;R2 and R3 are each independently aryl, heteroaryl, cycloalkyl or heterocycloalkyl;R4 and R5 are each independently alkyl, alkenyl, alkynyl, -CN; -OH, -O-alkyl, -NH2, -NH- alkyl, -NH-(CO)-alkyl, or -NH-(SO2)-alkyl;R6 is H, alkyl, alkenyl, alkynyl, -OH, -O-alkyl, -NH2, -NH-alkyl, -NH-(CO)-alkyl, or -NH- (SO2)-alkyl;254935-6729-8938v. 1R7 is aryl or heteroaryl;Rs is cycloalkyl, heterocycloalkyl, alkenyl, alkynyl, aryl, or heteroaryl;R9 and Rio are each independently H, alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, or heteroaryl;Y is CR17 or N; andR11, R12, Ri3, Ri4, R15, R16, and R17 are each independently H, alkyl, alkenyl, alkynyl, aryl, heterocycloalkyl, alkyl-heterocycloalkyl, heteroaryl, -(CC>2)-alkyl, -(CO2)-aryl -NH2, -NH- alkyl, -NH-alkenyl, -NH-alkynyl, -NH-aryl, -NH-heteroaryl, -OH, -OAc, -O-C(O)alkyl,, halo, -CF3. -OCF3-O-alkyl, -O-alkenyl, -O-aryl, -S-(alkyl), -S-(alkenyl), -S-(alkynyl), - S(O)-(alkyl), -SO2-(alkyl), -SO2-(aryl), -SO2-(heteroaryl), alkyl-OH, alkyl-O-alkyl, alkyl- NH2, alkyl-NH-alkyl, or alkyl-N(alkyl)2.

[0147] The present invention provides a method of inhibiting proliferation of tumor or cancer cells comprising contacting ID2 protein or suppressor complex CRLVHLwith the compound having the following structure:whereinA is a ring;X is NH, O, or S;Ri is an aryl or heteroaryl;R2 and R3 are each independently aryl, heteroaryl, cycloalkyl or heterocycloalkyl;R4 and R5 are each independently alkyl, alkenyl, alkynyl, -CN; -OH, -O-alkyl, -NH2, -NH- alkyl, -NH-(CO)-alkyl, or -NH-(SO2)-alkyl;R6 is H, alkyl, alkenyl, alkynyl, -OH, -O-alkyl, -NH2, -NH-alkyl, -NH-(CO)-alkyl, or -NH- (SO2)-alkyl;264935-6729-8938v. 1R.7 is aryl or heteroaryl;Rs is cycloalkyl, heterocycloalkyl, alkenyl, alkynyl, aryl, or heteroaryl;R9 and Rio are each independently H, alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, or heteroaryl;Y is CR17 or N; andR11, R12, Ri3, Ri4, R15, R16, and R17 are each independently H, alkyl, alkenyl, alkynyl, aryl, heterocycloalkyl, alkyl-heterocycloalkyl, heteroaryl, -(CC>2)-alkyl, -(CO2)-aryl -NH2, -NH- alkyl, -NH-alkenyl, -NH-alkynyl, -NH-aryl, -NH-heteroaryl, -OH, -OAc, -O-C(O)alkyl,, halo, -CF3. -OCF3-O-alkyl, -O-alkenyl, -O-aryl, -S-(alkyl), -S-(alkenyl), -S-(alkynyl), - S(O)-(alkyl), -SO2-(alkyl), -SO2-(aryl), -SO2-(heteroaryl), alkyl-OH, alkyl-O-alkyl, alkyl- NH2, alkyl-NH-alkyl, or alkyl-N(alkyl)2.

[0148] The present invention provides a method of suppressing the activity of tumor or cancer cells comprising contacting ID2 protein or suppressor complex CRLVHLwith the compound having the following structure:whereinA is a ring;X is NH, O, or S;Ri is an aryl or heteroaryl;R2 and R3 are each independently aryl, heteroaryl, cycloalkyl or heterocycloalkyl;R4 and R5 are each independently alkyl, alkenyl, alkynyl, -CN; -OH, -O-alkyl, -NH2, -NH- alkyl, -NH-(CO)-alkyl, or -NH-(SO2)-alkyl;R6 is H, alkyl, alkenyl, alkynyl, -OH, -O-alkyl, -NH2, -NH-alkyl, -NH-(CO)-alkyl, or -NH- (SO2)-alkyl;274935-6729-8938v. 1R.7 is aryl or heteroaryl;Rs is cycloalkyl, heterocycloalkyl, alkenyl, alkynyl, aryl, or heteroaryl;R9 and Rio are each independently H, alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, or heteroaryl;Y is CR17 or N; andR11, R12, Ri3, Ri4, R15, R16, and R17 are each independently H, alkyl, alkenyl, alkynyl, aryl, heterocycloalkyl, alkyl-heterocycloalkyl, heteroaryl, -(CC>2)-alkyl, -(CO2)-aryl -NH2, -NH- alkyl, -NH-alkenyl, -NH-alkynyl, -NH-aryl, -NH-heteroaryl, -OH, -OAc, -O-C(O)alkyl,, halo, -CF3. -OCF3-O-alkyl, -O-alkenyl, -O-aryl, -S-(alkyl), -S-(alkenyl), -S-(alkynyl), - S(O)-(alkyl), -SO2-(alkyl), -SO2-(aryl), -SO2-(heteroaryl), alkyl-OH, alkyl-O-alkyl, alkyl- NH2, alkyl-NH-alkyl, or alkyl-N(alkyl)2.

[0149] The present invention provides a method of treating tumor or cancer in a subject comprising administering the compound having the following structure:A is a ring;XisNH, O, or S;Ri is an aryl or heteroaryl;R2 and R3 are each independently aryl, heteroaryl, cycloalkyl or heterocycloalkyl;R4andRs are each independently alkyl, alkenyl, alkynyl, -CN; -OH, -O-alkyl, -NH2, -NH- alkyl, -NH-(CO)-alkyl, or -NH-(SO2)-alkyl;Re is H, alkyl, alkenyl, alkynyl, -OH, -O-alkyl, -NH2, -NH-alkyl, -NH-(CO)-alkyl, or -NH- (SO2)-alkyl;R7 is aryl or heteroaryl;284935-6729-8938v. 1Rs is cycloalkyl, heterocycloalkyl, alkenyl, alkynyl, aryl, or heteroaryl;R9 and Rio are each independently H, alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, or heteroaryl;Y is CR17 or N; andR11, R12, Ri3, Ri4, R15, R16, and R17 are each independently H, alkyl, alkenyl, alkynyl, aryl, heterocycloalkyl, alkyl-heterocycloalkyl, heteroaryl, -(CC>2)-alkyl, -(CO2)-aryl -NH2, -NH- alkyl, -NH-alkenyl, -NH-alkynyl, -NH-aryl, -NH-heteroaryl, -OH, -OAc, -O-C(O)alkyl,, halo, -CF3. -OCF3-O-alkyl, -O-alkenyl, -O-aryl, -S-(alkyl), -S-(alkenyl), -S-(alkynyl), - S(O)-(alkyl), -SO2-(alkyl), -SO2-(aryl), -SO2-(heteroaryl), alkyl-OH, alkyl-O-alkyl, alkyl- NH2, alkyl-NH-alkyl, or alkyl-N(alkyl)2.

[0150] In some embodiments of the method, A is a carbocycle.

[0151] In some embodiments of the method, A is cycloalkyl.

[0152] In some embodiments of the method, A is C5-12 cycloalkyl.

[0153] In some embodiments of the method, A is a C5-10 cycloalkyl.

[0154] In some embodiments of the method, A is cycloheptane or cyclopentane.

[0155] In some embodiments of the method, A is cycloheptane.

[0156] In some embodiments of the method, A is cyclopentane.

[0157] In some embodiments of the method, the compound has the following structure:

[0158] In some embodiments of the method, Ri is heteroaryl.

[0159] In some embodiments of the method, Ri is a 5-membered heterocycle.

[0160] In some embodiments of the method, Ri is furan, oxazole, 1,3,4-oxadiazole, thiophene, or thiazole.

[0161] In some embodiments of the method, Ri is furan or oxazole.294935-6729-8938v. 1

[0162] In some embodiments of the method, Ri is furan.

[0163] In some embodiments of the method, Ri is.

[0164] In some embodiments of the method, R2 is cycloalkyl or heterocycloalkyl.

[0165] In some embodiments of the method, R2 is heterocycloalkyl.

[0166] In some embodiments of the method, R2 is a 6-membered heterocycle.

[0167] In some embodiments of the method, R2 is a substituted or unsubstituted piperidine, piperazine, dioxane, or morpholine.

[0168] In some embodiments of the method, R2 is a substituted or unsubstituted piperazine or piperidine.

[0169] In some embodiments of the method, R2 has the following structure:wherein Z is CHR22 or NR22; andR18, R19, R20, R21, and R22 are each, independently H, halo, -CN, -CF3, -OCF3, -alkyl, - alkenyl, -alkynyl, -aryl, -heteroaryl, -NH2, -NH-alkyl, -NH-alkenyl, -NH-alkynyl, -NH- aryl, -NH-heteroaryl, -OH, -OAc, -O-C(O)alkyl, -O-alkyl, -O-alkenyl, -S-(alkyl), -S- (alkenyl), -S-(alkynyl), -S(O)-(alkyl), -SO2-(alkyl), -SO2-(aryl), -SO2-(heteroaryl), alkyl- OH, alkyl-O-alkyl, alkyl-NH2, alkyl-NH-alkyl, or alkyl-N(alkyl)2.

[0170] In some embodiments of the method, Z is NR22, wherein R22 is H, halo, -CN, -CF3, -OCF3, -alkyl, -alkenyl, -alkynyl, -aryl, -heteroaryl, -NH2, -NH-alkyl, -NH-alkenyl, -NH-alkynyl, -NH-aryl, -NH-heteroaryl, -OH, -OAc, -O-C(O)alkyl, -O-alkyl, -O-alkenyl, -S-(alkyl), -S-(alkenyl), -S-(alkynyl), -S(O)-(alkyl), -SO2-(alkyl), -SO2-(aryl), -SO2-(heteroaryl), alkyl-OH, alkyl-O-alkyl, alkyl-NH2, alkyl-NH-alkyl, or alkyl-N(alkyl)2.

[0171] In some embodiments of the method, R22 is H, halo, -CN, -CF3, -OCF3, or -alkyl.

[0172] In some embodiments of the method, R22 is alkyl.304935-6729-8938v. 1

[0173] In some embodiments of the method, R22 is C1-6 alkyl.

[0174] In some embodiments of the method, R22 is methyl.

[0175] In some embodiments of the method, one of Ris, R19, R20, and R21 is H.

[0176] In some embodiments of the method, two of Ris, R19, R20, and R21 are H.

[0177] In some embodiments of the method, three of Ris, R19, R20, and R21 are H.

[0178] In some embodiments of the method, Ris, R19, R20, and R21 are H.

[0179] In some embodiments of the method, R3 is aryl.

[0180] In some embodiments of the method, R3 is a 5-8-membered aromatic ring.

[0181] In some embodiments of the method, R3 is a 6-membered aromatic ring.

[0182] In some embodiments of the method, R3 is a substituted or unsubstituted benzene.

[0183] In some embodiments of the method, R3 has the following structure:R-23\^.JR24wherein R23, R24, R25, R26, R27 are each, independently H, halo, -CN, -CF3, -OCF3, -alkyl, -alkenyl, -alkynyl, -aryl, -heteroaryl, -NH2, -NH-alkyl, -NH-alkenyl, -NH-alkynyl, -NH- aryl, -NH-heteroaryl, -OH, -OAc, -O-C(O)alkyl, -O-alkyl, -O-alkenyl, -S-(alkyl), -S- (alkenyl), -S-(alkynyl), -S(O)-(alkyl), -SO2-(alkyl), -SO2-(aryl), -SO2-(heteroaryl), alkyl- OH, alkyl-O-alkyl, alkyl-NH2, alkyl-NH-alkyl, or alkyl-N(alkyl)2.

[0184] In some embodiments of the method, R25 is H, halo, -CN, -CF3, -OCF3, -alkyl, -alkenyl, -alkynyl, -aryl, -heteroaryl, -NH2, -NH-alkyl, -NH-alkenyl, -NH-alkynyl, -NH-aryl, -NH-heteroaryl, -OH, -OAc, -O-C(O)alkyl, -O-alkyl, -O-alkyl-haloalkyl, -O-alkenyl, -S-(alkyl), -S-(alkenyl), -S-(alkynyl), -S(O)-(alkyl), -SO2-(alkyl), -SO2-(aryl), -SO2-(heteroaryl), alkyl-OH, alkyl-O-alkyl, alkyl-NH2, alkyl-NH-alkyl, or alkyl-N(alkyl)2.

[0185] In some embodiments of the method, R25 is H, halo, -CN, -CF3, -OCF3, -alkyl or -O-alkyl-halo.314935-6729-8938v. 1

[0186] In some embodiments of the method, R25 is alkyl or -O-alkyl-halo.

[0187] In some embodiments of the method, R25 is C1-6 alkyl or -O-C1-6 alkyl-halo.

[0188] In some embodiments of the method, R25 is methyl or -O-CHF2.

[0189] In some embodiments of the method, one of R23, R24, R26, and R27 is H.

[0190] In some embodiments of the method, two of R23, R24, R26, and R27 are H.

[0191] In some embodiments of the method, three of R23, R24, R26, and R27 are H.

[0192] In some embodiments of the method, R23, R24, R26, and R27 are H.

[0193] In some embodiments of the method, R24 is H, halo, -CN, -CF3, -OCF3, -alkyl, -alkenyl, -alkynyl, -aryl, -heteroaryl, -NH2, -NH-alkyl, -NH-alkenyl, -NH-alkynyl, -NH-aryl, -NH-heteroaryl, -OH, -OAc, -O-C(O)alkyl, -O-alkyl, -O-alkyl-haloalkyl, -O-alkenyl, -S-(alkyl), -S-(alkenyl), -S-(alkynyl), -S(O)-(alkyl), -SO2-(alkyl), -SO2-(aryl), -SO2-(heteroaryl), alkyl-OH, alkyl-O-alkyl, alkyl-NH2, alkyl-NH-alkyl, or alkyl-N(alkyl)2.

[0194] In some embodiments of the method, one of R23, R25, R26, and R27 is H.

[0195] In some embodiments of the method, two of R23, R25, R26, and R27 are H.

[0196] In some embodiments of the method, three of R23, R25, R26, and R27 are H.

[0197] In some embodiments of the method, R23, R25, R26, and R27 are H.

[0198] In some embodiments of the method, R4 is alkyl, alkenyl, or alkynyl.

[0199] In some embodiments of the method, R4 is alkyl.

[0200] In some embodiments of the method, R4 is Ci-6 alkyl.

[0201] In some embodiments of the method, R4 is methyl.

[0202] In some embodiments of the method, the compound has the following structure:O X-R7

[0203] In some embodiments of the method, R5 is alkynyl or -CN.324935-6729-8938v. 1

[0204] In some embodiments of the method, Rs is -CN.

[0205] In some embodiments of the method, Re is H, halo, -CN, -CF3, -OCF3, -alkyl, -alkenyl, -alkynyl, -aryl, -heteroaryl, -NH2, -NH-alkyl, -NH-alkenyl, -NH-alkynyl, -NH-aryl, -NH-heteroaryl, -OH, -OAc, -O-C(O)alkyl, -O-alkyl, -O-alkyl-haloalkyl, -O-alkenyl, -S-(alkyl), -S-(alkenyl), -S-(alkynyl), -S(O)-(alkyl), -SO2-(alkyl), -SO2-(aryl), -SO2-(heteroaryl), alkyl-OH, alkyl-O-alkyl, alkyl-NH2, alkyl-NH-alkyl, or alkyl-N(alkyl)2.

[0206] In some embodiments of the method, Re is H or alkyl.

[0207] In some embodiments of the method, Re is H or methyl.

[0208] In some embodiments of the method, R7 is aryl.

[0209] In some embodiments of the method, R7 is Cs-Cs aryl.

[0210] In some embodiments of the method, R7 is Ce aryl.

[0211] In some embodiments of the method, R7 is a substituted or unsubstituted benzene.

[0212] In some embodiments of the method, R7 has the following structure:wherein R28, R29, R30, R31, R32 are each, independently H, halo, -CN, -CF3, -OCF3, -alkyl, -alkenyl, -alkynyl, -aryl, -heteroaryl, -NH2, -NH-alkyl, -NH-alkenyl, -NH-alkynyl, -NH- aryl, -NH-heteroaryl, -OH, -OAc, -O-C(O)alkyl, -O-alkyl, -O-alkenyl, -S-(alkyl), -S- (alkenyl), -S-(alkynyl), -S(O)-(alkyl), -SO2-(alkyl), -SO2-(aryl), -SO2-(heteroaryl), alkyl- OH, alkyl-O-alkyl, alkyl-NH2, alkyl-NH-alkyl, or alkyl-N(alkyl)2.

[0213] In some embodiments of the method, R30 is H, halo, -CN, -CF3, -OCF3, -alkyl, -alkenyl, -alkynyl, -aryl, -heteroaryl, -NH2, -NH-alkyl, -NH-alkenyl, -NH-alkynyl, -NH-aryl, -NH-heteroaryl, -OH, -OAc, -O-C(O)alkyl, -O-alkyl, -O-alkyl-haloalkyl, -O-alkenyl, -S-(alkyl), -S-(alkenyl), -S-(alkynyl), -S(O)-(alkyl), -SO2-(alkyl), -SO2-(aryl), -SO2-(heteroaryl), alkyl-OH, alkyl-O-alkyl, alkyl-NH2, alkyl-NH-alkyl, or alkyl-N(alkyl)2.334935-6729-8938v. 1

[0214] In some embodiments of the method, R30 is H, halo, -CN, -CF3, -OCF3, -alkyl or -O-alkyl-halo.

[0215] In some embodiments of the method, R30 is alkyl.

[0216] In some embodiments of the method, R30 is C1-6 alkyl.

[0217] In some embodiments of the method, R30 is propyl.

[0218] In some embodiments of the method, one of R28, R29, R31, and R32 is H.

[0219] In some embodiments of the method, two of R28, R29, R31, and R32 are H.

[0220] In some embodiments of the method, three of R28, R29, R31, and R32 are H.

[0221] In some embodiments of the method, R28, R29, R31, and R32 are H.

[0222] In some embodiments of the method, X is O.

[0223] The current invention provides a compound having the following structure:whereinReis H, alkyl, -O-alkyl, -NH-alkyl, -NH-(CO)-alkyl, or -NH-(SC>2)-alkyl; andR25 is -alkyl, -NH-alkyl, -O-alkyl, -O-alkyl-haloalkyl, -S(O)-(alkyl), -SO2-(alkyl), alkyl- OH, -alkyl-O-alkyl, alkyl-NH2, alkyl-NH-alkyl, or alkyl-N(alkyl)2.

[0224] In some embodiments of the method, the compound has the following structure:344935-6729-8938v. 1O

[0225] In some embodiments of the method, the compound has the following structure:

[0226] In some embodiments of the method, the compound has the following structure:N O

[0227] In some embodiments of the method, Rs has the following structure:wherein R33, R34, R35, R36, R37 are each, independently H, halo, -CN, -CF3, -OCF3, -alkyl, -alkenyl, -alkynyl, -aryl, -heteroaryl, -NH2, -NH-alkyl, -NH-alkenyl, -NH-alkynyl, -NH- aryl, -NH-heteroaryl, -OH, -OAc, -O-C(O)alkyl, -O-alkyl, -O-alkenyl, -S-(alkyl), -S-354935-6729-8938v. 1(alkenyl), -S-(alkynyl), -S(O)-(alkyl), -SO2-(alkyl), -SO2-(aryl), -SO2-(heteroaryl), alkyl-OH, alkyl-O-alkyl, alkyl-NH2, alkyl-NH-alkyl, or alkyl-N(alkyl)2.

[0228] In some embodiments of the method, R34 is H, halo, -CN, -CF3, -OCF3, -alkyl, -alkenyl, -alkynyl, -aryl, -heteroaryl, -NH2, -NH-alkyl, -NH-alkenyl, -NH-alkynyl, -NH-aryl, -NH-heteroaryl, -OH, -OAc, -O-C(O)alkyl, -O-alkyl, -O-alkyl-haloalkyl, -O-alkenyl, -S-(alkyl), -S-(alkenyl), -S-(alkynyl), -S(O)-(alkyl), -SO2-(alkyl), -SO2-(aryl), -SO2-(heteroaryl), alkyl-OH, alkyl-O-alkyl, alkyl-NH2, alkyl-NH-alkyl, or alkyl-N(alkyl)2i and R33, R35, R36, and R37 are H.

[0229] In some embodiments of the method, R34 is H, halo, -CN, -CF3, -OCF3, -alkyl or -O-alkyl-halo.

[0230] In some embodiments of the method, R34 is alkyl or halo.

[0231] In some embodiments of the method, R34 is C1-6 alkyl or halo.

[0232] In some embodiments of the method, R34 is methyl or F.

[0233] In some embodiments of the method, R9 is H or alkyl.

[0234] In some embodiments of the method, R9 is H or Ci-6 alkyl.

[0235] In some embodiments of the method, R9 is H or ethyl.

[0236] In some embodiments of the method, Rio has the following structure:wherein n is 0, 1, 2, or 3;R38, R39, R40, R41, R42 are each, independently H, halo, -CN, -CF3, -OCF3, -alkyl, -alkenyl, -alkynyl, -aryl, -heteroaryl, -NH2, -NH-alkyl, -NH-alkenyl, -NH-alkynyl, -NH-aryl, -NH- heteroaryl, -OH, -OAc, -O-C(O)alkyl, -O-alkyl, -O-alkenyl, -S-(alkyl), -S-(alkenyl), -S- (alkynyl), -S(O)-(alkyl), -SO2-(alkyl), -SO2-(aryl), -SO2-(heteroaryl), alkyl-OH, alkyl-O- alkyl, alkyl-NH2, alkyl-NH-alkyl, or alkyl-N(alkyl)2.

[0237] In some embodiments of the method, n is 0 or 1.364935-6729-8938v. 1

[0238] In some embodiments of the method, n is 0.

[0239] In some embodiments of the method, n is 1.

[0240] In some embodiments of the method, R38 and R41 are each independently H, halo, -CN, -CF3, -OCF3, -alkyl, -alkenyl, -alkynyl, -aryl, -heteroaryl, -NH2, -NH-alkyl, -NH-alkenyl, -NH-alkynyl, -NH-aryl, -NH-heteroaryl, -OH, -OAc, -O-C(O)alkyl, -O-alkyl, -O-alkyl-haloalkyl, -O-alkenyl, -S-(alkyl), -S-(alkenyl), -S-(alkynyl), -S(O)-(alkyl), -SO2-(alkyl), -SO2-(aryl), -SO2-(heteroaryl), alkyl-OH, alkyl-O-alkyl, alkyl-NH2, alkyl-NH-alkyl, or alkyl-N(alkyl)2; and R39, R40, and R42 are H.

[0241] In some embodiments of the method, R38 and R41 are each independently H, halo, -alkyl or -O-alkyl.

[0242] In some embodiments of the method, R38 is -O-alkyl and R41 is -alkyl.

[0243] In some embodiments of the method, R38 is -O-C1-6 alkyl and R41 is -C1-6 alkyl.

[0244] In some embodiments of the method, R38 is -O-methyl and R41 is methyl.

[0245] In some embodiments of the method, R38 and R41 are each independently halo.

[0246] In some embodiments of the method, R38 and R41 are F.

[0247] In some embodiments of the method, R38, R39, R40, R41 and R42 are H.

[0248] In some embodiments of the method, the compound has the following structure:NH

[0249] In some embodiments of the method, the compound has the following structure:n*13374935-6729-8938v. 1

[0250] In some embodiments of the method, Rn is H, alkenyl, aryl, substituted styrenyl or heterocycloalkyl-aryl.

[0251] In some embodiments of the method, Rn is H, styrenyl-O-alkyl, or N-containing heterocycloalkyl-aryl.

[0252] In some embodiments of the method, Rn is H, styrene-O-Ci-6 alkyl, or N-containing heterocycloalkyl-aryl, wherein the heterocylcoalkyl is piperidine.

[0253] In some embodiments of the method, Rn is H, dimethoxy styrene or N-phenyl-piperidine.

[0254] In some embodiments of the method, R12 is H or -(CO2)-alkyl.

[0255] In some embodiments of the method, R12 is H or -(CO2)-Ci-6 alkyl.

[0256] In some embodiments of the method, R12 is H or -(CO2)-ethyl.

[0257] In some embodiments of the method, R13 is H or alkyl.

[0258] In some embodiments of the method, R13 is H or alkyl-O-alkyl.

[0259] In some embodiments of the method, R13 is H or C1-6 alkyl-O-Ci-6 alkyl.

[0260] In some embodiments of the method, R13 is H or -CFh-O-isopropyl.

[0261] In some embodiments of the method, R14 is H or alkyl.

[0262] In some embodiments of the method, R14 is H or C1-6 alkyl.

[0263] In some embodiments of the method, R14 is H or methyl.

[0264] In some embodiments of the method, R15 is H, alkyl, alkyl-heterocycloalkyl-aryl, or -CF3.

[0265] In some embodiments of the method, R15 is H, C1-6 alkyl, C1-6 alkyl-N-containing heterocycloalkyl-benzene, or -CF3.

[0266] In some embodiments of the method, R15 is H, methyl, -CFF-piperidine- / ??-trifluoromethylbenzene, or -CF3.

[0267] In some embodiments of the method, Ri6 is H or -OH.

[0268] In some embodiments of the method, R17 is H or -NH-aryl.384935-6729-8938v. 1

[0269] In some embodiments of the method, Ri? is H or -NH-benzene.

[0270] In some embodiments of the method, Ri? is H or -NH-p-aniline.

[0271] In some embodiments of the method, Ri? is H or -NH-p-N, N-dimethylaniline.

[0272] In some embodiments of the method, the compound has the following structure:

[0273] The present invention provides a method of selecting one or more compounds that inhibit interaction between ID2 protein and tumor suppressor complex CRLVHLfrom a pool of compounds using an assay; wherein the assay comprises donor beads and acceptor beads, wherein the method comprises the following steps:a) adding, in any order:i) glutathione S-transferase tagged von Hippel-Lindau tumor suppressor protein and Elongin C protein complex (GST-VHL / EloC);ii) one or more compounds from the pool of compounds;iii) FLAG tags;iv) the acceptor beads; andv) the donor beads;b) exciting a luminescent signal at a wavelength between 670-690 nm, preferably at 680 nm; andc) obtaining an emission signal at a wavelength between 610-625 nm, preferably at 617 nm.394935-6729-8938v. 1

[0274] In some embodiments of the method, the addition is in the order of i), ii), iii), iv) and v).

[0275] In some embodiments of the method, the method further comprises incubating the donor beads before adding the acceptor beads.

[0276] In some embodiments of the method, the method further comprises incubating the donor beads after adding the acceptor beads.

[0277] In some embodiments of the method, the emission signal in step (c) is used to calculate the inhibition of the interaction between the ID2 protein and the suppressor complex CRLVHL.

[0278] In some embodiments of the method, the emission signals were used to calculate a Z-score to validate the assay; preferably, the Z-score is >0.85.

[0279] In some embodiments of the method, the method further comprises:(a) calculating an inhibition threshold of the compounds from the obtained emission signal in step (c); and(b) identifying the compounds in step (a) having at least 62% inhibition threshold; (c) identifying the compounds in step (b) that are soluble, are not singlet oxygen quenchers, or do not alter reactive oxygen species (ROS) levels;(d) conducting a surface plasmon resonance (SPR) binding analysis on the compounds identified in step (c); and(e) identifying and selecting the compounds having a 1:1 binding mode with the suppressor complex CRLVHL;wherein the compounds selected in step (e) are the compounds that inhibit interaction between the ID2 protein and the tumor suppressor complex CRIHL.

[0280] In some embodiments of the method, the method further comprises using Tanimoto similarity scores of the compounds from step (e) to generate a pool of compounds that inhibit interaction between the ID2 protein and the tumor suppressor complex CRIHL.

[0281] In some embodiments of the method, the method further comprises a calculation of PAMPA permeability scores to select compounds that inhibit interaction between the ID2 protein and the tumor suppressor complex CRIHL.

[0282] In some embodiments of the method, the compound has the following structure:404935-6729-8938v. 1O

[0283] In some embodiments of the method, the compound has the following structure:414935-6729-8938v. 1Definitions

[0284] Unless otherwise defined, all technical and / or scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the invention pertains. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of embodiments of the invention, exemplary methods and / or materials are described below. In case of conflict, the patent specification, including definitions, will control. In addition, the materials, methods, and examples are illustrative only and are not intended to be necessarily limiting.

[0285] In the discussion unless otherwise stated, adjectives such as “substantially” and “about” modifying a condition or relationship characteristic of a feature or features of an embodiment of the invention, are understood to mean that the condition or characteristic is defined to within tolerances that are acceptable for operation of the embodiment for an application for which it is intended. In embodiments, about means within a standard deviation using measurements generally acceptable in the art. In embodiments, about means a range extending to + / - 10% of the specified 424935-6729-8938v. 1value. In embodiments, about includes the specified value. Unless otherwise indicated, the word “or” in the specification and claims is considered to be the inclusive “or” rather than the exclusive “or” and indicates at least one of and any combination of items it conjoins.

[0286] It should be understood that the terms “a” and “an” as used above and elsewhere herein refer to “one or more” of the enumerated components. It will be clear to one of ordinary skill in the art that the use of the singular includes the plural unless specifically stated otherwise. Therefore, the terms “a,” “an” and “at least one” are used interchangeably in this application.

[0287] For purposes of better understanding the present teachings and in no way limiting the scope of the teachings, unless otherwise indicated, all numbers expressing quantities, percentages or proportions, and other numerical values used in the specification and claims, are to be understood as being modified in all instances by the term “about.” Accordingly, unless indicated to the contrary, the numerical parameters set forth in the following specification and attached claims are approximations that may vary depending upon the desired properties sought to be obtained. At the very least, each numerical parameter should at least be construed in light of the number of reported significant digits and by applying ordinary rounding techniques.

[0288] In the description and claims of the present application, each of the verbs, “comprise,” “include” and “have” and conjugates thereof, are used to indicate that the object or objects of the verb are not necessarily a complete listing of components, elements or parts of the subject or subjects of the verb. Other terms as used herein are meant to be defined by their well-known meanings in the art.General

[0289] For the foregoing embodiments, each embodiment disclosed herein is contemplated as being applicable to each of the other disclosed embodiment.

[0290] As used herein, all headings are simply for organization and are not intended to limit the disclosure in any manner. The content of any individual section may be equally applicable to all sections. All combinations of the various elements disclosed herein are within the scope of the invention.

[0291] Additional objects, advantages, and novel features of the present invention will become apparent to one ordinarily skilled in the art upon examination of the following examples, which434935-6729-8938v. 1are not intended to be limiting. Additionally, each of the various embodiments and aspects of the present invention as delineated hereinabove and as claimed in the claims section below finds experimental support in the following examples.

[0292] It is appreciated that certain features of the invention, which are, for clarity, described in the context of separate embodiments, may also be provided in combination in a single embodiment. Conversely, various features of the invention, which are, for brevity, described in the context of a single embodiment, may also be provided separately or in any suitable subcombination or as suitable in any other described embodiment of the invention. Certain features described in the context of various embodiments are not to be considered essential features of those embodiments, unless the embodiment is inoperative without those elements.

[0293] Examples are provided below to facilitate a more complete understanding of the invention. The following examples illustrate the exemplary modes of making and practicing the invention. However, the scope of the invention is not limited to specific embodiments disclosed in these Examples, which are for purposes of illustration only.

[0294] In some embodiments, a pharmaceutically acceptable salt of any of the above compounds is provided. In some embodiments, an ester of any of the above compounds is provided. In some embodiments, an enantiomer of any of the above compounds is provided.

[0295] In some embodiments, a package or composition comprising any of the above compounds is provided.

[0296] In some embodiments, a salt of the compound of the present invention is used in any of the above methods, uses, packages or compositions. In some embodiments, a pharmaceutically acceptable salt of the compound of the present invention is used in any of the above methods, uses, packages or compositions. In some embodiments, an ester of the compound of the present invention is used in any of the above methods, uses, packages or compositions. Any of the above compounds may be used in any of the disclosed methods, uses, packages or pharmaceutical compositions.

[0297] Any of the compounds used in the disclosed methods, uses, packages or pharmaceutical compositions may be replaced with any other compound disclosed in the present invention. Any444935-6729-8938v. 1of the above generic compounds may be used in any of the disclosed methods, uses, packages or compositions.

[0298] As used herein, "treating" means inhibiting, slowing, halting, or reversing the progression of a disease or infection. Treating may also mean improving one or more symptoms of a disease or infection.

[0299] The compounds of the present invention include all hydrates, solvates, and complexes of the compounds used by this invention. If a chiral center or another form of an isomeric center is present in a compound of the present invention, all forms of such isomer or isomers, including enantiomers and diastereomers, are intended to be covered herein. Compounds containing a chiral center may be used as a racemic mixture, an enantiomerically enriched mixture, or the racemic mixture may be separated using well-known techniques and an individual enantiomer may be used alone. The compounds described in the present invention are in racemic form or as individual enantiomers. The enantiomers can be separated using known techniques, such as those described in Pure and Applied Chemistry 69, 1469-1474, (1997) IUPAC. In cases in which compounds have unsaturated carbon-carbon double bonds, both the cis (Z) and trans (E) isomers are within the scope of this invention.

[0300] The compounds of the subject invention may have spontaneous tautomeric forms. In cases wherein compounds may exist in tautomeric forms, such as keto-enol tautomers, each tautomeric form is contemplated as being included within this invention whether existing in equilibrium or predominantly in one form.

[0301] In the compound structures depicted herein, hydrogen atoms are not shown for carbon atoms having less than four bonds to non-hydrogen atoms. However, it is understood that enough hydrogen atoms exist on said carbon atoms to satisfy the octet rule.

[0302] It is understood that the structures described in the embodiments of the methods hereinabove can be the same as the structures of the compounds described hereinabove.

[0303] It is understood that where a numerical range is recited herein, the present invention contemplates each integer between, and including, the upper and lower limits, unless otherwise stated.454935-6729-8938v. 1

[0304] Except where otherwise specified, if the structure of a compound of this invention includes an asymmetric carbon atom, it is understood that the compound occurs as a racemate, racemic mixture, and isolated single enantiomer. All such isomeric forms of these compounds are expressly included in this invention. Except where otherwise specified, each stereogenic carbon may be of the R or S configuration. It is to be understood accordingly that the isomers arising from such asymmetry (e.g., all enantiomers and diastereomers) are included within the scope of this invention, unless indicated otherwise. Such isomers can be obtained in substantially pure form by classical separation techniques and by stereochemically controlled synthesis, such as those described in " Enantiomers, Racemates and Resolutions" by J. Jacques, A. Collet and S. Wilen, Pub. John Wiley & Sons, NY, 1981. For example, the resolution may be carried out by preparative chromatography on a chiral column.

[0305] The subject invention is also intended to include all isotopes of atoms occurring on the compounds disclosed herein. Isotopes include those atoms having the same atomic number but different mass numbers. By way of general example and without limitation, isotopes of hydrogen include tritium and deuterium. Isotopes of carbon include C-13 and C-14.

[0306] It will be noted that any notation of a carbon in structures throughout this application, when used without further notation, are intended to represent all isotopes of carbon, such as12C,13C, or14C. Furthermore, any compounds containing13C or14C may specifically have the structure of any of the compounds disclosed herein.

[0307] It will also be noted that any notation of a hydrogen in structures throughout this application, when used without further notation, are intended to represent all isotopes of hydrogen, such as1H,2H, or3H. Furthermore, any compounds containing2H or3H may specifically have the structure of any of the compounds disclosed herein.

[0308] This invention also provides isotopic variants of the compounds disclosed herein, including wherein the isotopic atom is2H and / or wherein the isotopic atom13C. Accordingly, in the compounds provided herein hydrogen can be enriched in the deuterium isotope. It is to be understood that the invention encompasses all such isotopic forms.

[0309] Isotopically-labeled compounds can generally be prepared by conventional techniques known to those skilled in the art using appropriate isotopically-labeled reagents in place of the non-labeled reagents employed.464935-6729-8938v. 1

[0310] In the compounds used in the method of the present invention, the substituents may be substituted or unsubstituted, unless specifically defined otherwise.

[0311] In the compounds used in the method of the present invention, alkyl, heteroalkyl, monocycle, bicycle, aryl, heteroaryl, carbocycle, and heterocycle groups can be further substituted by replacing one or more hydrogen atoms with alternative non-hydrogen groups. These include, but are not limited to, halo, hydroxy, mercapto, amino, carboxy, cyano, carbamoyl and aminocarbonyl and aminothiocarbonyl.

[0312] It is understood that substituents and substitution patterns on the compounds used in the method of the present invention can be selected by one of ordinary skill in the art to provide compounds that are chemically stable and that can be readily synthesized by techniques known in the art from readily available starting materials. If a substituent is itself substituted with more than one group, it is understood that these multiple groups may be on the same carbon or on different carbons, so long as a stable structure results.

[0313] As used herein, "alkyl" is intended to include both branched and straight-chain saturated aliphatic hydrocarbon groups having the specified number of carbon atoms. Thus, Ci-Cn as in “Ci-Cn alkyl" is defined to include groups having 1,2, n-1 or n carbons in a linear or branched arrangement, and specifically includes methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, isopropyl, isobutyl, sec-butyl and so on. An embodiment can be C1-C12 alkyl, C2-C12 alkyl, C3-C12 alkyl, C4-C12 alkyl and so on. “Alkoxy" represents an alkyl group as described above attached through an oxygen bridge.

[0314] The term "alkenyl" refers to a non-aromatic hydrocarbon radical, straight or branched, containing at least 1 carbon to carbon double bond, and up to the maximum possible number of non-aromatic carbon-carbon double bonds may be present. Thus, C2-Cnalkenyl is defined to include groups having 1, 2...., n-1 or n carbons. For example, " C2-C6 alkenyl" means an alkenyl radical having 2, 3, 4, 5, or 6 carbon atoms, and at least 1 carbon-carbon double bond, and up to, for example, 3 carbon-carbon double bonds in the case of a Ce alkenyl, respectively. Alkenyl groups include ethenyl, propenyl, butenyl and cyclohexenyl. As described above with respect to alkyl, the straight, branched or cyclic portion of the alkenyl group may contain double bonds and may be substituted if a substituted alkenyl group is indicated. An embodiment can be C2-C12 alkenyl, C3-C12 alkenyl, C4-C12 alkenyl and so on.474935-6729-8938v. 1

[0315] The term "alkynyl" refers to a hydrocarbon radical straight or branched, containing at least 1 carbon to carbon triple bond, and up to the maximum possible number of non-aromatic carbon-carbon triple bonds may be present. Thus, C2-Cnalkynyl is defined to include groups having 1, 2...., n-1 or n carbons. For example, " C2-C6 alkynyl" means an alkynyl radical having 2 or 3 carbon atoms, and 1 carbon-carbon triple bond, or having 4 or 5 carbon atoms, and up to 2 carbon-carbon triple bonds, or having 6 carbon atoms, and up to 3 carbon-carbon triple bonds. Alkynyl groups include ethynyl, propynyl and butynyl. As described above with respect to alkyl, the straight or branched portion of the alkynyl group may contain triple bonds and may be substituted if a substituted alkynyl group is indicated. An embodiment can be a C2-Cnalkynyl. An embodiment can be C2-C12 alkynyl, C3-C12 alkynyl, C4-C12 alkynyl and so on.

[0316] “Alkylene”, “alkenylene” and “alkynylene” shall mean, respectively, a divalent alkane, alkene and alkyne radical, respectively. It is understood that an alkylene, alkenylene, and alkynylene may be straight or branched. An alkylene, alkenylene, and alkynylene may be unsubstituted or substituted.

[0317] As used herein, "heteroalkyl" includes both branched and straight-chain saturated aliphatic hydrocarbon groups having the specified number of carbon atoms and at least 1 heteroatom within the chain or branch.

[0318] As herein, "cycloalkyl" shall mean cyclic rings of alkanes of three to eight total carbon atoms, or any number within this range (i.e., cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl or cyclooctyl).

[0319] As used herein, "monocycle" includes any stable polyatomic carbon ring of up to 10 atoms and may be unsubstituted or substituted. Examples of such non-aromatic monocycle elements include but are not limited to: cyclobutyl, cyclopentyl, cyclohexyl, and cycloheptyl. Examples of such aromatic monocycle elements include but are not limited to: phenyl.

[0320] As used herein, "bicycle" includes any stable polyatomic carbon ring of up to 10 atoms that is fused to a polyatomic carbon ring of up to 10 atoms with each ring being independently unsubstituted or substituted. Examples of such non-aromatic bicycle elements include but are not limited to: decahydronaphthalene. Examples of such aromatic bicycle elements include but are not limited to: naphthalene.484935-6729-8938v. 1

[0321] As used herein, "aryl" is intended to mean any stable monocyclic, bicyclic or polycyclic carbon ring of up to 10 atoms in each ring, wherein at least one ring is aromatic, and may be unsubstituted or substituted. Examples of such aryl elements include but are not limited to, phenyl, p-toluenyl (4-methylphenyl), naphthyl, tetrahydro-naphthyl, indanyl, phenanthryl, anthryl or acenaphthyl. In cases where the aryl substituent is bicyclic and one ring is non-aromatic, it is understood that attachment is via the aromatic ring.

[0322] As used herein, the term “polycyclic” refers to unsaturated or partially unsaturated multiple fused ring structures, which may be unsubstituted or substituted.

[0323] The term “arylalkyl” refers to alkyl groups as described above wherein one or more bonds to hydrogen contained therein are replaced by a bond to an aryl group as described above. It is understood that an “arylalkyl” group is connected to a core molecule through a bond from the alkyl group and that the aryl group acts as a substituent on the alkyl group. Examples of arylalkyl moieties include, but are not limited to, benzyl (phenylmethyl), p-trifluoromethylbenzyl (4-trifluoromethylphenylmethyl), 1 -phenyl ethyl, 2 -phenyl ethyl, 3-phenylpropyl, 2-phenylpropyl and the like.

[0324] The term "heteroaryl", as used herein, represents a stable monocyclic, bicyclic or polycyclic ring of up to 10 atoms in each ring, wherein at least one ring is aromatic and contains from 1 to 4 heteroatoms selected from the group consisting of O, N and S. Bicyclic aromatic heteroaryl groups include phenyl, pyridine, pyrimidine or pyridizine rings that are (a) fused to a 6-membered aromatic (unsaturated) heterocyclic ring having one nitrogen atom; (b) fused to a 5- or 6-membered aromatic (unsaturated) heterocyclic ring having two nitrogen atoms; (c) fused to a 5-membered aromatic (unsaturated) heterocyclic ring having one nitrogen atom together with either one oxygen or one sulfur atom; or (d) fused to a 5-membered aromatic (unsaturated) heterocyclic ring having one heteroatom selected from O, N or S. Heteroaryl groups within the scope of this definition include but are not limited to: benzimidazolyl, benzofuranyl, benzofurazanyl, benzopyrazolyl, benzotri azolyl, benzothiophenyl, benzoxazolyl, carbazolyl, carbolinyl, quinolyl, furanyl, indolinyl, indolyl, indolazinyl, indazolyl, isobenzofuranyl, isoindolyl, isoquinolyl, isothiazolyl, isoxazolyl, naphthpyridinyl, oxadiazolyl, oxazolyl, isoxazoline, pyranyl, pyrazinyl, pyrazolyl, pyridazinyl, pyridopyridinyl, pyridazinyl, pyridyl, pyrimidyl, pyrrolyl, quinazolinyl, quinolyl, quinoxalinyl, tetrazolyl, tetrazolopyridyl, thiadiazolyl, thiazolyl, thienyl, triazolyl,494935-6729-8938v. 1acridinyl, carbazolyl, quinoxalinyl, pyrrazolyl, indolyl, benzotri azolyl, benzothiazolyl, benzoxazolyl, isoxazolyl, isothiazolyl, furanyl, thienyl, benzothienyl, benzofuranyl, quinolinyl, isoquinolinyl, oxazolyl, isoxazolyl, indolyl, pyrazinyl, pyridazinyl, pyridinyl, pyrimidinyl, or pyrrolyl. In cases where the heteroaryl substituent is bicyclic and one ring is non-aromatic or contains no heteroatoms, it is understood that attachment is via the aromatic ring or via the heteroatom containing ring, respectively. If the heteroaryl contains nitrogen atoms, it is understood that the corresponding N-oxides thereof are also encompassed by this definition.

[0325] The term “heteroarylalkyl” refers to alkyl groups as described above wherein one or more bonds to hydrogen contained therein are replaced by a bond to an heteroaryl group as described above. It is understood that an “heteroarylalkyl” group is connected to a core molecule through a bond from the alkyl group and that the heteroaryl group acts as a substituent on the alkyl group. Examples of heteroarylalkyl moieties include, but are not limited to, -CH2-(C5H4N), -CH2-CH2-(C5H4N) and the like.

[0326] The term "heterocycle", “heterocyclyl”, or “heterocyclic” refers to a mono- or polycyclic ring system which can be saturated or contains one or more degrees of unsaturation and contains one or more heteroatoms. Preferred heteroatoms include N, O, and / or S, including N-oxides, sulfur oxides, and dioxides. Preferably the ring is three to ten-membered and is either saturated or has one or more degrees of unsaturation. More preferably the ring is three to fourmembered and has one or more degrees of unsaturation. The heterocycle may be unsubstituted or substituted, with multiple degrees of substitution being allowed. Such rings may be optionally fused to one or more of another "heterocyclic" ring(s), heteroaryl ring(s), aryl ring(s), or cycloalkyl ring(s). Examples of heterocycles include, but are not limited to, aziridine, azirine, diazirine, oxirane, thiirane, azetidine, oxetane, thetane, tetrahydrofuran, pyran, 1,4-dioxane, 1,3-dioxane, piperidine, piperazine, pyrrolidine, morpholine, thiomorpholine, tetrahydrothiopyran, tetrahydrothiophene, 1,3-oxathiolane, and the like.

[0327] The term “carbocycle” or “carbocyclic” refers to a monocyclic ring system which can be saturated or contains one or more degrees of unsaturation and contains exclusively carbon atoms within the ring. However, it is understood that enough hydrogen atoms exist on said carbon atoms to satisfy the octet rule. Preferably the ring is three to ten-membered and is either saturated or has one or more degrees of unsaturation. More preferably the ring is five to six-membered and has one504935-6729-8938v. 1or more degrees of unsaturation. The carbocycle may be unsubstituted or substituted, with multiple degrees of substitution being allowed. Such rings may be optionally fused to one or more of another "carbocyclic" ring(s), heteroaryl ring(s), aryl ring(s), or cycloalkyl ring(s). Examples of carbocycles include, but are not limited to, cyclopropane, cyclobutane, cyclopentane, cyclohexane, cycloheptane, cyclooctane, benzene, and the like.

[0328] The alkyl, alkenyl, alkynyl, aryl, heteroaryl, and heterocyclyl substituents may be substituted or unsubstituted, unless specifically defined otherwise. In the compounds of the present invention, alkyl, alkenyl, alkynyl, aryl, heterocyclyl, cycloalkyl, and heteroaryl groups can be further substituted by replacing one or more hydrogen atoms with alternative non-hydrogen groups. These include, but are not limited to, halo, hydroxy, mercapto, amino, carboxy, cyano and carbamoyl.

[0329] As used herein, the term “halogen” or “halo” refers to F, Cl, Br, and I.

[0330] The term “ester” is intended to a mean an organic compound containing the R-O-CO-R’ group.

[0331] The terms “substitution”, “substituted” and “substituent” refer to a functional group as described above in which one or more bonds to a hydrogen atom contained therein are replaced by a bond to non-hydrogen or non-carbon atoms, provided that normal valencies are maintained and that the substitution results in a stable compound. Substituted groups also include groups in which one or more bonds to a carbon(s) or hydrogen(s) atom are replaced by one or more bonds, including double or triple bonds, to a heteroatom. Examples of substituent groups include the functional groups described above, and halogens (i.e., F, Cl, Br, and I); alkyl groups, such as methyl, ethyl, n-propyl, isopropryl, n-butyl, tert-butyl, and trifluoromethyl; hydroxyl; alkoxy groups, such as methoxy, ethoxy, n-propoxy, and isopropoxy; aryloxy groups, such as phenoxy; arylalkyloxy, such as benzyloxy (phenylmethoxy) and p-trifluorom ethylbenzyl oxy (4-trifluoromethylphenylmethoxy); heteroaryloxy groups; sulfonyl groups, such as trifluoromethanesulfonyl, methanesulfonyl, and p-toluenesulfonyl; nitro, nitrosyl; mercapto; sulfanyl groups, such as methylsulfanyl, ethylsulfanyl and propyl sulfanyl; cyano; amino groups, such as amino, methylamino, dimethylamino, ethylamino, and diethylamino; and carboxyl. Where multiple substituent moieties are disclosed or claimed, the substituted compound can be independently substituted by one or more of the disclosed or claimed substituent moieties, singly514935-6729-8938v. 1or pluraly. By independently substituted, it is meant that the (two or more) substituents can be the same or different.

[0332] In choosing the compounds of the present invention, one of ordinary skill in the art will recognize that the various substituents, i.e. Ri, R2, etc. are to be chosen in conformity with well-known principles of chemical structure connectivity.

[0333] The compounds used in the method of the present invention may be prepared by techniques known in organic synthesis and familiar to a practitioner ordinarily skilled in the art. However, these may not be the only means by which to synthesize or obtain the desired compounds.

[0334] The compounds used in the method of the present invention may be prepared by techniques described in Vogel’s Textbook of Practical Organic Chemistry, A. I. Vogel, A. R. Tatchell, B. S. Fumis, A. J. Hannaford, P. W. G. Smith, (Prentice Hall) 5th Edition (1996), March's Advanced Organic Chemistry: Reactions, Mechanisms, and Structure, Michael B. Smith, Jerry March, (Wiley-Interscience) 5th Edition (2007), and references therein, which are incorporated by reference herein. However, these may not be the only means by which to synthesize or obtain the desired compounds.

[0335] The various R groups attached to the aromatic rings of the compounds disclosed herein may be added to the rings by standard procedures, for example those set forth in Advanced Organic Chemistry: Part B: Reactions and Synthesis, Francis Carey and Richard Sundberg, (Springer) 5th ed. Edition. (2007), the content of which is hereby incorporated by reference.

[0336] Another aspect of the invention comprises a compound used in the method of the present invention as a pharmaceutical composition.

[0337] As used herein, the term “pharmaceutically active agent” means any substance or compound suitable for administration to a subject and furnishes biological activity or other direct effect in the treatment, cure, mitigation, diagnosis, or prevention of disease, or affects the structure or any function of the subject. Pharmaceutically active agents include, but are not limited to, substances and compounds described in the Physicians’ Desk Reference (PDR Network, LLC; 64th edition; November 15, 2009) and “Approved Drug Products with Therapeutic Equivalence Evaluations” (U. S. Department Of Health And Human Services, 30th edition, 2010), which are524935-6729-8938v. 1hereby incorporated by reference. Pharmaceutically active agents which have pendant carboxylic acid groups may be modified in accordance with the present invention using standard esterification reactions and methods readily available and known to those having ordinary skill in the art of chemical synthesis. Where a pharmaceutically active agent does not possess a carboxylic acid group, the ordinarily skilled artisan will be able to design and incorporate a carboxylic acid group into the pharmaceutically active agent where esterification may subsequently be carried out so long as the modification does not interfere with the pharmaceutically active agent’s biological activity or effect.

[0338] The compounds used in the method of the present invention may be in a salt form. As used herein, a “salt” is a salt of the instant compounds which has been modified by making acid or base salts of the compounds. In the case of compounds used to treat an infection or disease caused by a pathogen, the salt is pharmaceutically acceptable. Examples of pharmaceutically acceptable salts include, but are not limited to, mineral or organic acid salts of basic residues such as amines; alkali or organic salts of acidic residues such as phenols; alkali or organic salts of acidic residues such as carboxylic acids. The salts can be made using an organic or inorganic acid. Such acid salts are chlorides, bromides, sulfates, nitrates, phosphates, sulfonates, formates, tartrates, maleates, malates, citrates, benzoates, salicylates, ascorbates, and the like. Phenolate salts are alkali metal salts, sodium, potassium or lithium. The term "pharmaceutically acceptable salt" in this respect, refers to the relatively non-toxic, inorganic and organic acid or base addition salts of compounds of the present invention. These salts can be prepared in situ during the final isolation and purification of the compounds of the invention, or by separately reacting a purified compound of the invention in its free base or free acid form with a suitable organic or inorganic acid or base, and isolating the salt thus formed. Representative salts include the hydrobromide, hydrochloride, sulfate, bisulfate, phosphate, nitrate, acetate, valerate, oleate, palmitate, stearate, laurate, benzoate, lactate, phosphate, tosylate, citrate, maleate, fumarate, succinate, tartrate, napthylate, mesylate, glucoheptonate, lactobionate, and laurylsulphonate salts and the like. (See, e.g., Berge et al. (1977) " Pharmaceutical Salts", J. Pharm. Sci. 66:1-19).

[0339] The compounds of the present invention may also form salts with basic amino acids such a lysine, arginine, etc. and with basic sugars such as N-methylglucamine, 2-amino-2-deoxyglucose, etc. and any other physiologically non-toxic basic substance.534935-6729-8938v. 1

[0340] As used herein, “administering” an agent may be performed using any of the various methods or delivery systems well known to those skilled in the art. The administering can be performed, for example, orally, parenterally, intraperitoneally, intravenously, intraarterially, transdermally, sublingually, intramuscularly, rectally, transbuccally, intranasally, liposomally, via inhalation, vaginally, intraoccularly, via local delivery, subcutaneously, intraadiposally, intraarticularly, intrathecally, into a cerebral ventricle, intraventicularly, intratumorally, into cerebral parenchyma or intraparenchchymally.

[0341] The compounds used in the method of the present invention may be administered in various forms, including those detailed herein. The treatment with the compound may be a component of a combination therapy or an adjunct therapy, i.e. the subject or patient in need of the drug is treated or given another drug for the disease in conjunction with one or more of the instant compounds. This combination therapy can be sequential therapy where the patient is treated first with one drug and then the other or the two drugs are given simultaneously. These can be administered independently by the same route or by two or more different routes of administration depending on the dosage forms employed.

[0342] As used herein, a "pharmaceutically acceptable carrier" is a pharmaceutically acceptable solvent, suspending agent or vehicle, for delivering the instant compounds to the animal or human. The carrier may be liquid or solid and is selected with the planned manner of administration in mind. Liposomes are also pharmaceutically acceptable carriers as are slow-release vehicles.

[0343] The dosage of the compounds administered in treatment will vary depending upon factors such as the pharmacodynamic characteristics of a specific chemotherapeutic agent and its mode and route of administration; the age, sex, metabolic rate, absorptive efficiency, health and weight of the recipient; the nature and extent of the symptoms; the kind of concurrent treatment being administered; the frequency of treatment with; and the desired therapeutic effect.

[0344] A dosage unit of the compounds used in the method of the present invention may comprise a single compound or mixtures thereof with additional antibacterial agents. The compounds can be administered in oral dosage forms as tablets, capsules, pills, powders, granules, elixirs, tinctures, suspensions, syrups, and emulsions. The compounds may also be administered in intravenous (bolus or infusion), intraperitoneal, subcutaneous, or intramuscular form, or544935-6729-8938v. 1introduced directly, e.g. by injection, topical application, or other methods, into or onto a site of infection, all using dosage forms well known to those of ordinary skill in the pharmaceutical arts.

[0345] The compounds used in the method of the present invention can be administered in admixture with suitable pharmaceutical diluents, extenders, excipients, or in carriers such as the novel programmable sustained-release multi-compartmental nanospheres (collectively referred to herein as a pharmaceutically acceptable carrier) suitably selected with respect to the intended form of administration and as consistent with conventional pharmaceutical practices. The unit will be in a form suitable for oral, nasal, rectal, topical, intravenous or direct injection or parenteral administration. The compounds can be administered alone or mixed with a pharmaceutically acceptable carrier. This carrier can be a solid or liquid, and the type of carrier is generally chosen based on the type of administration being used. The active agent can be co-administered in the form of a tablet or capsule, liposome, as an agglomerated powder or in a liquid form. Examples of suitable solid carriers include lactose, sucrose, gelatin and agar. Capsule or tablets can be easily formulated and can be made easy to swallow or chew; other solid forms include granules, and bulk powders. Tablets may contain suitable binders, lubricants, diluents, disintegrating agents, coloring agents, flavoring agents, flow-inducing agents, and melting agents. Examples of suitable liquid dosage forms include solutions or suspensions in water, pharmaceutically acceptable fats and oils, alcohols or other organic solvents, including esters, emulsions, syrups or elixirs, suspensions, solutions and / or suspensions reconstituted from non-effervescent granules and effervescent preparations reconstituted from effervescent granules. Such liquid dosage forms may contain, for example, suitable solvents, preservatives, emulsifying agents, suspending agents, diluents, sweeteners, thickeners, and melting agents. Oral dosage forms optionally contain flavorants and coloring agents. Parenteral and intravenous forms may also include minerals and other materials to make them compatible with the type of injection or delivery system chosen.

[0346] Techniques and compositions for making dosage forms useful in the present invention are described in the following references: 7 Modem Pharmaceutics, Chapters 9 and 10 (Banker & Rhodes, Editors, 1979); Pharmaceutical Dosage Forms: Tablets (Lieberman et al., 1981); Ansel, Introduction to Pharmaceutical Dosage Forms 2nd Edition (1976); Remington's Pharmaceutical Sciences, 17th ed. (Mack Publishing Company, Easton, Pa., 1985); Advances in Pharmaceutical Sciences (David Ganderton, Trevor Jones, Eds., 1992); Advances in Pharmaceutical Sciences Vol.7. (David Ganderton, Trevor Jones, James McGinity, Eds., 1995); Aqueous Polymeric Coatings 554935-6729-8938v. 1for Pharmaceutical Dosage Forms (Drugs and the Pharmaceutical Sciences, Series 36 (James McGinity, Ed., 1989); Pharmaceutical Particulate Carriers: Therapeutic Applications: Drugs and the Pharmaceutical Sciences, Vol 61 (Alain Rolland, Ed., 1993); Drug Delivery to the Gastrointestinal Tract (Ellis Horwood Books in the Biological Sciences. Series in Pharmaceutical Technology; J. G. Hardy, S. S. Davis, Clive G. Wilson, Eds.); Modem Pharmaceutics Drugs and the Pharmaceutical Sciences, Vol 40 (Gilbert S. Banker, Christopher T. Rhodes, Eds.). All of the aforementioned publications are incorporated by reference herein.

[0347] Tablets may contain suitable binders, lubricants, disintegrating agents, coloring agents, flavoring agents, flow-inducing agents, and melting agents. For instance, for oral administration in the dosage unit form of a tablet or capsule, the active drug component can be combined with an oral, non-toxic, pharmaceutically acceptable, inert carrier such as lactose, gelatin, agar, starch, sucrose, glucose, methyl cellulose, magnesium stearate, dicalcium phosphate, calcium sulfate, mannitol, sorbitol and the like. Suitable binders include starch, gelatin, natural sugars such as glucose or beta-lactose, com sweeteners, natural and synthetic gums such as acacia, tragacanth, or sodium alginate, carboxymethylcellulose, polyethylene glycol, waxes, and the like. Lubricants used in these dosage forms include sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride, and the like. Disintegrators include, without limitation, starch, methyl cellulose, agar, bentonite, xanthan gum, and the like.

[0348] The compounds used in the method of the present invention may also be administered in the form of liposome delivery systems, such as small unilamellar vesicles, large unilamallar vesicles, and multilamellar vesicles. Liposomes can be formed from a variety of phospholipids, such as cholesterol, stearylamine, or phosphatidylcholines. The compounds may be administered as components of tissue-targeted emulsions.

[0349] The compounds used in the method of the present invention may also be coupled to soluble polymers as targetable drug carriers or as a prodrug. Such polymers include polyvinylpyrrolidone, pyran copolymer, polyhydroxylpropylmethacrylamide-phenol, polyhydroxyethylasparta-midephenol, or polyethyleneoxide-polylysine substituted with palmitoyl residues. Furthermore, the compounds may be coupled to a class of biodegradable polymers useful in achieving controlled release of a drug, for example, polylactic acid, polyglycolic acid, copolymers of polylactic and polyglycolic acid, polyepsilon caprolactone, polyhydroxy butyric564935-6729-8938v. 1acid, polyorthoesters, polyacetals, polydihydropyrans, polycyanoacylates, and crosslinked or amphipathic block copolymers of hydrogels.

[0350] Gelatin capsules may contain the active ingredient compounds and powdered carriers, such as lactose, starch, cellulose derivatives, magnesium stearate, stearic acid, and the like. Similar diluents can be used to make compressed tablets. Both tablets and capsules can be manufactured as immediate release products or as sustained release products to provide for continuous release of medication over a period of hours. Compressed tablets can be sugar coated or fdm coated to mask any unpleasant taste and protect the tablet from the atmosphere, or enteric coated for selective disintegration in the gastrointestinal tract.

[0351] For oral administration in liquid dosage form, the oral drug components are combined with any oral, non-toxic, pharmaceutically acceptable inert carrier such as ethanol, glycerol, water, and the like. Examples of suitable liquid dosage forms include solutions or suspensions in water, pharmaceutically acceptable fats and oils, alcohols or other organic solvents, including esters, emulsions, syrups or elixirs, suspensions, solutions and / or suspensions reconstituted from non-effervescent granules and effervescent preparations reconstituted from effervescent granules. Such liquid dosage forms may contain, for example, suitable solvents, preservatives, emulsifying agents, suspending agents, diluents, sweeteners, thickeners, and melting agents.

[0352] Liquid dosage forms for oral administration can contain coloring and flavoring to increase patient acceptance. In general, water, a suitable oil, saline, aqueous dextrose (glucose), and related sugar solutions and glycols such as propylene glycol or polyethylene glycols are suitable carriers for parenteral solutions. Solutions for parenteral administration preferably contain a water-soluble salt of the active ingredient, suitable stabilizing agents, and if necessary, buffer substances. Antioxidizing agents such as sodium bisulfite, sodium sulfite, or ascorbic acid, either alone or combined, are suitable stabilizing agents. Also used are citric acid and its salts and sodium EDTA. In addition, parenteral solutions can contain preservatives, such as benzalkonium chloride, methyl- or propyl-paraben, and chlorobutanol. Suitable pharmaceutical carriers are described in Remington's Pharmaceutical Sciences, Mack Publishing Company, a standard reference text in this field.

[0353] The compounds used in the method of the present invention may also be administered in intranasal form via use of suitable intranasal vehicles, or via transdermal routes, using those574935-6729-8938v. 1forms of transdermal skin patches well known to those of ordinary skill in that art. To be administered in the form of a transdermal delivery system, the dosage administration will generally be continuous rather than intermittent throughout the dosage regimen.

[0354] Parenteral and intravenous forms may also include minerals and other materials such as solutol and / or ethanol to make them compatible with the type of injection or delivery system chosen.

[0355] The compounds and compositions of the present invention can be administered in oral dosage forms as tablets, capsules, pills, powders, granules, elixirs, tinctures, suspensions, syrups, and emulsions. The compounds may also be administered in intravenous (bolus or infusion), intraperitoneal, subcutaneous, or intramuscular form, or introduced directly, e.g. by topical administration, injection or other methods, to the afflicted area, such as a wound, including ulcers of the skin, all using dosage forms well known to those of ordinary skill in the pharmaceutical arts.

[0356] Specific examples of pharmaceutically acceptable carriers and excipients that may be used to formulate oral dosage forms of the present invention are described in U. S. Pat. No.3,903,297 to Robert, issued Sept. 2, 1975. Techniques and compositions for making dosage forms useful in the present invention are described-in the following references: 7 Modem Pharmaceutics, Chapters 9 and 10 (Banker & Rhodes, Editors, 1979); Pharmaceutical Dosage Forms: Tablets (Lieberman et al., 1981); Ansel, Introduction to Pharmaceutical Dosage Forms 2nd Edition (1976); Remington's Pharmaceutical Sciences, 17th ed. (Mack Publishing Company, Easton, Pa., 1985); Advances in Pharmaceutical Sciences (David Ganderton, Trevor Jones, Eds., 1992); Advances in Pharmaceutical Sciences Vol 7. (David Ganderton, Trevor Jones, James McGinity, Eds., 1995); Aqueous Polymeric Coatings for Pharmaceutical Dosage Forms (Drugs and the Pharmaceutical Sciences, Series 36 (James McGinity, Ed., 1989); Pharmaceutical Particulate Carriers: Therapeutic Applications: Drugs and the Pharmaceutical Sciences, Vol 61 (Alain Rolland, Ed., 1993); Drug Delivery to the Gastrointestinal Tract (Ellis Horwood Books in the Biological Sciences. Series in Pharmaceutical Technology; J. G. Hardy, S. S. Davis, Clive G. Wilson, Eds.); Modem Pharmaceutics Drugs and the Pharmaceutical Sciences, Vol 40 (Gilbert S. Banker, Christopher T. Rhodes, Eds.). All of the aforementioned publications are incorporated by reference herein.

[0357] The active ingredient can be administered orally in solid dosage forms, such as capsules, tablets, powders, and chewing gum; or in liquid dosage forms, such as elixirs, syrups,584935-6729-8938v. 1and suspensions, including, but not limited to, mouthwash and toothpaste. It can also be administered parentally, in sterile liquid dosage forms.

[0358] Solid dosage forms, such as capsules and tablets, may be enteric-coated to prevent release of the active ingredient compounds before they reach the small intestine. Materials that may be used as enteric coatings include, but are not limited to, sugars, fatty acids, proteinaceous substances such as gelatin, waxes, shellac, cellulose acetate phthalate (CAP), methyl acrylatemethacrylic acid copolymers, cellulose acetate succinate, hydroxy propyl methyl cellulose phthalate, hydroxy propyl methyl cellulose acetate succinate (hypromellose acetate succinate), polyvinyl acetate phthalate (PVAP), and methyl methacrylate-methacrylic acid copolymers.

[0359] The compounds and compositions of the invention can be coated onto stents for temporary or permanent implantation into the cardiovascular system of a subject.

[0360] Each embodiment disclosed herein is contemplated as being applicable to each of the other disclosed embodiments. Thus, all combinations of the various elements described herein are within the scope of the invention.

[0361] This invention will be better understood by reference to the Experimental Details which follow, but those skilled in the art will readily appreciate that the specific experiments detailed are only illustrative of the invention as described more fully in the claims which follow thereafter.

[0362] “And / or” as used herein, for example, with option A and / or option B, encompasses the separate embodiments of (i) option A, (ii) option B, and (iii) option A plus option B.

[0363] All combinations of the various elements described herein are within the scope of the invention unless otherwise indicated herein or otherwise clearly contradicted by context.

[0364] Definitions: The terms used in this specification generally have their ordinary meanings in the art, within the context of this invention and the specific context where each term is used. Certain terms are discussed below, or elsewhere in the specification, to provide additional guidance to the practitioner in describing the methods of the invention and how to use them. Moreover, it will be appreciated that the same thing can be said in more than oneway. Consequently, alternative language and synonyms may be used for any one or more of the terms discussed herein, nor is any special significance to be placed upon whether or not a term is elaborated or discussed herein. Synonyms for certain terms are provided. A recital of one or more synonyms does not exclude the594935-6729-8938v. 1use of the other synonyms. The use of examples anywhere in the specification, including examples of any terms discussed herein, is illustrative only, and in no way limits the scope and meaning of the invention or any exemplified term. Likewise, the invention is not limited to its preferred embodiments.

[0365] The term “subject” as used in this application means a mammal. Mammals include canines, felines, rodents, bovine, equines, porcines, ovines, and primates including humans. Thus, the invention can be used in human medicine or also in veterinary medicine, e.g., to treat companion animals, farm animals, laboratory animals in zoological parks, and animals in the wild. The invention is particularly desirable for human medical applications. In a preferred embodiment the subject is a human.

[0366] The terms “therapeutically effective amount” or "amount effective to" encompasses, unless otherwise indicated, an amount sufficient to ameliorate or inhibit a symptom or sign of the medical condition. An effective amount for a particular subject may vary depending on factors such as the condition being treated, the overall health of the patient, the method route and dose of administration and the severity of side effects. An effective amount can be the maximal dose or dosing protocol that avoids significant side effects or toxic effects.EXAMPLESExample 1

[0367] AlphaScreen® donor beads coupled with Glutathione were used to capture the GST-VHL / EloC complex and acceptor beads (AlphaLISA®) that bind the Flag tag to immobilize Flag-H)2. Binding between VHL / EloC and H)2 elicits a strong luminescent signal at 617 nm upon excitation at 680 nm, which was be recorded by an EnVision™ Multilabel Plate Reader (PerkinElmer), instrument was used at the HTS facility of the Irving Cancer Center at Columbia University, CU-HTS (Figure 1).

[0368] Donor and acceptor beads were purchased from PerkinElmer. The CRLVHLcomplex expressed in Sf21 insect cells was obtained through EMD Millipore (Cat # 23-044). It included N-terminal GST tagged human VHL amino acid 54-end, A-terminal GST tagged human EloC, untagged human EloB and RBX1, and A-terminal 6xHis tagged human Cul-2. This recombinant complex was used the past in several different PPI assays with reproducible results in Lee et al.604935-6729-8938v. 12016. Flag tagged human ID2 was expressed in E. Coli and purified on Flag affinity matrix. High-quality Flag-ID2 protein was obtained. In preliminary experiments, the AlphaScreen® assay components were titrated to determine the optimal binding partners and bead concentration.

[0369] Each component of the binding reaction was titrated such that the final concentration of each is near the top of the linear response range and in the low nanomolar range. This binding reaction was stable at ambient temperature (>3 hours) and insensitive to concentrations of DMSO up to 10% (not shown). Preliminary tests were conducted in 96-well plates in 40pl volume. A prescreening was implemented using approximately 10,000 compounds on robotics and ssessed statistical factors for the HTS assay in a miniaturized binding reaction in a final volume of 25 pl in a 384-well micro plate format assay. The pre-screening campaign was performed in a 384-well micro plate format (test compound concentration = 10 pM, n = 1) with a final volume of 25 pL per well as follows:(1) dispensation of 10 pL of 2.5 x concentrated GST-VHL / EloC into a microplate using a robotic liquid handler;(2) transfer of 0.25 pL of test compounds in DMSO (1 mM stock) into each well using a compound transfer station with a nanoliter head, yielding a final assay concentration of each compound of 10 pM;(3) dispensation of 5 pL of 6X concentrated Flag-ID2 into the respective assay micro plates using a robotic liquid handler;(4) incubation of the assay micro plates for 1 hour at room temperature;(5) addition of 5 pL of AlphaScreen® acceptor beads for 3 hours followed by 5 pL of AlphaLISA® donor beads (20 pg / mL) for a further incubation of 2 hours at room temperature in the dark; and(6) reading of the assay microplates using laser excitation at 680 nm, with emission detected at 615 nm using the EnVision 2114 Multilabel Reader (PerkinElmer).

[0370] Small molecules in the screening library were selected to assure a high degree of diversity in the collection.

[0371] Screening assay was performed in buffer composed of 50 mM Tris pH 7.5, 120 mM NaCl, 0.05% Triton X-100. Each assay micro plate included the following controls: 16 wells 614935-6729-8938v. 1containing 1% v / v DMSO (positive control); 16 wells containing Flag-ID2 protein only and 16 wells containing only GST-VHL / EloC complex.General synthetic approaches to generate compoundsExample 2

[0372] Compounds of type Z can be synthesized from commercially available building blocks of types A and B using standard amide coupling reagents and conditions.Example 3

[0373] Compounds of type Y can be synthesized from building blocks of type C and 2-furoic acid (D), where Ri is hydrogen, methyl, or difluoromethoxy and R2 is hydrogen or methoxy. Compounds of type C can be synthesized according to US Patent 9,499,563. Compound C is joined with D using standard amide coupling reagents and conditions to produce intermediate of type E which is then treated with N-methylpiperazine under standard reductive amination conditions to produce Y.Rj =H, CHj, OCHF2R-, — 11. OCHj Example 4

[0374] Compound of type X can be synthesized from commercially available building blocks of types F and G using standard amide coupling reagents and conditions.624935-6729-8938v. 1RbR3= CH3, F R2= F, OCH3R4= H, CH2CH3n = 0, 1Example 5

[0375] Compounds of type W can be synthesized from commercially available building blocks types H and I using standard reductive amination conditions.Example 6

[0376] Compounds of type V can be synthesized from commercially available building blocks of type J and 1 -phenylpiperazine (K) using Pd-catalyzed Buchwald coupling conditions. Compounds of type J can be synthesized according to US Patent Publication 2008 / 0292626A1, but trans-esterifying with n-propanol.Example 7

[0377] Compounds of type U can be synthesized from commercially available building blocks of type L and 1 -phenylpiperazine (M) using Pd-catalyzed Buchwald coupling conditions.634935-6729-8938v. 1Example 8

[0378] Compounds of type T can be synthesized from commercially available building blocks types N and O. Building block N is appropriately protected using standard protecting reagents and conditions and then joined to O using Pd-catalyzed cross-coupling conditions. The protecting group is removed using standard reagents and conditions to produce compounds T.P = Protecting group Example 9

[0379] Compounds of type ZZ can be synthesized from commercially available building blocks AA, BB, and o-toluic acid (CC). Compound AA is joined with BB using standard amide coupling reagents and conditions to produce intermediate amid DD. DD is then deprotected using standard conditions and followed by joining with CC using standard amide coupling reagents and conditions to produce ZZ.tBoc =Example 10

[0380] Compounds of type YY can be synthesized from commercially available building blocks methyl cyanoacetate (EE) and 4-(4-Fluorobenzyloxy)benzaldehyde (FF). EE and FF are644935-6729-8938v. 1subjected to Knoevenagel condensation conditions to produce intermediate ester GG, which is then transaminated to intermediate amide HH. Amide HH is then treated with Lawesson’s reagent to produce YY.NEE HH Example 11

[0381] Compounds of type XX can be synthesized from commercially available compounds II and 4-fluorophenylacetonitrile JJ by using standard condensation conditions.oExample 12

[0382] Compound of type WW can be synthesized from commercially available building blocks KK and 4-chlorophenyl acetic acid (LL) using standard amide coupling reagents and conditions.654935-6729-8938v. 1Example 13

[0383] Compound of type VV can be synthesized from commercially available building blocks MM and NN using standard amide coupling reagents and conditions.Example 14

[0384] Compounds of type UU can be synthesized from commercially available building blocks 00 and 4-Bromoanisole (PP). 00 and PP are joined using Pd-catalyzed cross-coupling conditions to produce intermediate QQ which is treated with a strong base followed by a methylating agent to produce UU.00 PP Example 15

[0385] Compounds of type TT can be synthesized from building blocks RR and commercially available 2,3,4-trimethoxybenzaldehyde (SS) using standard condensation conditions. RR can be synthesized according to US Patent Publication 2015 / 0065482A1 using commercially available 2-bromohippuric acid.Example 16664935-6729-8938v. 1

[0386] Compounds of type ZZZ can be synthesized from commercially available building blocks AAA and type BBB using Pd-catalyzed Buchwald coupling conditions.Example 17

[0387] Compounds of type YYY can be synthesized from commercially available building blocks CCC and DDD using standard esterification reagents and conditions.Example 18

[0388] Compound of type XXX can be synthesized from commercially available building blocks EEE and FFF. EEE and FFF are joined using standard condensation conditions to produce intermediate GGG which is then reduced using standard hydrogenation conditions to produce XXX.Example 19

[0389] Compound of type WWW can be synthesized from commercially available building blocks HHH and III. HHH and III are joined using standard aldol reaction conditions to produce intermediate alcohol JJJ. JJJ is then protected with an appropriate protective group using standard conditions followed by treatment with Lawesson’s Reagent to produce intermediate thione KKK. KKK is then deprotected using standard conditions followed by standard oxidation conditions to produce WWW.674935-6729-8938v. 1o o OH O OP S O SDiscussion

[0390] The present invention provides inhibitors of the VHL / ID2 interaction. A high-throughput assay was developed to screen for compounds that show potential as pre-therapeutic leads that may be optimized into highly effective anti-tumor agents. A bead-based non-radioactive Amplified Luminescent Proximity Homogeneous Assay (AlphaScreen®) was developed to characterize the interaction between VHL-EloC and ID2, as depicted in Figure 1. This assay typically involves two types of beads (donor and acceptor beads) designed to associate with respective binding partners (Figure 1). The cascade of chemical reactions that occur when donor and acceptor beads are brought in close proximity (within -200 nm) is initiated by the interaction between the binding partners.

[0391] The statistical parameters of this assay were evaluated using the Z-factor, which is a statistical measure of the assay quality / reproducibility. After verifying the reproducibility of the assay, the present invention proceeded to perform the larger scale screening of library compounds. Z-score for both ID2 and VHL were >0.85, confirming the high quality of the screening campaign. The 224 plates that contained any hit were re-assayed as a confirmation screening, including 71,680 compounds of which 527 produced an inhibition threshold of - 62%.

[0392] Tanimoto similarity scores of the 391 hits were calculated, as was the Quantitative Estimation of Drug-likeness (QED) for each. The distance from the ID2 docking site on VHL was calculated for each hit according to the published model (Nature 2016). Hit compounds with both desirable PAMPA scores and calculated distances from the ID2 docking site were advanced.

[0393] AlphaLISA® TruHits kit and ROS-Glow were used to identify light scatterers (insoluble compounds), singlet oxygen quenchers or compounds that alter ROS levels and thus can interfere with the AlphaLISA® signal. Fifty-seven out of 391 compounds passed these filters and were intercrossed with results from 12-concentration dose response curves (performed in triplicate using 384 well format).684935-6729-8938v. 1

[0394] 65 compounds were selected based on dose response curves providing some reasonable indication of activity, minimal or no structural concerns, and similarity to other hits. For U-shaped and cinnamate-like compounds, which had the potential to be reactive, the glutathione reactivity assay was performed to confirm that their activity in blocking ID2 interaction with VHL in AlphaLISA® assay was not nonspecific.

[0395] Table 1 reports results of GSH-adducts and T1 / 2 of 20 compounds with U-shaped or cinnamate structure. Based on activity in AlphaLISA® assay, absence of interfering features and structural characteristics, 23 compounds were selected for compound binding to the target by surface plasmon resonance (SPR). 20 compounds showed steady state affinity whereas 3 compounds exhibited 1:1 binding mode (D411-0 16, D411-0511, 3112-5273; Figure 2).

[0396] Table 1. Summary of GSH adduct assay and T1 / 2 of 20 selected compounds characterized by cinnamate or U-shaped structure:Compound ID GSH-adducts Structure Tl / 2 < 30 GST pull-down min0086-0119 Y U-shaped Y Not tested0928-0127 U-shaped Not tested1989-9400 U-shaped Not tested2188-2186 U-shaped Y-binding2192-1096 Y U-shaped Not tested3512-0371 U-shaped Not tested3828-0523 Y U-shaped Not tested8012-7379 Y U-shaped Y Not tested D330-1156 U-shaped Y Not testedD411-0003 U-shaped Not testedD411-0511 U-shaped Y Not tested D411-0516 U-shaped Not tested 6246087 U-shaped Y- reduced binding 7472894 Y Cinnamate Y- reduced binding F1012-0068 Cinnamate Y Not tested5511-0457 Y Cinnamate Not tested694935-6729-8938v. 17391723 Cinnamate Y Not tested 5477354 Cinnamate Not tested D465-0478 Cinnamate Not tested

[0397] An additional molecule of the D411 series (D411-003) showed steady state affinity. These results were reproduced with newly synthesized molecule lots. D411 compounds are novel molecules. For example, the Tanimoto distance of D411-0516 from known VCB binding molecule = 0.59. 1875 molecules having D411 common scaffold are included in the Enamine REAL database. The pharmacophore elements of D411 molecules are shown in Figure 3. Independently of the SPR mode of compound binding to VCB, all three D411 molecules significantly reduced the ID2 interaction with VHL in vitro using GST pull-down assay (Figure 4).

[0398] 3112-5273 is also a novel compound (Tanimoto distance from known VCB binding molecule = 0.63), which is predicted to dock to the ID2-Thr-27 binding site on the VCB complex that was modeled in the work published in Lee et al. Nature 2016 (Figure 5). This is a particularly relevant feature of 3112-5273 that prompted the search for nearest-neighbor compounds in the HTS. 186 molecules were identified within 0.2 chemical distance in the Enamine REAL database. Twenty-seven nearest -neighbor compounds were included in the primary assay, but most of these molecules had shown low activity score, suggesting a weak structure-activity relationship (SAR). Among these nearest neighbors, 2 of the best performing candidates were subjected to SPR assays. One of these molecules (8010-3485, Tanimoto distance = 0.0369) exhibited 1:1 SPR binding mode (Figure 2, bottom right panel, and Table 2). Another near-neighbor molecule of 3112-5273 tested by SPR is 8012-3697, a molecule with Tanimoto distance = 0.09386 (Table 2). However, 8012-3697 showed the steady state affinity binding mode.

[0399] Table 2. Neighbor compounds of 3112-5273, 8010-3485 and 8012-3697 were tested by SPR for binding to VCB complex. 8010-3485 and 8012-3697 exhibited 1:1 and steady state affinity binding mode, respectively:704935-6729-8938v. 1Compound Structure Tanimoto Drug- Normalized Steady- ID Distance Likeness Drug- State from Score Likeness Binding 3112- Score by SPR 5273 (Y / N) 3112-5273 0 0.774 48.7 Yes8010-3485 0.0369 0.856 101.8 Yes2889-6051 0.04135 0.8654 21.6 NoS N o oNZM oX $°= r°X\\xzIZjTo cF0336- 0.06439 0.4359 7.7 No 03058012-3697 0.09386 0.325 1.6 No

[0400] Based on these results, several of the following compounds have performed as in vitro inhibitors of the ID2 / VHL interaction and with the expectation to serve as starting points and potential lead compounds for novel chemical probes and oncology therapeutics.

[0401] Based on in silico docking experiments, a plausible binding site for these compounds has been identified, as shown in Figure 5. This binding site is supported by the data demonstrating effective inhibition of the ID2-VHL interaction as this is the same site where ID2 binds to VHL.714935-6729-8938v. 1

[0402] The present invention identified small molecule inhibitors of the interaction between the oncogenic ID2 protein and the tumor suppressor complex CRLVHL, an interaction that leads to inactivation of the ubiquitin ligase activity of the complex. CRLVHLis the complex that targets HIF-alpha oncogenic proteins for degradation, limiting excessive HIF-alpha activity.

[0403] Small molecules that are able to interfere with ID2-CRLVHLcomplex thus restore the tumor suppression function of CRLVLH. These molecules are expected to be selectively active in cellular contexts such as the cancer cells and cancer stem cells, in which CRLVLHactivity is blunted by highly active ID2, thus potentially leaving normal cells, which mostly have low / inactive ID2, unaffected.

[0404] The present invention performed a cell free HT small molecule screening campaign testing the disruption of the binding of ID2 to the VCB ubiquitin ligase complex. Following confirmation assay and elimination of potential non-specific small molecule disruptors, the present invention selected a small number of candidates to confirm affinity and selectivity of biomolecular interactions using SPR. The outcome of this effort is to find 4 novel inhibitors of the ID2-VHL interaction including two different chemotypes with lead-like properties to support future research. These compounds (compounds D411-0516, D411-0511, and 3112-5273, 8010-3485) achieved validation of compound binding to VHL with orthogonal confirmation in SPR and are a priority for further exploration. Of equal interest is the possibility that these molecules may be of general utility in the field of VCB complex research.

[0405] X-ray crystallography and NMR will be used to locate and rationalize the binding of the small molecules identified in the present invention. This will open the path for future optimization of these molecules to tune the effect and ultimately demonstrate a therapeutically useful function in multiple cancer types with high expression of oncogenic HIF-a caused by ID2 interaction with and inactivation of the VCB complex.724935-6729-8938v. 1REFERENCES1. Soeda A, Park M, Lee D, et al. Hypoxia promotes expansion of the CD 133-positive glioma stem cells through activation of HIF-1 alpha. Oncogene. 2009; 28(45):3949- 3959; Li Z, Bao S, Wu Q, et al. Hypoxia-inducible factors regulate tumorigenic capacity of glioma stem cells. Cancer Cell. 2009; 15(6): 501— 513.2. Maxwell PH, Wiesener MS, Chang GW, et al. The tumor suppressor protein VHL targets hypoxia- inducible factors for oxygen-dependent proteolysis. Nature. 1999;399(6733):271-275; Ivan M, Kondo K, Yang HF, et al. HIF alpha targeted for VHL- mediated destruction by proline hydroxylation: implications for 02 sensing. Science. 2001; 292(5516):464-468.3. Lee SB, et al. An ID2 dependent mechanism for VHL inactivation in Cancer. Nature.2016; 529: 172-177.4. Lasorella, A., Benezra, R. & lavarone, A. The ID proteins: master regulators of cancer stem cells and tumour aggressiveness. Nature Rev. Cancer 14, 77-91(2014).5. Perk, J., lavarone, A. & Benezra, R. Id family of helix-loop-helix proteins in cancer. Nature Rev. Cancer 5, 603-614 (2005).6. Lasorella, A., Rothschild, G., Yokota, Y., Russell, R. G. & lavarone, A. Id2 mediates tumor initiation, proliferation, and angiogenesis in Rb mutant mice. Mol. Cell. Biol. 25, 3563-3574 (2005).7. Niola, F. et al. Mesenchymal high-grade glioma is maintained by the 1D-RAP1 axis. J. Clin. Invest. 123, 405-417 (2013).8. Vandeputte, D. A. et al. Expression and distribution of id helix-loop-helix proteins in human astrocytic tumors. Glia 38, 329-338 (2002).734935-6729-8938v. 1

Claims

CLAIMS1. A compound having the following structure:whereinA is a ring;X is NH, O, or S;Ri is an aryl or heteroaryl;R2 and R3 are each independently aryl, heteroaryl, cycloalkyl or heterocycloalkyl;R4andRs are each independently alkyl, alkenyl, alkynyl, -CN; -OH, -O-alkyl, -NH2, -NH- alkyl, -NH-(CO)-alkyl, or -NH-(SO2)-alkyl;R6 is H, alkyl, alkenyl, alkynyl, -OH, -O-alkyl, -NH2, -NH-alkyl, -NH-(CO)-alkyl, or -NH- (SO2)-alkyl;R7 is aryl or heteroaryl;Rs is cycloalkyl, heterocycloalkyl, alkenyl, alkynyl, aryl, or heteroaryl;R9 and Rio are each independently H, alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, or heteroaryl;Y is CR17 or N; andR11, R12, R13, R14, R15, Rie, and R17 are each independently H, alkyl, alkenyl, alkynyl, aryl, heterocycloalkyl, alkyl-heterocycloalkyl, heteroaryl, -(CO2)-alkyl, -(CO2)-aryl -NH2, -NH- alkyl, -NH-alkenyl, -NH-alkynyl, -NH-aryl, -NH-heteroaryl, -OH, -OAc, -O-C(O)alkyl,, halo, -CF3, -OCFs-O-alkyl, -O-alkenyl, -O-aryl, -S-(alkyl), -S-(alkenyl), -S-(alkynyl), - S(O)-(alkyl), -SO2-(alkyl), -SO2-(aryl), -SO2-(heteroaryl), alkyl-OH, alkyl-O-alkyl, alkyl- NH2, alkyl-NH-alkyl, or alkyl-N(alkyl)2.744937-1152-7546v.

2. The compound of claim 1, wherein A is a carbocycle; preferably, A is cycloalkyl; more preferably, A is a C5-12 cycloalkyl; more preferably, A is a C5-10 cycloalkyl; more preferably, A is cycloheptane or cyclopentane; more preferably, A is cycloheptane.

3. The compound of any one of claims 1-2 having the structure:preferably,(a) Ri is heteroaryl; more preferably Ri is a 5-membered aromatic heterocycle; more preferably, Ri is furan, oxazole, 1,3,4-oxadiazole, thiophene, or thiazole; more preferably Ri is furan or oxazole; more preferably Ri is furan; more preferably, Ri(b) R2 is cycloalkyl or heterocycloalkyl; preferably, R2 is heterocycloalkyl; more preferably, R2 is a 6-membered heterocycle; more preferably, R2 is a substituted or unsubstituted piperidine, piperazine, dioxane, or morpholine; more preferably, R2 is a substituted or unsubstituted piperazine or piperidine; more preferably, R2 is » / w Ivwherein Z is CHR22 or NR22; and Ris, R19, R20, R21, and R22 are each, independently H, halo, -CN, -CF3, -OCF3, -alkyl, -alkenyl, -alkynyl, -aryl, -heteroaryl, -NFh, -NH-alkyl, -NH-alkenyl, -NH-alkynyl, -NH-aryl, -NH- heteroaryl, -OH, -OAc, -O-C(O)alkyl, -O-alkyl, -O-alkenyl, -S-(alkyl), -S- (alkenyl), -S-(alkynyl), -S(O)-(alkyl), -SO2-(alkyl), -SO2-(aryl), -SO2- (heteroaryl), alkyl-OH, alkyl-O-alkyl, alkyl-NH2, alkyl-NH-alkyl, or alkyl- N(alkyl)2;754937-1152-7546v.2preferably, Z is NR22;(c) R3 is aryl; preferably R3 is Cs-Cs aryl; more preferably, R3 is Ce aryl; more preferably, R3 is a substituted or unsubstituted benzene; more preferably, R3 has the following structure:wherein R23, R24, R25, R26, R27 are each, independently H, halo, -CN, -CF3, - OCF3, -alkyl, -alkenyl, -alkynyl, -aryl, -heteroaryl, -NH2, -NH-alkyl, -NH- alkenyl, -NH-alkynyl, -NH-aryl, -NH-heteroaryl, -OH, -OAc, -O-C(O)alkyl, -O- alkyl, -O-alkyl-haloalkyl, -O-alkyl-halo, -O-alkenyl, -S-(alkyl), -S-(alkenyl), -S- (alkynyl), -S(O)-(alkyl), -SO2-(alkyl), -SO2-(aryl), -SO2-(heteroaryl), alkyl-OH, alkyl-O-alkyl, alkyl-NH2, alkyl-NH-alkyl, or alkyl-N(alkyl)2;(d) RHs alkyl, alkenyl, or alkynyl; preferably R4 is alkyl; more preferably, R4 is Ci- 6 alkyl; more preferably R4is methyl;(e) R5 is alkynyl or -CN; more preferably Rs is -CN;(f) Re is H, alkyl, alkenyl, alkynyl, -OH, -O-alkyl, -NH2, -NH-alkyl, -NH-(CO)- alkyl, or -NH-(SO2)-alkyl; preferably Re is H or alkyl; more preferably Re is H or methyl; and / or(g) R7 is aryl; preferably R7 is Cs-Cg aryl; more preferably, R7 is a Ce aryl; more preferably, R7 is a substituted or unsubstituted benzene; more preferably, R7 has the following structure:764937-1152-7546v.2wherein R28, R29, R30, R31, R32 are each, independently H, halo, -CN, -CF3, -OCF3, -alkyl, -alkenyl, -alkynyl, -aryl, -heteroaryl, -NH2, -NH-alkyl, -NH- alkenyl, -NH-alkynyl, -NH-aryl, -NH-heteroaryl, -OH, -OAc, -O- C(O)alkyl, -O-alkyl, -O-alkyl-haloalkyl, -O-alkyl-halo, -O-alkenyl, -S- (alkyl), -S-(alkenyl), -S-(alkynyl), -S(O)-(alkyl), -SO2-(alkyl), -SO2-(aryl), -SO2-(heteroaryl), alkyl-OH, alkyl-O-alkyl, alkyl-NH2, alkyl-NH-alkyl, or alkyl-N(alkyl)2.

4. The compound of claim 3, wherein R22 is H, halo, -CN, -CF3, -OCF3, -alkyl, -alkenyl, - alkynyl, -aryl, -heteroaryl, -NH2, -NH-alkyl, -NH-alkenyl, -NH-alkynyl, -NH-aryl, -NH- heteroaryl, -OH, -OAc, -O-C(O)alkyl, -O-alkyl, -O-alkenyl, -S-(alkyl), -S-(alkenyl), -S- (alkynyl), -S(O)-(alkyl), -SO2-(alkyl), -SO2-(aryl), -SO2-(heteroaryl), alkyl-OH, alkyl-O- alkyl, alkyl-NH2, alkyl-NH-alkyl, or alkyl-N(alkyl)2j and one of Ris, R19, R20, and R21 is H; preferably, two of Ris, R19, R20, and R21 are H; more preferably, three of Ris, R19, R20, and R21 are H; more preferably, Ris, R19, R20, and R21 are H;preferably, R22 is H, halo, -CN, -CF3, -OCF3, or -alkyl; more preferably R22 is alkyl; more preferably R22 is Ci-6 alkyl; more preferably R22 is methyl.

5. The compound of claim 3, wherein R25 is H, halo, -CN, -CF3, -OCF3, -alkyl, -alkenyl, - alkynyl, -aryl, -heteroaryl, -NH2, -NH-alkyl, -NH-alkenyl, -NH-alkynyl, -NH-aryl, -NH- heteroaryl, -OH, -OAc, -O-C(O)alkyl, -O-alkyl, -O-alkyl-haloalkyl, -O-alkyl-halo, -O- alkenyl, -S-(alkyl), -S-(alkenyl), -S-(alkynyl), -S(O)-(alkyl), -SO2-(alkyl), -SO2-(aryl), - SO2-(heteroaryl), alkyl-OH, alkyl-O-alkyl, alkyl-NH2, alkyl-NH-alkyl, or alkyl-N(alkyl)2; and one of R23, R24, R26, and R27 is H; preferably, two of R23, R24, R26, and R27 are H; more preferably, three of R23, R24, R26, and R27 are H; more preferably, R23, R24, R26, and R27 are H;preferably, R25 is H, halo, -CN, -CF3, -OCF3, -alkyl or -O-alkyl-halo; preferably R25 is alkyl or -O-alkyl-halo; more preferably R25 is C1-6 alkyl or -O-C1-6 alkyl-halo; more preferably R25 is methyl or -O-CHF2.

6. The compound of claim 3, wherein R24 is H, halo, -CN, -CF3, -OCF3, -alkyl, -alkenyl, - alkynyl, -aryl, -heteroaryl, -NH2, -NH-alkyl, -NH-alkenyl, -NH-alkynyl, -NH-aryl, -NH-774937-1152-7546v.2heteroaryl, -OH, -OAc, -O-C(O)alkyl, -O-alkyl, -O-alkyl-haloalkyl, -O-alkyl-halo, -O- alkenyl, -S-(alkyl), -S-(alkenyl), -S-(alkynyl), -S(O)-(alkyl), -SO2-(alkyl), -SO2-(aryl), - SO2-(heteroaryl), alkyl-OH, alkyl-O-alkyl, alkyl-NH2, alkyl-NH-alkyl, or alkyl-N(alkyl)2; and one of R23, R25, R26, and R27 is H; preferably, two of R23, R25, R26, and R27 are H; more preferably, three of R23, R25, R26, and R27 are H; more preferably R23, R25, R26, and R27 are H;preferably, R24 is H, halo, -CN, -CF3, -OCF3, -alkyl or -O-alkyl-halo; preferably R24 is CF3.

7. The compound of claim 3, wherein R30 is H, halo, -CN, -CF3, -OCF3, -alkyl, -alkenyl, - alkynyl, -aryl, -heteroaryl, -NH2, -NH-alkyl, -NH-alkenyl, -NH-alkynyl, -NH-aryl, -NH- heteroaryl, -OH, -OAc, -O-C(O)alkyl, -O-alkyl, -O-alkyl-haloalkyl, -O-alkyl-halo, -O- alkenyl, -S-(alkyl), -S-(alkenyl), -S-(alkynyl), -S(O)-(alkyl), -SO2-(alkyl), -SO2-(aryl), - SO2-(heteroaryl), alkyl-OH, alkyl-O-alkyl, alkyl-NH2, alkyl-NH-alkyl, or alkyl-N(alkyl)2; and one of R28, R29, R31, and R32 is H; preferably, two of R28, R29, R31, and R32 are H; more preferably, three of R28, R29, R31, and R32 are H; more preferably, R28, R29, R31, and R32 are H;preferably, R30 is H, halo, -CN, -CF3, -OCF3, -alkyl or -O-alkyl-halo; preferably R30 is alkyl; more preferably R30 is Ci-6 alkyl; more preferably R30 is propyl.

8. The compound of any one of claims 1-7, wherein X is O.

9. The compound of claim 3 having the structure:whereinReis H, alkyl, -O-alkyl, -NH-alkyl, -NH-(CO)-alkyl, or -NH-(SO2)-alkyl; and784937-1152-7546v.2R25 is -alkyl, -NH-alkyl, -O-alkyl, -O-alkyl-halo, -S(O)-(alkyl), -SO2-(alkyl), alkyl- OH, -alkyl-O-alkyl, alkyl-NTb, alkyl-NH-alkyl, or alkyl-N(alkyl)2.

10. The compound of claim 9 having the structure:

11. The compound of claim 1 having the structure:

12. The compound of claim 1 having the structure:

13. The compound of claim 12, wherein(a) Rs has the following structure:794937-1152-7546v.2wherein R33, R34, R35, R36, R37 are each, independently H, halo, -CN, -CF3, -OCF3, -alkyl, -alkenyl, -alkynyl, -aryl, -heteroaryl, -NH2, -NH-alkyl, -NH-alkenyl, -NH- alkynyl, -NH-aryl, -NH-heteroaryl, -OH, -OAc, -O-C(O)alkyl, -O-alkyl, -O- alkenyl, -S-(alkyl), -S-(alkenyl), -S-(alkynyl), -S(O)-(alkyl), -SO2-(alkyl), -SO2- (aryl), -SO2-(heteroaryl), alkyl-OH, alkyl-O-alkyl, alkyl-NH2, alkyl-NH-alkyl, or alkyl-N(alkyl)2;(b) R9 is H or alkyl; preferably R9 is H or Ci-6 alkyl; more preferably R9 is H or ethyl; (c) Rio has the following structure:wherein n is 0, 1, 2, or 3; preferably n is 0 or 1; andR38, R39, R40, R41, R42 are each, independently H, halo, -CN, -CF3, -OCF3, -alkyl, -alkenyl, -alkynyl, -aryl, -heteroaryl, -NH2, -NH-alkyl, -NH-alkenyl, -NH-alkynyl, -NH-aryl, -NH-heteroaryl, -OH, -OAc, -O-C(O)alkyl, -O-alkyl, -O-alkenyl, -S- (alkyl), -S-(alkenyl), -S-(alkynyl), -S(O)-(alkyl), -SO2-(alkyl), -SO2-(aryl), -SO2- (heteroaryl), alkyl-OH, alkyl-O-alkyl, alkyl-NH2, alkyl-NH-alkyl, or alkyl- N(alkyl)2.

14. The compound of claim 13, wherein(a) R34 is H, halo, -CN, -CF3, -OCF3, -alkyl, -alkenyl, -alkynyl, -aryl, -heteroaryl, -NH2, - NH-alkyl, -NH-alkenyl, -NH-alkynyl, -NH-aryl, -NH-heteroaryl, -OH, -OAc, -O- C(O)alkyl, -O-alkyl, -O-alkyl-haloalkyl, -O-alkenyl, -S-(alkyl), -S-(alkenyl), -S- (alkynyl), -S(O)-(alkyl), -SO2-(alkyl), -SO2-(aryl), -SO2-(heteroaryl), alkyl-OH, alkyl-804937-1152-7546v.2O-alkyl, alkyl-NH2, alkyl-NH-alkyl, or alkyl-N(alkyl)2; and R33, R35, R36, and R37 are H;preferably, R34 is H, halo, -CN, -CF3, -OCF3, -alkyl or -O-alkyl-halo; more preferably R34 is alkyl or halo; more preferably R34 is C1-6 alkyl or halo; more preferably R34 is methyl or F; and(b) R38 and R41 are each independently H, halo, -CN, -CF3, -OCF3, -alkyl, -alkenyl, - alkynyl, -aryl, -heteroaryl, -NH2, -NH-alkyl, -NH-alkenyl, -NH-alkynyl, -NH-aryl, - NH-heteroaryl, -OH, -OAc, -O-C(O)alkyl, -O-alkyl, -O-alkyl-haloalkyl, -O-alkenyl, - S-(alkyl), -S-(alkenyl), -S-(alkynyl), -S(O)-(alkyl), -SO2-(alkyl), -SO2-(aryl), -SO2- (heteroaryl), alkyl-OH, alkyl-O-alkyl, alkyl-NH2, alkyl-NH-alkyl, or alkyl-N(alkyl)2; and R39, R40, and R42 are H;preferably, R38 and R41 are each independently H, halo, -alkyl or -O-alkyl; more preferably, R38 is -O-alkyl and R41 is -alkyl; more preferably R38 is -O- Ci-6 alkyl and R41 is -C1-6 alkyl; more preferably R38 is -O-methyl and R41 is methyl; more preferably, R38 and R41 are each independently halo; more preferably R38 and R41 are F; more preferably, R38, R39, R40, R41 and R42 are H.

15. The compound of claim 12 having the structure:

16. The compound of claim 12, wherein(a) Rn is H, alkenyl, aryl, substituted styrenyl or heterocycloalkyl-aryl; preferably Ru is H, styrenyl-O-alkyl, or N-containing heterocycloalkyl-aryl; more preferably Rn is H, styrene-O-Ci-6 alkyl, or N-containing heterocycloalkyl-aryl, wherein the814937-1152-7546v.2heterocylcoalkyl is piperidine; more preferably Rn is H, dimethoxy styrene or N- phenyl-piperidine;(b) R12 is H or -(CC>2)-alkyl; preferably R12 is H or -(CO2)-Ci-6 alkyl; more preferably R12 is H or -(CO2)-ethyl;(c) R13 is H or alkyl; preferably R13 is H or alkyl-O-alkyl; more preferably R13 is H or C1-6 alkyl-O-Ci-6 alkyl; more preferably R13 is H or -CH2-O-isopropyl;(d) R14 is H or alkyl; more preferably R14 is H or Ci-6 alkyl; more preferably R14 is H or methyl;(e) R15 is H, alkyl, alkyl-heterocycloalkyl-aryl, or -CF3; preferably R15 is H, C1-6 alkyl, Ci- 6 alkyl-N-containing heterocycloalkyl-benzene, or -CF3; more preferably R15 is H, methyl, -CFF-piperidine-zn-trifluoromethylbenzene, or -CF3;(f) Ri6 is H or -OH; and(g) R17 is H or -NH-aryl; preferably R17 is H or -NH-benzene; more preferably R17 is H or -NH- -aniline; more preferably R17 is H or -NH- / ?-N, N-dimethylaniline.

17. The compounds of claim 1 having the structure:

18. A compound having the structure:824937-1152-7546v.2whereinA is a ring;X is CH2, N, or S;Ri is an aryl or heteroaryl;R2and R3 are each independently aryl, heteroaryl, cycloalkyl or heterocycloalkyl;R4 is alkenyl, alkynyl, -CN; -OH, -O-alkyl, -NH2, -NH-alkyl, -NH-(CO)-alkyl, or -NH- (SO2)-alkyl;Rs is alkyl, alkenyl, alkynyl, -CN; -OH, -O-alkyl, -NH2, -NH-alkyl, -NH-(CO)-alkyl, or - NH-(SO2)-alkyl;R(, is H, alkyl, alkenyl, alkynyl, -OH, -O-alkyl, -NH2, -NH-alkyl, -NH-(CO)-alkyl, or -NH- (SO2)-alkyl;R7 is aryl or heteroaryl;Rs is cycloalkyl, heterocycloalkyl, alkenyl, alkynyl, aryl, or heteroaryl;R9 and Rio are each independently alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, or heteroaryl;Yis CRn;R11, Ri2, R13, R14, RIS, and Rie are each independently H, alkyl, alkenyl, alkynyl, aryl, heterocycloalkyl, alkyl-heterocycloalkyl, heteroaryl, -(CO2)-alkyl, -(CO2)-aryl -NH2, -NH- alkyl, -NH-alkenyl, -NH-alkynyl, -NH-aryl, -NH-heteroaryl, -OH, -OAc, -O-C(O)alkyl,, halo, -CF3, -OCFs-O-alkyl, -O-alkenyl, -O-aryl, -S-(alkyl), -S-(alkenyl), -S-(alkynyl), - S(O)-(alkyl), -SO2-(alkyl), -SO2-(aryl), -SO2-(heteroaryl), alkyl-OH, alkyl-O-alkyl, alkyl- NH2, alkyl-NH-alkyl, or alkyl-N(alkyl)2; and834937-1152-7546v.2R17 is alkyl, alkenyl, alkynyl, aryl, heterocycloalkyl, alkyl-heterocycloalkyl, heteroaryl, - (CO2)-alkyl, -(CO2)-aryl -NH2, -NH-alkyl, -NH-alkenyl, -NH-alkynyl, -NH-heteroaryl, - OH, -OAc, -O-C(O)alkyl,, halo, -CF3. -OCF3-O-alkyl, -O-alkenyl, -O-aryl, -S-(alkyl), -S- (alkenyl), -S-(alkynyl), -S(O)-(alkyl), -SO2-(alkyl), -SO2-(aryl), -SO2-(heteroaryl), alkyl- OH, alkyl-O-alkyl, alkyl-NH2, alkyl-NH-alkyl, or alkyl-N(alkyl)2.

19. A compound having the following structure:whereinA is a ring;X is NH, O, or S;Ri is an aryl or heteroaryl;R2 and R3 are each independently aryl, heteroaryl, cycloalkyl or heterocycloalkyl; tandRs are each independently alkyl, alkenyl, alkynyl, -CN; -OH, -O-alkyl, -NH2, -NH- alkyl, -NH-(CO)-alkyl, or -NH-(SO2)-alkyl;Re is H, alkyl, alkenyl, alkynyl, -OH, -O-alkyl, -NH2, -NH-alkyl, -NH-(CO)-alkyl, or -NH- (SO2)-alkyl;R7 is aryl or heteroaryl;Rs is cycloalkyl, heterocycloalkyl, alkenyl, alkynyl, aryl, or heteroaryl;R9 and Rio are each independently H, alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, or heteroaryl;Y is CR17 or N; andR11, R12, Ri3, Ri4, RIS, R16, and R17 are each independently H, alkyl, alkenyl, alkynyl, aryl, heterocycloalkyl, alkyl-heterocycloalkyl, heteroaryl, -(CO2)-alkyl, -(CO2)-aryl -NH2, -NH-844937-1152-7546v.2alkyl, -NH-alkenyl, -NH-alkynyl, -NH-aryl, -NH-heteroaryl, -OH, -OAc, -O-C(O)alkyl,, halo, -CF3, -OCFs-O-alkyl, -O-alkenyl, -O-aryl, -S-(alkyl), -S-(alkenyl), -S-(alkynyl), - S(O)-(alkyl), -SO2-(alkyl), -SO2-(aryl), -SO2-(heteroaryl), alkyl-OH, alkyl-O-alkyl, alkyl- NH2, alkyl-NH-alkyl, or alkyl-N(alkyl)2;wherein the compound is other than854937-1152-7546v.

220. A pharmaceutical composition comprising the compound of any one of claims 1-19 and a pharmaceutically acceptable carrier.

21. A method of(a) inhibiting interaction between ID2 protein and tumor suppressor complex CRLVHLcomprising contacting the ID2 protein or the suppressor complex CRLVHLwith the compound of any one of claims 1-19 or the pharmaceutical composition of claim 20;(b) inhibiting proliferation of tumor or cancer cells comprising contacting ID2 protein or suppressor complex CRLVHLwith the compound of any one of claims 1-19 or the pharmaceutical composition of claim 20;(c) suppressing activity of tumor or cancer cells comprising contacting ID2 protein or suppressor complex CRLVHLwith the compound of any one of claims 1-19 or the pharmaceutical composition of claim 20; and / or(d) treating tumor or cancer in a subject comprising administering the compound of any one of claims 1-19 or the pharmaceutical composition of claim 20 to the subject thereby to treat the subject.

22. The method of claim 21, wherein the cancer is bladder cancer, breast cancer, colorectal cancer, kidney cancer, lung cancer, lymphoma, melanoma, oral and oropharyngeal cancer, pancreatic cancer, prostate cancer, thyroid cancer or uterine cancer; or wherein the subject is human.864937-1152-7546v.

223. An assay for selecting one or more compounds that inhibit interaction between ID2 protein and tumor suppressor complex CRIHLfrom a pool of compounds; wherein the assay comprises donor beads and acceptor beads.

24. The assay of claim 23, wherein the compound has the following structure:874937-1152-7546v.24937-1152-7546v.2O26. A method of selecting one or more compounds that inhibit interaction between ID2 protein and tumor suppressor complex CRLVHLfrom a pool of compounds using the assay of claim 23, wherein the method comprises the following steps:adding, in any order:i) a glutathione S-transferase tagged von Hippel-Lindau tumor suppressor protein and an Elongin C protein complex (GST-VHL / EloC);ii) one or more compounds from the pool of compounds;iii) FLAG tags;iv) acceptor beads; andv) donor beads;exciting a luminescent signal at a wavelength between 670-690 nm, preferably at 680 nm;and894937-1152-7546v.2obtaining an emission signal at a wavelength between 610-625 nm, preferably at 617 nm.

27. The method of claim 26, wherein the(a) addition is in the order of i), ii), iii), iv) and v);(b) method further comprises incubating the donor beads before adding the acceptor beads; or incubating the acceptor beads after adding the donor beads;(c) emission signal in step (c) is used to calculate the inhibition of the interaction between the ID2 protein and the suppressor complex CRLV HL; or(d) emission signal was used to calculate a Z-score to validate the assay; preferably, the Z-score is >0.85.

28. The method of claim 26, further comprising(a) calculating an inhibition threshold of the compounds from the obtained emission signal in step (c);(b) identifying the compounds in step (a) having at least 62% inhibition threshold; (c) identifying the compounds in step (b) that are soluble, are not singlet oxygen quenchers, or do not alter reactive oxygen species (ROS) levels;(d) conducting a surface plasmon resonance (SPR) binding analysis on the compounds identified in step (c); and / or(e) identifying and selecting the compounds having a 1:1 binding mode with the suppressor complex CRLVHL;wherein the compounds selected in step (e) are the compounds that inhibit the interaction between the ID2 protein and the tumor suppressor complex CRIHL.

29. The method of claim 28, further comprising use of Tanimoto similarity scores of the compounds from step (e) to generate a pool of compounds that inhibit interaction between the ID2 protein and the tumor suppressor complex CRLVHL; and / or further comprising a calculation of PAMPA permeability scores to select compounds that inhibit interaction between the 1D2 protein and the tumor suppressor complex CRLVHL.904937-1152-7546v.

230. The method of any one of claims 26-29, wherein the one or more compounds have the structure:

31. The method of any one of claims 26-29, wherein the one or more compounds have the structure:914937-1152-7546v.2F924937-1152-7546v.24937-1152-7546v.2

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