Compounds and formulations for restoring tumor suppressor functions

Abstract

Formulations, compounds, and methods that target p53 mutants for treatment of cancers with p53 mutations are described. Treatments include administration of a compound to bind with p53 mutants.

Description

COMPOUNDS AND FORMULATIONS FOR RESTORING TUMOR SUPPRESSOR FUNCTIONSCROSS-REFERNCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of priority to U.S. Provisional Appl. No. 63 / 560,520, entitled “Compounds and Formulations for Restoring Tumor Suppressor Functions,” filed March 1 , 2024, the contents of which is herein incorporated by reference.SEQUENCE LISTING

[0002] The instant application contains a Sequence Listing which has been submitted electronically in XML format and is hereby incorporated by reference in its entirety. Said XML copy, created on February 25, 2025, is named 08969. PCT.xml and is 4,096 bytes in size.STATEMENT OF FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT

[0003] This invention was made with government support under Grant No. CA267495- 01 A1 , awarded by the National Institutes of Health and under Grant No. CA-283462-01 , awarded by the National Institutes of Health. The government has certain rights in the invention.TECHNICAL FIELD

[0004] The invention is generally directed toward compounds, formulations, and methods for restoring tumor suppressor p53 functions.BACKGROUND

[0005] The TP53 gene provides instructions for making a protein called tumor protein p53 (or p53). This protein acts as a tumor suppressor, which means that it regulates cell division by keeping cells from growing and dividing (proliferating) too fast or in an uncontrolled way.

[0006] The p53 protein locates in the nucleus of cells throughout the body, where it attaches (binds) directly to DNA. When DNA in a cell becomes damaged by agents such as toxic chemicals, radiation, or ultraviolet rays, this protein plays an important role in determining whether the DNA will be repaired, or the damaged cell will self-destruct (undergo apoptosis). If the DNA is to be repaired, p53 activates cell cycle arrest and a repair mechanism to fix the damage. If the DNA cannot be repaired, p53 prevents the cell from dividing and signals to undergo apoptosis. By stopping cells with mutated or damaged DNA from dividing, p53 helps prevent the development of tumors. When the p53 protein itself is mutated, it can lose the ability to control tumorigenesis, resulting in cancer cell proliferation.

[0007] Several cancers are associated with mutated p53, including (but not limited to) ovarian cancers, breast cancers, esophageal cancers, colorectal cancers, head and neck cancers, larynx cancers, and lung cancers. The tumor suppressor p53 has long been a rationale target for therapeutic exploration for treating cancer. Targeting p53 mutants for cancer therapy requires novel solutions due to the difficulty of restoring the protein’s tumor suppressor function. Due to the fact that p53 is a transcription factor and requires a challenging task of restoring function, mutant p53 isoforms have been considered undruggable. A number of small molecules have been identified to restore certain tumor suppressor activity of select p53 mutants in cancer cell lines. However, most of these molecules act indirectly, or lack a comprehensive treatment strategy for the many mutant p53 isoforms. These molecules do not show discriminative efficacy on tumors with mutant p53.SUMMARY

[0008] Various embodiments are directed towards formulations, compounds and methods to restore tumor suppressor activity of p53 mutants in human cancer cell lines. The small molecules in accordance with many embodiments bind directly to mutant p53. In some embodiments, the small molecules form covalent bonds with the mutant p53. In some embodiments, the small molecules form non-covalent bonds with the mutant p53.Direct binding to mutant p53 can induce conformation changes in the mutant p53. The conformation changes can activate and restore p53 tumor suppressor activities.

[0009] Various aspects of the disclosure are related to a compound of Formula I:or a therapeutically acceptable salt thereof; wherein R1is H, alkyl, benzyl, aryl, heteroaryl, heterocycle, alkyl, or alkyl chain and heteroatom;R2is H or alkyl;R3is H or halogen; andR4is H or CH3.

[0010] Various aspects of the disclosure are related to a compound of Formula II:or a therapeutically acceptable salt thereof; wherein R1is H, alkyl, benzyl, aryl, heteroaryl, or alkyl chain and heteroatoms;R2is H, CH3, or alkyl;R3is H or halogen; andR4is H or CH3.

[0011] Various aspects of the disclosure are related to a compound of Formula III:or a therapeutically acceptable salt thereof; wherein R1is H, alkyl, benzyl, aryl, heteroaryl, or alkyl chain and heteroatoms;R2is H or alkyl;R3is H or halogen; andR4is H or CH3.

[0012] Various aspects of the disclosure are related to a compound of Formula IV:or a therapeutically acceptable salt thereof; wherein R1is H, alkyl, benzyl, aryl, heteroaryl, or alkyl chain and heteroatoms;R2is H or alkyl;R3is H or halogen; andR4is H or CH3.

[0013] Various aspects of the disclosure are related to a compound of Formula V:or a therapeutically acceptable salt thereof; wherein R1is H, alkyl, benzyl, aryl, heteroaryl, or alkyl chain and heteroatoms;R2is H or alkyl;R3is H or halogen;R4is H or CH3;X is CH or N; andY is S or 0.

[0014] Various aspects of the disclosure are related to a compound of Formula Vl-a or Vl-b:or a therapeutically acceptable salt thereof; wherein R1is H, alkyl, benzyl, aryl, heteroaryl, or alkyl chain and heteroatoms;R2is H or alkyl;R3is H or halogen;R4is H or CH3;X is CH or N; andX1, X2, X3, and X4are each independently C or N.

[0015] Various aspects of the disclosure are related to a compound of Formula VII:VII or a therapeutically acceptable salt thereof; wherein R2is H or alkyl;R3is H or halogen;R4is H or CH3; andX is 0 or S.

[0016] Various aspects of the disclosure are related to a compound of Formula VIII:or a therapeutically acceptable salt thereof; wherein R1and R2are H or halogen;R3is H, alkyl, aryl, benzyl, or acyl; and R4is OMe.

[0017] Various aspects of the disclosure are related to a compound of Formula IX:or a therapeutically acceptable salt thereof; wherein R1is H, Boc, benzoyl, p-trifuoromethylbenzyl, p-methoxybenzyl, pentyl, or 4- pentynyl; and R2is OMe.

[0018] Various aspects of the disclosure are related to a compound of Formula X:or a therapeutically acceptable salt thereof; wherein R1is H or alkyl; and R2is OMe.

[0019] Various aspects of the disclosure are related to a compound of Formula XI:or a therapeutically acceptable salt thereof; wherein R1is H or alkyl; andR2is amine, alkyl amine, or dialkyl amine.

[0020] Various aspects of the disclosure are related to a compound of Formula XII:or a therapeutically acceptable salt thereof; wherein R1is H or alkyl; andR2is acylamide, methacrylamide or NHC0NMe2.

[0021] Various aspects of the disclosure are related to a compound of Formula XIII:or a therapeutically acceptable salt thereof; wherein R1is H or alkyl;R2is Boc; andR3is H or alkyl.

[0022] Various aspects of the disclosure are related to a compound of Formula XIV:or a therapeutically acceptable salt thereof; wherein R1is alkyl or 4-pentynyl; and R2is acylamide or N-methylacrylamide.

[0023] Various aspects of the disclosure are related to a compound of Formula XV:or a therapeutically acceptable salt thereof; wherein R1is H or alkyl; andR2is H, acylamide, or OMe.

[0024] Various aspects of the disclosure are related to a compound of Formula XVI:XVI or a therapeutically acceptable salt thereof; wherein R1is H, alkyl, benzyl, aryl, heteroaryl, heterocycle, or alkyl chain with heteroatom; R2is H, or alkyl;R3is H, or halogen;R4is H, or CH3; andX1, X2, X3, and X4are each independently C or N.

[0025] Various aspects of the disclosure are related to a compound selected from any one of the compounds depicted in Tables 1 -6 or therapeutically acceptable salts thereof.

[0026] Various aspects of the disclosure are related to a pharmaceutical composition comprising a compound of any one of Formula I to Formula XVI, and a pharmaceutically acceptable carrier, adjuvant, or vehicle.

[0027] Various aspects of the disclosure are related to a method of treating cancer in a patient comprising administering to said patient a compound of any one of Formula I toFormula XVI, or a pharmaceutical composition of a compound of any one of Formula I to Formula XVI, wherein the cancer is associated with p53 mutagenesis.

[0028] Various aspects of the disclosure are related to a method of treating cancer in a patient in which the compound comprises a functional group that binds with and stabilizes a p53 mutant such that the stabilization of the p53 mutant reactivates gene expression of p53 wild type.

[0029] Various aspects of the disclosure are related to a method of treating cancer in a patient in which prior to administering the compound, assessing a sample derived from the patient for a presence of a p53 mutant within the cancer.

[0030] Various aspects of the disclosure are related to a method of treating cancer in a patient in which the p53 mutant comprises a mutation within one or more residue positions selected from the group consisting of: R175, G245, R248, R249, R273, R282, R280, Y220, A138, P151 , P152, G154, R158, V172, H193, H214, S215, Y234, P250, I255, V272, V274, E285, and E286.

[0031] Various aspects of the disclosure are related to a method of treating cancer in a patient in which the cancer is ovarian cancer, breast cancer, esophageal cancer, colorectal cancer, head and neck cancer, larynx cancers, and lung cancer.

[0032] Various aspects of the disclosure are related to a method of treating cancer in a patient in which the cancer is a human ovarian cancer or a human breast cancer.BRIEF DESCRIPTION OF THE DRAWINGS

[0033] The description and claims will be more fully understood with reference to the following figures and data graphs, which are presented as exemplary embodiments of the invention and should not be construed as a complete recitation of the scope of the invention.

[0034] Figure 1 illustrates a viability assay of compounds 11 and 60 in ovarian cancer cells with p53 mutations in accordance with various embodiments.

[0035] Figure 2 illustrates a cellular thermal shift assay of compounds 11 and 60 in ovarian cancer cells with p53 mutations in accordance with various embodiments.

[0036] Figure 3 illustrates a DNA binding assay for p53 in presence of compounds 11 and 60 in ovarian cancer cells with p53 mutations in vivo in accordance with various embodiments.

[0037] Figure 4 illustrates a chromatin immunoprecipitation analysis of compounds 11 and 60 in ovarian cancer cells with p53 mutations in accordance with various embodiments.

[0038] Figure 5 illustrates an RNA transcription induction analysis of compounds 11 and 60 in ovarian cancer cells with p53 mutations in accordance with various embodiments.

[0039] Figure 6A illustrates a viability assay of compounds in various cancer cells with p53 mutations in accordance with various embodiments.

[0040] Figure 6B illustrates the ICso values of various cancer cell lines in accordance with various embodiments.

[0041] Figure 7 illustrates compound 11 restores DNA binding of various p53 mutants in vivo in accordance with various embodiments.

[0042] Figure 8 is an immunoblotting assay that shows the effect of compound 11 on p21 protein levels.

[0043] Figure 9 is an assay to test covalent attachment of compounds 11 , 23, and 24 to p53 proteins.

[0044] Figure 10 provides experimental results that indicate peptides from p53G245Sare covalently bonded with compound 11 as identified by mass spectrometry.DETAILED DESCRIPTION

[0045] Turning now to the drawings and data, compounds, formulations and methods for restoring and / or reactivating tumor suppressor functions of p53 mutations are described. In some embodiments, compounds can bind directly to mutant destabilized p53. The compounds can bind with the p53 mutants covalently and / or non-covalently. The binding with the compounds can stabilize the p53 mutant conformations and restore the wild-type functions. The restoration of wild-type functions by the compounds can prevent growth and / or induce death in cancer cells. The compounds can bind andstabilize conformation of mutant p53 and induce its activation to prevent cancer cell growth and / or induce cancer cell death. The compounds can be used with a pharmaceutical formulation for treatment of various human cancers with p53 mutations such as (but not limited to) ovarian cancers, breast cancers, esophageal cancers, colorectal cancers, head and neck cancers, larynx cancers, and lung cancers.Terms of Art

[0046] “Acrylamide”, by itself or as part of another substituent means, unless otherwise stated, refers to an organic compound with the chemical formula CH2=CHC(O)NH2.

[0047] “Alkyl”, by itself or as part of another substituent means, unless otherwise stated, refers to a straight or branched hydrocarbon chain having the number of carbon atoms designated (e.g. C1-10 means one to ten carbon atoms) and including straight, branched chain, or cyclic substituent groups. Examples include methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, pentyl, neopentyl, hexyl, and cyclopropylmethyl. In some embodiments, a straight chain or a branched chain alkyl can have less than or equal to 30 carbon atoms in its backbone.

[0048] “Alkane”, by itself or as part of another substituent means, unless otherwise stated, refers to saturated aliphatic groups, including a compound of carbon (C) and hydrogen (H) that contains only carbon hydrogen single bonds.

[0049] “Alkyne”, by itself or as part of another substituent means, unless otherwise stated, refers to a hydrocarbon structure that contains a carbon-carbon triple bond.

[0050] “Aromatic”, by itself or as part of another substituent means, unless otherwise stated, refers to a carbocycle or heterocycle with one or more polyunsaturated rings and having aromatic character, e.g. having (4n+2) delocalized IT (pi) electrons, where n is an integer.

[0051] “Aryl”, by itself or in combination with other terms, means, unless otherwise stated, refers to a carbocyclic aromatic system containing one or more rings (typically one, two or three rings), wherein such rings may be attached together in a pendent manner, such as a biphenyl, or may be fused, such as naphthalene. Examples of aryl groups include phenyl, anthracyl, and naphthyl.

[0052] “Benzimidazole”, by itself or in combination with other terms, means, unless otherwise stated, refers to a heterocyclic aromatic organic compound with fused rings of the aromatic compounds benzene and imidazole.

[0053] “Benzyl”, by itself or as part of another substituent means, unless otherwise stated, refers to a benzene ring attached to a methylene group, R-CH2-C6H5.

[0054] “Enone”, by itself or as part of another substituent means, unless otherwise stated, refers to a chemical structure where a ketone is conjugated with an alkene.

[0055] “Halogen” or “halo”, by itself or as part of another substituent means, unless otherwise stated, refers to a fluorine (F), chlorine (Cl), bromine (Br), or iodine (I) atom.

[0056] “Heteroaryl”, by itself or as part of another substituent means, unless otherwise stated, refers to an aromatic molecule containing at least one heteroatom as part of the aromatic ring.

[0057] “Heteroatom”, by itself or as part of another substituent means, unless otherwise stated, refers to an atom that is not a carbon atom or a hydrogen atom.

[0058] “Heterocycle”, by itself or as part of another substituent means, unless otherwise stated, refers to a cyclic compound that has atoms of at least two different elements as members of its ring(s). A heterocycle may be aromatic or non-aromatic in nature.

[0059] “Hydrocarbon”, by itself or as part of another substituent means, unless otherwise stated, refers to an organic chemical compound that consists entirely of the elements carbon (C) and hydrogen (H).

[0060] “Isoxazole”, by itself or as part of another substituent means, unless otherwise stated, refers to an azole with an oxygen atom (O) next to a nitrogen atom (N).

[0061] “R” in the molecular formula above and throughout are meant to indicate any suitable organic molecule.

[0062] “Small molecules”, by itself or as part of another substituent means, unless otherwise stated, refers to organic molecules and / or organic compounds with molecular weight less than or equal to about 1000 Daltons.Compounds

[0063] Several embodiments are directed towards formulations and compounds and their use as therapeutics to treat cancers. A number of compounds have been identified that can target the p53 mutants by binding and restabilizing un-stabilized p53 mutant conformations. In some embodiments, a compound is a small molecule. In some embodiments, a compound is a small molecule comprising a benzimidazole. In some embodiments, a compound is a small molecule comprising a benzimidazole and an acrylamide. The small molecules can bind p53 mutants covalently.

[0064] In some embodiments, a compound is a small molecule comprising a benzimidazole, an acrylamide, and an enone. In some embodiments, a formula to bind p53 mutant comprises a compound of Formula I:wherein R1is H, alkyl, benzyl, aryl, heteroaryl, heterocycle, or alkyl chain with heteroatom; R2is H, or alkyl;R3is H, or halogen; andR4is H, or CH3.

[0065] In some embodiments, R1is an alkyl chain with a heteroatom at the end. The examples of heteroatoms include (but are not limited to) N, O, S, OH, alcohol, ether, and amine. In certain embodiments, R1is an alkyl chain with a heteroatom OH. In certain embodiments, R1is an alkyl chain with a heteroatom amine. In some embodiments, R1is an alkyl chain with a heterocyclic ring at the end. In certain embodiments, R1is an alkyl chain with a morpholine.

[0066] In some embodiments, the alkyl or alkyl chain with heteroatom of R1is 1 to 12 carbons (C1-12). In some embodiments, the alkyl or alkyl chain with heteroatom of R1is 1 to 10 carbons (C1-10). In some embodiments, the alkyl or alkyl chain with heteroatom of R1is 1 to 8 carbons (C1 -8). In some embodiments, the alkyl or alkyl chain with heteroatom of R1is 1 to 6 carbons (C1-6). In some embodiments, the alkyl or alkyl chain with heteroatom of R1is 1 to 4 carbons (C1-4).

[0067] In some embodiments, the benzyl, aryl, heteroaryl, or heterocycle of R1is a 6- membered ring. In some embodiments, the benzyl, aryl, heteroaryl, or heterocycle of R1is a 5-membered ring. In some embodiments, the heteroaryl or heterocycle of R1comprises one or more nitrogen atoms (N). In some embodiments, the heteroaryl or heterocycle of R1comprises one or more oxygen atoms (O).

[0068] In some embodiments, the alkyl of R2is 1 to 8 carbons (C1-8). In some embodiments, the alkyl of R2is 1 to 6 carbons (C1-6). In some embodiments, the alkyl of R2is 1 to 4 carbons (C1-4). In some embodiments, the alkyl of R2is an ethyl. In some embodiments, the alkyl of R2is a methyl.

[0069] In some embodiments, the halogen of R3is fluoro, chloro, bromo, or iodo. In some embodiments, the halogen of R3is fluoro, chloro, or bromo. In some embodiments, the halogen of R3is fluoro or chloro. In some embodiments, a cyano group is utilized in place of the halogen. In some embodiments, a hydroxyl group is utilized in place of the halogen. In some embodiments, a methoxy group is utilized in place of the halogen.

[0070] In many embodiments, covalent linkers including (but not limited to) acrylamide can be implemented in the small molecules. In some embodiments, the acrylamide can form a covalent linkage with p53. In some embodiments, other covalent linkers can replace the acrylamide of the small molecules, such as (for example) propargylamide . In several embodiments, the covalent linkers can be chemical groups that contain one or more reactive ends that can chemically attach to specific functional groups on proteins, polypeptides, or other molecules. In certain embodiments, the covalent linkers can form covalent bonds with the p53 mutants. As can be readily appreciated, small molecules in accordance with various embodiments can utilize any of a variety of covalent linkers asappropriate to the requirements of specific applications. In some embodiments, a formula to bind p53 mutant comprises a compound of Formula II:II wherein R1is H, alkyl, benzyl, aryl, heteroaryl, heterocycle, or alkyl chain with heteroatom; R2is H, or alkyl;R3is H, or halogen; andR4is H, or CH3.

[0071] In some embodiments, R1is H, alkyl, benzyl, aryl, heteroaryl, or alkyl chain with heteroatoms.

[0072] In some embodiments, R1is an alkyl chain with a heteroatom at the end. The examples of heteroatoms include (but are not limited to) N, 0, S, OH, alcohol, ether, and amine. In certain embodiments, R1is an alkyl chain with a heteroatom OH. In certain embodiments, R1is an alkyl chain with a heteroatom amine. In some embodiments, R1is an alkyl chain with a heterocyclic ring at the end. In certain embodiments, R1is an alkyl chain with a morpholine.

[0073] In some embodiments, the alkyl or alkyl chain with heteroatom of R1is 1 to 12 carbons (C1-12). In some embodiments, the alkyl or alkyl chain with heteroatom of R1is 1 to 10 carbons (C1-10). In some embodiments, the alkyl or alkyl chain with heteroatom of R1is 1 to 8 carbons (C1 -8). In some embodiments, the alkyl or alkyl chain with heteroatom of R1is 1 to 6 carbons (C1-6). In some embodiments, the alkyl or alkyl chain with heteroatom of R1is 1 to 4 carbons (C1-4).

[0074] In some embodiments, the benzyl, aryl, heteroaryl, or heterocycle of R1is a 6- membered ring. In some embodiments, the benzyl, aryl, heteroaryl, or heterocycle of R1is a 5-membered ring. In some embodiments, the heteroaryl or heterocycle of R1comprises one or more nitrogen atoms (N). In some embodiments, the heteroaryl or heterocycle of R1comprises one or more oxygen atoms (0).

[0075] In some embodiments, the alkyl of R2is 1 to 8 carbons (C1-8). In some embodiments, the alkyl of R2is 1 to 6 carbons (C1-6). In some embodiments, the alkyl of R2is 1 to 4 carbons1-4 In some embodiments, the alkyl of R2is an ethyl. In some embodiments, the alkyl of R2is a methyl.

[0076] In some embodiments, the halogen of R3is fluoro, chloro, bromo, or iodo. In some embodiments, the halogen of R3is fluoro, chloro, or bromo. In some embodiments, the halogen of R3is fluoro or chloro. In some embodiments, a cyano group is utilized in place of the halogen. In some embodiments, a hydroxyl group is utilized in place of the halogen. In some embodiments, a methoxy group is utilized in place of the halogen.

[0077] In some embodiments, a compound is a small molecule comprising a benzimidazole, an acrylamide, and an isoxazole. In some embodiments, a formula to bind p53 mutant comprises a compound of Formula III:wherein R1is H, alkyl, benzyl, aryl, heteroaryl, heterocycle, or alkyl chain with heteroatom;R2is H, or alkyl;R3is H, or halogen; andR4is H, or CH3.

[0078] In some embodiments, R1is H, alkyl, benzyl, aryl, heteroaryl, or alkyl chain with heteroatoms.

[0079] In some embodiments, the alkyl or alkyl chain with heteroatom of R1is 1 to 12 carbons (C1-12). In some embodiments, the alkyl or alkyl chain with heteroatom of R1is 1 to 10 carbons (C1-10). In some embodiments, the alkyl or alkyl chain with heteroatom of R1is 1 to 8 carbons (C1-8). In some embodiments, the alkyl or alkyl chain with heteroatom of R1is 1 to 6 carbons (Ci-e). In some embodiments, the alkyl or alkyl chain with heteroatom of R1is 1 to 4 carbons (C1-4).

[0080] In some embodiments, the benzyl, aryl, heteroaryl, or heterocycle of R1is a 6- membered ring. In some embodiments, the benzyl, aryl, heteroaryl, or heterocycle of R1is a 5-membered ring. In some embodiments, the heteroaryl or heterocycle of R1comprises one or more nitrogen atoms (N). In some embodiments, the heteroaryl or heterocycle of R1comprises one or more oxygen atoms (O).

[0081] In some embodiments, the alkyl of R2is 1 to 8 carbons (C1-8). In some embodiments, the alkyl of R2is 1 to 6 carbons (C1-6). In some embodiments, the alkyl of R2is 1 to 4 carbons (C1-4). In some embodiments, the alkyl of R2is an ethyl. In some embodiments, the alkyl of R2is a methyl.

[0082] In some embodiments, the halogen of R3is fluoro, chloro, bromo, or iodo. In some embodiments, the halogen of R3is fluoro, chloro, or bromo. In some embodiments, the halogen of R3is fluoro or chloro. In some embodiments, a cyano group is utilized in place of the halogen. In some embodiments, a hydroxyl group is utilized in place of the halogen. In some embodiments, a methoxy group is utilized in place of the halogen.

[0083] In some embodiments, a compound is a small molecule comprising and benzimidazole, an acrylamide, and an alkyne. In some embodiments, a formula to bind p53 mutant comprises a compound of Formula IV:IV wherein R1is H, alkyl, benzyl, aryl, heteroaryl, heterocycle, or alkyl chain with heteroatom; R2is H, or alkyl;R3is H, or halogen; andR4is H, or CH3.

[0084] In some embodiments, R1is H, alkyl, benzyl, aryl, heteroaryl, or alkyl chain with heteroatoms.

[0085] In some embodiments, R1is an alkyl chain with a heteroatom at the end. The examples of heteroatoms include (but are not limited to) N, O, S, OH, alcohol, ether, and amine. In certain embodiments, R1is an alkyl chain with a heteroatom OH. In certain embodiments, R1is an alkyl chain with a heteroatom amine. In some embodiments, R1is an alkyl chain with a heterocyclic ring at the end. In certain embodiments, R1is an alkyl chain with a morpholine.

[0086] In some embodiments, the alkyl or alkyl chain with heteroatom of R1is 1 to 12 carbons (C1-12). In some embodiments, the alkyl or alkyl chain with heteroatom of R1is 1 to 10 carbons (C1-10). In some embodiments, the alkyl or alkyl chain with heteroatom of R1is 1 to 8 carbons (C1-8). In some embodiments, the alkyl or alkyl chain with heteroatomof R1is 1 to 6 carbons (C1-6). In some embodiments, the alkyl or alkyl chain with heteroatom of R1is 1 to 4 carbons (C1-4).

[0087] In some embodiments, the benzyl, aryl, heteroaryl, or heterocycle of R1is a 6- membered ring. In some embodiments, the benzyl, aryl, heteroaryl, or heterocycle of R1is a 5-membered ring. In some embodiments, the heteroaryl or heterocycle of R1comprises one or more nitrogen atoms (N). In some embodiments, the heteroaryl or heterocycle of R1comprises one or more oxygen atoms (0).

[0088] In some embodiments, the alkyl of R2is 1 to 8 carbons (C1-8). In some embodiments, the alkyl of R2is 1 to 6 carbons (C1-6). In some embodiments, the alkyl of R2is 1 to 4 carbons (C1-4). In some embodiments, the alkyl of R2is an ethyl. In some embodiments, the alkyl of R2is a methyl.

[0089] In some embodiments, the halogen of R3is fluoro, chloro, bromo, or iodo. In some embodiments, the halogen of R3is fluoro, chloro, or bromo. In some embodiments, the halogen of R3is fluoro or chloro. In some embodiments, a cyano group is utilized in place of the halogen. In some embodiments, a hydroxyl group is utilized in place of the halogen. In some embodiments, a methoxy group is utilized in place of the halogen.

[0090] In some embodiments, a compound is a small molecule comprising a benzimidazole, an acrylamide, and a central heterocycle. In some embodiments, a formula to bind p53 mutant comprises a compound of Formula V:wherein R1is H, alkyl, benzyl, aryl, heteroaryl, heterocycle, or alkyl chain with heteroatom;R2is H, or alkyl;R3is H, or halogen;R4is H, or CH3;X is CH, or N; andY is S, or 0.

[0091] In some embodiments, R1is H, alkyl, benzyl, aryl, heteroaryl, or alkyl chain with heteroatoms.

[0092] In some embodiments, R1is an alkyl chain with a heteroatom at the end. The examples of heteroatoms include (but are not limited to) N, O, S, OH, alcohol, ether, and amine. In certain embodiments, R1is an alkyl chain with a heteroatom OH. In certain embodiments, R1is an alkyl chain with a heteroatom amine. In some embodiments, R1is an alkyl chain with a heterocyclic ring at the end. In certain embodiments, R1is an alkyl chain with a morpholine.

[0093] In some embodiments, the alkyl or alkyl chain with heteroatom of R1is 1 to 12 carbons (C1-12). In some embodiments, the alkyl or alkyl chain with heteroatom of R1is 1 to 10 carbons (C1-10). In some embodiments, the alkyl or alkyl chain with heteroatom of R1is 1 to 8 carbons (C1 -8). In some embodiments, the alkyl or alkyl chain with heteroatom of R1is 1 to 6 carbons (C1-6). In some embodiments, the alkyl or alkyl chain with heteroatom of R1is 1 to 4 carbons (C1-4).

[0094] In some embodiments, the benzyl, aryl, heteroaryl, or heterocycle of R1is a 6- membered ring. In some embodiments, the benzyl, aryl, heteroaryl, or heterocycle of R1is a 5-membered ring. In some embodiments, the heteroaryl or heterocycle of R1comprises one or more nitrogen atoms (N). In some embodiments, the heteroaryl or heterocycle of R1comprises one or more oxygen atoms (O).

[0095] In some embodiments, the alkyl of R2is 1 to 8 carbons (C1-8). In some embodiments, the alkyl of R2is 1 to 6 carbons (C1-6). In some embodiments, the alkyl of R2is 1 to 4 carbons (C1-4). In some embodiments, the alkyl of R2is an ethyl. In some embodiments, the alkyl of R2is a methyl.

[0096] In some embodiments, the halogen of R3is fluoro, chloro, bromo, or iodo. In some embodiments, the halogen of R3is fluoro, chloro, or bromo. In some embodiments, the halogen of R3is fluoro or chloro. In some embodiments, a cyano group is utilized in place of the halogen. In some embodiments, a hydroxyl group is utilized in place of the halogen. In some embodiments, a methoxy group is utilized in place of the halogen.

[0097] In some embodiments, a compound is a small molecule comprising a benzimidazole, an acrylamide, and a central heterocycle. In some embodiments, a formula to bind p53 mutant comprises a compound of Formula VI:wherein R1is H, alkyl, benzyl, aryl, heteroaryl, heterocycle, or alkyl chain with heteroatom; R2is H, or alkyl;R3is H, or halogen;R4is H, or CH3;X is CH or N; andX1, X2, X3, and X4are each independently C, N, or 0.

[0098] In some embodiments, R1is H, alkyl, benzyl, aryl, heteroaryl, or alkyl chain with heteroatoms.

[0099] In some embodiments, R1is an alkyl chain with a heteroatom at the end. The examples of heteroatoms include (but are not limited to) N, O, S, OH, alcohol, ether, and amine. In certain embodiments, R1is an alkyl chain with a heteroatom OH. In certain embodiments, R1is an alkyl chain with a heteroatom amine. In some embodiments, R1isan alkyl chain with a heterocyclic ring at the end. In certain embodiments, R1is an alkyl chain with a morpholine.

[0100] In some embodiments, the alkyl or alkyl chain with heteroatom of R1is 1 to 12 carbons (C1-12). In some embodiments, the alkyl or alkyl chain with heteroatom of R1is 1 to 10 carbons (C1-10). In some embodiments, the alkyl or alkyl chain with heteroatom of R1is 1 to 8 carbons (C1 -8). In some embodiments, the alkyl or alkyl chain with heteroatom of R1is 1 to 6 carbons (C1-6). In some embodiments, the alkyl or alkyl chain with heteroatom of R1is 1 to 4 carbons (C1-4).

[0101] In some embodiments, the benzyl, aryl, heteroaryl, or heterocycle of R1is a 6- membered ring. In some embodiments, the benzyl, aryl, heteroaryl, or heterocycle of R1is a 5-membered ring. In some embodiments, the heteroaryl or heterocycle of R1comprises one or more nitrogen atoms (N). In some embodiments, the heteroaryl or heterocycle of R1comprises one or more oxygen atoms (O).

[0102] In some embodiments, the alkyl of R2is 1 to 8 carbons (C1-8). In some embodiments, the alkyl of R2is 1 to 6 carbons (C1-6). In some embodiments, the alkyl of R2is 1 to 4 carbons (C1-4). In some embodiments, the alkyl of R2is an ethyl. In some embodiments, the alkyl of R2is a methyl.

[0103] In some embodiments, the halogen of R3is fluoro, chloro, bromo, or iodo. In some embodiments, the halogen of R3is fluoro, chloro, or bromo. In some embodiments, the halogen of R3is fluoro or chloro. In some embodiments, a cyano group is utilized in place of the halogen. In some embodiments, a hydroxyl group is utilized in place of the halogen. In some embodiments, a methoxy group is utilized in place of the halogen.

[0104] In some embodiments, X1and X3are nitrogen atoms (N). In some embodiments, X2and X4are nitrogen atoms (N).

[0105] In some embodiments, a compound is a non-benzimidazole based small molecule comprising an acrylamide and an enone. In some embodiments, a formula to bind p53 mutant comprises a compound of Formula VII:VII wherein R2is H, or alkyl;R3is H, or halogen;R4is H, or CH3; andX is 0, or S.

[0106] In some embodiments, the alkyl of R2is 1 to 8 carbons (C1-8). In some embodiments, the alkyl of R2is 1 to 6 carbons (C1-6). In some embodiments, the alkyl of R2is 1 to 4 carbons (C1-4). In some embodiments, the alkyl of R2is an ethyl. In some embodiments, the alkyl of R2is a methyl.

[0107] In some embodiments, the halogen of R3is fluoro, chloro, bromo, or iodo. In some embodiments, the halogen of R3is fluoro, chloro, or bromo. In some embodiments, the halogen of R3is fluoro or chloro. In some embodiments, a cyano group is utilized in place of the halogen. In some embodiments, a hydroxyl group is utilized in place of the halogen. In some embodiments, a methoxy group is utilized in place of the halogen.

[0108] In some embodiments, a formula to bind p53 mutant comprises a compound of Formula VIII:wherein R1and R2are H or halogen;R3is H, alkyl, aryl, benzyl, or acyl; andR4is OMe.

[0109] In some embodiments, a formula to bind p53 mutant comprises a compound ofFormula IX:wherein R1is H, Boc, benzoyl, p-trifuoromethylbenzyl, p-methoxybenzyl, pentyl, or 4- pentynyl; and R2is OMe.

[0110] In some embodiments, a formula to bind p53 mutant comprises a compound ofFormula X:wherein R1is H or alkyl; and R2is OMe.

[0111] In some embodiments, a formula to bind p53 mutant comprises a compound ofFormula XI:wherein R1is H or alkyl; andR2is amine, alkyl amine, or dialkyl amine.

[0112] In some embodiments, a formula to bind p53 mutant comprises a compound of Formula XII:wherein R1is H or alkyl; andR2is acylamide, methacrylamide or NHC0NM62.

[0113] In some embodiments, a formula to bind p53 mutant comprises a compound of Formula XIII:wherein R1is H or alkyl;R2is Boc; andR3is H or alkyl.

[0114] In some embodiments, a formula to bind p53 mutant comprises a compound of Formula XIV:XIV wherein R1is alkyl or 4-pentynyl; andR2is acylamide or N-methylacrylamide.

[0115] In some embodiments, a formula to bind p53 mutant comprises a compound of Formula XV:wherein R1is H or alkyl; andR2is H, acylamide or OMe.

[0116] In some embodiments, a compound is a small molecule comprising a benzimidazole, an acrylamide, an enone, and a heteroaryl. In some embodiments, a formula to bind p53 mutant comprises a compound of Formula XVI:wherein R1is H, alkyl, benzyl, aryl, heteroaryl, heterocycle, or alkyl chain with heteroatom;R2is H, or alkyl;R3is H, or halogen;R4is H, or CH3; andX1, X2, X3, and X4are each independently C or N.

[0117] In some embodiments, R1is an alkyl chain with a heteroatom at the end. The examples of heteroatoms include (but are not limited to) N, 0, S, OH, alcohol, ether, and amine. In certain embodiments, R1is an alkyl chain with a heteroatom OH. In certain embodiments, R1is an alkyl chain with a heteroatom amine. In some embodiments, R1is an alkyl chain with a heterocyclic ring at the end. In certain embodiments, R1is an alkyl chain with a morpholine.

[0118] In some embodiments, the alkyl or alkyl chain with heteroatom of R1is 1 to 12 carbons (C1-12). In some embodiments, the alkyl or alkyl chain with heteroatom of R1is 1 to 10 carbons (C1-10). In some embodiments, the alkyl or alkyl chain with heteroatom of R1is 1 to 8 carbons (C1 -8). In some embodiments, the alkyl or alkyl chain with heteroatom of R1is 1 to 6 carbons (C1-6). In some embodiments, the alkyl or alkyl chain with heteroatom of R1is 1 to 4 carbons (CM).

[0119] In some embodiments, the benzyl, aryl, heteroaryl, or heterocycle of R1is a 6- membered ring. In some embodiments, the benzyl, aryl, heteroaryl, or heterocycle of R1is a 5-membered ring. In some embodiments, the heteroaryl or heterocycle of R1comprises one or more nitrogen atoms (N). In some embodiments, the heteroaryl or heterocycle of R1comprises one or more oxygen atoms (O).

[0120] In some embodiments, the alkyl of R2is 1 to 8 carbons (C1-8). In some embodiments, the alkyl of R2is 1 to 6 carbons (C1-6). In some embodiments, the alkyl of R2is 1 to 4 carbons (C1-4). In some embodiments, the alkyl of R2is an ethyl. In some embodiments, the alkyl of R2is a methyl.

[0121] In some embodiments, the halogen of R3is fluoro, chloro, bromo, or iodo. In some embodiments, the halogen of R3is fluoro, chloro, or bromo. In some embodiments, the halogen of R3is fluoro or chloro. In some embodiments, a cyano group is utilized in place of the halogen. In some embodiments, a hydroxyl group is utilized in place of the halogen. In some embodiments, a methoxy group is utilized in place of the halogen.

[0122] In some embodiments, X1and X3are nitrogen atoms (N). In some embodiments, X2and X4are nitrogen atoms (N).

[0123] In some embodiments, a compound is a small molecule comprising a benzimidazole, an enone, and a phenol ether. In some embodiments, a compound isselected from Table 1 . In some embodiments, a formula to bind p53 mutant comprises a compound of Table 1.Table 1

[0124] In some embodiments, a compound is a small molecule comprising a benzimidazole, alkynone, and a phenol ester. In some embodiments, a compound is selected from Table 2. In some embodiments, a formula to bind p53 mutant comprises a compound of Table 2.Table 2

[0125] In some embodiments, a compound is a small molecule comprising a benzimidazole, an enone, and an aniline. In some embodiments, a compound is selected from Table 3. In some embodiments, a formula to bind p53 mutant comprises a compound of Table 3.Table 3

[0126] In some embodiments, a compound is a small molecule comprising a benzimidazole, an enone, and an aryl acrylamide. In some embodiments, a compound is selected from Table 4. In some embodiments, a formula to bind p53 mutant comprises a compound of Table 4.Table 4

[0127] In some embodiments, a compound is a small molecule comprising a benzimidazole and an isoxazole. In some embodiments, a compound is selected from Table 5. In some embodiments, a formula to bind p53 mutant comprises a compound of Table 5.Table 5

[0128] In some embodiments, a compound is selected from Table 6. In some embodiments, a formula to bind p53 mutant comprises a compound of Table 6.Table 6Pharmaceutical Formulae

[0129] Provided herein are various embodiments of pharmaceuticals and / or supplements for use in a treatment for cancer, together with one or more pharmaceutically acceptable carriers thereof and optionally one or more other active ingredients. Proper formulation is dependent upon the route of administration chosen. In some embodiments, pharmaceutical formulae are utilized within a therapeutic for treatment of a condition such as cancer.

[0130] The term "active ingredient" refers to a compound, which is administered, alone or in combination with one or more pharmaceutically acceptable excipients or carriers, to a subject for treating, preventing, or ameliorating one or more symptoms of a disorder. In various embodiments, active ingredients include any one of the compounds listed herein,including those described within Tables 1 -6. In some embodiments, an active ingredient is benzimidazole based small molecules. In some embodiments, an active ingredient is benzimidazole based small molecules comprising acrylamide. In some embodiments, an active ingredient is benzimidazole based small molecules comprising acrylamide and enone. In some embodiments, an active ingredient is benzimidazole based small molecules comprising acrylamide and isoxazole. In some embodiments, an active ingredient is benzimidazole based small molecules comprising acrylamide and alkyne. In some embodiments, an active ingredient is benzimidazole based small molecules comprising acrylamide and heterocycle. In some embodiments, an active ingredient is non-benzimidazole based small molecules comprising acrylamide. In some embodiments, an active ingredient is non-benzimidazole based small molecules comprising acrylamide and enone.

[0131] The compounds disclosed herein can exist as therapeutically acceptable salts. The term "therapeutically acceptable salt," as used herein, represents salts or zwitterionic forms of the compounds disclosed herein which are therapeutically acceptable as defined herein. The salts can be prepared during the final isolation and purification of the compounds or separately by reacting the appropriate compound with a suitable acid or base. Therapeutically acceptable salts include acid and basic addition salts. For a more complete discussion of the preparation and selection of salts, refer to "Handbook of Pharmaceutical Salts, Properties, and Use," Stah and Wermuth, Ed., (Wiley-VCH and VHCA, Zurich, 2002) or S. M. Berge, L. D. Bighley and D. C. Monkhouse, J. Pharm. Sci. 1977, 66, 1-19.

[0132] Modes of administration, in accordance with multiple embodiments, include, but are not limited to intratumoral, oral, intravenous, subcutaneous, intramuscular, intrauterine (e.g., an implantable depot such as a hormone releasing intrauterine device), intraperitoneal, transdermal (e.g., patch), transmucosal (e.g., sublingual, nasal, vaginal or rectal) or a combination thereof. The actual amount of drug needed will depend on factors such as the size, age and severity of disease in the afflicted individual. The actual amount of drug needed will also depend on the effective concentration ranges of the various active ingredients. Vehicles of administration, in accordance with variousembodiments, include ointments, solutions, gels, creams, suppositories, implants, tablets, or capsules, as appropriate.

[0133] The compounds disclosed herein can be dried (e.g., precipitated) or kept solubilized within a solution, suspension, emulsion, colloid, gel, etc. Whether dried or kept solubilized, the components can be formulated for storage and / or delivery, which can be inclusive of a buffer (e.g., saline, lactate, acetate, gluconate), salts, minerals, or other components for storage such as preservatives.

[0134] For oral (or other gastrointestinal) administration, numerous coating agents can be used in accordance with various embodiments of the invention. In some embodiments, the coating agent is one which acts as a coating agent in conventional delayed release oral formulations, including polymers for enteric coating. Examples include hypromellose phthalate (hydroxy propyl methyl cellulose phthalate; HPMCP); hydroxypropylcellulose (HPC; such as KLUCEL®); ethylcellulose (such as ETHOCEL®); and methacrylic acid and methyl methacrylate (MAA / MMA; such as EUDRAGIT®).

[0135] Various embodiments of formulations also include at least one disintegrating agent. In some embodiments, a disintegrating agent is a super disintegrant agent. In many embodiments, disintegrants are combined with a resin. Additional disintegrating agents include, but are not limited to, agar, calcium carbonate, maize starch, potato starch, tapioca starch, alginic acid, alginates, certain silicates, and sodium carbonate. Suitable super disintegrating agents include, but are not limited to crospovidone, croscarmellose sodium, AMBERLITE (Rohm and Haas, Philadelphia, Pa.), and sodium starch glycolate.

[0136] Several embodiments of a formulation further utilize other components and excipients. For example, sweeteners, flavors, buffering agents, and flavor enhancers to make the dosage form more palatable. Sweeteners include, but are not limited to, fructose, sucrose, glucose, maltose, mannose, galactose, lactose, sucralose, saccharin, aspartame, acesulfame K, and neotame. Common flavoring agents and flavor enhancers that may be included in the formulation of the present invention include, but are not limited to, maltol, vanillin, ethyl vanillin, menthol, citric acid, fumaric acid, ethyl maltol and tartaric acid.

[0137] Multiple embodiments of a formulation also include a surfactant. In certain embodiments, surfactants are selected from the group consisting of Tween 80, sodium lauryl sulfate, and docusate sodium.Various embodiments of a formulation also include a lubricant. In certain embodiments, lubricants are selected from the group consisting of, but are not limited to, magnesium stearate, stearic acid, sodium stearyl fumarate, calcium stearate, hydrogenated vegetable oil, mineral oil, fish oil, castor oil, sesame oil, polyethylene glycol, polyethylene glycol 4000-6000, talc, and glyceryl behenate.

[0138] In some embodiments, active ingredients are administered in a therapeutically effective amount as part of a course of treatment. As used in this context, to "treat" means to ameliorate at least one symptom of a disorder to be treated or to provide a beneficial physiological effect. For example, one such amelioration of a symptom could be reduction of tumor sizes of various cancers.

[0139] A therapeutically effective amount can be an amount sufficient to prevent, reduce, ameliorate, or eliminate the symptoms of cancer complications susceptible to such treatment.

[0140] Dosage, toxicity and therapeutic efficacy of the compounds can be determined, e.g., by standard pharmaceutical procedures in cell cultures or experimental animals, e.g., for determining the LDso (the dose lethal to 50% of the population) and the EDso (the dose therapeutically effective in 50% of the population). The dose ratio between toxic and therapeutic effects is the therapeutic index and it can be expressed as the ratio LDso / EDso. Compounds that exhibit high therapeutic indices are preferred. While compounds that exhibit toxic side effects may be used, care should be taken to design a delivery system that targets such compounds to the site of affected tissue in order to minimize potential damage to other tissue and organs and, thereby, reduce side effects.

[0141] Data obtained from cell culture assays or animal studies can be used in formulating a range of dosage for use in humans. If the pharmaceutical is provided systemically, the dosage of such compounds lies preferably within a range of circulating concentrations that include the EDso with little or no toxicity. The dosage may vary within this range depending upon the dosage form employed and the route of administrationutilized. For any compound used in the method of the invention, the therapeutically effective dose can be estimated initially from cell culture assays. A dose may be formulated in animal models to achieve a circulating plasma concentration or within the local environment to be treated in a range that includes the EDso as determined in cell culture or animal models. Such information can be used to more accurately determine useful doses in humans. Levels in plasma may be measured, for example, by mass spectrometry.

[0142] An "effective amount" is an amount sufficient to effect beneficial or desired results. For example, a therapeutic amount is one that achieves the desired therapeutic effect. This amount can be the same or different from a prophy lactically effective amount, which is an amount necessary to prevent onset of disease or disease symptoms. An effective amount can be administered in one or more administrations, applications or dosages. A therapeutically effective amount of a composition depends on the composition selected. The compositions can be administered from one or more times per day to one or more times per week; including once every other day, as determined to be beneficial. The skilled artisan will appreciate that certain factors may influence the dosage and timing required to effectively treat a subject, including but not limited to the severity of the disease or disorder, previous treatments, the general health and / or age of the subject, and other diseases present. Moreover, treatment of a subject with a therapeutically effective amount of the compositions described herein can include a single treatment or a series of treatments. For example, several divided doses may be administered daily, one dose, or cyclic administration of the compounds to achieve the desired therapeutic result.

[0143] Pharmaceutical compositions may be formulated as a modified release dosage form, including delayed-, extended-, prolonged-, sustained, pulsatile-, controlled-, accelerated- and fast-, targeted-, programmed-release, and gastric retention dosage forms. These dosage forms can be prepared utilizing the various method embodiments as described herein.

[0144] Preservatives and other additives, like antimicrobial, antioxidant, chelating agents, and inert gases, can also be present. One common preservative is benzyl alcohol.(See generally, Remington: The Science and Practice of Pharmacy, 21 st Edition;Lippincott Williams & Wilkins: Philadelphia, PA, 2005.)Methods of Treatment

[0145] Several embodiments are directed towards treatments of individuals with benzimidazole based small molecules and related compounds. In some embodiments, pharmaceutical compounds are administered to an individual or patient having cancer. In some embodiments, pharmaceutical compounds are administered to an individual having cancer with p53 mutations. Generally, any p53 protein having a mutation resulting an oncogenic effect (e.g., partial or full loss of tumor suppression) that retains in-frame encoding can be targeted by the pharmaceutical compounds. In some implementations, pharmaceutical compounds target can target p53 proteins having one or more missense mutations that results in an oncogenic effect. In some implementations, pharmaceutical compounds target can target p53 proteins having a small in-frame deletion or insertion (e.g., between 1 and 5 amino acids) that results in an oncogenic effect. Examples of residues that are commonly mutated within p53 mutants that can be targeted with the small molecules include (but are not limited to) R175, G245, R248, R249, R273, R282, R280, Y220, A138, K139, P151 , P152, G154, R158, V172, H193, H214, S215, Y234, P250, 1255, V272, V274, E285, and E286. Many positions have several known mutations. For example, R175 mutations include (but are not limited to) R175H, R175G, R175D, R175A, R175Y, R175F, R175K, R175E, R175Q, R175del, and R175_C176del. Table 7 lists various residue positions that are commonly mutated within p53 that pharmaceutical compounds of the disclosure can target. Various p53 mutants that can be targeted are described within The TP53 Database maintained by the National Cancer Institute of the U.S. National Institutes of Health and several manuscript publications (see The TP53 Database (R20, Jul 2019; R21 , Jan 2025): tp53[dot]cancer[dot]gov; J. S. Funk, et al., Nat Genet. 2025 Jan;57(1 ):140-153; K. C. de Andrade, et al., Cell Death Differ. 2022 May;29(5): 1071 -1073; and M. Olivier, et al., Cold Spring Harb Perspect Biol. 2010 Jan;2(1 ):a001008; the disclosures of which are hereby incorporated by reference).Table 7. p53 Mutants

[0146] Various embodiments are directed to diagnostic procedures and / or treatments related to cancer. In several embodiments, an individual can be diagnosed as having a cancer associated with p53 mutagenesis. Examples of cancers associated with p53 mutagenesis include (but are not limited to) (but not limited to) ovarian cancers, breast cancers, esophageal cancers, colorectal cancers, head and neck cancers, larynx cancers, and lung cancers. An individual or cancer patient of the one of the cancers associated with p53 mutagenesis can be administered a therapy regimen inclusive of a therapeutic comprising one or more pharmaceutical compounds. In many embodiments, an individual can be diagnosed as having a cancer associated with p53 and thenadministered a pharmaceutical compound. In some embodiments, prior to administration of a pharmaceutical compound a sample of the patient is assessed for p53 mutagenesis. Upon detection of a p53 mutation, the individual an individual can be administered a therapy regimen inclusive of a therapeutic comprising one or more pharmaceutical compounds. Mutagenesis of p53 mutation can be assessed by a number of means, including (but not limited to) nucleic acid sequencing, nucleic acid hybridization assay, and immunohistochemistry. Assessment can be performed on a sample derived from a collection of cancer cells (e.g., tumor tissue for solid cancers, circulating white blood cells for blood cancer, lymph tissue for metastatic cancer, etc.). Alternatively (or additionally), assessment can be performed on cell-free nucleic acids from a noncellular source (e.g., plasma, lymph, cerebral spinal fluid, urine, stool, saliva, etc.).

[0147] In some embodiments, a cancer cell with a p53 mutation is contacted with a p53-targeting compound, which can result in inhibition of proliferation or inducement of cell death. In some embodiments, pharmaceutical compounds are administered to a recipient having a cancer, which can allow the compounds contact cancer cells with a p53 mutation. In some embodiments, pharmaceutical compounds are administered to a recipient having a cancer, which can allow the compounds to contact cancer cells and to bind to p53 mutants.

[0148] Mutant p53 isoforms are often localized to the cytoplasm, whereas activated wild-type p53 localizes to chromatin in cancer cells. The pharmaceutical compounds can induce chromatin localization of mutant p53. In some embodiments, pharmaceutical compounds are administered to a recipient, which can induce chromatin translocation of mutant p53 within cancer cells. Chromatin translocation can be assessed by various techniques, such as (for example) immunohistochemistry of a tissue biopsy.

[0149] Mutant p53 isoforms lose the function to induce expression of p53 target genes. In some embodiments, pharmaceutical compounds are administered to a recipient, which can restore the expression of p53 target genes within cancer cells. Expression of p53 target genes can be assessed by various techniques, such as (for example) RNA- sequencing or RT-PCR.

[0150] Mutant p53 isoforms lose the function to bind to target gene promoters. In some embodiments, pharmaceutical compounds are administered to an individual, which can restore p53 promoter binding within cancer cells. Association of p53 with promoters can be assessed by various techniques, such as (for example) chromatinimmunoprecipitation.

[0151] Administration of pharmaceutical compounds can be combined with standards of care for the complication. Administration of compounds can be combined with surgery, radiation therapy, chemotherapy, immunotherapy, targeted therapy, cell replacement therapy, hormone therapy, or other therapies, which can be determined as appropriate to the cancer type. The compounds can be administered as an adjuvant treatment or a neoadjuvant treatment.

[0152] In various embodiment, an individual is administered a pharmaceutical compound described herein, including benzimidazole based small molecules, or a related compound thereof, for a period of 2 to 4 weeks, for a period of 4 to 6 weeks, for a period of 6 to 8 weeks, for a period of 8 to 10 weeks, for a period of 10 to 12 weeks, for a period of 12 to 14 weeks, for a period of 14 to 19 weeks, for a period of 20 weeks, for a period of 21 weeks, for a period of 22 weeks, for a period of 23 weeks, for a period of 25 weeks, for a period of 26 weeks, for a period of 27 weeks, for a period of 28 weeks, for a period of 29 weeks, for a period of 30 weeks, for a period of 35 weeks, for a period of 37 weeks, for a period of 38 weeks, for a period of 39 weeks, for a period of 40 weeks, or for a period of more than 50 weeks.

[0153] In various embodiments, the effective amount of a pharmaceutical compound described herein, including benzimidazole based small molecules, or a related compound thereof is 0.5-1 mg / day, 1 -5mg / day, 5-10mg / day, 10-15mg / day, 15-20mg / day, 20- 25mg / day, 25-30mg / day, 30-35mg / day, 35-40mg / day, 40-45mg / day, 45- 50mg / day, 50- 55mg / day, 55-60mg / day, 60-65mg / day, 65-70mg / day, 70-75mg / day, 75-80mg / day, 80- 85mg / day, 85-90mg / day, 90-95mg / day or 95-100mg / day, 100-200mg / day, 200- 300mg / day, 300-400mg / day, 400-500mg / day, 500-600mg / day, 600-700mg / day, 700- 800mg / day, 800-900mg / day, 900-1 OOOmg / day, 1000-1100mg / day, 1100-1200mg / day, 1200-1300mg / day, 1300-1400mg / day, 1400-1500mg / day, 1500-1600mg / day, 1600-1700mg / day, 1700-1800mg / day, 1800-1900mg / day, 1900-2000mg / day, 2000-2100mg / day, 2100-2200mg / day, 2200-2300mg / day, 2300-2400mg / day, 2400-2500mg / day, 2500-2600mg / day, 2600-2700mg / day, 2700-2800mg / day, 2800-2900mg / day, 2900-3000mg / day, 3000-3100mg / day, 3100-3200mg / day, 3200-3300mg / day, 3300-3400mg / day, 3400-3500mg / day, 3500-3600mg / day, 3600-3700mg / day, 3700-3800mg / day, 3800-3900mg / day, 3900-4000mg / day, 4000-4200mg / day, 4200-4400mg / day, 4400-4600mg / day, 4600-4800mg / day or 4800-5000mg / day.NUMBERED EMBODIMENTSEmbodiment 1 . A medicament for the treatment of an individual having a cancer with a p53 mutation, the medicament comprising a compound selected from the group consisting of: a benzimidazole-related compound, a benzimidazole and acrylamide-related compound, a benzimidazole, enone, and acrylamide-related compound, a benzimidazole, acrylamide, isoxazole-related compound, a benzimidazole, acrylamide, and alkyne-related compound, a benzimidazole, acrylamide, and heterocycle-related compound, a non-benzimidazole and acrylamide-related compound, a benzimidazole, enone, and phenol-related compound, a benzimidazole and phenol-related compound, a benzimidazole, enone, and aniline-related compound, a benzimidazole, enone, benzene, and amide-related compound, and a benzimidazole and isoxazole-related compound.Embodiment 2. The medicament of embodiment 1 , wherein the medicament comprising a compound having the structure:wherein R1is H, CH3, alkyl, benzyl, aryl, heteroaryl, or alkyl chain and heteroatom;R2is H, CH3, or alkyl;R3is H, F, Cl, or halogen; andR4isH,orCH3.Embodiments. The medicament of embodiment 1, wherein the medicament comprising a compound having the structure:wherein R1is H, CH3, alkyl, benzyl, aryl, heteroaryl, or alkyl chain and heteroatoms;R2is H, CH3, or alkyl;R3is H, F, Cl, or halogen; andR4isH,orCH3.Embodiment 4. The medicament of embodiment 1, wherein the medicament comprising a compound having the structure:wherein R1is H, CH3, alkyl, benzyl, aryl, heteroaryl, or alkyl chain and heteroatoms;R2is H, CH3, or alkyl;R3is H, F, Cl, or halogen; andR4isH,orCH3.Embodiments. The medicament of embodiment 1, wherein the medicament comprising a compound having the structure:wherein R1is H, CH3, alkyl, benzyl, aryl, heteroaryl, or alkyl chain and heteroatoms;R2is H, CH3, or alkyl;R3is H, F, Cl, or halogen; andR4is H, or CH3.Embodiment 6. The medicament of embodiment 1 , wherein the medicament comprising a compound having the structure:wherein R1is H, CH3, alkyl, benzyl, aryl, heteroaryl, or alkyl chain and heteroatoms;R2is H, CH3, or alkyl;R3is H, F, Cl, or halogen;R4is H, or CH3;X is CH, or N; andY is S, or 0.Embodiment 7. The medicament of embodiment 1 , wherein the medicament comprising a compound having the structure:wherein R1is H, CH3, alkyl, benzyl, aryl, heteroaryl, or alkyl chain and heteroatoms;R2is H, CH3, or alkyl;R3is H, F, Cl, or halogen;R4is H, or CH3; andX is CH, or N.Embodiment s. The medicament of embodiment 1, wherein the medicament comprising a compound having the structure:wherein R2is H, CH3, or alkyl;R3is H, F, Cl, or halogen;R4is H, or CH3; andX is 0, or S.Embodiment 9. The medicament of embodiment 1 , wherein the compound binds with and stabilizes a p53 mutant such that the stabilization of the p53 mutant reactivates gene expression of p53 wild type.Embodiment 10. The medicament of embodiment 9, wherein the p53 mutant is selected from the group consisting of: R175, G245, R248, R249, R273, R282, Y220, A138, P151 , P152, G154, R158, V172, H193, H214, S215, Y234, P250, I255, V272, V274, E285, and E286.Embodiment 11. The medicament of embodiment 9, wherein the p53 mutant is present in a human ovarian cancer or a human breast cancer.Embodiment 12. A method for the treatment of an individual having a cancer with a p53 mutation, comprising: administering a medicament comprising a compound selected from the group consisting of: a benzimidazole-related compound, a benzimidazole and acrylamide- related compound, a benzimidazole, enone, and acrylamide-related compound, a benzimidazole, acrylamide, isoxazole-related compound, a benzimidazole, acrylamide, and alkyne-related compound, a benzimidazole, acrylamide, and heterocycle-related compound, a non-benzimidazole and acrylamide-related compound, a benzimidazole, enone, and phenol-related compound, a benzimidazole and phenol-related compound, a benzimidazole, enone, and aniline-related compound, a benzimidazole, enone, benzene, and amide-related compound, and a benzimidazole and isoxazole-related compound; wherein the compound binds with and stabilizes a p53 mutant such that the stabilization of the p53 mutant reactivates gene expression of p53 wild type.EXAMPLES

[0154] Biological data support the formulations, compounds, and methods to inhibit cancer cell proliferation with p53 mutants. In the ensuing sections, examples of formulations, compounds, and methods to stabilize p53 mutants are provided, indicating that the compounds described herein can be utilized to bind p53 mutants and / or treat an individual having cancer cells with p53 mutations.Example 1: Synthesis of Exemplary Provided CompoundsRoute A. (E)-3-(3-methoxyphenyl)-1-(1-methyl-1H-benzo[c / ]imidazol-2-yl)prop-2-en- 1-one (5)Step 1. (E)-1-(1H-benzo[d]imidazol-2-yl)-3-(3-methoxyphenyl)prop-2-en-1-one 60)

[0155] To a scintillation vial equipped with a magnetic stir bar, (50 mg, 0.31 mmol, 1 equiv.) of 2-acetylbenzimidazole were dissolved in 0.62 mL of EtOH at 20 °C and open to air. To this solution, (41 pL, 0.34 mmol, 1.1 equiv.) of 3-methoxybenzaldehyde followed by (0.31 mL, 1 .25 mmol, 4 equiv.) 4 M aq. NaOH were added, and the solution was stirred for 12 hours. The reaction was quenched with sat. aq. NH4CI and the mixture was extracted with EtOAc three times. The combined organic layers were then dried with Na2SO4 and concentrated in vacuo to yield a yellow solid. The material was further purified by chromatography (15% EtOAc / hexanes) to give enone 60 as a yellow powder (62 mg, 71 % yield).

[0156] 1H NMR: (500 MHz, K2CO3 buffered CDCI3) δ 11 .02 (bs, 1 H), 8.13 (d, J = 15.9 Hz, 1 H), 8.05 (d, J = 15.9 Hz, 1 H), 7.95 (d, J = 8.0 Hz, 1 H), 7.58 (d, J = 8.0 Hz, 1 H), 7.43 (t, J = 7.5 Hz, 1 H), 7.39 - 7.35 (m, 1 H), 7.33 (m, 2H), 7.26 (s, 1 H), 6.99 (m, 1 H), 3.85 (s, 3H).

[0157] 13C{1H} NMR: (125 MHz, K2CO3 buffered CDCI3) δ 181.8, 160.1 , 149.0, 146.2,143.8, 136.0, 134.1 , 130.1 , 126.7, 124.0, 122.3, 122.0, 121.3, 117.7, 113.5, 112.4, 55.6.

[0158] HRMS (ESI-TOF) m / z: [M+Na]+Calcd for C17H14N2O2Na 301.0953; Found 301.0946.Step 2. (E)-3-(3-methoxyphenyl)-1 -(1 -methyl-1 H-benzo[d]limidazol-2-yl)prop-2-en- 1-one (5)

[0159] In a scintillation vial equipped with a magnetic stir bar under an inert atmosphere of argon (100 mg, 0.36 mmol, 1 equiv.) of 60 was dissolved in DMF (2 mL). NaH (22 mg, 60% dispersion in mineral oil, 0.54 mmol, 1.5 equiv.) was added. After stirring at rt for 15 min, Mel was added (35 pL, 0.54 mmol, 1.5 equiv.), and the reaction was allowed to stir overnight. The reaction was quenched with water and diluted with EtOAc. The organic layers were washed with water three times followed by once with brine, dried with MgSCk, and concentrated in vacuo. The crude material was purified by chromatography (15% EtOAc / hexanes) to give 5 as a yellow solid (62 mg, 56% yield).

[0160] 1H NMR: (500 MHz, CDCI3) δ 8.24 (d, J = 16.0 Hz, 1 H), 7.94 (d, J = 8.2 Hz, 1 H), 7.89 (d, J = 16.0 Hz, 1 H), 7.46 (app. d, J = 3.9 Hz, 2H), 7.39 (ddd, J = 8.2, 4.5, 3.6 Hz, 1 H), 7.33 (app. d, J = 4.9 Hz, 2H), 7.27 (app. s, 1 H), 6.99 - 6.96 (m, 1 H), 4.22 (s, 3H), 3.87 (s, 3H).

[0161] 13C{1H} NMR: (125 MHz, CDCI3) δ 183.0, 160.1 , 147.3, 144.8, 141.8, 137.2,136.2, 130.0, 126.1 , 124.0, 123.3, 122.2, 121.9, 117.3, 113.4, 110.7, 55.6, 32.6.

[0162] HRMS (ESI-TOF) m / z: [M+Na]+Calcd for Ci8Hi6N2O2Na 315.1110; Found 315.1113.Intermediate S1. 1-(5,6-dibromo-1H-benzo[d]imidazol-2-yl)ethan-1-oneStep 1. 1-(5,6-dibromo-1H-benzo[cf]imidazol-2-yl)ethan-1-ol

[0163] To 4, 5-dibromobenzene-1 ,2-diamine (2.50 g, 9.40 mmol, 1 equiv.) and DL- lactic acid (85% v / v in water, 1.2 mL, 14.1 mmol, 1.5 equiv.) was added 10 mL of 4M HCI. The reaction was stirred at reflux overnight, after which the pH was adjusted to 4. The resulting precipitate was filtered and washed with cold water to give 2.58 g (86% yield) of the title compound.Step 2. 1-(5,6-dibromo-1H-benzo[cflimidazol-2-yl)ethan-1-one (S1)

[0164] To 1 -(5,6-dibromo-1 / - / -benzo[d]imidazol-2-yl)ethan-1 -ol (1.00 g, 3.13 mmol, 1 equiv.) in DCM (30 mL) was added sodium bicarbonate (5.30 g, 62.5 mmol, 20 equiv.) and 3-Oxo-1 A5,2-benziodoxole-1 , 1 , 1 (3 / 7)-triyl triacetate (Dess-Martin periodinane, 1.99 g, 4.69 mmol, 1 .5 equiv.). The reaction was stirred at rt for 1 h and then diluted with sat. aq. NaHCO3 and sat. aq. NaS2O3. The aqueous layer was extracted twice with DCM, and the combined organic layers were dried over Na2SO4, concentrated under vacuum, and subjected to chromatography (0-30% EtOAc in hexanes) to give 710 mg (71 % yield) of the title compound as an orange solid.Intermediate S2. 1 -(5,6-difluoro-1 W-benzo[d]imidazol-2-yl)ethan-1 -one

[0165] The title compound was made in a manner analogous to 1 -(5,6-dibromo-1 H- benzo[d]imidazol-2-yl)ethan-1 -one (S1 ) using 4,5-difluorobenzene-1 ,2-diamine instead of 4,5-dibromobenzene-1 ,2-diamine.(E)-1 -(5,6-dibromo-1 H-benzo[cflimidazol-2-yl)-3-(3-methoxyphenyl)prop-2-en-1 -one (3)

[0166] The title compound was made in a manner analogous to (E)-1 -(1H- benzo[d]imidazol-2-yl)-3-(3-methoxyphenyl)prop-2-en-1 -one (60) except using ketone 1 - (5,6-dibromo-1H-benzo[d]imidazol-2-yl)ethan-1 -one (S1) instead of 2- acetylbenzimidazole. Orange solid.

[0167] 1H NMR: (500 MHz, DMSO-d6) δ 13.75 (bs, 1 H), 8.12 (bs, 2H), 8.02 (d, J = 16.1 Hz, 1 H), 7.96 (d, J = 16.1 Hz, 1 H), 7.48 - 7.35 (m, 3H), 7.08 - 7.07 (m, 1 H), 3.84 (s, 3H).

[0168] 13C{1H} NMR: (125 MHz, DMSO-d6) δ 180.8, 159.7, 150.6, 145.1 , 135.6, 130.2,121.5, 117.5, 113.7, 55.3. [N-H Benzimidazoles are known to have very broad13C peaks that are not observed under normal conditions.]

[0169] HRMS (ESI-TOF) m / z: [M+H]+Calcd for Ci7Hi2Br2N2O2H 434.9344 (79Br,79Br); Found 434.9342.(E)-1-(5,6-dichloro-1H-benzo[d]imidazol-2-yl)-3-(3-methoxyphenyl)prop-2-en-1-one (2)

[0170] The title compound was made in a manner analogous to (E)-1 -(1 / - / - benzo[d]imidazol-2-yl)-3-(3-methoxyphenyl)prop-2-en-1 -one (60) except using ketone 1 - (5,6-dichloro-1 H-benzo[d]imidazol-2-yl)ethan-1 -one, which is commercially available or synthetically accessible, instead of 2-acetylbenzimidazole. Yellow solid.

[0171] 1H NMR: (500 MHz, DMSO-d6) δ 13.78 (s, 1 H), 8.06 - 7.93 (m, 4H), 7.47 - 7.38 (m, 3H), 7.08 (d, J = 7.9 Hz, 1 H), 3.84 (s, 3H).

[0172] 13C{1H} NMR: (125 MHz, DMSO-d6) δ 180.7, 159.7, 150.9, 145.1 , 135.6, 130.2,121.5 (2), 117.5, 113.7, 55.3. [N-H Benzimidazoles are known to have very broad13C peaks that are not observed under normal conditions.](£)-1-(5,6-difluoro-1H-benzo[cflimidazol-2-yl)-3-(3-methoxyphenyl)prop-2-en-1-one

[0173] The title compound was made in a manner analogous to (E)-1 -(1 / - / - benzo[d]imidazol-2-yl)-3-(3-methoxyphenyl)prop-2-en-1 -one (60) except using ketone 1 - (5,6-difluoro-1 H-benzo[d]imidazol-2-yl)ethan-1 -one (S2) instead of 2- acetylbenzimidazole. Yellow solid.

[0174] 1H NMR: (500 MHz, 1 :1 CD3OD:CD2Cl2) δ 8.04 (d, J = 15.9 Hz, 1 H), 7.95 (J =15.9 Hz, 1 H), 7.54 (bs, 2H), 7.38 - 7.33 (m, 2H), 7.31 (s, 1 H), 7.02 (ddd, J = 7.8, 2.1 , 1.3Hz, 1 H), 3.87 (s, 3H).

[0175] 13C{1H} NMR: (125 MHz, 1 :1 CD3OD:CD2Cl2) δ 181.2, 160.7, 146.1 , 136.5,130.5, 122.4, 121.6, 117.9, 113.6, 55.7. [N-H Benzimidazoles are known to have very broad13C peaks that are not observed under normal conditions.]

[0176] HRMS (ESI-TOF) m / z: [M + H]+Calcd for C17H12F2N2O2H 315.0945; Found315.0934.(E)-1-(1H-benzo[d]imidazol-2-yl)-3-(6-methoxypyridin-2-yl)prop-2-en-1-one (46)

[0177] The title compound was made in a manner analogous to (E)-1 -(1H- benzo[d]imidazol-2-yl)-3-(3-methoxyphenyl)prop-2-en-1 -one (60) except using aldehyde 6-methoxypicolinaldehyde instead of 3-methoxybenzaldehyde. Yellow solid.

[0178] 1H NMR: (500 MHz, DMSO-d6) δ 13.50 (bs, 1 H), 8.48 (d, J = 15.7 Hz, 1 H), 7.92 (d, J = 15.7 Hz, 1 H), 7.82 (t, J = 7.5 Hz, 1 H), 7.73 (bs, 2H), 7.65 (d, J = 7.5 Hz, 1 H), 7.38 (bs, 2H), 7.36 (d, J = 7.8 Hz, 1 H), 2.59 (s, 3H).

[0179] 13C{1H} NMR: (150 MHz, DMSO-d6) δ 181 .3, 158.7, 151.5, 149.0, 142.9, 137.6,124.9, 124.5, 123.6, 24.2. [N-H Benzimidazoles are known to have very broad13C peaks that are not observed under normal conditions.](E)-1-(1H-benzo[c / ]imidazol-2-yl)-3-(2-methoxypyridin-4-yl)prop-2-en-1-one (47)

[0180] The title compound was made in a manner analogous to (E)-1 -(1 / - / - benzo[d]imidazol-2-yl)-3-(3-methoxyphenyl)prop-2-en-1 -one (60) except using aldehyde 2-methoxyisonicotinaldehyde instead of 3-methoxybenzaldehyde. Yellow solid.

[0181] 1H NMR: (500 MHz, DMSO-d6) δ 8.28 (d, J = 15.6 Hz, 1 H), 8.26 (d, J = 4.4 Hz, 1 H), 7.83 (d, J = 16.2 Hz, 1 H), 7.71 - 7.70 (m, 2H), 7.45 (d, J = 5.2 Hz, 1 H), 7.32 - 7.30 (m, 2H), 7.21 (s, 1 H), 3.90 (s, 3H).

[0182] 13C{1H} NMR: (150 MHz, DMSO-de) 3 181 .5, 164.5, 147.7, 144.7, 140.4, 126.5,123.9, 114.9, 110.4, 53.4. [N-H Benzimidazoles are known to have very broad13C peaks that are not observed under normal conditions.](E)-1-(1H-benzo[d]imidazol-2-yl)-3-(4-methoxypyridin-2-yl)prop-2-en-1-one (49)

[0183] The title compound was made in a manner analogous to (E)-1-(1 / 7- benzo[d]imidazol-2-yl)-3-(3-methoxyphenyl)prop-2-en-1 -one (60) except using aldehyde 4-methoxypicolinaldehyde instead of 3-methoxybenzaldehyde. Yellow solid.

[0184] 1H NMR: (500 MHz, DMSO-d6) δ 8.61 (d, J = 5.9 Hz, 1 H), 8.55 (d, J = 15.9 Hz, 1 H), 7.92 (d, J = 15.8 Hz, 1 H), 7.74 (app. s, 2H), 7.68 (s, 1 H), 7.40 - 7.39 (m, 2H), 7.21 - 7.20 (m, 1 H), 3.98 (s, 3H).(E)-1-(5,6-difluoro-1-methyl-1H-benzo[c / ]imidazol-2-yl)-3-(3-methoxyphenyl)prop-2- en-1-one (6)

[0185] (E)-1 -(5,6-difluoro-1 / - / -benzo[c / ]imidazol-2-yl)-3-(3-methoxyphenyl)prop-2-en-1-one (1) was subjected to Step 2 of Route A, with (1) replacing (E)-1-(1 H- benzo[d]imidazol-2-yl)-3-(3-methoxyphenyl)prop-2-en-1 -one (). Yellow solid.

[0186] 1H NMR: (500 MHz, CDCI3) δ 8.14 (d, J = 15.9 Hz, 1 H), 7.88 (d, J = 15.9 Hz, 1 H), 7.68 (dd, J = 9.9, 7.5 Hz, 1 H), 7.36 - 7.32 (m, 2H), 7.26 - 7.22 (m, 2H), 6.99 (dt, J = 7.2, 2.1 Hz, 1 H), 4.18 (s, 3H), 3.88 (s, 3H).

[0187] 13C{1H} NMR: (125 MHz, CDCI3) δ 182.3, 160.1 , 151.5, 150.1 , 149.7, 145.2,137.1 , 136.1 , 132.9, 130.1 , 122.9, 122.1 , 117.3, 113.6, 108.8 (d, J = 19.5 Hz), 98.3 (d, J = 22.3 Hz), 55.6, 33.0.19F{1H} NMR: (375 MHz, CDCI3) δ -135.3 (d, J = 20.2 Hz), -140.2 (d, J = 20.3 Hz).Intermediate S5. 1 -(1 -phenyl-1 H-benzo[d]imidazol-2-yl)ethan-1 -one

[0188] The title compound was made in a manner analogous to 1 -(5,6-dibromo-1 / - / - benzo[d]imidazol-2-yl)ethan-1-one (S1) using A / 1-phenylbenzene-1 ,2-diamine instead of 4,5-dibromobenzene-1 ,2-diamine.(E)-3-(3-methoxyphenyl)-1-(1 -phenyl-1 H-benzo[d]irnidazol-2-yl)prop-2-en-1 -one (7)

[0189] The title compound was made in a manner analogous to (E)-3-(3- methoxyphenyl)-1-(1-methyl-1 / - / -benzo[c / ]imidazol-2-yl)prop-2-en-1 -one (5) except using ketone 1-(1 -phenyl-1 H-benzo[d]imidazol-2-yl)ethan-1 -one (S5) instead of 2- acetylbenzimidazole. Yellow solid.

[0190] 1H NMR: (600 MHz, CDCI3) δ 8.20 (d, J = 16.0 Hz, 1 H), 8.01 (d, J = 8.4 Hz,1 H), 7.81 (d, J = 15.9 Hz, 1 H), 7.60 - 7.53 (m, 3H), 7.47 - 7.37 (m, 4H), 7.32 (t, J = 7.7Hz, 1 H), 7.29 (d, J = 7.5 Hz, 1 H), 7.24 (s, 1 H), 7.20 (d, J = 8.3 Hz, 1 H), 6.98 (dd, J = 7.9, 1.7 Hz, 1 H), 3.88 (s, 3H).

[0191] 13C{1H} NMR: (150 MHz, CDCI3) δ 181.5, 160.0, 147.4, 145.2, 141.9, 138.1 ,137.1 , 136.2, 130.0, 129.6, 128.9, 127.2, 126.6, 124.4, 123.0, 122.1 , 121.8, 117.2, 113.5, 111.8, 55.6.

[0192] HRMS (ESI-TOF) m / z: [M+Na]+Calcd for C23Hi8N2O2Na 377.1269; Found 377.1266.tert-butyl (E)-(3-(3-(1 -methyl-1 H-benzo[d]imidazol-2-yl)-3-oxoprop-1 -en-1 - yl)benzyl)carbamate (35)

[0193] The title compound was made in a manner analogous to (E)-3-(3- methoxyphenyl)-1-(1-methyl-1 / - / -benzo[rt]imidazol-2-yl)prop-2-en-1 -one (5) except using aldehyde tert-butyl (3-formylbenzyl)carbamate instead of 3-methoxybenzaldehyde. Yellow solid.

[0194] 1H NMR: (500 MHz, CDCI3) δ 8.25 (d, J = 16.0 Hz, 1 H), 7.94 - 7.89 (m, 2H), 7.65 - 7.63 (m, 2H), 7.48 - 7.46 (m, 2H), 7.41 - 7.35 (m, 3H), 4.38 (d, J = 4.8 Hz, 2H), 4.24 (s, 3H), 1.49 (s, 9H).

[0195] 13C{1H} NMR: (150 MHz, CDCI3) δ 183.0, 156.0, 147.3, 144.3, 141.9, 139.8,137.2, 135.1 , 130.0, 129.3, 128.2, 127.7, 126.0, 123.8, 123.3, 121.9, 110.6, 79.8, 44.5, 32.5, 28.5.tert-butyl (E)-(4-(3-(1 -methyl-1 H-benzo[rt]imidazol-2-yl)-3-oxoprop-1 -en-1 - yl)phenyl)carbamate (36)

[0196] The title compound was made in a manner analogous to (E)-3-(3- methoxyphenyl)-1-(1-methyl-1 / - / -benzo[d]imidazol-2-yl)prop-2-en-1 -one (5) except using aldehyde tert-butyl (4-formylphenyl)carbamate instead of 3-methoxybenzaldehyde. Yellow solid.

[0197] 1H NMR: (500 MHz, CDCI3) δ 8.15 (d, J = 15.9 Hz, 1 H), 7.93 (d, J = 8.2 Hz, 1 H), 7.88 (d, J = 15.9 Hz, 1 H), 7.69 (d, J = 8.2 Hz, 2H), 7.48 - 7.43 (m, 4H), 7.40 - 7.37 (m, 1 H), 6.66 (s, 1 H), 4.23 (s, 3H), 1.53 (s, 9H).

[0198] 13C{1H} NMR: (150 MHz, CDCI3) δ 183.0, 152.3, 147.5, 144.4, 141.9, 141.3,141.0, 137.2, 130.5, 130.3, 129.4, 125.8, 123.8, 121.9, 121.4, 118.3, 110.5, 81.2, 32.5, 28.3.

[0199] HRMS (ESI-TOF) m / z\ [M+Na]+Calcd for Cz^NsCteNa 400.1637; Found 400.1648.(E)-3-(6-meth oxy py ri di n -2-y I )-1 -(1 -methyl-1 H-benzo[c / ]imidazol-2-yl)prop-2-en-1 - one (50)

[0200] The title compound was made in a manner analogous to (E)-3-(3- methoxyphenyl)-1-(1 -methyl-1 H-benzo[c / ]imidazol-2-yl)prop-2-en-1 -one (5) except using aldehyde 6-methoxypicolinaldehyde instead of 3-methoxybenzaldehyde. Yellow solid.

[0201] 1H NMR: (500 MHz, CDCI3) δ 8.56 (d, J = 15.5 Hz, 1 H), 7.94 - 7.89 (m, 2H), 7.62 (t, J = 7.50 Hz, 1 H), 7.49 - 7.36 (m, 4H), 7.15 (d, J = 7.33 Hz, 1 H), 4.21 (s, 3H), 2.61 (s, 3H).

[0202] 13C{1H} NMR: (150 MHz, CDCI3) δ 183.6, 159.4, 153.1 , 147.4, 144.0, 142.2,137.5, 137.1 , 127.0, 126.3, 124.5, 124.1 , 122.3, 121.9, 110.8, 32.7, 24.9.(E)-3-(4-methoxypyridin-2-yl)-1 -(1 -methyl-1 H-benzo[d]imidazol-2-yl)prop-2-en-1 - one (52)

[0203] The title compound was made in a manner analogous to (E)-3-(3- methoxyphenyl)-1-(1 -methyl-1 H-benzo[c / ]imidazol-2-yl)prop-2-en-1 -one (5) except using aldehyde 4-methoxypicolinaldehyde instead of 3-methoxybenzaldehyde. Yellow solid.

[0204] 1H NMR: (500 MHz, CDCI3) δ 8.57 - 8.53 (m, 2H), 7.93 - 7.87 (m, 2H), 7.47 (m, 2H), 7.38 (m, 1 H), 7.21 (s, 1 H), 6.83 - 6.82 (m, 1 H), 4.23 (s, 3H), 3.91 (s, 3H).

[0205] 13C{1H} NMR: (150 MHz, CDCI3) δ 183.4, 166.7, 155.2, 151.5, 147.3, 143.5,142.3, 137.5, 127.7, 126.4, 124.1 , 122.4, 110.9, 110.8, 110.6, 55.8, 32.7.(E)-3-(2-methoxypyridin-4-yl)-1 -(1 -methyl-1 H-benzo[c / ]imidazol-2-yl)prop-2-en-1 - one (51)

[0206] The title compound was made in a manner analogous to (E)-3-(3- methoxyphenyl)-1-(1 -methyl-1 H-benzo[d]imidazol-2-yl)prop-2-en-1 -one (5) except using 2-methoxyisonicotinaldehyde instead of 3-methoxybenzaldehyde. Yellow solid.

[0207] 1H NMR: (500 MHz, CDCI3) δ 8.34 (d, J = 16.0 Hz, 1 H), 8.21 (d, J = 5.5 Hz, 1 H), 7.93 (d, J = 8.5 Hz, 1 H), 7.75 (d, J = 16.0 Hz, 1 H), 7.47 (d, J = 4.0 Hz, 2H), 7.41 - 7.38 (m, 1 H), 7.21 - 7.20 (m, 1 H), 6.98 (s, 1 H), 4.23 (s, 3H), 3.97 (s, 3H).

[0208] 13C{1H} NMR: (150 MHz, CDCI3) δ 182.1 , 164.6, 147.0, 146.3, 144.1 , 141.4,140.8, 136.7, 126.5, 125.8, 123.5, 121.6, 114.6, 110.2, 110.1 , 53.2, 32.0.(E)-1-(1-benzyl-1H-benzo[d]imidazol-2-yl)-3-(3-methoxyphenyl)prop-2-en-1-one (4)

[0209] To intermediate (E)-1-(1 / - / -benzo[cf]imidazol-2-yl)-3-(3-methoxyphenyl)prop-2- en-1-one (60), 200 mg, 0.72 mmol, 1 equiv.) in 3.6 mL of DMF under argon was addedK2CO3 (147 mg, 1.07 mmol, 1.5 equiv.). The reaction was stirred for 30 minutes at rt, afterwhich BnBr (130 pL, 1 .07 mmol, 1 .5 equiv.) was added. The reaction was heated at 80 °C for 2.5 h and then diluted with water and then EtOAc. The aqueous layer was extracted with EtOAc three times and then the combined organic layers were washed with water three times. The combined organic layers were dried over Na2SO4, concentrated under vacuum, and subjected to chromatography (10-25% EtOAc in hexanes) to give 73 mg (28% yield) of the title compound as a yellow solid.

[0210] 1H NMR: (500 MHz, CDCI3) δ 8.27 (d, J = 15.9 Hz, 1 H), 7.97 (d, J = 7.3 Hz, 1 H), 7.88 (d, J = 15.9 Hz, 1 H), 7.46 - 7.38 (m, 3H), 7.35 - 7.23 (m, 6H), 7.19 (d, J = 7.3 Hz, 2H), 6.99 - 6.97 (m, 1 H), 5.98 (s, 2H), 3.87 (s, 3H).

[0211] 13C{1H} NMR: (125 MHz, CDCI3) δ 182.7, 160.0, 146.9, 144.9, 142.0, 136.9,136.8, 136.1 , 129.9, 128.8, 127.8, 126.9, 126.3, 124.1 , 123.2, 122.1 , 122.0, 117.2, 113.4, 111.3, 55.5, 48.9.

[0212] HRMS (ESI-TOF) m / z: [M + Na]+Calcd for C24H2oN202Na 391.1422; Found 391.1424.(£)-1 -(1 -acetyl-1 H-benzo[d]imidazol-2-yl)-3-(3-methoxyphenyl)prop-2-en-1 -one (8)

[0213] To a scintillation vial equipped with a magnetic stir bar (E)-1 -(1 / - / - benzo[d]imidazol-2-yl)-3-(3-methoxyphenyl)prop-2-en-1 -one (60, 100 mg, 0.36 mmol, 1 equiv.) was dissolved in DCM (3.6 mL) at 20 °C and open to air. To this solution, triethylamine (300 pL, 2.15 mmol, 6 equiv.) was added, then acetic anhydride (160 pL, 1.4 mmol, 4 equiv.). The reaction was stirred for 16 hours before quenching with sat. aq. NH4CI and extracting with DCM three times. The organic extracts were dried with Na2SO4 and concentrated in vacuo to give a yellow solid. The material was purified using flash column chromatography (20% EtOAc / Hexanes) to give 8 as a yellow solid (47 mg, 40%).

[0214] 1H NMR: (600 MHz, CDCI3) δ 7.98 (d, J = 8.4 Hz, 1 H), 7.93 (d, J = 16.0 Hz, 1 H), 7.91 (s, 1 H), 7.80 (d, J = 16.0 Hz, 1 H), 7.52 (t, J = 7.8 Hz, 1 H), 7.46 (t, J = 8.0 Hz, 1 H), 7.36 (t, J = 7.6 Hz, 1 H), 7.31 (d, J = 7.6 Hz, 1 H), 7.23 (s, 1 H), 7.02 (dd, J = 8.2, 2.2 Hz, 1 H), 3.87 (s, 3H), 2.69 (s, 3H).

[0215] 13C{1H} NMR: (150 MHz, CDCI3) δ 183.0, 170.4, 160.2, 149.4, 147.4, 141.8,135.7, 134.1 , 130.2, 128.1 , 125.6, 123.3, 122.2, 121.9, 117.7, 114.5, 113.7, 55.6, 27.5.tert-butyl (E)-2-(3-(3-methoxyphenyl)acryloyl)-1 H-benzo[d]imidazole-1 -carboxylate (13)

[0216] To a scintillation vial equipped with a magnetic stir bar (E)-1 -(1 H- benzo[d]imidazol-2-yl)-3-(3-methoxyphenyl)prop-2-en-1 -one (60, 40 mg, 0.14 mmol, 1 equiv.) was dissolved in DCM (5.6 mL) at 20 °C and open to air. To this solution, triethylamine (200 pL, 1.44 mmol, 10 equiv.) and DMAP (1.7 mg, 0.014 mmol, 0.1 equiv.) were added, then di-fert-butyl dicarbonate (157 mg, 0.72 mmol, 5 equiv.). The reaction was stirred for 16 hours before quenching with sat. aq. NH4CI and extracting with DCM three times. The organic extracts were dried with Na2SO4 and concentrated in vacuo to give a yellow solid. The material was purified using flash column chromatography (30% EtOAc / Hexanes) to give 8 as a yellow solid (26 mg, 67%).

[0217] 1H NMR: (600 MHz, CDCI3) δ 8.02 (d, J = 7.7 Hz, 1 H), 7.86 (d, J = 8.2 Hz, 1 H), 7.70 (d, J = 16.0 Hz, 1 H), 7.51 (t, J = 7.4 Hz, 1 H), 7.43 (t, J = 7.8 Hz, 1 H), 7.34 (d, J = 15.9 Hz, 1 H), 7.32 (d, J = 15.6 Hz, 1 H), 7.22 (d, J = 7.7 Hz, 1 H), 7.13 (app. s, 1 H), 6.98 (dd, J = 8.3, 2.8 Hz, 1 H), 3.84 (s, 3H), 1 .62 (s, 9H).

[0218] 13C{1H} NMR: (150 MHz, CDCI3) δ 184.8, 160.1 , 149.8, 147.7 (2), 142.0, 135.7,133.2, 130.1 , 126.8, 125.2, 125.1 , 121.8, 121.5, 117.5, 114.5, 113.5, 86.7, 55.5, 27.8.

[0219] HRMS (ESI-TOF) m / z: [M+Na]+Calcd for C22H22N2O4Na 401.1477; Found 401.1481.(E)-1-(1-benzoyl-1H-benzo[d]imidazol-2-yl)-3-(3-methoxyphenyl)prop-2-en-1-one (14)

[0220] The title compound was prepared in a manner analogous to (E)-1 -(1 -acetyl-1 / - / - benzo[d]imidazol-2-yl)-3-(3-methoxyphenyl)prop-2-en-1 -one (8) except with benzoyl chloride instead of acetic anhydride. Yellow solid.

[0221] 1H NMR: (500 MHz, CDCI3) δ 8.02 - 8.00 (m, 1 H), 7.90 (d, J = 16.0 Hz, 1 H), 7.80 (d, J = 16.0 Hz, 1 H), 7.75 - 7.73 (m, 2H), 7.63 (t, J = 7.4 Hz, 1 H), 7.51 - 7.45 (m, 5H), 7.32 (t, J = 8.0 Hz, 1 H), 7.27 (d, J = 7.2 Hz, 1 H), 7.20 (app. t, J = 1 .4 Hz, 1 H), 6.99 (dd, J = 8.1 , 2.5 Hz, 1 H), 3.86 (s, 3H).

[0222] 13C{1H} NMR: (125 MHz, CDCI3) δ 180.9, 160.1 , 149.9, 146.6, 142.3, 135.9,135.5, 134.5, 130.3, 130.1 , 130.0, 129.2, 128.6, 127.5, 125.2, 122.3, 122.2, 122.0, 117.6,113.6, 112.8, 55.6.

[0223] HRMS (ESI-TOF) m / z\ [M+Na]+Calcd for C24Hi8N2O3Na 405.1215; Found 405.1204(E)-3-(3-methoxyphenyl)-1 -(1 -(4-(trifluoromethyl)benzyl)-1 H-benzo[d]imidazol-2- yl)prop-2-en-1-one (15)

[0224] To an oven-dried scintillation vial equipped with a magnetic stir bar under argon was added (E)-1-(1 / - / -benzo[d]imidazol-2-yl)-3-(3-methoxyphenyl)prop-2-en-1-one (60, 24 mg, 0.09 mmol, 1 equiv.) in 0.43 mL of DMF. K2CO3 (18 mg, 0.13 mmol, 1.5 equiv.) and 1 -(bromomethyl)-4-(trifluoromethyl)benzene (31 mg, 0.13 mmol, 1.5 equiv.) were then added in succession. The reaction was stirred at 80 °C for 16 h and then quenched after addition of water. The mixture was extracted with EtOAc three times, then the combined organic extracts were washed with water three times. The combined organic extracts were then dried over Na2SO4 and concentrated in vacuo. The material was purified using flash column chromatography (10% EtOAc / Hexanes) to give 15 as a yellow solid (12 mg, 22%).

[0225] 1H NMR: (600 MHz, CDCI3) δ 8.26 (d, J = 15.9 Hz, 1 H), 7.99 (dd, J = 7.6, 1 .5 Hz, 1 H), 7.88 (d, J = 15.9 Hz, 1 H), 7.56 (d, J = 8.1 Hz, 2H), 7.47 - 7.40 (m, 3H), 7.36 - 7.32 (m, 2H), 7.29 - 7.25 (m, 3H), 6.99 (dt, J = 6.9, 2.1 Hz, 1 H) δ.62 (s, 2H), 3.89 (s, 3H).

[0226] 13C{1H} NMR: (150 MHz, CDCI3) δ 182.8, 160.1 , 146.9, 145.3, 142.2, 140.8,136.8, 136.1 , 130.0, 127.1 , 126.7, 125.9 (q, J = 3.9 Hz), 124.4, 124.1 , 123.1 , 122.4, 122.2, 117.3, 113.6, 110.9, 55.6, 48.6.

[0227] HRMS (ESI-TOF) m / z\ [M+Na]+Calcd for C25Hi9F3N2O2Na 459.1296; Found 459.1298.(E)-3-(3-methoxyphenyl)-1-(1-pentyl-1H-benzo[tf]imidazol-2-yl)prop-2-en-1-one(16)

[0228] The title compound was synthesized in an analogous manner to (E)-3-(3- methoxyphenyl)-1-(1-(4-(trifluoromethyl)benzyl)-1 / - / -benzo[d]imidazol-2-yl)prop-2-en-1- one (15) except using 1 -bromopropane instead of 1 -(bromomethyl)-4- (trifluoromethyl)benzene. Yellow solid (38 mg, 40%).

[0229] 1H NMR: (500 MHz, CDCI3) δ 8.26 (d, J = 16.0 Hz, 1 H), 7.94 (d, J = 8.2 Hz, 1 H), 7.89 (d, J = 16.0 Hz, 1 H), 7.48 (d, J = 8.3 Hz, 1 H), 7.44 (td, J = 6.9, 0.9 Hz, 1 H), 7.38 (td, J = 6.9, 1.5 Hz, 1 H), 7.34 (d, J = 5.1 Hz, 2H), 7.28 - 7.26 (m, 1 H), 7.00 - 6.96 (m, 1 H), 4.69 (t, J = 7.8 Hz, 2H), 3.88 (s, 3H), 1.92 - 1 .86 (m, 2H), 1.42 - 1 .36 (m, 4H), 0.91 (t, J = 6.9 Hz, 3H).

[0230] 13C{1H} NMR: (150 MHz, CDCI3) δ 182.8, 159.9, 146.9, 144.6, 141.9, 136.7,136.1 , 129.9, 125.9, 123.8, 123.3, 122.1 , 121.9, 117.1 , 113.3, 110.9, 55.5, 45.8, 30.2,29.1 , 22.5, 14.1.

[0231] HRMS (ESI-TOF) m / z: [M+H]+Calcd for C22H24N2O2H 349.1908; Found 349.1916.(E)-1-(1-(4-methoxybenzyl)-1H-benzo[cf]imidazol-2-yl)-3-(3-methoxyphenyl)prop-2- en-1-one (22)

[0232] The title compound was synthesized in an analogous manner to (E)-3-(3- methoxyphenyl)-1-(1-(4-(trifluoromethyl)benzyl)-1 / - / -benzo[d]imidazol-2-yl)prop-2-en-1- one (15) except using 1 -(bromomethyl)-4-methoxybenzene instead of l-(bromomethyl)- 4-(trifluoromethyl)benzene. Yellow solid (30 mg, 21 %).

[0233] 1H NMR: (500 MHz, CDCI3) δ 8.25 (d, J = 16.0 Hz, 1 H), 7.95 (app. d, J = 7.7 Hz, 1 H), 7.88 (d, J = 16.0 Hz, 1 H), 7.49 (d, J = 7.9 Hz, 1 H), 7.24 (td, J = 7.1 , 1.3 Hz, 1 H), 7.33 (td, J = 8.2, 1.3 Hz, 1 H), 7.34 - 7.32 (m, 2H), 7.26 - 7.25 (m, 1 H), 7.18 (d, J = 8.8 Hz, 2H), 7.00 - 6.97 (m, 1 H), 7.81 (app. d, J = 9.0 Hz, 2H), 5.91 (s, 2H), 3.88 (s, 3H), 3.75 (s, 3H).

[0234] 13C{1H} NMR: (150 MHz, CDCI3) δ 182.8, 160.0, 159.2, 146.9, 144.8, 142.0,136.8, 136.1 , 129.9, 128.9, 128.4, 126.2, 124.0, 123.3, 122.1 , 122.0, 117.1 , 114.2, 113.4, 111.3, 55.5, 55.3, 48.4.

[0235] HRMS (ESI-TOF) m / z\ [M+Na]+Calcd for C25H22N2O3Na 421.1528; Found 421.1522.(E)-3-(3-methoxyphenyl)-1-(1-(pent-4-yn-1-yl)-1H-benzo[d]imidazol-2-yl)prop-2-en- 1-one(23)

[0236] The title compound was synthesized in an analogous manner to (E)-3-(3- methoxyphenyl)-1-(1-(4-(trifluoromethyl)benzyl)-1 / - / -benzo[d]imidazol-2-yl)prop-2-en-1- one (15) except using 5-bromo-1 -pentyne instead of 1 -(bromomethyl)-4- (trifluoromethyl)benzene and 10 mol% Nal was added. Yellow solid.

[0237] 1H NMR: (600 MHz, CDCI3) δ 8.25 (d, J = 16.0 Hz, 1 H), 7.94 (dt, J = 8.3, 0.9 Hz, 1 H), 7.89 (d, J = 16.0 Hz, 1 H), 7.59 (dt, J = 8.3, 0.9 Hz, 1 H), 7.47 (ddd, J = 8.2, 7.1 ,1.1 Hz, 1 H), 7.40 (ddd, J = 8.2, 7.1 , 1.1 Hz, 1 H), 7.35 - 7.33 (m, 2H), 7.28 - 7.26 (m, 1 H), 7.00 - 6.97 (m, 1 H), 4.82 (app. t, J = 7.2 Hz, 2H), 3.88 (s, 3H), 2.31 (td, J = 6.9, 2.7 Hz, 2H), 2.14 (app. p, J = 6.9 Hz, 2H), 2.08 (t, J = 2.7 Hz, 1 H).

[0238] 13C{1H} NMR: (150 MHz, CDCI3) δ 182.8, 160.0, 146.9, 144.7, 141.9, 136.8,136.1 , 129.9, 126.2, 123.9, 123.3, 122.1 , 121.9, 117.1 , 113.4, 110.8, 83.1 , 69.5, 55.5, 44.5, 29.1 , 16.1.

[0239] HRMS (ESI-TOF) m / z\ [M+Na]+Calcd for C22H2oN202Na 367.1422; Found 367.1424.Route B. (E)- / V-(3-(3-(1 -methyl-1 H-benzo[cf]imidazol-2-yl)-3-oxoprop-1 -en-1 - yl)phenyl)acrylamide (11)Step 1. tert-butyl (E)-(3-(3-(1H-benzo[d]imidazol-2-yl)-3-oxoprop-1-en-1- yl)phenyl)carbamate (S3)

[0240] To a dram vial equipped with a magnetic stir bar, (88 mg, 0.55 mmol, 1 equiv.) of 2-acetylbenzimidazole, commercially available or synthetically accessible, were dissolved in 0.55 mL of EtOH at rt and open to air. To this solution, (122 mg, 0.55 mmol, 1 equiv.) of tert-butyl (3-formylphenyl)carbamate followed by (0.55 mL, 2.2 mmol, 4 equiv.) 4 M aq. NaOH were added and the solution was allowed to stir overnight. The reaction was quenched with sat. aq. NH4CI and the resulting yellow solid was filtered and then dried in a vacuum desiccator over P2Os overnight to give enone S3 as a yellow solid (180 mg, 90% yield).

[0241] 1H NMR: (500 MHz, CDCI3) δ 10.41 (br s, 1 H), 8.10 (d, J = 16.0 Hz, 1 H), 8.05 (d, J = 16.0 Hz, 1 H), 7.96 (d, J = 8.1 Hz, 1 H), 7.71 (s, 1 H), 7.58 (d, J = 8.1 Hz, 1 H), 7.51 (dd, J = 8.1 , 1.2 Hz, 1 H), 7.47 (d, J = 6.8 Hz, 1 H), 7.44 (td, J = 8.1 , 1.2 Hz, 1 H), 7.42 - 7.36 (m, 1 H), 7.36 (t, J = 8.0 Hz, 1 H), 6.68 (br s, 1 H), 1 .56 (s, 9H).

[0242] 13C{1H} NMR: (125 MHz, THF-d8) 5 181.6, 153.4, 150.3, 144.7 (2), 141.5,136.2, 135.7, 129.9, 126.1 , 123.5, 123.3, 122.5, 122.3, 121.0, 118.2, 112.8, 79.8, 28.4.

[0243] HRMS (ESI-TOF) m / z: [M+Na]+Calcd for C2i H2i N3O3Na 386.1481 ; Found 386.1484.Step 2. tert-butyl (E)-(3-(3-(1-methyl-1H-benzo[d]imidazol-2-yl)-3-oxoprop-1-en-1- yl)phenyl)carbamate (19)

[0244] To a flask equipped with a magnetic stir bar, NaH (60% dispersion in mineral oil, 33 mg, 0.83 mmol, 1.0 equiv.) was added to benzimidazole S3 (300 mg, 0.83 mmol, 1 equiv.) in 2.0 mL of DMF under Ar at rt. After stirring for 5 minutes, methyl iodide (80 pL, 1.24 mmol, 1.5 equiv.) was added. The reaction was allowed to stir overnight and then was quenched with water and diluted with EtOAc. The organic layer was washed with water three times, and then with brine once. The organic layer was then dried with MgSO4 and concentrated in vacuo. The material was further purified by chromatography (80:15:5 EtOAc: CHCl3:N Eta) to give methylated benzimidazole 19 as a yellow foam (200 mg, 64% yield).

[0245] 1H NMR: (600 MHz, CDCI3) δ 8.22 (d, J = 16.0 Hz, 1 H), 7.92 (d, J = 8.2 Hz, 1 H), 7.87 (d, J = 16.0 Hz, 1 H), 7.68 (s, 1 H), 7.50 (d, J = 6.7 Hz, 1 H), 7.47 - 7.45 (m, 2H), 7.42 (d, J = 7.6 Hz, 1 H), 7.39 - 7.37 (m, 1 H), 7.24 (d, J = 7.9 Hz, 1 H), 6.69 (s, 1 H), 4.22 (s, 3H), 1.54 (s, 9H).

[0246] 13C{1H} NMR: (150 MHz, CDCI3) δ 183.1 , 152.8, 147.3, 144.5, 142.0, 139.2,137.3, 135.7, 129.6, 126.1 , 123.9, 123.8, 123.6, 122.0, 121.0, 118.8, 110.6, 80.9, 32.5, 28.5.

[0247] HRMS (ESI-TOF) m / z\ [M+Na]+Calcd for (z^NsOsNa 400.1637; Found 400.1636.Step 3. (E)-3-(3-aminophenyl)-1-(1-methyl-1H-benzo[d]imidazol-2-yl)prop-2-en-1- one (10)

[0248] To a flask equipped with a magnetic stir bar (210 pL, 2.7 mmol, 10 equiv.) of TFA was added to (100 mg, 0.27 mmol, 1 equiv.) of Boc-protected aniline 19 dissolved in 2.7 mL of CH2CI2 at rt. The reaction was allowed to run overnight and then quenched with sat. aq. NaHCOs, and the mixture was extracted with CH2CI2 three times. The combined organic layers were then dried with MgSO4 and concentrated in vacuo to give aniline 10 as a yellow solid (64 mg, 85% yield).

[0249] 1H NMR: (500 MHz, CDCI3) δ 8.19 (J = 16.0 Hz, 1 H), 7.93 (d, J = 8.1 Hz, 1 H), 7.84 (d, J = 15.9 Hz, 1 H), 7.48 - 7.44 (m, 2H), 7.38 (ddd, J = 8.1 , 5.8, 2.4 Hz, 1 H), 7.21 (t, J = 7.7 Hz, 1 H), 7.13 (d, J = 7.6 Hz, 1 H), 7.08 (s, 1 H), 6.75 (dd, J = 7.9, 1.6 Hz, 1 H), 4.23 (s, 3H), 3.79 (bs, 2H).

[0250] 13C{1H} NMR: (125 MHz, CDCI3) δ 183.2, 147.6, 147.0, 145.1 , 142.1 , 137.3,136.0, 129.9, 126.0, 123.8, 123.1 , 122.1 , 120.2, 117.9, 114.7, 110.6, 32.5.

[0251] HRMS (ESI-TOF) m / z: [M+H]+Calcd for C17H15N3OH 278.1293; Found 278.1282.Step 4. (E)- / V-(3-(3-(1 -methyl-1 H-benzo[d]imidazol-2-yl)-3-oxoprop-1 -en-1 - yl)phenyl)acrylamide (11)

[0252] To a flask equipped with a magnetic stir bar of aniline 10 (89 mg, 0.32 mmol, 1 equiv.) and Et3N (30 pL, 0.38 mmol, 3 equiv.) were dissolved in 3.0 mL of CH2CI2 at rt under Ar. After 1 hour, the reaction was quenched with water and extracted with EtOAc three times. The combined organic layers were then dried with MgSCk, 0.2 mg of 4- methoxyphenol was added as a solution in CH2CI2 and concentrated in vacuo. The material was further purified by chromatography (20% EtOAc / CHCI3) to give acrylamide 11 as an orange solid (20 mg, 19% yield).

[0253] 1H NMR: (600 MHz, DMSO-d6) δ 10.35 (s, 1 H), 8.21 (s, 1 H), 8.18 (d, J = 16.2 Hz, 1 H), 7.91 (d, J = 8.2 Hz, 1 H), 7.83 (d, J = 16.0 Hz, 1 H), 7.80 (d, J = 8.0 Hz, 1 H), 7.76 (d, J = 8.3 Hz, 1 H), 7.53 - 7.44 (m, 3H), 7.40 (t, J = 7.6 Hz, 1 H), 6.47 (dd, J = 10.2, 6.8 Hz, 1 H), 6.32 (d, J = 17.2 Hz, 1 H), 5.81 (d, J = 10.1 Hz, 1 H), 4.19 (s, 3H).

[0254] 13C{1H} NMR: (150 MHz, DMSO-d6) δ 182.4, 163.9, 147.4, 143.8, 141.7, 140.2,137.5, 135.3, 132.2, 130.2, 127.8, 126.3, 125.7, 124.1 , 123.6, 122.4, 121.7, 118.3, 112.1 , 32.8.

[0255] HRMS (ESI-TOF) m / z: [M+Na]+Calcd for C2oHi7N302Na 354.1219; Found 354.121.tert-butyl (E)-methy l(3-(3-(1-methy 1-1 H-benzo[d]i midazol -2-yl )-3-oxoprop-1-en-1- yl)phenyl)carbamate (20)

[0256] The title compound was another product of Step 2, Route B. tert-butyl (E)-(3- (3-(1-methyl-1 / - / -benzo[d]imidazol-2-yl)-3-oxoprop-1 -en-1 -yl)phenyl)carbamate (19). 79 mg (16% yield) of yellow foam was obtained.

[0257] 1H NMR: (500 MHz, CDCI3) δ 8.23 (d, J = 16.0 Hz, 1 H), 7.92 (d, J = 8.2 Hz, 1 H), 7.89 (d, J = 16.0 Hz, 1 H), 7.61 (s, 1 H), 7.56 (d, J = 7.5 Hz, 1 H), 7.48 - 7.44 (m, 2H), 7.40 - 7.36 (m, 2H), 7.31 (d, J = 7.8 Hz, 1 H), 4.23 (s, 3H), 3.30 (s, 3H), 1 .47 (s, 9H).

[0258] 13C{1H} NMR: (150 MHz, CDCI3) δ 183.0, 154.7, 147.3, 144.6, 144.2, 142.0,137.3, 135.5, 129.2, 128.2, 126.1 (2), 125.9, 123.9, 123.6, 122.0, 110.7, 80.7, 37.4, 32.5, 28.5.

[0259] HRMS (ESI-TOF) m / z\ [M+Na]+Calcd for C23H25N3O3Na 414.1794; Found 414.1794.tert-butyl (E)-methyl(2-(3-(1-methyl-1H-benzo[cflimidazol-2-yl)-3-oxoprop-1-en-1- yl)pyridin-4-yl)carbamate (53)

[0260] The title compound was made in a manner analogous to fert-butyl (E)-(3-(3-(1 - methyl-1 / - / -benzo[cf]imidazol-2-yl)-3-oxoprop-1-en-1 -yl)phenyl)carbamate (19) except aldehyde tert-butyl (2-formylpyridin-4-yl)carbamate was used instead of tert-butyl (3- formylphenyl)carbamate. Yellow solid.

[0261] 1H NMR: (400 MHz, CDCI3) δ 8.43 (d, J = 4.6 Hz, 1 H), 8.36 (d, J = 16.1 Hz, 1 H), 7.96 - 7.91 (m, 2H), 7.81 (d, J = 16.0 Hz, 1 H), 7.47 (app. s, 2H), 7.42 - 7.38 (m, 1 H), 7.34 (d, J = 4.4 Hz, 1 H), 4.23 (s, 3H), 3.42 (s, 3H), 1 .56 (s, 9H).(E)-1-(1-methyl-1H-benzo[tf]imidazol-2-yl)-3-(3-(methylamino)phenyl)prop-2-en-1- one (21)

[0262] The title compound was made in a manner analogous to Step 3, Route B, using tert-butyl (E)-methyl(3-(3-(1 -methyl-1 H-benzo[d]imidazol-2-yl)-3-oxoprop-1 -en-1 - yl)phenyl)carbamate (20) instead of (E)-(3-(3-(1-methyl-1H-benzo[d]imidazol-2-yl)-3- oxoprop-1 -en-1 -yl)phenyl)carbamate (19).

[0263] 1H NMR: (500 MHz, CDCI3) δ 8.20 (d, J = 16.0 Hz, 1 H), 7.93 (d, J = 8.2 Hz, 1 H), 7.88 (d, J = 16.0 Hz, 1 H), 7.47 - 7.43 (m, 2H), 7.38 (ddd, J = 8.3, 5.7, 2.6 Hz, 1 H), 7.23 (t, J = 7.7 Hz, 1 H), 7.10 (d, J = 7.6 Hz , 1 H), 6.96 (s, 1 H), 6.68 (dd, J = 8.0, 2.1 Hz, 1 H), 4.22 (s, 3H), 2.89 (s, 3H).

[0264] 13C{1H} NMR: (125 MHz, CDCI3) δ 183.2, 149.8, 147.5, 145.7, 142.0, 137.2,135.7, 129.7, 126.0, 123.8, 122.7, 121.9, 118.9, 115.5, 111.9, 110.6, 32.5, 30.8.

[0265] HRMS (ESI-TOF) m / z: [M+H]+Calcd for C18H17N3OH 292.1450; Found 292.1461.(E)- / V-methyl- / V-(3-(3-(1-methyl-1H-benzo[d]imidazol-2-yl)-3-oxoprop-1-en-1- yl)phenyl)acrylamide (25)

[0266] The title compound was made in a manner analogous to Step 4, Route B, using(E)-1-(1-methyl-1H-benzo[c(]imidazol-2-yl)-3-(3-(methylamino)phenyl)prop-2-en-1-one(21)

[0267] instead of (E)-3-(3-aminophenyl)-1-(1-methyl-1H-benzo[d]imidazol-2-yl)prop- 2-en-1-one (10).

[0268] 1H NMR: (500 MHz, CDCI3) δ 8.26 (d, J = 16.1 Hz, 1H), 7.92 (d, J = 8.1 Hz, 1H), 7.88 (d, J = 16.1 Hz, 1H), 7.70 (d, J = 7.6 Hz, 1H), 7.56 (s, 1H), 7.49-7.45 (m, 3H), 7.39 (ddd, J = 8.3, 5.3, 3.0 Hz, 1H), 7.23 (d, J = 6.9 Hz, 1H), 6.40 (dd, J= 16.8, 1.8 Hz, 1H), 6.09 (dd, J= 16.1, 10.7 Hz, 1H), 5.55 (dd, J= 10.3, 1.4 Hz, 1H), 4.23 (s, 3H), 3.40 (s, 3H).

[0269] 13C{1H} NMR: (125 MHz, CDCI3) δ 182.8, 165.8, 147.1, 144.3, 143.0, 142.0,137.3, 136.6, 130.3, 129.5, 128.4, 128.2, 128.1, 127.4, 126.3, 124.5, 124.1, 122.1, 110.7, 37.6, 32.6.

[0270] HRMS (ESI-TOF) m / z: [M+H]+Calcd for C21H19N3O2H 346.1555; Found 346.1545.(E)-W-(3-(3-(1 -methyl -1 H-benzo[d]imidazol-2-yl)-3-oxoprop-1 -en-1 -yl)phenyl)but-2- ynamide (29)

[0271] Using intermediate (E)-3-(3-aminophenyl)-1-(1-methyl-1H-benzo[c / ]imidazol-2- yl)prop-2-en-1 -one (10, 20 mg, 0.072 mmol) in 0.25 mL of DCM, 2,5-dioxopyrrolidin-1 -yl but-2-ynoate (150 mg, 0.83 mmol) was added. The mixture was stirred overnight at room temperature. The mixture was quenched with sat. aq. NaHCOs, extracted with EA, dried over MgSCM, and concentrated under vacuum. This resulted in 6 mg of (E)-A / -(3-(3-(1- methyl-1 H-benzo[c / ]imidazol-2-yl)-3-oxoprop-1 -en-1 -yl)phenyl)but-2-ynam ide (29) as a yellow solid.

[0272] 1H NMR: (600 MHz, CDCI3) δ 8.22 (d, J = 15.6 Hz, 1 H), 7.92 (d, J = 8.3 Hz, 1 H), 7.86 (d, J = 15.9 Hz, 1 H), 7.75 (s, 1 H), 7.70 - 7.67 (m, 2H), 7.51 (d, J = 7.8 Hz, 1 H), 7.47 - 7.46 (m, 2H), 7.39 - 7.36 (m, 2H), 4.23 (s, 3H), 2.01 (s, 3H).

[0273] HRMS (ESI-TOF) m / z: [M+H]+Calcd for C21H17N3O2H 344.1399; Found 344.1411.

[0274] (E)- / V-(3-(3-(1 -methyl -1 W-benzo[cflimidazol-2-yl)-3-oxoprop-1 -en-1 -y I )pheny I )- ethenesulfonamide (44)

[0275] To intermediate (E)-3-(3-aminophenyl)-1-(1-methyl-1H-benzo[d]imidazol-2- yl)prop-2-en-1 -one (10), 20 mg, 0.072 mmol) in 0.7 mL of DCM under an inert atmosphere of argon, 2-chloroethane-1 -sulfonyl chloride (9 pL, 0.087 mmol) and triethylamine (30 pL,0.22 mmol) were added. The mixture was stirred overnight at room temperature. The mixture was quenched with water, extracted with DCM, dried over MgSO4, and concentrated under vacuum. This resulted in 24 mg of (E)-A / -(3-(3-(1 -methyl-1 / - / - benzo[d]imidazol-2-yl)-3-oxoprop-1 -en-1 -yl)phenyl)-ethenesulfonamide (44) as a yellow solid.

[0276] 1H NMR: (500 MHz, CDCI3) δ 8.21 (d, J = 15.8 Hz, 1 H), 7.93 (d J = 7.9 Hz, 1 H), 7.83 (d, J = 15.8 Hz, 1 H), 7.52 (d, J = 7.9 Hz, 1 H), 7.48 - 7.47 (m, 3H), 7.41 - 7.35 (m, 2H), 7.30 (d, J = 8.8 Hz, 1 H), 7.09 (s, 1 H), 6.60 (dd, J = 16.6, 9.8 Hz, 1 H), 6.32 (d, J = 16.5 Hz, 1 H), 5.99 (d, J = 10.2 Hz, 1 H), 4.23 (s, 3H).

[0277] 13C{1H} NMR: (150 MHz, CDCI3) δ 182.6, 146.9, 143.6, 141.5, 137.4, 137.1 ,136.1 , 135.3, 130.0, 128.6, 126.2, 125.7, 124.1 , 124.0, 123.0, 121.7, 120.8, 110.7, 32.5.

[0278] HRMS (ESI-TOF) m / z\ [M+Na]+Calcd for CigHnNsOsSNa 390.0888; Found 390.0906.(E)- / V-(3-(3-(1 -methyl -1 H-benzo[d]imidazol-2-yl)-3-oxoprop-1 -en-1 - yl)phenyl)methacrylamide (12)

[0279] Intermediate (E)-3-(3-aminophenyl)-1-(1-methyl-1 H-benzo[d]imidazol-2- yl)prop-2-en-1 -one (10), 20 mg, 0.072 mmol, 1 equiv.) was dissolved in 0.72 mL of DMF under an inert atmosphere of argon. Then triethylamine (30 pL, 0.21 mmol, 3 equiv.) and then methacryloyl chloride (8 pL, 0.086 mmol, 1.2 equiv.) were added. The reaction was quenched with water and diluted with EtOAc. The organic layer was washed with water three times, and then with brine once. The organic layer was dried with MgSCh and concentrated in vacuo. The material was further purified by chromatography (80:20 CHCI3: EtOAc, 1 % NEts) to give the title compound (4 mg, 16%) as a yellow solid.

[0280] 1H NMR: (600 MHz, CDCI3) δ 8.24 (d, J = 15.9 Hz, 1 H), 7.93 (d, J = 8.4 Hz, 1 H), 7.89 (d, J = 16.0 Hz, 1 H), 7.84 (s, 1 H), 7.80 (d, J = 8.2 Hz, 1 H), 7.69 (s, 1 H), 7.50 - 7.45 (m, 3H), 7.41 - 7.38 (m, 2H), 5.85 (s, 1 H), 5.51 (s, 1 H), 4.23 (s, 3H), 2.10 (s, 3H).

[0281] 13C{1H} NMR: (150 MHz, CDCI3) δ 183.0, 166.8, 147.3, 144.1 , 142.0, 140.9,138.6, 137.3, 135.7, 129.8, 126.1 , 125.4, 124.0, 123.7, 122.6, 122.0, 120.4, 119.8, 110.7,32.6, 18.9.

[0282] HRMS (ESI-TOF) m / z\ [M+Na]+Calcd for C2i Hi9N3O2Na 368.1375; Found 368.1375.(£)-1 -(3-(3-( 1 -methyl-1 H-benzo[cflimidazol-2-yl)-3-oxoprop-1 -en-1 -y I )ph eny I )-1 H- pyrrole-2, 5-dione (54)

[0283] Aniline (E)-3-(3-aminophenyl)-1 -(1 -methyl-1 H-benzo[c^imidazol-2-yl)prop-2 - en-1-one (10, 50 mg, 0.18 mmol, 1 equiv.), maleic anhydride (18 mg, 0.18 mmol, 1 equiv.) in CHCh (0.18 mL) was stirred for 1 hour at rt and then concentrated under vacuum. Then, under an atmosphere of argon, acetic anhydride (60 pL, 0.90 mmol, 5 equiv.) and NaOAc (14 mg, 0.09 mmol, 0.5 equiv.) was added, and the reaction was heated at 100 °C for 6 hours. The reaction was then diluted with water, extracted with DCM, dried over MgSO4, and concentrated under vacuum to give the title compound as a yellow solid.

[0284] 1H NMR: (400 MHz, CDCI3) δ 8.27 (d, J = 15.9 Hz, 1 H), 7.94 - 7.90 (m, 2H), 7.76 (d, J = 8.1 Hz, 1 H), 7.73 (s, 1 H), 7.54 (t, J = 7.8 Hz, 1 H), 7.48 - 7.46 (m, 2H), 7.42 - 7.37 (m, 2H), 6.90 (s, 2H), 4.24 (s, 3H).

[0285] 13C{1H} NMR: (150 MHz, CDCI3) δ 182.9, 169.4, 147.2, 143.3, 142.0, 137.3,136.1 , 134.5, 132.1 , 129.8, 128.4, 128.2, 126.5, 126.2, 124.3, 124.0, 122.1 , 110.7, 32.6.

[0286] HRMS (ESI-TOF) m / z: [M+H]+Calcd for C21H15N3O3H 358.1192; Found 358.1196.(E)-W-(3-(3-(5,6-difluoro-1 -methyl-1 H-benzo[d]imidazol-2-yl)-3-oxoprop-1 -en-1 - yl)phenyl)acrylamide (43)

[0287] The title compound was made in a manner analogous to (E)- / V-(3-(3-(1 -methyl- 1 / - / -benzo[d]imidazol-2-yl)-3-oxoprop-1 -en-1 -yl)phenyl)acrylam ide (11) except using ketone 1 -(5,6-difluoro-1 / - / -benzo[cf|imidazol-2-yl)ethan-1 -one S2 instead of 2- acetylbenzimidazole. Yellow solid.

[0288] 1H NMR: (500 MHz, DMSO-d6) δ 10.35 (s, 1 H), 8.21 (s, 1 H), 8.09 (d, J = 15.6 Hz, 1 H), 8.02 - 7.98 (m, 2H), 7.84 - 7.77 (m, 2H), 7.50 - 7.43 (m, 2H), 6.46 (dd, J = 16.2, 10.0 Hz, 1 H), 6.30 (d, J = 16.2 Hz, 1 H), 5.81 (d, J = 10.0 Hz, 1 H), 4.15 (s, 3H).(E)-W-(3-(3-oxo-3-(1 -(4-(trifluoromethyl)benzyl)-1 H-benzo[d]imidazol-2-yl)prop-1 - en-1-yl)phenyl)acrylamide (26)

[0289] te / t-buty I (E)-(3-(3-oxo-3-(1 -(4-(trifluoromethyl)benzyl)-1 H-benzo[c / ]imidazol-2- yl)prop-1 -en-1 -yl)phenyl)carbamate (S6) was made in a manner analogous to (E)-3-(3- methoxyphenyl)-1-(1-(4-(trifluoromethyl)benzyl)-1H-benzo[tf|imidazol-2-yl)prop-2-en-1- one (15) except using tert-butyl (E)-(3-(3-(1H-benzo[c / ]imidazol-2-yl)-3-oxoprop-1-en-1- yl)phenyl)carbamate (S3) instead of (E)-1-(1H-benzo[d]imidazol-2-yl)-3-(3- methoxyphenyl)prop-2-en-1-one 60. The intermediate (S6) was then subjected to steps 3-4 from Route B, with (S6) replacing tert-butyl (E)-(3-(3-(1 -methyl-1 H-benzo[d]imidazol-2-yl)-3-oxoprop-1-en-1-yl)phenyl)carbamate (19) to give the title compound as a yellow solid.

[0290] 1H NMR: (600 MHz, CDCI3) δ 8.23 (d, J = 15.9 Hz, 1 H), 7.98 (d, J = 7.7 Hz, 1 H), 7.89 (s, 1 H), 7.83 (app. d, J = 16.4 Hz, 2H), 7.55 (d, J = 8.0 Hz, 2H), 7.47-7.40 (m, 4H), 7.37 (t, J = 7.8 Hz, 1 H), 7.29-7.25 (m, 2H), 6.48 (d, J = 16.8 Hz, 1 H), 6.35 (dd, J = 16.9, 10.4 Hz, 1 H), 6.01 (s, 2H), 5.81 (d, J = 10.2 Hz, 1 H).

[0291] HRMS (ESI-TOF) m / z: [M+Na]+Calcd for C27H2oF3N302Na 498.1405; Found 498.1416.(E)- / V-(3-(3-oxo-3-(1-(pent-4-yn-1-yl)-1H-benzo[d]imidazol-2-yl)prop-1-en-1- yl)phenyl)acrylamide (24)

[0292] tert-butyl (E)-(3-(3-oxo-3-(1 -(pent-4-yn-1 -yl)-1 H-benzo[rt]imidazol-2-yl)prop-1 - en-1-yl)phenyl)carbamate (S7) was made in a manner analogous to (E)-3-(3- methoxyphenyl)-1 -(1 -(pent-4-yn-1 -yl)-1 A7-benzo[c / ]imidazol-2-yl)prop-2-en-1 -one (23) except using tert-butyl (E)-(3-(3-(1 / - / -benzo[d]imidazol-2-yl)-3-oxoprop-1-en-1- yl)phenyl)carbamate (S3) instead of (E)-1-(1H-benzo[d]imidazol-2-yl)-3-(3- methoxyphenyl)prop-2-en-1-one 60. The intermediate (S7) was then subjected to steps 3-4 from Route B, with (S7) replacing tert-butyl (E)-(3-(3-(1-methyl-1H-benzo[d]imidazol- 2-yl)-3-oxoprop-1-en-1-yl)phenyl)carbamate (19) to give the title compound as a yellow solid.

[0293] 1H NMR: (600 MHz, CDCI3) δ 8.22 (app. d, J = 16.0 Hz, 2H), 7.92 (d, J = 8.1 Hz, 1 H), 7.83 (app. d, J = 16.0 Hz, 2H), 7.78 (s, 1 H), 7.60 (d, J = 8.2 Hz, 1 H), 7.47 (t, J = 7.6 Hz, 1 H), 7.44 (d, J = 7.7 Hz, 1 H), 7.40 (t, J = 7.7 Hz, 1 H), 7.36 (t, J = 7.9 Hz, 1 H), 6.50 (d, J = 16.9 Hz, 1 H), 6.40 (dd, J = 16.8, 10.0 Hz, 1 H), 5.80 (d, J = 10.0 Hz, 1 H), 4.81(t, J = 7.2 Hz, 2H), 2.31 (td, J = 7.0, 2.6 Hz, 2H), 2.14 (p, J = 7.0 Hz, 2H), 2.08 (t, J = 2.6 Hz, 1 H).

[0294] 13C{1H} NMR: (150 MHz, CDCI3) δ 182.6, 164.1 , 146.9, 144.2, 141.9, 138.7,136.9, 135.6, 131.2, 129.8, 128.5, 126.4, 125.5, 124.1 , 123.6, 122.6, 121.9, 119.6, 111.0,83.2, 69.7, 44.6, 29.2, 16.2.

[0295] HRMS (ESI-TOF) m / z\ [M+Na]+Calcd for C24H2i N3O2Na 406.1531 ; Found 406.1546.(£)-W-(4-(3-(1 -methyl -1 W-benzo[cflimidazol-2-yl)-3-oxoprop-1 -en-1 - yl)phenyl)acrylamide (37)

[0296] The title compound was made in a manner analogous to (E)- / V-(3-(3-(1 -methyl- 1 / - / -benzo[d]imidazol-2-yl)-3-oxoprop-1 -en-1 -yl)phenyl)acrylam ide (11) except using tert- butyl (4-formylphenyl)carbamate instead of tert-butyl (3-formylphenyl)carbamate. Yellow solid.

[0297] 1H NMR: (600 MHz, CDCI3) δ 8.17 (d, J = 16.4 Hz, 1 H), 7.92 (d, J = 8.1 Hz, 1 H), 7.87 (d, J = 15.8 Hz, 1 H), 7.80 (s, 1 H), 7.72 - 7.68 (m, 3H), 7.47 - 7.46 (m, 2H), 7.40 - 7.37 (m, 1 H), 6.46 (d, J = 16.6 Hz, 1 H), 6.27 (dd, J = 17.0, 11.0 Hz, 1 H), 5.79 (d, J =10.2, 1 H), 4.22 (s, 3H).

[0298] 13C{1H} NMR: (150 MHz, CDCI3) δ 182.9, 163.6, 147.4, 144.0, 141.8, 140.3,137.2, 130.9, 130.9, 130.2, 128.5, 126.0, 123.9, 122.1 , 121.8, 119.9, 110.6, 32.5.Route C. (E)-3-(3-methoxyphenyl)-1 -(1 -(3-morpholinopropyl)-1 H-benzo[cf]imidazol- 2-yl)prop-2-en-1-one (28)Step 1. (E)-1-(1-(3-((tert-butyldimethylsilyl)oxy)propyl)-1H-benzo[d]imidazol-2-yl)- 3-(3-methoxyphenyl)prop-2-en-1-one

[0299] To a solution of (E)-1 -(1 H-benzo[c(]imidazol-2-yl)-3-(3-methoxyphenyl)prop-2- en-1-one (60) (280 mg, 1.01 mmol) in 5 mL of DMF, fert-butyl(3- chloropropoxy)dimethylsilane (422 mg, 2.02 mmol), which is commercially available or synthetically accessible, Nal (152 mg, 1 .01 mmol), and CS2CO3 (988 mg, 3.03 mmol) was added. The mixture was stirred overnight at 80 °C. The mixture was washed with water and brine, dried over MgSO4, and concentrated under vacuum. The residue was applied onto a silica gel column (90:10 Hex / EA). This resulted in 200 mg of (E)-1 -(1 -(3-((tert- butyldimethylsilyl)oxy)propyl)-1H-benzo[d]imidazol-2-yl)-3-(3-methoxyphenyl)prop-2-en- 1-one as a yellow solid.Step 2. (E)-1 -(1 -(3-hydroxypropyl)-1 H-benzo[d]imidazol-2-yl)-3-(3- methoxyphenyl)prop-2-en-1 -one (27)

[0300] To a solution of (E)-1 -(1 -(3-((tert-butyldimethylsilyl)oxy)propyl)-1 H- benzo[d]imidazol-2-yl)-3-(3-methoxyphenyl)prop-2-en-1-one (196 mg, 0.43 mmol) in 4.3 mL of THF at 0 °C, TBAF (1 .3 mL, 1 M in THF, 1 .3 mmol) was added. The mixture waswarmed to room temperature and stirred for 1 hour. The mixture was quenched with aqueous NH4CI, extracted with EA, dried over MgSCU, and concentrated under vacuum. The residue was applied onto a silica gel column (60:40 Hex / EA). This resulted in 118 mg of (E)-1-(1-(3-hydroxypropyl)-1 H-benzo[c / ]imidazol-2-yl)-3-(3-methoxyphenyl)prop-2-en-1-one as a yellow solid.

[0301] 1H NMR: (600 MHz, CDCI3) δ 8.27 (d, J = 15.9 Hz, 1 H), 7.94 (d, J = 8.1 Hz, 1 H), 7.91 (d, J = 15.9 Hz, 1 H), 7.55 (d, J = 8.2 Hz, 1 H), 7.46 (t, J = 7.1 Hz, 1 H), 7.40 (t, J = 7.6 Hz, 1 H), 7.24 (d, J = 4.8 Hz, 2H), 7.27 (s, 1 H), 7.00 - 6.98 (m, 1 H), 4.84 (t, J = 6.5 Hz, 2H), 3.88 (s, 3H), 3.61 (t, J = 5.2 Hz, 2H), 2.79 (s, 1 H), 2.16 (p, J = 6.1 Hz, 2H).

[0302] 13C{1H} NMR: (150 MHz, CDCI3) δ 183.5, 160.1 , 147.2, 145.5, 142.1 , 136.8,136.1 , 130.1 , 126.4, 124.2, 123.4, 122.3, 122.1 , 117.4, 113.6, 111.1 , 58.8, 55.6, 42.2, 33.0.

[0303] HRMS (ESI-TOF) m / z\ [M+H]+Calcd for C20H20N2O3H 337.1552; Found 337.1552.Step 3. (E)-3-(2-(3-(3-methoxyphenyl)acryloyl)-1H-benzo[d]imidazol-1-yl)propanal

[0304] To a solution of (E)-1-(1-(3-hydroxypropyl)-1H-benzo[d]imidazol-2-yl)-3-(3- methoxyphenyl)prop-2-en-1-one (27 mg, 0.080 mmol) in 0.4 mL of DMSO under an inert atmosphere of argon, IBX (67.5 mg, 0.240 mmol) was added. The solution was stirred at room temperature for 1 hour. Et20 was added to the mixture and the resulting solid was filtered through celite. This resulted in 8 mg of (E)-3-(2-(3-(3-methoxyphenyl)acryloyl)-1 / - / - benzo[d]imidazol-1 -yl)propanal as a yellow solid.Step 4. (E)-3-(3-methoxyphenyl)-1-(1-(3-morpholinopropyl)-1H-benzo[d]imidazol-2-yl)prop-2-en-1 -one

[0305] To a solution of (E)-3-(2-(3-(3-methoxyphenyl)acryloyl)-1 H-benzo[c / ]imidazol- 1-yl)propanal (8 mg, 0.024 mmol) in 0.05 mL of DCM under an inert atmosphere of argon, NaBH(OAc)3 (9.7 mg, 0.046 mmol) and morpholine (3.8 pL, 0.043 mmol) were added. The mixture was stirred at room temperature overnight. The mixture was quenched withNaHCOs, extracted with EA, dried over MgSCh, and concentrated under vacuum. This resulted in 3 mg of (E)-3-(3-methoxyphenyl)-1 -(1-(3-morpholinopropyl)-1 / - / - benzo[d]imidazol-2-yl)prop-2-en-1 -one as a yellow solid.

[0306] 1H NMR: (600 MHz, CDCI3) δ 8.26 (d, J = 16.1 Hz, 1 H), 7.93 (d, J = 8.2 Hz, 1 H), 7.87 (d, J = 15.9 Hz, 1 H), 7.57 (d, J = 8.3 Hz, 1 H), 7.44 (t, J = 7.2 Hz, 1 H), 7.39 (t, J = 7.73 Hz, 1 H), 7.35 - 7.34 (m, 2H), 7.27 - 7.26 (m, 1 H), 7.00 - 6.98 (m, 1 H), 4.77 (t, J = 7.0 Hz, 2H), 3.88 (s, 3H), 3.71 (app. s, 4H), 2.39 (t, J = 6.7 Hz, 6H), 2.09 (p, J = 6.8 Hz, 2H).

[0307] 13C{1H} NMR: (150 MHz, CDCI3) δ 182.8, 160.0, 147.1 , 144.7, 141.9, 136.9,136.1 , 130.0, 126.0, 123.9, 123.2, 122.2, 122.0, 117.2, 113.3, 111.1 , 67.0, 55.6 (2), 53.6, 43.7, 27.0.

[0308] HRMS (ESI-TOF) n? / z: [M+H]+Calcd for C24H27N3O3H 406.2131 ; Found 406.2122.(E)-N-(3-(3-(1 -(3-morpholinopropyl)-1H-benzo[c / ]imidazol-2-yl)-3-oxoprop-1 -en-1 - yl)phenyl)acrylamide (31)

[0309] Using fe / t-butyl (E)-(3-(3-(1H-benzo[d]imidazol-2-yl)-3-oxoprop-1-en-1- yl)phenyl)carbamate (S3) instead of (E)-1 -(1 H-benzo[d]imidazol-2-yl)-3-(3- methoxyphenyl)prop-2-en-1-one (60) steps 1 -4 of Route C were performed followed by steps 3-4 of route B. Yellow solid.

[0310] 1H NMR: (500 MHz, CDCI3) δ 8.25 (d, J = 15.9 Hz, 1 H), 7.93 (d, J = 7.8 Hz, 1 H), 7.87 - 7.80 (m, 3H), 7.57 (d, J = 8.3 Hz, 1 H), 7.49 - 7.37 (m, 4H), 6.48 (d, J = 16.2Hz, 1 H), 6.34 (dd, J = 16.2, 10.2 Hz, 1 H), 5.81 (d, J = 10.2 Hz, 1 H), 4.77 (t, J = 7.12 Hz, 2H), 3.71 (t, J = 4.3 Hz, 4H), 2.41 (t, J = 5.8 Hz, 6H), 2.10 (m, 2H).Route D. 3-(3-methoxyphenyl)-1 -(1 -methyl-1 H-benzo[cf]imidazol-2-yl)prop-2-yn-1 - one (9)Step 1. / V-methoxy- / V,1 -dimethyl-1 H-benzo[d]imidazole-2-carboxamide

[0311] To 1-methyl-2-(trichloromethyl)-1 / - / -benzo[d]imidazole (synthetically accessible, 100 mg, 0.43 mmol, 1 equiv.) dissolved in 5.5 ml_ of 3:1 MeCN:water was added A / ,O-dimethylhydroxylamine hydrochloride (83 mg, 0.85 mmol, 2 equiv.) and then K2CO3 (188 mg, 1.36 mmol, 3.2 equiv.). The reaction was allowed to stir at rt for 6 h, after which the reaction was diluted with sat. aq. NH4CI and then EtOAc. The organic layer was washed with water and then brine, dried with MgSCh, concentrated in vacuo, and then subjected at chromatography (0-60% EtOAc in hexanes) to give 78 mg (89% yield) of the title compound as a white crystalline solid.Step 2. 3-(3-methoxyphenyl)-1 -(1 -methyl-1 H-benzo[d]imidazol-2-yl)prop-2-yn-1 - one (9)

[0312] To 1-ethynyl-3-methoxybenzene (200 pL, 1 .0 equiv. dissolved in 15 mL of THF under argon at -78 °C was added n-BuLi (2.5 M in hexanes, 690 pL, 1.73 mmol, 1.1 equiv.). The reaction was warmed to 0 °C and stirred at that temperature for 30 minutes. The reaction was then cooled back to -78 °C and / V-methoxy-A / , 1 -dimethyl-1 H- benzo[d]imidazole-2-carboxamide (414 mg, 1.89 mmol, 1.2 equiv.) was added in a minimal amount of THF dropwise. The reaction was warmed to rt, stirred for 30 minutes at that temperature, and then quenched with sat. aq. NH4CI. The aqueous layer was extracted with EtOAc three times, dried with Na2SO4, and concentrated under vacuum.The crude material was subjected to chromatography (0-40% EtOAc in hexanes) to give 215 mg (47% yield) of the title compound as a yellow oil.

[0313] 1H NMR: (500 MHz, CDCI3) δ 7.99 (d, J = 7.7 Hz, 1 H), 7.50 - 7.45 (m, 2H), 7.42 - 7.38 (m, 2H), 7.33 (d, J = 8.2 Hz, 1 H), 7.31 - 7.28 (m, 1 H), 7.04 (dd, J = 8.3, 2.5, 1 H), 4.19 (s, 3H), 3.85 (s, 3H).

[0314] 13C{1H} NMR: (125 MHz, CDCI3) δ 170.4, 159.4, 146.6, 142.4, 137.1 , 129.7,126.7, 126.2, 124.1 , 122.6, 121.0, 118.2, 117.7, 110.6, 94.6, 88.1 , 55.5, 32.3.

[0315] HRMS (ESI-TOF) m / z: [M+H]+Calcd for C18H14N2O2H 291.1133; Found 291.1134.fert-butyl (3-(3-(1-methyl-1H-benzo[c / ]imidazol-2-yl)-3-oxoprop-1-yn-1- yl)phenyl)carbamate (38)

[0316] The title compound was made in a manner analogous to 3-(3-methoxyphenyl)- 1-(1 -methyl-1 / - / -benzo[d]imidazol-2-yl)prop-2-yn-1 -one (9) except using 1 ,1 - dimethylethyl A / -(3-ethynylphenyl)carbamate (commercially available or synthetically accessible) instead of 1 -ethynyl-3-methoxybenzene and 2.2 equivalents of n-BuLi.

[0317] 1H NMR: (500 MHz, CDCI3) δ 8.00 (d, J = 8.3 Hz, 1 H), 7.74 (s, 1 H), 7.56 (d, J = 8.0 Hz, 1 H), 7.49 - 7.44 (m, 3H), 7.40 (ddd, J = 8.3, 6.0, 2.0 Hz, 1 H), 7.24 (t, J = 8.1 Hz, 1 H), 6.53 (s, 1 H), 4.20 (s, 3H), 1.53 (s, 9H).

[0318] HRMS (ESI-TOF) m / z\ [M+Na]+Calcd for C22H2i N3O3Na 398.1481 ; Found 398.1484.Intermediate S4. / V-(3-ethynylphenyl)acrylamide

[0319] The title compound was made in a manner analogous to Step 4 of Route B, except using 3-ethynylaniline instead of (E)-3-(3-aminophenyl)-1-(1-methyl-1H- benzo[d]imidazol-2-yl)prop-2-en-1 -one (10).

[0320] 1H NMR: (500 MHz, CDCI3) δ 7.70 (bs, 1 H), 7.63 (d, J = 7.0 Hz, 1 H), 7.51 (s, 1 H), 7.28 (t, J = 7.7 Hz, 1 H), 7.24 (d, J = 7.6 Hz, 1 H), 6.43 (d, J = 16.9 Hz, 1 H), 6.26 (dd, J = 16.9, 10.2 Hz, 1 H), 5.77 (d, J = 10.2 Hz, 1 H), 3.06 (s, 1 H). / V-(3-(3-(1 -methyl -1 H-benzo[d]imidazol-2-yl)-3-oxoprop-1 -yn-1 - yl)phenyl)acrylamide (56)

[0321] The title compound was made in a manner analogous to 3-(3-methoxyphenyl)-1-(1-methyl-1H-benzo[d]imidazol-2-yl)prop-2-yn-1-one (9) except using N-(3- ethynylphenyl)acrylamide (S4) instead of 1-ethynyl-3-methoxybenzene and 2.2 equivalents of n-BuLi.

[0322] 1H NMR: (500 MHz, DMSO-d6) δ 10.41 (s, 1 H), 8.17 (s, 1 H), 7.93 (d, J = 8.3 Hz, 1 H), 7.80 (d, J = 7.9 Hz, 1 H), 7.77 (d, J = 8.0 Hz, 1 H), 7.53 - 7.46 (m, 3H), 7.41 (t, J = 7.6 Hz, 1 H), 6.45 (dd, J = 17.0, 10.2 Hz, 1 H), 6.31 (d, J = 17.1 Hz, 1 H), 5.82 (d, J = 10.1Hz, 1 H), 4.13 (s, 3H).

[0323] 13C{1H} NMR: (150 MHz, DMSO-d6) δ 170.1 , 164.0, 146.9, 142.1 , 140.1 , 137.5,132.0, 130.4, 128.4, 128.1 , 126.9, 124.4, 123.4, 122.7, 122.1 , 119.9, 112.3, 93.0, 88.5,32.6.

[0324] HRMS (ESI-TOF) m / z\ [M+H]+Calcd C20H15N3O2H 330.1242; Found 330.1245.(Z)-3-methoxy-3-(3-methoxyphenyl)-1-(1 -methyl-1 H-benzo[c / |imidazol-2-yl)prop-2- en-1-one (55)

[0325] 3-(3-methoxyphenyl)-1-(1 -methyl-1 H-benzo[c(]imidazol-2-yl)prop-2-yn-1 -one(9) dissolved in 280 pL of 0.25 M NaOH in MeOH (1 equiv.) was stirred at rt for 3 h. The reaction was then diluted with water and extracted with EtOAc, dried over MgSO4, concentrated under vacuum, and subjected to chromatography (75:25 hexanes / EtOAc) to give 8 mg (36% yield) of the title compound as a yellow solid.

[0326] 1H NMR: (500 MHz, CDCI3) δ 7.89 (d, J = 8.0 Hz, 1 H), 7.42 - 7.41 (m, 2H), 7.38 - 7.31 (m, 2H), 7.12 - 7.11 (m, 2H), 7.05 (s, 1 H), 6.99 (dd, J = 8.3, 1.6 Hz, 1 H), 4.08 (s, 3H), 4.05 (s, 3H), 3.82 (s, 3H).

[0327] LCMS: m / z = 323 [M+H]+5-(3-methoxyphenyl)-3-(1 -methyl-1 W-benzo[d]imidazol-2-yl)isoxazole 18)

[0328] To (Z)-1 -methyl-1 / - / -benzo[d]imidazole-2-carbaldehyde oxime (50 mg, 0.29 mmol, 1 equiv.) in 1.4 mL DMF was added / V-chlorosuccinimide (42 mg, 0.31 mmol, 1.1 equiv.) at 40 °C under argon. After 3 h at 40 °C, 1-ethynyl-3-methoxybenzene (38 pL, 0.30 mmol, 1.05 equiv.) was added and the reaction was heated at 40 °C overnight. The reaction was diluted with 2 mL of ice water and then extracted with Et20 three times. The combined organic layers were dried with Na2SO4, concentrated under vacuum, and subjected to chromatography (80:20 hexanes: EtOAc) to give the title compound as a white solid (25 mg, 28% yield).

[0329] 1H NMR: (500 MHz, CDCI3) δ 7.85 (d, J = 7.9 Hz, 1 H), 7.46 - 7.31 (m, 7H), 7.00 (d, J = 7.9 Hz, 1 H), 4.22 (s, 3H), 3.87 (s, 3H).

[0330] 13C{1H} NMR: (125 MHz, CDCI3) δ 170.1 , 160.2, 157.2, 143.2, 143.0, 136.8,130.4, 128.1 , 124.3, 123.1 , 120.5, 118.5, 116.8, 111.0, 110.1 , 100.8, 55.5, 32.3.

[0331] HRMS (ESI-TOF) m / z: [M+H]+Calcd for C18H15N3O2H 306.1242; Found 306.1230.3-(1 -methyl-1 H-benzo[cf]imidazol-2-yl)-5-phenylisoxazole (17)

[0332] The title compound was made in a manner analogous to 5-(3-methoxyphenyl)- 3-(1 -methyl-1 / - / -benzo[c / ]imidazol-2-yl)isoxazole (18), except using ethynylbenzene instead of 1 -ethynyl-3-methoxybenzene. White solid.

[0333] 1H NMR: (500 MHz, CDCI3) δ 7.88 - 7.86 (m, 3H), 7.54 - 7.47 (m, 4H), 7.41 (td, J = 7.0, 1.1 Hz, 1 H), 7.37 - 7.34 (m, 2H), 4.26 (s, 3H).

[0334] 13C{1H} NMR: (150 MHz, CDCI3) δ 170.2, 157.2, 143.2, 142.9, 136.8, 130.8,129.3, 127.0, 126.1 , 124.3, 123.1 , 120.5, 110.2, 100.6, 32.5.

[0335] HRMS (ESI-TOF) m / z: [M+H]+Calcd for C17H13N3OH 276.1137; Found 276.1128.fert-butyl (3-(3-(1 -methyl-1 H-benzo[c / ]imidazol-2-yl)isoxazol-5-yl)phenyl)carbamate (32)

[0336] The title compound was made in a manner analogous to 5-(3-methoxyphenyl)- 3-(1 -methyl-1 / - / -benzo[c / ]imidazol-2-yl)isoxazole (18), except using 1 ,1 -dimethylethyl N-(3-ethynylphenyl)carbamate (commercially available or synthetically accessible) instead of 1-ethynyl-3-methoxybenzene. White solid.

[0337] 1H NMR: (600 MHz, CDCI3) δ 7.94 (s, 1 H), 7.89 (d, J = 8.0 Hz, 1 H), 7.53 (d, J = 7.6 Hz, 1 H), 7.49 - 7.47 (m, 2H), 7.44 - 7.41 (m, 3H), 7.37 (t, J = 7.5 Hz, 1 H), 6.65 (s, 1 H), 4.26 (s, 3H), 1.55 (s, 9H). / V-(3-(3-(1 -methyl -1 H-benzo[d]imidazol-2-yl)isoxazol-5-yl)phenyl)acrylamide (33)

[0338] The title compound was made in a manner analogous to (E)- / V-(3-(3-(1-methyl- 1 / - / -benzo[d]imidazol-2-yl)-3-oxoprop-1 -en-1-yl)phenyl)acrylamide (11) using steps 3-4 of route B, except that terf-butyl (3-(3-(1-methyl-1 H-benzo[d]imidazol-2-yl)isoxazol-5- yl)phenyl)carbamate (32) was used instead of tert-butyl (E)-(3-(3-(1 -methyl-1 H- benzo[d]imidazol-2-yl)-3-oxoprop-1 -en-1 -yl)phenyl)carbamate (19).

[0339] 1H NMR: (500 MHz, DMSO-d6) δ 10.43 (s, 1 H), 8.36 (s, 1 H), 7.81 - 7.73 (m, 4H), 7.67 (s, 1 H), 7.55 (t, J = 8.0 Hz, 1 H), 7.41 (ddd, J = 8.2, 7.2, 1 .0 Hz, 1 H), 7.33 (ddd, J = 8.1 , 7.4, 1.0 Hz, 1 H), 6.50 (dd, J = 17.1 , 10.1 Hz, 1 H), 6.32 (dd, J = 17.0, 1.7 Hz, 1 H), 5.82 (dd, J = 10.1 , 1.8 Hz, 1 H), 4.19 (s, 3H).

[0340] 13C{1H} NMR: (150 MHz, DMSO-d6) δ 169.3, 163.5, 157.0, 142.4 (2), 139.9,136.6, 131.6, 130.0, 127.5, 126.7, 123.9, 122.7, 121.6, 121.4, 119.7, 116.0, 111.1 , 101.3, 32.1.

[0341] HRMS (ESI-TOF) m / z\ [M+Na]+Calcd for C2oHi6N402Na 367.1171 ; Found 367.1188.2-(4-(3-methoxyphenyl)-1H-1, 2, 3-triazol-1-yl)-1 -methyl-1 H-benzo[d]imidazole (39)

[0342] 2 -azido-1 -methyl-1 H-benzo[c / ]imidazole (synthetically accessible, 20 mg, 0.12 mmol, 1 equiv.), Cui (22 mg, 0.12 mmol, 1 equiv.), and sodium L-ascorbate (33 mg, 0.12 mmol, 1 equiv.) were added to a flask. The flask was backfilled with argon and 1 .2 mL of degassed 1 :2 t-BuOH:H2O and 1-ethynyl-3-methoxybenzene (30 pL, .23 mmol, 2 equiv.) were added. The reaction was allowed to stir overnight at rt. EtOAc and water were added, and the organic layer was washed with water three times. The organic layer was dried with MgSO4, concentrated under vacuum, and then subjected to chromatography (75:25 hexanes: EtOAc) to give 11 mg (32% yield) of the title compound as a white solid.

[0343] 1H NMR: (500 MHz, CDCI3) δ 7.80 (d, J = 7.6 Hz, 1 H), 7.54 (s, 1 H), 7.48 (t, J = 6.6 Hz, 2H), 7.44 - 7.35 (m, 3H), 6.95 (dd, J = 8.1 , 2.2 Hz, 1 H), 4.19 (s, 3H), 3.89 (s, 3H).

[0344] 13C{1H} NMR: (150 MHz, CDCI3) δ 160.3, 147.7, 140.6, 142.9 (b), 136.0 (b),130.8, 130.3, 124.2, 123.7, 120.7, 120.2, 118.5, 115.2, 111.1 , 110.3, 55.5, 32.6. Peaks indicated as broad (b) were confirmed using HMBC.

[0345] HRMS (ESI-TOF) m / z: [M+Na]+Calcd for CizHisNsONa 328.1174, found 328.1175.1-methyl-2-(4-phenyl-1H-1,2,3-triazol-1-yl)-1H-benzo[d]imidazole (34)

[0346] The title compound was made in a manner analogous to 2-(4-(3- methoxyphenyl)-1 / - / -1 ,2,3-triazol-1 -yl)-1 -methyl-1 / - / -benzo[c / ]imidazole (39) except using ethynylbenzene instead of 1 -ethynyl-3-methoxybenzene. White solid.

[0347] 1H NMR: (500 MHz, CDCI3) δ 8.72 (s, 1 H), 7.94 (d, J = 7.8 Hz, 2H), 7.79 (d, J = 7.7 Hz, 1 H), 7.50 - 7.36 (m, 6H), 4.19 (s, 3H).tert-butyl (3-(1 -(1 -methyl-1 H-benzo[d]imidazol-2-yl)-1 H-1 ,2,3-triazol-4- yl)phenyl)carbamate (40)

[0348] The title compound was made in a manner analogous to 2-(4-(3- methoxyphenyl)-1H-1 ,2,3-triazol-1-yl)-1 -methyl-1 H-benzo[d]imidazole (39) except using terf-buty I (3-ethynylphenyl)carbamate instead of 1 -ethynyl-3-methoxybenzene.

[0349] 1H NMR: (600 MHz, CDCI3) δ 8.74 (s, 1 H), 7.97 (s, 1 H), 7.80 (d, J = 7.4 Hz, 1 H), 7.61 (d, J = 7.3 Hz, 1 H), 7.50 (d, J = 7.8 Hz, 1 H), 7.42 - 7.36 (m, 4H), 6.69 (s, 1 H), 4.17 (s, 3H), 1.54 (s, 9H).

[0350] 13C{1H} NMR: (150 MHz, CDCI3) δ 152.8, 147.5, 142.9 (b), 140.4, 139.3, 135.9(b), 130.2, 129.9, 124.2, 123.7, 120.9, 120.7, 120.2, 119.0, 116.1 , 110.2, 80.9, 32.5, 28.5. Peaks indicated as broad (b) were confirmed using HMBC.

[0351] HRMS (ESI-TOF) m / z: [M+Na]+Calcd for C2i H22N6O2Na 413.1702; Found 413.1702.2-(6-(3-methoxy phenyl )pyri di n -2-yl )-1 -methyl-1 H-benzo[d]imidazole (45)Step 1. 6-(3-methoxyphenyl)picolinaldehyde

[0352] Using degassed solvents under an atmosphere of argon, 6- bromopicolinaldehyde (99 mg, 0.55 mmol, 1.0 equiv.), (3-methoxyphenyl)boronic acid (100 mg, 0.66 mmol, 1 .2 equiv.), Pd(PPhi3)4 (32 mg, 0.028 mmol, 0.05 equiv.), and K2CO3 (228 mg, 1.65 mmol, 3.0 equiv.) were stirred in 4 mL of 1 :1 DME:water at 100 °C overnight. The reaction was then diluted with water, extracted with EtOAc, dried overMgS04, concentrated under vacuum, and subjected to chromatography (95:5 hexanes: EtOAc) to give 63 mg (45% yield) of the title compound as a white solid.Step 2. 2-(6-(3-methoxyphenyl)pyridin-2-yl)-1-methyl-1H-benzo[d]imidazole (45)

[0353] 6-(3-methoxyphenyl)picolinaldehyde (50 mg, 0.23 mmol, 1.0 equiv.) and A / -1 - methylbenzene-1 ,2-diamine (29 mg, 0.23 mmol, 1.0 equiv.) were heated in 0.5 mL DMF in a sealed vial at 120 °C overnight. The reaction was then diluted with water, extracted with EtOAc, dried over MgSO4, concentrated under vacuum, and subjected to chromatography (0-30% EtOAc in hexanes) to give 49 mg (66% yield) of the title compound as a yellow oil.

[0354] 1H NMR: (500 MHz, CDCI3) δ 8.36 (d, J = 7.8 Hz, 1 H), 7.91 (t, J = 7.9 Hz, 1 H), 7.87 (d, J = 7.5 Hz, 1 H), 7.79 (d, J = 7.9 Hz, 1 H), 7.68 (app. t, J = 1 .7 Hz, 1 H), 7.64 (d, J = 7.8 Hz, 1 H), 7.46 (d, J = 7.7 Hz, 1 H), 7.42 (t, J = 8.1 Hz, 1 H), 7.38 - 7.31 (m, 2H), 7.01 (dd, J = 8.1 , 2.3 Hz, 1 H), 4.38 (s, 3H), 3.90 (s, 3H).

[0355] 13C{1H} NMR: (150 MHz, CDCI3) δ 160.2, 156.0, 150.2, 149.7, 141.8, 140.3,137.9, 137.0, 129.9, 123.7, 123.5, 123.0, 120.6, 119.8, 119.3, 114.9, 112.6, 110.0, 55.4, 33.1.Example 2: Viability Assay of compound treatment in ovarian cancer and pancreatic cancer cells with p53 mutations

[0356] Provided in Fig. 1 is a viability assay of compound treatment in ovarian cancer cells with p53 mutations. Fig. 1 shows the effect of compounds 60 and 11 on p53R175Hcancer mutant. The TOV112D-p53R175Hovarian cancer (TOV) cells are used in the viability assay. TOV cells were cultured in the presence of different concentrations of 60 and 11 for about 72 hours. DMSO was used as the control (vehicle). Numbers of alive cells were measured using the CellTiter-Glo reagent. Data are presented as standard deviation (SD) of mean (n=3).

[0357] Table 8, below, provides the results of the viability assay in TOV112D-p53R175Hcells. The letter codes for the I C50 of the viability assay in TOV112D-p53R175Hcells include: A (<1 .5 μM), B (≥1 .5-6.2 μM), C (>6.2 μM).Table 8: Viability assay in TOV112D-p53R175H

[0358] Table 9, below, provides the results of the viability assay in BxPC3-p53Y220Ccells. The letter codes for the IC50 of the viability assay in BxPC3-p53Y220Ccells include: A (<1 .5 μM), B (>1 .5-6.2 μM), C (>6.2 μM).Table 9: Viability assay in BxPC3-p53Y220CExample 3: Cellular Thermal Shift Assay (CETSA) of compound treatment in ovarian cancer cells with p53 mutations.

[0359] Provided in Fig. 2 is a cellular thermal shift assay (CETSA) of compound treatment in ovarian cancer cells with a p53R175Hmutation. Fig. 2 shows the effect of compounds 60 and 11 on p53R175Hcancer mutant within TOV112D-p53R175Hovarian cancer cells. The CETSA was performed with the TOV cells treated with 30 μM 60, 30 μM 11, or DMSO (vehicle) respectively for 3 hours. Following treatment, cells can be aliquoted, heated at indicated temperatures, lysed and analyzed by Western blot. Data are represented as SD of mean (n = 2). Compound 11 stabilized the p53 mutant as temperatures increased better than DMSO, suggesting the compound interacts with the protein to prevent unfolding.Example 4: Chromatin binding assay in ovarian cancer cells with p53 mutations

[0360] Provided in Fig. 3 is a chromatin binding assay in ovarian cancer cells with a p53Ri 75Hmutation. Fig. 3 shows the effect of compounds 60 and 11 on promoter binding of the p53R175Hmutant within TOV112D-p53R175Hovarian cancer cells. The TOV cells were treated with 30 μM 60, 30 μM 11, or DMSO (vehicle) respectively for 3 hours and fractionated into cytosolic and nuclear soluble (C) as well as chromatin bound (N) fractions. GAPDH can be used as a cytosolic protein control and Histone 3 can be used as a chromatin bound protein control. 11 compounds can restore DNA binding of p53R175Hin vivo.Example 5: Chromatin Immunoprecipitation (ChIP) analysis of compound treatment in ovarian cancer cells with p53 mutations

[0361] Provided in Fig. 4 is a chromatin immunoprecipitation (ChIP) analysis of compound treatment in ovarian cancer cells with a p53R175Hmutation. Fig. 4 shows the effect of 60 and 11 on the p53R175Hmutant on association with the puma promoter within TOV112D-p53R175Hovarian cancer cells. Cells were treated with 30 μM 60, 30 μM 11, or DMSO (vehicle) respectively for 30 minutes, then cross-liked, quenched and harvested. ChIP was performed using a p53 specific antibody and eluted DNA was examined by quantitative PCR using primers that target the p53 binding site in the promoter region of puma. Data are represented as SD (n = 3). ChIP analysis shows induction of site-specific promoter binding of p53R175Hby 11 compounds in TOV cells.Example 6: RNA induction analysis of compounds in ovarian cancer cells with p53 mutations

[0362] Provided in Fig. 5 is RNA induction analysis of compounds in ovarian cancer cells with a p53R175Hmutation. Fig. 5 shows the effect of compounds 60 and 11 on the p53R175Hmutant on target gene expression within TOV112D-p53R175Hovarian cancer cells. Quantitative RT-PCR of p53 target genes NOXA and p21 is shown. TOV cells were treated with 30 μM 60, 30 μM 11, or DMSO (vehicle) for 3 hours. Data are represented as SD of mean (n = 3). Treatment with compound 11 robustly induced p53 target gene expression as compared to DMSO.Example 7: Viability assay in various cancer cells with p53 mutations

[0363] Provided in Fig. 6A is viability assay of compounds in various cancer cells each comprising a p53 mutation. Fig. 6A shows the effect of compounds 11 on the viability of the p53 cancer mutant cell lines. Cells such as TOV112D-p53R175Hovarian cancer cells, BXPC3-p53Y220Ccancer cells, SK-Mel-p53G245Scancer cells, MB468-p53R273Hcancer cells, MB231-p53R280Qcancer cells, and HCT116 wild type p53, were cultured in the presence of different concentrations of 11 for 72 hours. DMSO is used as the control (vehicle). Relative number of viable cells were measured using the CellTiter-Glo reagent.Data are presented as SD of mean (n=3). The 11 compounds reduced viable cell proliferation in each of the cancer cell lines. Fig. 6B shows the I Cso values of the cell lines. The letter codes for the I Cso include: A (<1 .5 μM), B (≥1 .5-6.2 μM), C (>6.2 μM).Example 8: p53 nuclear enrichment of various cancer cells

[0364] Provided in Fig. 7 shows compound 11 enhances chromatin association of various p53 mutants. Cells were treated with compounds (15 μM) or vehicle (DMSO) for 3 hours and were fractionated into a cytosolic soluble (C) fraction and a chromatin bound (N) fraction. Enrichment of p53 within the chromatin bound fractions was quantified with densitometric analysis and normalized to H3. Data are presented as SD of mean (n=2).

[0365] Several compounds have been tested for nuclear enrichment of p53R175Hwithin TOV112D cells. Table 10, below, provides the results of the nuclear enrichment assay in TOV112D-p53R175Hovarian cancer cells. Compounds that enriched the amount of mutant p53 in the nuclear fraction as compared to vehicle (DMSO) are marked “Enriched.” “No” indicates there was no difference between the amount of mutant p53 in the nuclear fraction as compared to vehicle (DMSO).Table 10: Nuclear Enrichment assay in TOV112D-p53R175HExample 9: Immunoblotting assay of compound 11 on p21 protein.

[0366] Provided in Fig. 8 is an immunoblot that shows the effect of compound 11 on p21 protein levels, which is an expressed from the CDKN1A gene targeted by p53. TOV112D-p53R175Hovarian cancer cells, BXPC3-p53Y220Ccancer cells, SK-Mel-p53G245Scancer cells, MB468-p53R273Hcancer cells, MB231-p53R280Qcancer cells, and HCT116 wild type p53, were cultured in the presence of 15μM of compound 11 for 3 hours. DMSO is used as the control (vehicle). Cell lysates were analyzed by immunoblotting after separation of proteins by SDS-PAGE. Compound 11 increased the amount of p21 protein present.Example 10: Covalent attachment assay of compounds on p53 protein

[0367] Provided in Figure 9 are results of an assay to assess covalent attachment of compounds to p53 proteins. A method for identifying and distinguishing covalent from non-covalent interactions between a test compound and recombinant p53 protein was developed using click chemistry-based pull-down technology.

[0368] Compounds were immobilized to beads via azide click chemistry and then blocked with bovine serum albumin. Recombinant biotinylated p53 mutants p53G245Sand p53R175Hwere incubated with beads for a period of X hours, and then washed to remove excess protein. The S. cerevisiae Met 30 protein was utilized as a negative control. The beads were then mixed with streptavidin-HRP, washed to remove excess HRP, and plated within a well of a 96-well plate. Detection of HRP by chemiluminescence indicates the relative level of conjugation between the compound and the p53 mutant. The results show that compounds 11 , 23, and 24 covalently attached to the p53 mutants.Example 10: Mass spectrometry-based covalent attachment assay

[0369] Provided in Fig. 10 are sequences of mutant p53 peptides that were identified to covalently bind with compound 11 , as determined by mass spectrometry. Recombinant 6x-His-tagged p53G245Swere immobilized onto Ni-NTA beads. The beads were mixed within a solution containing compound 11 (50 μM) or DMSO (control) and incubated for 30 minutes, then washed to removed excess compound. The beads are then reduced, alkylated, and digested with trypsin. Peptides within the trypsin product were purified through a C18 tip to remove contaminants and then analyzed via LC-MS. Peptides with non-specific modification corresponding to compound 11 were identified (Fig. 10). The peptides that showed a mass shift within the sample treated with compound 11 were p53 sequences: residues 121-132, 139-156, and 181 -196, indicating that compound 11 covalently bound recombinant p53 mutant at these peptides.

Claims

CLAIMSWHAT IS CLAIMED IS:

1. A compound of Formula I:or a therapeutically acceptable salt thereof; wherein R1is H, alkyl, benzyl, aryl, heteroaryl, heterocycle, alkyl, or alkyl chain and heteroatom;R2is H or alkyl;R3is H or halogen; andR4is H or CH3.

2. A compound of Formula II:or a therapeutically acceptable salt thereof; wherein R1is H, alkyl, benzyl, aryl, heteroaryl, or alkyl chain and heteroatoms;R2is H, CH3, or alkyl;R3is H or halogen; andR4is H or CH3.

3. A compound of Formula III:or a therapeutically acceptable salt thereof; wherein R1is H, alkyl, benzyl, aryl, heteroaryl, or alkyl chain and heteroatoms;R2is H or alkyl;R3is H or halogen; andR4is H or CH3.

4. A compound of Formula IV:or a therapeutically acceptable salt thereof; wherein R1is H, alkyl, benzyl, aryl, heteroaryl, or alkyl chain and heteroatoms;R2is H or alkyl;R3is H or halogen; andR4is H or CH3.

5. A compound of Formula V:or a therapeutically acceptable salt thereof; wherein R1is H, alkyl, benzyl, aryl, heteroaryl, or alkyl chain and heteroatoms;R2is H or alkyl;R3is H or halogen;R4is H or CH3;X is CH or N; andY is S or 0.

6. A compound of Formula Vl-a or Vl-b:or a therapeutically acceptable salt thereof; wherein R1is H, alkyl, benzyl, aryl, heteroaryl, or alkyl chain and heteroatoms;R2is H or alkyl;R3is H or halogen;R4is H or CH3;X is CH or N; andX1, X2, X3, and X4are each independently C or N.

7. A compound of Formula VII:VII or a therapeutically acceptable salt thereof; wherein R2is H or alkyl;R3is H or halogen;R4is H or CH3; andX is 0 or S.

8. A compound of Formula VIII:or a therapeutically acceptable salt thereof; wherein R1and R2are H or halogen;R3is H, alkyl, aryl, benzyl, or acyl; and R4is OMe.

9. A compound of Formula IX:or a therapeutically acceptable salt thereof; wherein R1is H, Boc, benzoyl, p-trifuoromethylbenzyl, p-methoxybenzyl, pentyl, or 4- pentynyl; and R2is OMe.

10. A compound of Formula X:or a therapeutically acceptable salt thereof; wherein R1is H or alkyl; and R2is OMe.

11. A compound of Formula XI:or a therapeutically acceptable salt thereof; wherein R1is H or alkyl; andR2is amine, alkyl amine, or dialkyl amine.

12. A compound of Formula XII:or a therapeutically acceptable salt thereof; wherein R1is H or alkyl; andR2is acylamide, methacrylamide or NHC0NMe2.-U S-13. A compound of Formula XIII:or a therapeutically acceptable salt thereof; wherein R1is H or alkyl;R2is Boc; andR3is H or alkyl.

14. A compound of Formula XIV:or a therapeutically acceptable salt thereof; wherein R1is alkyl or 4-pentynyl; and R2is acylamide or N-methylacrylamide.

15. A compound of Formula XV:or a therapeutically acceptable salt thereof; wherein R1is H or alkyl; andR2is H, acylamide, or OMe.

16. A compound of Formula XVI:XVI or a therapeutically acceptable salt thereof; wherein R1is H, alkyl, benzyl, aryl, heteroaryl, heterocycle, or alkyl chain with heteroatom;R2is H, or alkyl;R3is H, or halogen;R4is H, or CH3; andX1, X2, X3, and X4are each independently C or N.

17. A compound selected from any one of the compounds depicted in Tables 1 -6 or therapeutically acceptable salts thereof.

18. A pharmaceutical composition comprising a compound of any one of claims 1 -17, and a pharmaceutically acceptable carrier, adjuvant, or vehicle.

19. A method of treating cancer in a patient comprising administering to said patient a compound according to any of one claims 1 -17, or the pharmaceutical composition of claim 18, wherein the cancer is associated with p53 mutagenesis.

20. A method of claim 19, wherein the compound comprises a functional group that binds with and stabilizes a p53 mutant such that the stabilization of the p53 mutant reactivates gene expression of p53 wild type.21 . A method of claim 19 or 20 further comprising: prior to administering the compound, assessing a sample derived from the patient for a presence of a p53 mutant within the cancer.

22. A method of any one of claims 19-21 , wherein the p53 mutant comprises a mutation within one or more residue positions selected from the group consisting of: R175, G245, R248, R249, R273, R282, R280, Y220, A138, P151 , P152, G154, R158, V172, H193, H214, S215, Y234, P250, I255, V272, V274, E285, and E286.

23. A method of any one of claims 19-22, wherein the cancer is ovarian cancer, breast cancer, esophageal cancer, colorectal cancer, head and neck cancer, larynx cancers, and lung cancer.

24. A method of claim 23, wherein the cancer is a human ovarian cancer or a human breast cancer.

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