Histone acetyltransferase modulators and compositions and uses thereof

Abstract

Compounds and compositions comprising compounds that modulate histone acyl transferase (HAT). Methods for treating neurodegenerative disorders, conditions associated with accumulated amyloid-beta peptide deposits, Tau protein levels, and / or accumulations of alpha-synuclein as well as cancer by administering a compound that modulates HAT to a subject.

Description

Attorney Docket No.: 0019240.01318WO1 Date of Electronic Filing: July 29, 2024 HISTONE ACETYLTRANSFERASE MODULATORS AND COMPOSITIONS AND USES THEREOF CROSS REFERENCE TO RELATED APPLICATION

[0001] The application claims the benefit of and priority to U.S. Provisional Application No.63 / 516,069, filed on July 27, 2023, the content of which is hereby incorporated by reference its entirety. BACKGROUND

[0002] All patents, patent applications and publications cited herein are hereby incorporated by reference in their entirety. The disclosures of these publications in their entireties are hereby incorporated by reference into this application in order to more fully describe the state of the art as known to those skilled therein as of the date of the invention described and claimed herein.

[0003] This patent disclosure contains material that is subject to copyright protection. The copyright owner has no objection to the facsimile reproduction by anyone of the patent document or the patent disclosure as it appears in the U.S. Patent and Trademark Office patent file or records, but otherwise reserves any and all copyright rights.

[0004] Modulation of the acetylation state of histones, transcription factors, and other regulatory proteins is known to influence their activity within neuronal, cancer and inflammatory cells. The acetylation state of a protein is controlled by the activity of two main groups of enzymes, histone deacetylases (HDACs) and histone acetyl transferases (HATs). The HDAC removes acetyl-groups while the HATs transfer acetyl-groups to the protein of interest.

[0005] Classically, modulation of acetylation status is known to influence the condensation of chromatin. In cancer, histones are deacetylated maintaining a condensed chromatin structure, and a transcriptionally silenced state. This transcriptional inactivation is mediated by HDACs which remove acetyl groups from histone tails, maintain a condensed chromatic structure. Inhibitors of HDACs help maintain transcriptionally active chromatin, theoretically allowing for expression of tumor suppressor genes. One observation that has evolved is that histones are not the only targets of acetylation. It is now accepted that post- ActiveUS 205301239 1Attorney Docket No.: 0019240.01318WO1 Date of Electronic Filing: July 29, 2024 translational acetylation of intracellular proteins such as tumor suppressors (p53) and oncogenes (Bcl6) plays a critical role in influencing their activity. It has been established that there is a network of proteins and enzymes that can be modified by acetylation, now collectively referred to as the acetylome.

[0006] Histone Acetyltransferases (HATs) are involved in histone acetylation (leading to gene activation), chromosome decondensation, DNA repair and non-histone substrate modification. The post-translational acetylation status of chromatin is governed by the competing activities of two classes of enzymes, HATs and HDACs.

[0007] Alzheimer’s disease (AD) is an irreversible neurodegenerative disease characterized by memory loss, synaptic dysfunction and accumulation of extracellular amyloid β-peptides (Aβ), and intraneuronal tau protein. The pathogenesis of AD is believed to be caused by high levels and aggregation of amyloid-β (Aβ) and tau in the brain. Aβ and tau has been found to impair memory by reducing acetylation of specific histone lysines important for memory formation. Histones are proteins that closely associate with DNA molecules and play an important role in gene transcription.

[0008] Currently available therapies for AD are palliative and do not cure the disease. Cholinesterase inhibitors such as Razadyne® (galantamine), Exelon® (rivastigmine), Aricept® (donepezil), and Cognex® (tacrine) have been prescribed for early stages of Alzheimer’s disease, and may temporarily delay or prevent progression of symptoms related to AD. However, as AD progresses, the brain loses less acetylcholine, thereby rendering cholinesterase inhibitors unproductive as treatment for AD. Namenda® (memantine), an N- methyl D-aspartate (NMDA) antagonist, is also prescribed to treat moderate to severe Alzheimer’s disease; however only temporary benefits are realized. Namzaric (donepezil and memantine combination) is also prescribed but shares the same problems as the other therapies. Very recently, Leqembi (Lecanemab), slowing disease progression, has got full FDA approval for treatment of early AD. However, it is so far prescribed for early stages of the disease to a limited population due to possible serious side effects, and with unknown yet knowledge on long-term side effects and efficacy.

[0009] It would be beneficial to have additional HAT modulators, such as HAT activators, which can be used for treatments for a variety of disease states for which HAT activity is implicated. For example, additional HAT modulators would be useful to provide ActiveUS 205301239 2Attorney Docket No.: 0019240.01318WO1 Date of Electronic Filing: July 29, 2024 alternative methods for treating neurodegenerative diseases, neurological disorders and cancers. In particular, it would be beneficial to have additional HAT modulators for treatment of dementia and memory loss associated with Alzheimer’s disease and Alzheimer’s disease related dementia (ADRD). SUMMARY

[0010] The present disclosure is directed to compounds and compositions that modify HAT activity and their methods of use in treating various conditions, such as neurodegenerative disease or cancer. Thus, pharmaceutical compositions may comprise a compound enhancing HAT activity, and the methods may comprise administering to a subject a compound or composition that increases HAT activity.

[0011] Iin one aspect, the disclosure provides a compound of Formula (I),wherein, Rais H, OH, CN, C1-C6-alkyl, O-(C3-C8-cycloalkyl), O-(C3-C8-heterocycloalkyl), O-(C2-C6- alkenyl), O-(C1-C6-alkyl), O-(C2-C6-alkyl)-N(R1)2, halogen, or haloalkyl; Rbis H, C1-C6-alkyl, C2-C6-alkenyl, C3-C8-cycloalkyl, C2-C6-heteroalkyl, C3-C8- heterocycloalkyl, aryl, heteroaryl, O-(C1-C6-alkyl), O-(C3-C8-cycloalkyl), O-(C2-C6-alkenyl), or O-(C3-C8-heterocycloalkyl); Rcis H, C1-C6-alkyl, C1-C6-haloalkyl, O-(C1-C6-alkyl), O-(C1-C6-haloalkyl), halogen, CN, or NO2; Rdis H, OH, C1-C6-alkyl, O-(C3-C8-cycloalkyl), O-(C3-C8-heterocycloalkyl), O-(C2-C6- alkenyl), O-(C1-C6-alkyl), or O-(C2-C6-alkyl)-N(R1)2; ActiveUS 205301239 3Attorney Docket No.: 0019240.01318WO1 Date of Electronic Filing: July 29, 2024 R1is independently H, -(C1-C4-alkyl), -(C1-C4-haloalkyl), -(C3-C8-cycloalkyl), -(C3-C8- heterocycloalkyl), aryl or heteroaryl; W is CH or N; X is -CO-, -CON(R10)-, -CON(R1)(CH2)n-, -(CH2)nN(R1)-, or -C=N-; Z is CH or N, and n is an integer from 1-3, or a pharmaceutically acceptable salt or hydrate thereof.

[0012] In accordance with one embodiment, the present disclosure provides a compound of Formula (I) wherein, Rais OH, CN, C1-C6-alkyl, O-(C1-C6-alkyl), halogen, or haloalkyl; Rbis C1-C6-alkyl, C2-C6-heteroalkyl, aryl, heteroaryl, O-(C1-C6-alkyl); Rcis H, C1-C6-alkyl, C1-C6-haloalkyl, halogen, or CN; Rdis OH, C1-C6-alkyl, O-(C1-C6-alkyl), or O-(C2-C6-alkyl)-N(R1)2; R1is independently H, -(C1-C4-alkyl), or -(C1-C4-haloalkyl); W is CH;Z is CH, and n is an integer from 1-3, or a pharmaceutically acceptable salt or hydrate thereof.

[0013] In accordance with one embodiment, the present disclosure provides a compound of Formula(I) wherein, Rais OH, O-(C1-C6-alkyl), halogen, or haloalkyl; Rbis C1-C6-alkyl, O-(C1-C6-alkyl); Rcis C1-C6-alkyl, halogen, CF3or CN; Rdis OH, C1-C6-alkyl, O-(C1-C6-alkyl), or O-(C2-C6-alkyl)-N(R1)2; R1is independently H, -(C1-C4-alkyl), or -(C1-C4-haloalkyl); W is CH; ActiveUS 205301239 4Attorney Docket No.: 0019240.01318WO1 Date of Electronic Filing: July 29, 2024 X is -CON(R1)-; and Z is CH, or a pharmaceutically acceptable salt or hydrate thereof.

[0014] In accordance with one embodiment, the present disclosure provides a compound of Formula (I) wherein, wherein X is –C(O)N(R1)-.

[0015] In accordance with one embodiment, the present disclosure provides a compound of Formula (I) wherein, Rbis C1-C6-alkyl, O-(C1-C6-alkyl).

[0016] In accordance with one embodiment, the present disclosure provides a compound of Formula (I) wherein the compound of Formula (I) iswherein Rais OH, O-(C1-C6-alkyl), halogen, or haloalkyl; Rbis C1-C6-alkyl, O-(C1-C6-alkyl); Rcis C1-C6-alkyl, halogen, CF3 or CN; Rdis OH, C1-C6-alkyl, O-(C1-C6-alkyl), or O-(C2-C6-alkyl)-N(R1)2; R1is independently H, -(C1-C4-alkyl), or -(C1-C4-haloalkyl); W is CH; and Z is CH, or a pharmaceutically acceptable salt or hydrate thereof.

[0017] In accordance with one embodiment, the present disclosure provides a compound of Formula (I) wherein the compound of Formula (I) is ActiveUS 205301239 5Attorney Docket No.: 0019240.01318WO1 Date of Electronic Filing: July 29, 2024wherein Rais OH, O-(C1-C3-alkyl), halogen, or haloalkyl; Rbis C1-C3-alkyl, O-(C1-C3-alkyl); Rcis C1-C6-alkyl, halogen, CF3or CN; Rdis O-(C1-C6-alkyl), or O-(C2-C6-alkyl)-N(R1)2; and R1is independently H, -(C1-C2-alkyl), or -(C1-C4-haloalkyl), or a pharmaceutically acceptable salt or hydrate thereof.

[0018] In accordance with one embodiment, the present disclosure provides a compound of Formula (I-b), wherein Rais O-(C1-C3-alkyl); Rbis O-(C1-C3-alkyl); Rcis C1-C6-alkyl, halogen, CF3 or CN; Rdis O-(C2-C6-alkyl)-N(R1)2; and R1is independently H, -(C1-C2-alkyl), or a pharmaceutically acceptable salt or hydrate thereof.

[0019] In accordance with one embodiment, the present disclosure provides a compound having the structure: ActiveUS 205301239 6Attorney Docket No.: 0019240.01318WO1 Date of Electronic Filing: July 29, 2024or a pharmaceutically acceptable salt or solvate thereof.

[0020] In accordance with one embodiment, the present disclosure provides a compound that is a HAT activator.

[0021] In accordance with one aspect, the present disclosure provides pharmaceutical compositions disclosed herein containing a compound of Formula (I) and a pharmaceutically acceptable excipient.

[0022] In some aspects, the present disclosure provides a method of increasing histone acetylation in a subject, the method comprising administering to the subject a therapeutically effective amount of a compound of Formula (I) or a composition comprising a compound of Formula (I).

[0023] In some aspects, the present disclosure provides a method of treating a neurodegenerative disease or condition in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a compound of Formula (I) or a composition comprising a compound of Formula (I).

[0024] In some aspects, the present disclosure provides a method of treating cancer in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a compound of Formula (I) or a composition comprising a compound of Formula (I).

[0025] In some aspects, the present disclosure provides a enhancing learning or memory in a subject by administering to the subject a therapeutically effective amount of a compound of Formula (I) or a composition comprising a compound of Formula (I). ActiveUS 205301239 7Attorney Docket No.: 0019240.01318WO1 Date of Electronic Filing: July 29, 2024

[0026] In some embodiments, the present disclosure provides a method of treating a subject who has at least one mutant HAT enzyme gene.

[0027] The present disclosure is directed to compounds and compositions that modify HAT activity and their methods of use in treating various conditions, such as neurodegenerative disease or cancer. Thus, pharmaceutical compositions may comprise a compound enhancing HAT activity, and the methods may comprise administering to a subject a compound or composition that increases HAT activity.

[0028] The pharmaceutical compositions disclosed herein comprise a compound of Formula (I) and a pharmaceutically acceptable excipient.

[0029] In some embodiments, the present disclosure provides a method of increasing histone acetylation in a subject, the method comprising administering to the subject a therapeutically effective amount of a compound of Formula (I) or a composition comprising a compound of Formula (I).

[0030] In some embodiments, the present disclosure provides a method of treating a neurodegenerative disease or condition in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a compound of Formula (I) or a composition comprising a compound of Formula (I).

[0031] In some embodiments, the present disclosure provides a method of treating cancer in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a compound of Formula (I) or a composition comprising a compound of Formula (I). BRIEF DESCRIPTION OF THE FIGURES

[0032] FIG.1A, FIG.1B, FIG.1C and FIG.1D display pharmacokinetic concentration values for EZ115HAT in plasma and brain following 10mg / kg dose via intravenous (IV) and 20mg / kg dose via oral gavage (PO) administrations in Sprague Dawley rats. In addition, brain concentrations of EZ115HAT were evaluated at various times post dose up to 24 hours after dosing.

[0033] FIG.2A is a graph showing EZ115HAT biological activity (EC50) on p300 H3K18. ActiveUS 205301239 8Attorney Docket No.: 0019240.01318WO1 Date of Electronic Filing: July 29, 2024

[0034] FIG.2B is a graph showing EZ115HAT biological activity (EC50) on p300 H3K27.

[0035] FIG.3A is a schematic showing the preparation of EZ115HAT through a route of synthesis in accordance with one embodiment.

[0036] FIG.3B is a schematic showing the preparation of EZ115HAT through a route of synthesis in accordance with another embodiment.

[0037] FIG.4 is a plot of EZ115HAT concentration in brain and plasma following intravenous (IV) and oral (PO) administrations.

[0038] FIG.5A is a graph showing efficacy of EZ115HAT in the APP / PS1 mouse model of amyloid elevation as related to synaptic plasticity.

[0039] FIG.5B is a graph showing efficacy of EZ115HAT in the APP / PS1 mouse model of amyloid elevation as related to fear memory.

[0040] FIG.5C is a graph showing efficacy of EZ115HAT in the APP / PS1 mouse model of amyloid elevation as related to spatial memory.

[0041] FIG.6A is a graph showing efficacy of EZ115HAT in the hTau / Mapt-KO mouse model of tau elevation as related to synaptic plasticity.

[0042] FIG.6B is a graph showing efficacy of EZ115HAT in the hTau / Mapt-KO mouse model of tau elevation as related to fear memory.

[0043] FIG.6C is a graph showing efficacy of EZ115HAT in the hTau / Mapt-KO mouse model of tau elevation as related to spatial memory.

[0044] FIG.7A is a graph showing results for time to reach a visible platform task for the hTau / Mapt-KO mouse model of tau elevation for different treatment conditions.

[0045] FIG.7B is a graph showing results for average speed to reach a visible platform task for the hTau / Mapt-KO mouse model of tau elevation for different treatment conditions.

[0046] FIG.7C is a chart showing results for cued amygdala dependent memory for the hTau / Mapt-KO mouse model of tau elevation for different treatment conditions. ActiveUS 205301239 9Attorney Docket No.: 0019240.01318WO1 Date of Electronic Filing: July 29, 2024

[0047] FIG.7D is a chart showing results for the percentage of time spent in the center during an open field test for the hTau / Mapt-KO mouse model of tau elevation for different treatment conditions.

[0048] FIG.7E is a chart showing results for number of entries into the center during an open field test for the hTau / Mapt-KO mouse model of tau elevation for different treatment conditions.

[0049] FIG.7F is a chart showing results for capability of perceiving an electric shock for the hTau / Mapt-KO mouse model of tau elevation for different treatment conditions.

[0050] FIG.8 is a graph showing efficacy of EZ115HAT in AD relevant mouse models characterized by Aβ and tau oligomer elevation using LTP as a test of synaptic plasticity.

[0051] FIG.9 provides an EZ115HAT dose-response curve during assessment of LTP deficit due to oligomeric tau elevation. DETAILED DESCRIPTION

[0052] In the following description, certain specific details are set forth in order to provide a thorough understanding of various embodiments. However, one skilled in the art will understand that the invention can be practiced without these details. In other instances, well-known structures have not been shown or described in detail to avoid unnecessarily obscuring descriptions of the embodiments.

[0053] All definitions, as defined and used herein, should be understood to control over dictionary definitions, definitions in documents incorporated by reference, and / or ordinary meanings of the defined terms. Use of flow diagrams is not meant to be limiting with respect to the order of operations performed for all embodiments.

[0054] Reference throughout this specification to “one embodiment” or “an embodiment,” etc. means that a particular feature, structure or characteristic described in connection with the embodiment is included in at least one embodiment. Thus, the appearances of the phrases “in one embodiment” or “in an embodiment” in various places throughout this specification are not necessarily all referring to the same embodiment. Furthermore, the particular features, structures, or characteristics can be combined in any suitable manner in one or more embodiments. Also, as used in this specification and the ActiveUS 205301239 10Attorney Docket No.: 0019240.01318WO1 Date of Electronic Filing: July 29, 2024 appended claims, the singular forms “a,” “an,” and “the” include plural referents unless the content clearly dictates otherwise. It should also be noted that the term “or” is generally employed in its sense including “and / or” unless the content clearly dictates otherwise.

[0055] “Alkyl” or “alkyl group” refers to a fully saturated, straight or branched hydrocarbon chain radical, and which is attached to the rest of the molecule by a single bond. Alkyls comprising any number of carbon atoms from 1 to 12 are included. An alkyl comprising up to 12 carbon atoms is a C1-C12alkyl, an alkyl comprising up to 10 carbon atoms is a C1-C10 alkyl, an alkyl comprising up to 6 carbon atoms is a C1-C6 alkyl and an alkyl comprising up to 5 carbon atoms is a C1-C5alkyl. A C1-C5alkyl includes C5alkyls, C4alkyls, C3alkyls, C2alkyls and C1alkyl (i.e., methyl). A C1-C6alkyl includes all moieties described above for C1-C5 alkyls but also includes C6 alkyls. A C1-C10 alkyl includes all moieties described above for C1-C5 alkyls and C1-C6 alkyls, but also includes C7, C8, C9 and C10alkyls. Similarly, a C1-C12alkyl includes all the foregoing moieties, but also includes C11and C12 alkyls. Non-limiting examples of C1-C12 alkyl include methyl, ethyl, n-propyl, i- propyl, sec-propyl, n-butyl, i-butyl, sec-butyl, t-butyl, n-pentyl, t-amyl, n-hexyl, n-heptyl, n- octyl, n-nonyl, n-decyl, n-undecyl, and n-dodecyl. Unless stated otherwise specifically in the specification, an alkyl group can be optionally substituted.

[0056] “Alkylene” or “alkylene chain” refers to a fully saturated, straight or branched divalent hydrocarbon chain radical. Alkylenes comprising any number of carbon atoms from 1 to 12 are included. Non-limiting examples of C1-C12alkylene include methylene, ethylene, propylene, n-butylene, ethenylene, propenylene, n-butenylene, propynylene, n-butynylene, and the like. The alkylene chain is attached to the rest of the molecule through a single bond and to the radical group through a single bond. The points of attachment of the alkylene chain to the rest of the molecule and to the radical group can be through one carbon or any two carbons within the chain. Unless stated otherwise specifically in the specification, an alkylene chain can be optionally substituted.

[0057] “Alkenyl” or “alkenyl group” refers to a straight or branched hydrocarbon chain radical having from two to twelve carbon atoms, and having one or more carbon-carbon double bonds. Each alkenyl group is attached to the rest of the molecule by a single bond. Alkenyl group comprising any number of carbon atoms from 2 to 12 are included. An alkenyl group comprising up to 12 carbon atoms is a C2-C12 alkenyl, an alkenyl comprising up to 10 carbon atoms is a C2-C10alkenyl, an alkenyl group comprising up to 6 carbon atoms is a C2- ActiveUS 205301239 11Attorney Docket No.: 0019240.01318WO1 Date of Electronic Filing: July 29, 2024 C6 alkenyl and an alkenyl comprising up to 5 carbon atoms is a C2-C5 alkenyl. A C2-C5 alkenyl includes C5 alkenyls, C4 alkenyls, C3 alkenyls, and C2 alkenyls. A C2-C6 alkenyl includes all moieties described above for C2-C5alkenyls but also includes C6alkenyls. A C2- C10 alkenyl includes all moieties described above for C2-C5 alkenyls and C2-C6 alkenyls, but also includes C7, C8, C9 and C10 alkenyls. Similarly, a C2-C12 alkenyl includes all the foregoing moieties, but also includes C11and C12alkenyls. Non-limiting examples of C2-C12alkenyl include ethenyl (vinyl), 1-propenyl, 2-propenyl (allyl), iso-propenyl, 2-methyl-1- propenyl, 1-butenyl, 2-butenyl, 3-butenyl, 1-pentenyl, 2-pentenyl, 3-pentenyl, 4-pentenyl, 1- hexenyl, 2-hexenyl, 3-hexenyl, 4-hexenyl, 5-hexenyl, 1-heptenyl, 2-heptenyl, 3-heptenyl, 4- heptenyl, 5-heptenyl, 6-heptenyl, 1-octenyl, 2-octenyl, 3-octenyl, 4-octenyl, 5-octenyl, 6- octenyl, 7-octenyl, 1-nonenyl, 2-nonenyl, 3-nonenyl, 4-nonenyl, 5-nonenyl, 6-nonenyl, 7- nonenyl, 8-nonenyl, 1-decenyl, 2-decenyl, 3-decenyl, 4-decenyl, 5-decenyl, 6-decenyl, 7- decenyl, 8-decenyl, 9-decenyl, 1-undecenyl, 2-undecenyl, 3-undecenyl, 4-undecenyl, 5- undecenyl, 6-undecenyl, 7-undecenyl, 8-undecenyl, 9-undecenyl, 10-undecenyl, 1-dodecenyl, 2-dodecenyl, 3-dodecenyl, 4-dodecenyl, 5-dodecenyl, 6-dodecenyl, 7-dodecenyl, 8- dodecenyl, 9-dodecenyl, 10-dodecenyl, and 11-dodecenyl. Examples of C1-C3alkyl includes methyl, ethyl, n-propyl, and i-propyl. Examples of C1-C4alkyl includes methyl, ethyl, n- propyl, i-propyl, n-butyl, i-butyl, and sec-butyl. Unless stated otherwise specifically in the specification, an alkyl group can be optionally substituted.

[0058] “Alkenylene” or “alkenylene chain” refers to a straight or branched divalent hydrocarbon chain radical, having from two to twelve carbon atoms, and having one or more carbon-carbon double bonds. Non-limiting examples of C2-C12alkenylene include ethene, propene, butene, and the like. The alkenylene chain is attached to the rest of the molecule through a single bond and to the radical group through a single bond. The points of attachment of the alkenylene chain to the rest of the molecule and to the radical group can be through one carbon or any two carbons within the chain. Unless stated otherwise specifically in the specification, an alkenylene chain can be optionally substituted.

[0059] “Alkynyl” or “alkynyl group” refers to a straight or branched hydrocarbon chain radical having from two to twelve carbon atoms, and having one or more carbon-carbon triple bonds. Each alkynyl group is attached to the rest of the molecule by a single bond. Alkynyl groups comprising any number of carbon atoms from 2 to 12 are included. An alkynyl group comprising up to 12 carbon atoms is a C2-C12alkynyl, an alkynyl comprising up to 10 carbon ActiveUS 205301239 12Attorney Docket No.: 0019240.01318WO1 Date of Electronic Filing: July 29, 2024 atoms is a C2-C10 alkynyl, an alkynyl group comprising up to 6 carbon atoms is a C2-C6 alkynyl and an alkynyl comprising up to 5 carbon atoms is a C2-C5 alkynyl. A C2-C5 alkynyl includes C5alkynyls, C4alkynyls, C3alkynyls, and C2alkynyls. A C2-C6alkynyl includes all moieties described above for C2-C5 alkynyls but also includes C6 alkynyls. A C2-C10 alkynyl includes all moieties described above for C2-C5 alkynyls and C2-C6 alkynyls, but also includes C7, C8, C9and C10alkynyls. Similarly, a C2-C12alkynyl includes all the foregoing moieties, but also includes C11and C12alkynyls. Non-limiting examples of C2-C12alkenyl include ethynyl, propynyl, butynyl, pentynyl and the like. Unless stated otherwise specifically in the specification, an alkyl group can be optionally substituted.

[0060] “Alkoxy” refers to a radical of the formula -ORawhere Rais an alkyl, alkenyl or alkynyl radical as defined above containing one to twelve carbon atoms. Unless stated otherwise specifically in the specification, an alkoxy group can be optionally substituted.

[0061] “Alkylamino” refers to a radical of the formula -NHRaor -NRaRawhere each Rais, independently, an alkyl, alkenyl or alkynyl radical as defined above containing one to twelve carbon atoms. Unless stated otherwise specifically in the specification, an alkylamino group can be optionally substituted.

[0062] “Alkylcarbonyl” refers to the –C(=O)Ramoiety, wherein Rais an alkyl, alkenyl or alkynyl radical as defined above. A non-limiting example of an alkyl carbonyl is the methyl carbonyl (“acetal”) moiety. Alkylcarbonyl groups can also be referred to as “Cw-Cz acyl” where w and z depicts the range of the number of carbons in Ra, as defined above. For example, “C1-C10 acyl” refers to alkylcarbonyl group as defined above, where Ra is C1-C10 alkyl, C1-C10alkenyl, or C1-C10alkynyl radical as defined above. Unless stated otherwise specifically in the specification, an alkyl carbonyl group can be optionally substituted.

[0063] “Aryl” refers to a hydrocarbon ring system radical comprising hydrogen, 5 to 18 carbon atoms and at least one aromatic ring. For purposes of this invention, the aryl radical can be a monocyclic, bicyclic, tricyclic or tetracyclic ring system, which can include fused or bridged ring systems. Aryl radicals include, but are not limited to, aryl radicals derived from aceanthrylene, acenaphthylene, acephenanthrylene, anthracene, azulene, benzene, chrysene, fluoranthene, fluorene, as-indacene, s-indacene, indane, indene, naphthalene, phenalene, phenanthrene, pleiadene, pyrene, and triphenylene. Unless stated otherwise specifically in the specification, the term “aryl” is meant to include aryl radicals that are optionally substituted. ActiveUS 205301239 13Attorney Docket No.: 0019240.01318WO1 Date of Electronic Filing: July 29, 2024

[0064] “Aralkyl” refers to a radical of the formula -Rb-Rc where Rb is an alkylene, alkenylene or alkynylene group as defined above and Rc is one or more aryl radicals as defined above, for example, benzyl, diphenylmethyl and the like. Unless stated otherwise specifically in the specification, an aralkyl group can be optionally substituted.

[0065] “Carbocyclyl,” “carbocyclic ring” or “carbocycle” refers to a rings structure, wherein the atoms which form the ring are each carbon. Carbocyclic rings can comprise from 3 to 20 carbon atoms in the ring. Carbocyclic rings include aryls and cycloalkyl, cycloalkenyl and cycloalkynyl as defined herein. Unless stated otherwise specifically in the specification, a carbocyclyl group can be optionally substituted.

[0066] “Cycloalkyl” refers to a stable non-aromatic monocyclic or polycyclic fully saturated hydrocarbon radical consisting solely of carbon and hydrogen atoms, which can include fused or bridged ring systems, having from three to twenty carbon atoms, preferably having from three to ten carbon atoms, and which is attached to the rest of the molecule by a single bond. Monocyclic cycloalkyl radicals include, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. Polycyclic cycloalkyl radicals include, for example, adamantyl, norbornyl, decalinyl, 7,7-dimethyl-bicyclo[2.2.1]heptanyl, and the like. Unless otherwise stated specifically in the specification, a cycloalkyl group can be optionally substituted.

[0067] “Cycloalkenyl” refers to a stable non-aromatic monocyclic or polycyclic hydrocarbon radical consisting solely of carbon and hydrogen atoms, having one or more carbon-carbon double bonds, which can include fused or bridged ring systems, having from three to twenty carbon atoms, preferably having from three to ten carbon atoms, and which is attached to the rest of the molecule by a single bond. Monocyclic cycloalkenyl radicals include, for example, cyclopentenyl, cyclohexenyl, cycloheptenyl, cycloctenyl, and the like. Polycyclic cycloalkenyl radicals include, for example, bicyclo[2.2.1]hept-2-enyl and the like. Unless otherwise stated specifically in the specification, a cycloalkenyl group can be optionally substituted.

[0068] “Cycloalkynyl” refers to a stable non-aromatic monocyclic or polycyclic hydrocarbon radical consisting solely of carbon and hydrogen atoms, having one or more carbon-carbon triple bonds, which can include fused or bridged ring systems, having from three to twenty carbon atoms, preferably having from three to ten carbon atoms, and which is ActiveUS 205301239 14Attorney Docket No.: 0019240.01318WO1 Date of Electronic Filing: July 29, 2024 attached to the rest of the molecule by a single bond. Monocyclic cycloalkynyl radicals include, for example, cycloheptynyl, cyclooctynyl, and the like. Unless otherwise stated specifically in the specification, a cycloalkynyl group can be optionally substituted.

[0069] “Cycloalkylalkyl” refers to a radical of the formula -Rb-Rd where Rb is an alkylene, alkenylene, or alkynylene group as defined above and Rd is a cycloalkyl, cycloalkenyl, cycloalkynyl radical as defined above. Unless stated otherwise specifically in the specification, a cycloalkylalkyl group can be optionally substituted.

[0070] “Haloalkyl” refers to an alkyl radical, as defined above, that is substituted by one or more halo radicals, as defined above, e.g., trifluoromethyl, difluoromethyl, trichloromethyl, 2,2,2-trifluoroethyl, 1,2-difluoroethyl, 3-bromo-2-fluoropropyl, 1,2-dibromoethyl, and the like. Unless stated otherwise specifically in the specification, a haloalkyl group can be optionally substituted.

[0071] “Haloalkenyl” refers to an alkenyl radical, as defined above, that is substituted by one or more halo radicals, as defined above, e.g., 1-fluoropropenyl, 1,1-difluorobutenyl, and the like. Unless stated otherwise specifically in the specification, a haloalkenyl group can be optionally substituted.

[0072] “Haloalkynyl” refers to an alkynyl radical, as defined above that is substituted by one or more halo radicals, as defined above, e.g., 1-fluoropropynyl, 1-fluorobutynyl, and the like. Unless stated otherwise specifically in the specification, a haloalkenyl group can be optionally substituted.

[0073] “Heterocyclyl,” “heterocyclic ring” or “heterocycle” refers to a stable 3- to 20-membered non-aromatic ring radical which consists of two to twelve carbon atoms and from one to six heteroatoms selected from the group consisting of nitrogen, oxygen and sulfur. Heterocyclyl or heterocyclic rings include heteroaryls as defined below. Unless stated otherwise specifically in the specification, the heterocyclyl radical can be a monocyclic, bicyclic, tricyclic or tetracyclic ring system, which can include fused or bridged ring systems; and the nitrogen, carbon or sulfur atoms in the heterocyclyl radical can be optionally oxidized; the nitrogen atom can be optionally quaternized; and the heterocyclyl radical can be partially or fully saturated. Examples of such heterocyclyl radicals include, but are not limited to, dioxolanyl, thienyl[1,3]dithianyl, decahydroisoquinolyl, imidazolinyl, imidazolidinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl, octahydroindolyl, ActiveUS 205301239 15Attorney Docket No.: 0019240.01318WO1 Date of Electronic Filing: July 29, 2024 octahydroisoindolyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, oxazolidinyl, piperidinyl, piperazinyl, 4-piperidonyl, pyrrolidinyl, pyrazolidinyl, quinuclidinyl, thiazolidinyl, tetrahydrofuryl, trithianyl, tetrahydropyranyl, thiomorpholinyl, thiamorpholinyl, 1-oxo-thiomorpholinyl, and 1,1-dioxo-thiomorpholinyl. Unless stated otherwise specifically in the specification, a heterocyclyl group can be optionally substituted.

[0074] “N-heterocyclyl” refers to a heterocyclyl radical as defined above containing at least one nitrogen and where the point of attachment of the heterocyclyl radical to the rest of the molecule is through a nitrogen atom in the heterocyclyl radical. Unless stated otherwise specifically in the specification, a N-heterocyclyl group can be optionally substituted.

[0075] “Heterocyclylalkyl” refers to a radical of the formula -Rb-Rewhere Rbis an alkylene, alkenylene, or alkynylene chain as defined above and Re is a heterocyclyl radical as defined above, and if the heterocyclyl is a nitrogen-containing heterocyclyl, the heterocyclyl can be attached to the alkyl, alkenyl, alkynyl radical at the nitrogen atom. Unless stated otherwise specifically in the specification, a heterocyclylalkyl group can be optionally substituted.

[0076] “Heteroaryl” refers to a 5- to 20-membered ring system radical comprising hydrogen atoms, one to thirteen carbon atoms, one to six heteroatoms selected from the group consisting of nitrogen, oxygen and sulfur, and at least one aromatic ring. For purposes of this invention, the heteroaryl radical can be a monocyclic, bicyclic, tricyclic or tetracyclic ring system, which can include fused or bridged ring systems; and the nitrogen, carbon or sulfur atoms in the heteroaryl radical can be optionally oxidized; the nitrogen atom can be optionally quaternized. Examples include, but are not limited to, azepinyl, acridinyl, benzimidazolyl, benzothiazolyl, benzindolyl, benzodioxolyl, benzofuranyl, benzooxazolyl, benzothiazolyl, benzothiadiazolyl, benzo[b][1,4]dioxepinyl, 1,4-benzodioxanyl, benzonaphthofuranyl, benzoxazolyl, benzodioxolyl, benzodioxinyl, benzopyranyl, benzopyranonyl, benzofuranyl, benzofuranonyl, benzothienyl (benzothiophenyl), benzotriazolyl, benzo[4,6]imidazo[1,2-a]pyridinyl, carbazolyl, cinnolinyl, dibenzofuranyl, dibenzothiophenyl, furanyl, furanonyl, isothiazolyl, imidazolyl, indazolyl, indolyl, indazolyl, isoindolyl, indolinyl, isoindolinyl, isoquinolyl, indolizinyl, isoxazolyl, naphthyridinyl, oxadiazolyl, 2-oxoazepinyl, oxazolyl, oxiranyl, 1-oxidopyridinyl, 1-oxidopyrimidinyl, 1- oxidopyrazinyl, 1-oxidopyridazinyl, 1-phenyl-1H-pyrrolyl, phenazinyl, phenothiazinyl, phenoxazinyl, phthalazinyl, pteridinyl, purinyl, pyrrolyl, pyrazolyl, pyridinyl, pyrazinyl, ActiveUS 205301239 16Attorney Docket No.: 0019240.01318WO1 Date of Electronic Filing: July 29, 2024 pyrimidinyl, pyridazinyl, quinazolinyl, quinoxalinyl, quinolinyl, quinuclidinyl, isoquinolinyl, tetrahydroquinolinyl, thiazolyl, thiadiazolyl, triazolyl, tetrazolyl, triazinyl, and thiophenyl (i.e., thienyl). Unless stated otherwise specifically in this disclosure, a heteroaryl group can be optionally substituted.

[0077] “N-heteroaryl” refers to a heteroaryl radical as defined above containing at least one nitrogen and where the point of attachment of the heteroaryl radical to the rest of the molecule is through a nitrogen atom in the heteroaryl radical. Unless stated otherwise specifically in the specification, an N-heteroaryl group can be optionally substituted.

[0078] “Heteroarylalkyl” refers to a radical of the formula -Rb-Rfwhere Rbis an alkylene, alkenylene, or alkynylene chain as defined above and Rfis a heteroaryl radical as defined above. Unless stated otherwise specifically in the specification, a heteroarylalkyl group can be optionally substituted.

[0079] The term “substituted” used herein means any of the above groups (i.e., alkyl, alkylene, alkenyl, alkenylene, alkynyl, alkynylene, alkoxy, alkylamino, alkylcarbonyl, thioalkyl, aryl, aralkyl, carbocyclyl, cycloalkyl, cycloalkenyl, cycloalkynyl, cycloalkylalkyl, haloalkyl, heterocyclyl, N-heterocyclyl, heterocyclylalkyl, heteroaryl, N-heteroaryl and / or heteroarylalkyl) wherein at least one hydrogen atom is replaced by a bond to a non-hydrogen atoms such as, but not limited to: a halogen atom such as F, Cl, Br, I and CF3; an oxygen atom in groups such as hydroxyl groups, alkoxy groups, and ester groups; a sulfur atom in groups such as thiol groups, thioalkyl groups, sulfone groups, sulfonyl groups, and sulfoxide groups; a nitrogen atom in groups such as amines, amides, alkylamines, dialkylamines, arylamines, alkylarylamines, diarylamines, N-oxides, imides, and enamines; a silicon atom in groups such as trialkylsilyl groups, dialkylarylsilyl groups, alkyldiarylsilyl groups, and triarylsilyl groups; and other heteroatoms in various other groups. “Substituted” also means any of the above groups in which one or more hydrogen atoms are replaced by a higher-order bond (e.g., a double- or triple-bond) to a heteroatom such as oxygen in oxo, carbonyl, carboxyl, and ester groups; and nitrogen in groups such as imines, oximes, hydrazones, and nitriles. For example, “substituted” includes any of the above groups in which one or more hydrogen atoms are replaced with -NRgC(=O)ORh, -NRgSO2Rh, -OC(=O)NRgRh, -ORg, -SRg, -SORg, -SO2Rg, -OSO2Rg, -S O2ORg, =NSO2Rg, and -SO2NRgRh. “Substituted also means any of the above groups in which one or more hydrogen atoms are replaced ActiveUS 205301239 17Attorney Docket No.: 0019240.01318WO1 Date of Electronic Filing: July 29, 2024 with -C(=O)Rg, -C(=O)ORg, -C(=O)NRgRh, -CH2SO2Rg, -CH2SO2NRgRh. In the foregoing, Rg and Rh are the same or different and independently hydrogen, alkyl, alkenyl, alkynyl, alkoxy, alkylamino, thioalkyl, aryl, aralkyl, cycloalkyl, cycloalkenyl, cycloalkynyl, cycloalkylalkyl, haloalkyl, haloalkenyl, haloalkynyl, heterocyclyl, N-heterocyclyl, heterocyclylalkyl, heteroaryl, N-heteroaryl and / or heteroarylalkyl. “Substituted” further means any of the above groups in which one or more hydrogen atoms are replaced by a bond to an amino, cyano, hydroxyl, imino, nitro, oxo, thioxo, halo, alkyl, alkenyl, alkynyl, alkoxy, alkylamino, thioalkyl, aryl, aralkyl, cycloalkyl, cycloalkenyl, cycloalkynyl, cycloalkylalkyl, haloalkyl, haloalkenyl, haloalkynyl, heterocyclyl, N-heterocyclyl, heterocyclylalkyl, heteroaryl, N-heteroaryl and / or heteroarylalkyl group. In addition, each of the foregoing substituents can also be optionally substituted with one or more of the above substituents.

[0080] “Optional” or “optionally” means that the subsequently described event of circumstances can or cannot occur, and that the description includes instances where said event or circumstance occurs and instances in which it does not. For example, “optionally substituted aryl” means that the aryl radical can or cannot be substituted and that the description includes both substituted aryl radicals and aryl radicals having no substitution.

[0081] The compounds disclosed herein, or their pharmaceutically acceptable salts can contain one or more asymmetric centers and can thus give rise to enantiomers, diastereomers, and other stereoisomeric forms that can be defined, in terms of absolute stereochemistry, as (R)- or (S)- or, as (D)- or (L)- for amino acids. The present invention is meant to include all such possible isomers, as well as their racemic and optically pure forms whether or not they are specifically depicted herein. Optically active (+) and (-), (R)- and (S)-, or (D)- and (L)- isomers can be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques, for example, chromatography and fractional crystallization. Conventional techniques for the preparation / isolation of individual enantiomers include chiral synthesis from a suitable optically pure precursor or resolution of the racemate (or the racemate of a salt or derivative) using, for example, chiral high pressure liquid chromatography (HPLC). When the compounds described herein contain olefinic double bonds or other centers of geometric asymmetry, and unless specified otherwise, it is intended that the compounds include both E and Z geometric isomers. Likewise, all tautomeric forms are also intended to be included. ActiveUS 205301239 18Attorney Docket No.: 0019240.01318WO1 Date of Electronic Filing: July 29, 2024

[0082] A “stereoisomer” refers to a compound made up of the same atoms bonded by the same bonds but having different three-dimensional structures, which are not interchangeable. The present invention contemplates various stereoisomers and mixtures thereof and includes “enantiomers”, which refers to two stereoisomers whose molecules are nonsuperimposable mirror images of one another.

[0083] A “tautomer” refers to a proton shift from one atom of a molecule to another atom of the same molecule. The present disclosure includes tautomers of any said compounds.

[0084] “Pharmaceutically acceptable carrier, diluent or excipient” includes without limitation any adjuvant, carrier, excipient, glidant, sweetening agent, diluent, preservative, dye / colorant, flavor enhancer, surfactant, wetting agent, dispersing agent, suspending agent, stabilizer, isotonic agent, solvent, or emulsifier which has been approved by the United States Food and Drug Administration as being acceptable for use in humans or domestic animals.

[0085] “Pharmaceutically acceptable salt” includes both acid and base addition salts.

[0086] “Pharmaceutically acceptable acid addition salt” refers to those salts which retain the biological effectiveness and properties of the free bases, which are not biologically or otherwise undesirable, and which are formed with inorganic acids such as, but are not limited to, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid and the like, and organic acids such as, but not limited to, acetic acid, 2,2-dichloroacetic acid, adipic acid, alginic acid, ascorbic acid, aspartic acid, benzenesulfonic acid, benzoic acid, 4- acetamidobenzoic acid, camphoric acid, camphor-10-sulfonic acid, capric acid, caproic acid, caprylic acid, carbonic acid, cinnamic acid, citric acid, cyclamic acid, dodecylsulfuric acid, ethane-1,2-disulfonic acid, ethanesulfonic acid, 2-hydroxyethanesulfonic acid, formic acid, fumaric acid, galactaric acid, gentisic acid, glucoheptonic acid, gluconic acid, glucuronic acid, glutamic acid, glutaric acid, 2-oxo-glutaric acid, glycerophosphoric acid, glycolic acid, hippuric acid, isobutyric acid, lactic acid, lactobionic acid, lauric acid, maleic acid, malic acid, malonic acid, mandelic acid, methanesulfonic acid, mucic acid, naphthalene-1,5- disulfonic acid, naphthalene-2-sulfonic acid, 1-hydroxy-2-naphthoic acid, nicotinic acid, oleic acid, orotic acid, oxalic acid, palmitic acid, pamoic acid, propionic acid, pyroglutamic acid, pyruvic acid, salicylic acid, 4-aminosalicylic acid, sebacic acid, stearic acid, succinic acid, tartaric acid, thiocyanic acid, p-toluenesulfonic acid, trifluoroacetic acid, undecylenic acid, and the like. ActiveUS 205301239 19Attorney Docket No.: 0019240.01318WO1 Date of Electronic Filing: July 29, 2024

[0087] “Pharmaceutically acceptable base addition salt” refers to those salts which retain the biological effectiveness and properties of the free acids, which are not biologically or otherwise undesirable. These salts are prepared from addition of an inorganic base or an organic base to the free acid. Salts derived from inorganic bases include, but are not limited to, the sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum salts and the like. Preferred inorganic salts are the ammonium, sodium, potassium, calcium, and magnesium salts. Salts derived from organic bases include, but are not limited to, salts of primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines and basic ion exchange resins, such as ammonia, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, diethanolamine, ethanolamine, deanol, 2-dimethylaminoethanol, 2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, hydrabamine, choline, betaine, benethamine, benzathine, ethylenediamine, glucosamine, methylglucamine, theobromine, triethanolamine, tromethamine, purines, piperazine, piperidine, N-ethylpiperidine, polyamine resins and the like. Particularly preferred organic bases are isopropylamine, diethylamine, ethanolamine, trimethylamine, dicyclohexylamine, choline and caffeine.

[0088] As used herein, a “subject” can be a human, non-human primate, mammal, rat, mouse, cow, horse, pig, sheep, goat, dog, cat, insect and the like. The subject can be suspected of having or at risk for having a disease or condition treatable with a HAT modulator.

[0089] A “pharmaceutical composition” refers to a formulation of a compound of the invention and a medium generally accepted in the art for the delivery of the biologically active compound to subjects, such as mammals, e.g., humans. Such a medium includes all pharmaceutically acceptable carriers, diluents or excipients therefor.

[0090] An “effective amount” refers to a therapeutically effective amount or a prophylactically effective amount. A “therapeutically effective amount” refers to an amount effective, at dosages and for periods of time necessary, to achieve the desired therapeutic result, such as decreased symptoms, reduced tumor size, increased life span or increased life expectancy. A therapeutically effective amount of a compound can vary according to factors such as the disease state, age, sex, and weight of the subject, and the ability of the compound to elicit a desired response in the subject. Dosage regimens can be adjusted to provide the ActiveUS 205301239 20Attorney Docket No.: 0019240.01318WO1 Date of Electronic Filing: July 29, 2024 optimum therapeutic response. A therapeutically effective amount is also one in which any toxic or detrimental effects of the compound are outweighed by the therapeutically beneficial effects. A “prophylactically effective amount” refers to an amount effective, at dosages and for periods of time necessary, to achieve the desired prophylactic result, such as fewer symptoms, less severe symptoms, smaller tumors, increased life span, increased life expectancy or prevention of the progression of the disease or condition. Typically, a prophylactic dose is used in subjects prior to or at an earlier stage of disease, so that a prophylactically effective amount can be less than a therapeutically effective amount.

[0091] “Treating” or “treatment” as used herein covers the treatment of the disease or condition of interest in a subject, such as a mammal, preferably a human, having the disease or condition of interest, and includes (but is not limited to): 1. preventing the disease or condition from occurring in a subject, in particular, when such subject is predisposed to the condition but has not yet been diagnosed as having it; 2. inhibiting the disease or condition, e.g., arresting its development; 3. relieving the disease or condition, e.g., causing regression of the disease or condition (ranging from reducing the severity of the disease or condition to curing the disease of condition); or 4. relieving the symptoms resulting from the disease or condition, e.g., relieving pain without addressing the underlying disease or condition. As used herein, the terms “disease” and “condition” can be used interchangeably or can be different in that the particular malady or condition cannot have a known causative agent (so that etiology has not yet been worked out) and it is therefore not yet recognized as a disease but only as an undesirable condition or syndrome, wherein a more or less specific set of symptoms have been identified by clinicians.

[0092] Throughout the present specification, the terms “about” and / or “approximately” can be used in conjunction with numerical values and / or ranges. The term “about” is understood to mean those values near to a recited value. For example, “about 40 [units]” can mean within ± 25% of 40 (e.g., from 30 to 50), within ± 20%, ± 15%, ± 10%, ± 9%, ± 8%, ± ActiveUS 205301239 21Attorney Docket No.: 0019240.01318WO1 Date of Electronic Filing: July 29, 2024 7%, ± 6%, ± 5%, ± 4%, ± 3%, ± 2%, ± 1%, less than ± 1%, or any other value or range of values herein. Furthermore, the phrases “less than about [a value]” or “greater than about [a value]” should be understood in view of the definition of the term “about” provided herein. The terms “about” and “approximately” can be used interchangeably.

[0093] Throughout the present specification, numerical ranges are provided for certain quantities. It is to be understood that these ranges comprise all subranges therein. Thus, the range “from 50 to 80” includes all possible ranges therein (e.g., 51-79, 52-78, 53-77, 54-76, 55-75, 60-70, etc.). Furthermore, all values within a given range can be an endpoint for the range encompassed thereby (e.g., the range 50-80 includes the ranges with endpoints such as 55-80, 50-75, etc.).

[0094] Stimulation of memory associated gene expression is dependent upon cAMP response element-binding (CREB) protein phosphorylation (pCREB), which is required for CREB’s ability to bind to CREB binding protein (CBP). CBP belongs to one of 5 HAT families including 1) p300 / CBP (p300 and CBP), 2) GNAT [Gcn5 and p300 / CBP associated factor (PCAF), HAT1, and ELP3], 3) MYST (Tat-interactive protein-60 - Tip60, MYST1-4), 4) nuclear receptor coactivators (steroid receptor coactivator-1 - SRC-1, ACTR), 5) divers (TAF1, TFIIIC90, P160, CLOCK) (Sadoul K, et al., J Biomedicine & Biotechnology, 2011, 2011:970382). These families show high sequence similarity within each family, but poor to no sequence similarity between families and the size of the HAT domain of each family is different (Marmorstein R, et al., Biochimica et biophysica acta, 2009, 1789(1):58-68). Importantly, several HATs have been involved in numerous memory-related neural processes (Korzus E, et al., Neuron.2004, 42(6):961-72). For instance, transgenic dominant-negative mice with deficient CBP activity have stressed the importance of CBP HAT activity in memory processes. A conditional transgenic mouse expressing an inhibitory truncated form of p300, lacking its HAT domain, showed that p300 HAT activity is required for long-term recognition and contextual fear memory (Oliveira AM, et al., Learning & Memory 2007, 14(9):564-7). PCAF knock-out (KO) mice showed a defect in short-term memory, but not long-term memory, at 2 months of age, whereas at 6 months the KO mice also exhibited long-term memory impairment (Alarcon JM, et al., Neuron, 2004, 42(6):947-59). CBP, p300 and PCAF protein levels were increased during the formation of spatial memory in normal mice (Bousiges O, et al., Neuropsychopharmacology 2010, 35(13):2521-37). Tip60 and (Src- 3) are also involved in the transcription of memory-associated genes (Panikker P, et al., ActiveUS 205301239 22Attorney Docket No.: 0019240.01318WO1 Date of Electronic Filing: July 29, 2024 Journal of Neuroscience: the official journal of the Society for Neuroscience.2018; 38(19):4569-83). Accordingly, several studies showed changes in acetylation of histones, such as H2A, H2B, H3 and H4, in response to memory formation. Consistent with these findings, inhibitors of HDACs (HDACIs) enhance LTP, and fear memory, with HDAC2 inhibition reinstating learning and long-term memory in the CK-p25 transgenic mouse model of neurodegeneration (but see mice lacking HDAC1 in neurons and astrocytes which exhibit marked cognitive deficits as the brain ages, together with astrogliosis and impaired hippocampal LTP thus suggesting that HDAC1 inhibition might negatively affect memory). Altogether these findings suggest that increase in histone acetylation is involved in synaptic plasticity and memory formation.

[0095] Following below are more detailed descriptions of various concepts related to, and embodiments of the compounds and methods disclosed herein for the treatment of cancer and neurodegenerative diseases. It should be appreciated that various concepts introduced above and discussed in greater detail below may be implemented in any of numerous ways, as the disclosed concepts are not limited to any particular manner of implementation. Examples of specific implementations and applications are provided primarily for illustrative purposes. Compounds

[0096] In some aspects of the present disclosure, compounds of Formula (I) are providedwherein, Rais H, OH, CN, C1-C6-alkyl, O-(C3-C8-cycloalkyl), O-(C3-C8-heterocycloalkyl), O-(C2-C6- alkenyl), O-(C1-C6-alkyl), O-(C2-C6-alkyl)-N(R1)2, halogen, or haloalkyl; Rbis H, C1-C6-alkyl, C2-C6-alkenyl, C3-C8-cycloalkyl, C2-C6-heteroalkyl, C3-C8- heterocycloalkyl, aryl, heteroaryl, O-(C1-C6-alkyl), O-(C3-C8-cycloalkyl), O-(C2-C6-alkenyl), or O-(C3-C8-heterocycloalkyl); ActiveUS 205301239 23Attorney Docket No.: 0019240.01318WO1 Date of Electronic Filing: July 29, 2024 Rcis H, C1-C6-alkyl, C1-C6-haloalkyl, O-(C1-C6-alkyl), O-(C1-C6-haloalkyl), halogen, CN, NO2 or CF3; Rdis H, OH, C1-C6-alkyl, O-(C3-C8-cycloalkyl), O-(C3-C8-heterocycloalkyl), O-(C2-C6- alkenyl), O-(C1-C6-alkyl), or O-(C2-C6-alkyl)-N(R1)2; R1is independently H, -(C1-C4-alkyl), -(C1-C4-haloalkyl), -(C3-C8-cycloalkyl), -(C3-C8- heterocycloalkyl), aryl or heteroaryl; W is CH or N; X is -CO-, -CON(R10)-, -CON(R1)(CH2)n-, -(CH2)nN(R1)-, or -C=N-; Z is CH or N, and n is an integer from 1-3, or a pharmaceutically acceptable salt or hydrate thereof.

[0097] In some embodiments, the present disclosure provides compounds of Formula (I), wherein, Rais OH, CN, C1-C6-alkyl, O-(C1-C6-alkyl), halogen, or haloalkyl; Rbis C1-C6-alkyl, C2-C6-heteroalkyl, aryl, heteroaryl, O-(C1-C6-alkyl); Rcis H, C1-C6-alkyl, C1-C6-haloalkyl, halogen, CN or CF3; Rdis OH, C1-C6-alkyl, O-( C1-C6-alkyl), or O-( C2-C6-alkyl)-N(R1)2; R1is independently H, -( C1-C4-alkyl), or -( C1-C4-haloalkyl); W is CH; X is -CON(R1)-, -CON(R1)(CH2)n-; Z is CH, and n is an integer from 1-3, or a pharmaceutically acceptable salt or hydrate thereof.

[0098] In some embodiments, the present disclosure provides compounds of Formula (I), wherein ActiveUS 205301239 24Attorney Docket No.: 0019240.01318WO1 Date of Electronic Filing: July 29, 2024 Rais OH, O-(C1-C6-alkyl), halogen, or haloalkyl; Rbis C1-C6-alkyl, O-(C1-C6-alkyl); Rcis C1-C6-alkyl, or halogen or CF3; Rdis OH, C1-C6-alkyl, O-(C1-C6-alkyl), or O-(C2-C6-alkyl)-N(R1)2; R1is independently H, -(C1-C4-alkyl), or -(C1-C4-haloalkyl); W is CH; X is -CON(R1)-; and Z is CH, or a pharmaceutically acceptable salt or hydrate thereof.

[0099] In some embodiments of Formula (I), X is –C(O)N(R1)-.

[0100] In some embodiments of Formula (I), Rais OH, O-(C1-C6-alkyl), halogen, or haloalkyl.

[0101] In some embodiments of Formula (I), Rbis C1-C6-alkyl, O-(C1-C6-alkyl).

[0102] In some embodiments, the compound of Formula (I) is,wherein Rais OH, O-(C1-C6-alkyl), halogen, or haloalkyl; Rbis C1-C6-alkyl, O-(C1-C6-alkyl); Rcis C1-C6-alkyl, or halogen or CF3; Rdis OH, C1-C6-alkyl, O-(C1-C6-alkyl), or O-(C2-C6-alkyl)-N(R1)2; R1is independently H, -(C1-C4-alkyl), or -(C1-C4-haloalkyl); W is CH; and ActiveUS 205301239 25Attorney Docket No.: 0019240.01318WO1 Date of Electronic Filing: July 29, 2024 Z is CH, or a pharmaceutically acceptable salt or hydrate thereof.

[0103] In some embodiments, the compound of Formula (I) is,wherein Rais OH, O-(C1-C3-alkyl), halogen, or haloalkyl; Rbis C1-C3-alkyl, O-(C1-C3-alkyl); Rcis halogen or CF3Rdis O-(C1-C6-alkyl), or O-(C2-C6-alkyl)-N(R1)2; and R1is independently H, -(C1-C2-alkyl), or -(C1-C4-haloalkyl), or a pharmaceutically acceptable salt or hydrate thereof.

[0104] In some embodiments, the compound of Formula (I) is,wherein Rais O-(C1-C3-alkyl); Rbis O-(C1-C3-alkyl); Rcis Cl; F, and CF3; Rdis O-(C2-C6-alkyl)-N(R1)2; and R1is independently H, -(C1-C2-alkyl), or a pharmaceutically acceptable salt or hydrate thereof. ActiveUS 205301239 26Attorney Docket No.: 0019240.01318WO1 Date of Electronic Filing: July 29, 2024

[0105] In accordance with certain embodiments, the compounds have structures of:or a pharmaceutically acceptable salt or solvate thereof. Methods of Use

[0106] In an embodiment, a HAT activator compound can be used in combination with one or more HDAC inhibitors to treat a neurodegenerative disease in a subject in need thereof. Non-limiting examples of neurodegenerative diseases include Adrenoleukodystrophy (ALD), Alcoholism, Alexander's disease, Alper's disease, Alzheimer's disease, Alzheimer's disease Related Dementia (ADRD), argyrophilic grain disease (AGD), and globular glial tauopathy (GGT), the neurofibrillary tangle-predominant senile dementia (now included also in the category of primary age-related tauopathy, PART), Behavioral variant frontotemporal dementia; Semantic variant primary progressive aphasia, non- fluent / agrammatic variant primary progressive aphasia, logopenic variant primary progressive aphasia, Amyotrophic lateral sclerosis (Lou Gehrig's Disease), Ataxia telangiectasia, Batten disease (also known as Spielmeyer-Vogt-Sjögren-Batten disease), Bovine spongiform encephalopathy (BSE), Canavan disease, Cockayne syndrome, Corticobasal degeneration, Creutzfeldt-Jakob disease, Familial fatal insomnia, Frontotemporal lobar degeneration, Huntington's disease, HIV-associated dementia, Kennedy's disease, Krabbe's disease, Lewy body dementia, Neuroborreliosis, Machado- Joseph disease (Spinocerebellar ataxia type 3), Multiple System Atrophy, Multiple sclerosis, Narcolepsy, Niemann Pick disease, Parkinson's disease, Pelizaeus-Merzbacher Disease, Pick's disease, Primary lateral sclerosis, Prion diseases, Progressive Supranuclear Palsy, Refsum's disease, Rett’s syndrome, Tau-positive FrontoTemporal dementia, Tau-negative FrontoTemporal dementia, Sandhoff disease, Tauopathy, Schilder's disease, Subacute combined degeneration of spinal cord secondary to Pernicious Anaemia, Spielmeyer-Vogt- ActiveUS 205301239 27Attorney Docket No.: 0019240.01318WO1 Date of Electronic Filing: July 29, 2024 Sjogren-Batten disease (also known as Batten disease), Spinocerebellar ataxia (multiple types with varying characteristics), Spinal muscular atrophy, Steele-Richardson-Olszewski disease, Tabes dorsalis, and Toxic encephalopathy.

[0107] In some embodiments, the neurodegenerative disease is selected from Alzheimer’s Disease, ADRD, Tauopathy, ALS, Parkinson’s Disease, and Huntington’s Disease. In some embodiments, the neurodegenerative disease is Alzheimer’s Disease. In some embodiments, the neurodegenerative disease is ADRD. In some embodiments, the neurodegenerative disease is tauopathy. In some embodiments, the neurodegenerative disease is ALS. In some embodiments, the neurodegenerative disease is Parkinon’s Disease. In some embodiments, the neurodegenerative disease is Huntington’s Disease.

[0108] Non-limiting examples of cancers include B cell lymphoma, colon cancer, lung cancer, renal cancer, bladder cancer, T cell lymphoma, myeloma, leukemia, chronic myeloid leukemia, acute myeloid leukemia, chronic lymphocytic leukemia, acute lymphocytic leukemia, hematopoietic neoplasias, thymoma, lymphoma, sarcoma, lung cancer, liver cancer, non-Hodgkin’s lymphoma, Hodgkin’s lymphoma, uterine cancer, renal cell carcinoma, hepatoma, adenocarcinoma, breast cancer, pancreatic cancer, liver cancer, prostate cancer, head and neck carcinoma, thyroid carcinoma, soft tissue sarcoma, ovarian cancer, primary or metastatic melanoma, squamous cell carcinoma, basal cell carcinoma, brain cancer, angiosarcoma, hemangiosarcoma, bone sarcoma, fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma, osteogenic sarcoma, chordoma, angiosarcoma, endotheliosarcoma, lymphangiosarcoma, lymphangioendotheliosarcoma, synovioma, testicular cancer, uterine cancer, cervical cancer, gastrointestinal cancer, mesothelioma, Ewing's tumor, leiomyosarcoma, rhabdomyosarcoma, colon carcinoma, pancreatic cancer, breast cancer, ovarian cancer, prostate cancer, squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, sweat gland carcinoma, sebaceous gland carcinoma, papillary carcinoma, Waldenstroom's macroglobulinemia, papillary adenocarcinomas, cystadenocarcinoma, bronchogenic carcinoma, bile duct carcinoma, choriocarcinoma, seminoma, embryonal carcinoma, Wilms' tumor, lung carcinoma, epithelial carcinoma, cervical cancer, testicular tumor, glioblastoma, glioma, diffuse midline glioma, astrocytoma, medulloblastoma, craniopharyngioma, ependymoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma, meningioma, retinoblastoma, leukemia, melanoma, ActiveUS 205301239 28Attorney Docket No.: 0019240.01318WO1 Date of Electronic Filing: July 29, 2024 neuroblastoma, small cell lung carcinoma, bladder carcinoma, lymphoma, multiple myeloma, follicular lymphoma and medullary carcinoma.

[0109] In some embodiments, the cancer is colon cancer, lung cancer, renal cancer, leukemia, CNS cancer, melanoma, ovarian cancer, breast cancer, or prostate cancer.

[0110] In some embodiments, the cancer is colon cancer, renal cancer, T cell leukemia, myeloma, leukemia, acute myeloid leukemia, acute lymphocytic leukemia, renal cell carcinoma, adenocarcinoma, glioblastoma, breast carcinoma, prostate carcinoma, or lung carcinoma.

[0111] In some embodiments, the cancer is Hodgkin’s lymphoma, non-Hodgkin’s lymphoma, B cell lymphoma, T cell lymphoma, or follicular lymphoma. In other embodiments, the B cell lymphoma is diffuse large B-cell lymphoma. In further embodiments, the diffuse large B-cell lymphoma is a germinal center-derived diffuse large B cell lymphoma, an activated B-cell-derived (ABC) diffuse large B-cell lymphoma, or a non- germinal center diffuse large B cell lymphoma.

[0112] In some embodiments, the cancer is glioblastoma, or diffuse midline glioma.

[0113] Epigenetic modifications including acetylation of histones may contribute to gene expression changes important to learning and memory (Science 2010: 328(5979), 701-702; herein incorporated by reference in its entirety). Addition of acetyl groups to histones by enhances gene expression, while their removal by HDACs reduces gene expression. Reduction in histone acetylation has been linked to age-induced memory impairment and various neurodegenerative diseases (Science 2010: 328(5979), 701-702; herein incorporated by reference in its entirety). HDAC inhibitors have been shown to enhance memory in mice (Nature 459, 55-60 (7 May 2009); herein incorporated by reference in its entirety). Although clinical trials of several HDAC inhibitors are currently underway to try to prevent deacetylation, the alternative strategy of increasing histone acetylation by activating HAT has not been significantly explored.

[0114] In some embodiments, the present disclosure provides for compounds with histone acetyltransferase activity which can be used in combination with one or more HDAC inhibitors to treat patients with cancers or neurodegenerative diseases. In some embodiments, the compounds are HAT activators. In some embodiments, the compounds are HDAC ActiveUS 205301239 29Attorney Docket No.: 0019240.01318WO1 Date of Electronic Filing: July 29, 2024 inhibitors. In some embodiments, the compounds have good HAT activation potency, high selectivity, reasonable pharmacokinetics and / or good permeability across the blood-brain- barrier (BBB). In some embodiments, these compounds can be used as therapy with decreased side effects for AD patients. In some embodiments, the compounds improve cognition or memory in AD and Alzheimer’s-like pathologies, as well as minimize the side effects for subjects afflicted with other neurodegenerative diseases. In some embodiments, the compounds disclosed herein can also be developed as anti-cancer therapies. In some embodiments, acetylation of histone proteins increases gene expression in a subject resulting in enhanced memory and cognition.

[0115] In some embodiments, the present disclosure provides a method for reducing amyloid beta (Aβ) protein deposits in a subject in need thereof, the method comprising administering to the subject a HAT activator and, optionally, a HDAC inhibitor. In some embodiments, the subject exhibits abnormally elevated levels of amyloid beta plaques. In some embodiments, the subject is afflicted with Alzheimer's disease, ADRD, Tauopathy, Lewy body dementia, inclusion body myositis, or cerebral amyloid angiopathy.

[0116] In some embodiments, the present disclosure provides a method for reducing tau protein deposits in a subject in need thereof, the method comprising administering to the subject a HAT activator and, optionally, a HDAC inhibitor. In some embodiments, the subject exhibits abnormally elevated levels of neurofibrillary tangles. In some embodiments, the subject is afflicted with Alzheimer's disease, ADRD, Tauopathy.

[0117] In further embodiments, the present disclosure provides for the utilization of HAT agonists optionally in combination with one or more HDAC inhibitors as memory enhancers in normal subjects (for example, a subject not afflicted with a neurodegenerative disease). In further embodiments, the present disclosure provides for the utilization of HAT agonists, optionally in combination with one or more HDAC inhibitors as memory enhancers in aging subjects (for example, a subject who is >55 years old). In further embodiments, the present disclosure provides for the utilization of HAT agonists optionally in combination with one or more HDAC inhibitors as memory enhancers for other conditions associated with cognitive decrease / impairment. Non-limiting examples of conditions associated with cognitive decrease / impairment include a variety of syndromes associated with mental retardation and syndromes associated with learning disabilities, Parkinson’s disease, Pick’s disease, argyrophilic grain disease (AGD), and globular glial tauopathy (GGT), the neurofibrillary ActiveUS 205301239 30Attorney Docket No.: 0019240.01318WO1 Date of Electronic Filing: July 29, 2024 tangle-predominant senile dementia (now included also in the category of primary age-related tauopathy, PART), Behavioral variant frontotemporal dementia; Semantic variant primary progressive aphasia, non-fluent / agrammatic variant primary progressive aphasia, logopenic variant primary progressive aphasia, a Lewy body disease, amyotrophic lateral sclerosis, Huntington’s disease, Creutzfeld-Jakob disease, Down syndrome, multiple system atrophy, neuronal degeneration with brain iron accumulation type I (Hallervorden-Spatz disease), pure autonomic failure, REM sleep behavior disorder, mild cognitive impairment (MCI), cerebral amyloid angiopathy (CAA), mild cognitive deficits, aging, vascular dementias mixed with Alzheimer’s disease, a neurodegenerative disease characterized by abnormal amyloid deposition, and any combination thereof.

[0118] In some embodiments, the present disclosure provides methods for identifying a one or more HAT activators and, optionally, one or more HDAC inhibitors that can acetylate histone proteins thus increasing gene expression in a subject resulting in enhanced memory and cognition. In some embodiments, the present disclosure provides methods for identifying one or more HAT activators and, optionally, one or more HDAC inhibitors can acetylate histone proteins thus increasing gene expression in a subject resulting in enhanced memory and cognition.

[0119] To shrink the candidate pool of HAT modulator and, optionally, HDAC modulator combinations to be tested in animal models of neurodegenerative diseases, such as animals that exhibit elevated levels of inclusion bodies, for example Aβ accumulation animal models (e.g., animal models of AD), or, for example, or tau accumulation animal models (e.g. animal model of tauopathy), or a mouse model for Huntington’s disease, HAT activators or HDAC inhibitors can first be screened or selected based on their possession of certain characteristics, such as having one or more of: an EC50 no greater than about 500 nM; a histone acetylation activity in vitro; and the ability to penetrate the BBB. HAT activator and HDAC inhibitor combinations can first be screened or selected based on their possession of certain characteristics, such as having a histone acetylation activity in vitro or resulting in increased histone acetylation in vitro compared to histone acetylation in vitro of the HAT activator or HDAC inhibitor alone.

[0120] In some embodiments, the candidate pool of HAT activator and, optionally, HDAC inhibitor combinations can be tested in animal models of neurodegenerative diseases, such as, but not limited to, animals that exhibit elevated levels of inclusion bodies, for ActiveUS 205301239 31Attorney Docket No.: 0019240.01318WO1 Date of Electronic Filing: July 29, 2024 example Aβ accumulation animal models (e.g., animal models of AD), or tau accumulation animal models (e.g. animal model of tauopathy), or a mouse model for Huntington’s disease to determine whether they increase gene expression in a subject resulting in enhanced memory and cognition. As used herein, a HAT activator compound does not necessarily preclude the possibility that the compound may also be able to inhibit other HATs. As used herein, a HDAC inhibitor compound does not necessarily preclude the possibility that the compound may also be able to activate other HDACs.

[0121] A HAT activator compound can be a compound that increases the activity and / or expression of a HAT molecule (e.g., p300, CBP, GCN5, GCN5L, PCAF, or HAT1) in vivo and / or in vitro. HAT activator compounds can be compounds that exert their effect on the activity of a HAT protein via the expression, via post-translational modifications, or by other means. In some embodiments, a HAT activator compound increases HAT protein or mRNA expression, or acetyltransferase activity by at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 97%, at least about 99%, or 100%.

[0122] A HDAC inhibitor compound can be a compound that decreases the activity and / or expression of a HDAC molecule in vivo and / or in vitro. HDAC modulator compounds can be compounds that exert their effect on the activity of a HDAC protein via the expression, via post-translational modifications, or by other means. In some embodiments, a HDAC inhibitor compound decreases HDAC protein or mRNA expression, or deacetyltransferase activity by at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 97%, at least about 99%, or 100%.

[0123] Test compounds or agents that bind to a HAT molecule (such as p300, CBP, GCN5, GCN5L, PCAF, or HAT1), and / or have a stimulatory effect on the activity or the expression of a HAT molecule, can be identified by various assays. The assay can be a binding assay comprising direct or indirect measurement of the binding of a test compound or a known HAT ligand to the active site of a HAT protein. The assay can also be an activity assay comprising direct or indirect measurement of the activity of a HAT molecule. The assay can also be an expression assay comprising direct or indirect measurement of the ActiveUS 205301239 32Attorney Docket No.: 0019240.01318WO1 Date of Electronic Filing: July 29, 2024 expression of a HAT mRNA or protein. The various screening assays can be combined with an in vivo assay comprising measuring the effect of the test compound on cognitive and synaptic function in an animal model for neurodegenerative disorders, such as, but not limited to, AD, ADRD, tauopathy, Parkinson’s Disease, ALS, or Huntington’s Disease. The assay can be an assay comprising measuring the effect of the test compounds on cell viability. In one embodiment, the cells are cancer cells, such as, but not limited to B-cell lymphoma cell lines, T-cell lymphoma cell lines (e.g. Ly1, Ly7, Ly10, SU-DHL2, HH, or H9 cell lines), or glioblastoma.

[0124] Activators of the expression of a HAT molecule can also be identified via contacting a HAT-positive cell or tissue with a test compound and determining the expression of a HAT protein or HAT mRNA in the cell. The protein or mRNA expression level of a HAT molecule in the presence of the test compound can be compared to the protein or mRNA expression level of a HAT protein in the absence of the test compound. The test compound can then be identified as an activator of expression of a HAT protein (such as p300, CBP, PCAF, TIP60, Src-3, GCN5, GCN5L, or HAT1) based on this comparison. For example, when expression of HAT protein or mRNA is statistically or significantly more in the presence of the test compound than in its absence, the compound is identified as an activator of the expression of a HAT protein or mRNA. In other words, the test compound can also be a HAT activator compound (such as an agonist). The expression level of a HAT protein or mRNA in cells can be determined by methods described herein.

[0125] Determining the ability of a test compound to bind to a HAT molecule, a HDAC molecule or a variant thereof can be accomplished using real-time Bimolecular Interaction Analysis (BIA) [McConnell, (1992); Sjolander, S., and Urbaniczky, C. Integrated fluid handling system for biomolecular interaction analysis. Anal. Chem.1991, 63, 2338-2345; herein incorporated by reference in its entirety]. BIA is a technology for studying biospecific interactions in real time, without labeling any of the interactants (e.g., BIA-core™). Changes in optical phenomenon surface plasmon resonance (SPR) can be used as an indication of real- time reactions between biological molecules.

[0126] In some embodiments, the present disclosure provides for compounds that bind to a HAT activator protein, such as p300, CBP, GCN5, PCAF, TIP60, Src-3, GCN5, GCN5L, or HAT1. These compounds can be identified by the screening methods and assays described herein, and enhance the activity or expression of HAT activator proteins. ActiveUS 205301239 33Attorney Docket No.: 0019240.01318WO1 Date of Electronic Filing: July 29, 2024

[0127] Test compounds or agents that bind to a HAT molecule and / or have a stimulatory effect on the activity or the expression of a HAT molecule, can be combined with one or more test compounds or agents that bind to a HDAC molecule. The assay can be an activity assay comprising direct or indirect measurement of the activity of a HAT molecule and / or a HDAC molecule. The assay can also be an expression assay comprising direct or indirect measurement of the expression of a HAT mRNA or protein and / or a HDAC mRNA or protein. The various screening assays can be combined with an in vivo assay comprising measuring the effect of a HAT activator and a HDAC inhibitor on cognitive and synaptic function in an animal model for neurodegenerative disorders, such as, but not limited to, AD, ADRD, tauopathy, Parkinson’s Disease, ALS, or Huntington’s Disease. The assay can be an assay comprising measuring the effect of the test compounds on cell viability. In one embodiment, the cells are cancer cells, such as, but not limited to B-cell lymphoma cell lines, T-cell lymphoma cell lines, or glioblastoma. In one embodiment, the effect of a HAT activator and one or more HDAC inhibitors in combination is compared to the effect of a HAT activator or HDAC inhibitor alone. Pharmaceutical Compositions

[0128] In some embodiments, the present disclosure provides pharmaceutical compositions comprising an effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof. In some embodiments, the pharmaceutical compositions provided herein comprise one or more pharmaceutically acceptable carriers or excipients.

[0129] In various embodiments, the pharmaceutical compositions of the present disclosure can be formulated for administration by a variety of means including orally, parenterally, by inhalation spray, topically, or rectally in formulations containing pharmaceutically acceptable carriers, adjuvants and vehicles. The term parenteral as used here includes subcutaneous, intravenous, intramuscular, and intraarterial injections with a variety of infusion techniques. Intraarterial and intravenous injection as used herein includes administration through catheters.

[0130] The effective amount of a compound of Formula (I), pharmaceutically acceptable salts, esters, prodrugs, hydrates, solvates and isomers thereof, or a pharmaceutical ActiveUS 205301239 34Attorney Docket No.: 0019240.01318WO1 Date of Electronic Filing: July 29, 2024 composition comprising a compound of Formula (I) or a pharmaceutically acceptable salt thereof may be determined by one skilled in the art based on known methods.

[0131] In one embodiment, a pharmaceutical composition or a pharmaceutical formulation of the present disclosure comprises a compound of Formula (I) or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier, diluent, and / or excipient. Pharmaceutically acceptable carriers, diluents or excipients include without limitation any adjuvant, carrier, excipient, glidant, sweetening agent, diluent, preservative, dye / colorant, flavor enhancer, surfactant, wetting agent, dispersing agent, suspending agent, stabilizer, isotonic agent, solvent, or emulsifier which has been approved by the United States Food and Drug Administration as being acceptable for use in humans or domestic animals.

[0132] In one embodiment, suitable pharmaceutically acceptable carriers include, but are not limited to, inert solid fillers or diluents and sterile aqueous or organic solutions. Pharmaceutically acceptable carriers are well known to those skilled in the art and include, but are not limited to, from about 0.01 to about 0.1 M and preferably 0.05M phosphate buffer or 0.8% saline. Such pharmaceutically acceptable carriers can be aqueous or non-aqueous solutions, suspensions and emulsions. Examples of non-aqueous solvents suitable for use in the present application include, but are not limited to, propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable organic esters such as ethyl oleate.

[0133] Aqueous carriers suitable for use in the present application include, but are not limited to, water, ethanol, alcoholic / aqueous solutions, glycerol, emulsions or suspensions, including saline and buffered media. Oral carriers can be elixirs, syrups, capsules, tablets and the like.

[0134] Liquid carriers suitable for use in the present application can be used in preparing solutions, suspensions, emulsions, syrups, elixirs and pressurized compounds. The active ingredient can be dissolved or suspended in a pharmaceutically acceptable liquid carrier such as water, an organic solvent, a mixture of both or pharmaceutically acceptable oils or fats. The liquid carrier can contain other suitable pharmaceutical additives such as solubilizers, emulsifiers, buffers, preservatives, sweeteners, flavoring agents, suspending agents, thickening agents, colors, viscosity regulators, stabilizers or osmo-regulators.

[0135] Liquid carriers suitable for use in the present application include, but are not limited to, water (partially containing additives as above, e.g. cellulose derivatives, preferably ActiveUS 205301239 35Attorney Docket No.: 0019240.01318WO1 Date of Electronic Filing: July 29, 2024 sodium carboxymethyl cellulose solution), alcohols (including monohydric alcohols and polyhydric alcohols, e.g. glycols) and their derivatives, and oils (e.g. fractionated coconut oil and arachis oil). For parenteral administration, the carrier can also include an oily ester such as ethyl oleate and isopropyl myristate. Sterile liquid carriers are useful in sterile liquid form comprising compounds for parenteral administration. The liquid carrier for pressurized compounds disclosed herein can be halogenated hydrocarbon or other pharmaceutically acceptable propellant.

[0136] Solid carriers suitable for use in the present application include, but are not limited to, inert substances such as lactose, starch, glucose, methyl-cellulose, magnesium stearate, dicalcium phosphate, mannitol and the like. A solid carrier can further include one or more substances acting as flavoring agents, lubricants, solubilizers, suspending agents, fillers, glidants, compression aids, binders or tablet-disintegrating agents; it can also be an encapsulating material. In powders, the carrier can be a finely divided solid which is in admixture with the finely divided active compound. In tablets, the active compound is mixed with a carrier having the necessary compression properties in suitable proportions and compacted in the shape and size desired. The powders and tablets preferably contain up to 99% of the active compound. Suitable solid carriers include, for example, calcium phosphate, magnesium stearate, talc, sugars, lactose, dextrin, starch, gelatin, cellulose, polyvinylpyrrolidine, low melting waxes and ion exchange resins. A tablet may be made by compression or molding, optionally with one or more accessory ingredients. Compressed tablets may be prepared by compressing in a suitable machine the active ingredient in a free flowing form such as a powder or granules, optionally mixed with a binder (e.g., povidone, gelatin, hydroxypropylmethyl cellulose), lubricant, inert diluent, preservative, disintegrant (e.g., sodium starch glycolate, cross-linked povidone, cross-linked sodium carboxymethyl cellulose) surface active or dispersing agent. Molded tablets may be made by molding in a suitable machine a mixture of the powdered compound moistened with an inert liquid diluent. The tablets may optionally be coated or scored and may be formulated so as to provide slow or controlled release of the active ingredient therein using, for example, hydroxypropyl methylcellulose in varying proportions to provide the desired release profile. Tablets may optionally be provided with an enteric coating, to provide release in parts of the gut other than the stomach. ActiveUS 205301239 36Attorney Docket No.: 0019240.01318WO1 Date of Electronic Filing: July 29, 2024

[0137] Parenteral carriers suitable for use in the present application include, but are not limited to, sodium chloride solution, Ringer's dextrose, dextrose and sodium chloride, lactated Ringer's and fixed oils. Intravenous carriers include fluid and nutrient replenishers, electrolyte replenishers such as those based on Ringer's dextrose and the like. Preservatives and other additives can also be present, such as, for example, antimicrobials, antioxidants, chelating agents, inert gases and the like.

[0138] Carriers suitable for use in the present application can be mixed as needed with disintegrants, diluents, granulating agents, lubricants, binders and the like using conventional techniques known in the art. The carriers can also be sterilized using methods that do not deleteriously react with the compounds, as is generally known in the art.

[0139] Diluents may be added to the formulations of the present disclosure. Diluents increase the bulk of a solid pharmaceutical composition and / or combination and may make a pharmaceutical dosage form containing the composition and / or combination easier for the patient and care giver to handle. Diluents for solid compositions and / or combinations include, for example, microcrystalline cellulose (e.g., AVICEL), microfine cellulose, lactose, starch, pregelatinized starch, calcium carbonate, calcium sulfate, sugar, dextrates, dextrin, dextrose, dibasic calcium phosphate dihydrate, tribasic calcium phosphate, kaolin, magnesium carbonate, magnesium oxide, maltodextrin, mannitol, polymethacrylates (e.g., EUDRAGIT®), potassium chloride, powdered cellulose, sodium chloride, sorbitol, and talc.

[0140] The pharmaceutical composition of the present invention may be prepared into any type of formulation and drug delivery system by using any of the conventional methods well-known in the art. The inventive pharmaceutical composition may be formulated into injectable formulations, which may be administered by routes including intrathecal, intraventricular, intravenous, intraperitoneal, intranasal, intraocular, intramuscular, subcutaneous or intraosseous. Also, it may also be administered orally, or parenterally through the rectum, the intestines or the mucous membrane in the nasal cavity (see Gennaro, A. R., ed. (1995) Remington's Pharmaceutical Sciences). The composition may be administered topically, instead of enterally. For instance, the composition may be injected, or delivered via a targeted drug delivery system such as a reservoir formulation or a sustained release formulation. ActiveUS 205301239 37Attorney Docket No.: 0019240.01318WO1 Date of Electronic Filing: July 29, 2024

[0141] The pharmaceutical formulation of the present invention may be prepared by any well-known methods in the art, such as mixing, dissolving, granulating, dragee-making, levigating, emulsifying, encapsulating, entrapping, or lyophilizing processes. As mentioned above, the compositions of the present invention may include one or more physiologically acceptable carriers such as excipients and adjuvants that facilitate processing of active molecules into preparations for pharmaceutical use.

[0142] Proper formulation is dependent upon the route of administration chosen. For injection, for example, the composition may be formulated in an aqueous solution, preferably in physiologically compatible buffers such as Hank's solution, Ringer's solution, or physiological saline buffer. For transmucosal or nasal administration, penetrants appropriate to the barrier to be permeated are used in the formulation. Such penetrants are generally known in the art. In a one embodiment, the compound may be prepared in an oral formulation. For oral administration, the compounds can be formulated readily by combining the active compounds with pharmaceutically acceptable carriers known in the art. Such carriers enable the disclosed compound to be formulated as tablets, pills, dragees, capsules, liquids, gels, syrups, slurries, suspensions and the like, for oral ingestion by a subject. The compounds may also be formulated in rectal compositions such as suppositories or retention enemas, e.g., containing conventional suppository bases such as cocoa butter or other glycerides.

[0143] Pharmaceutical preparations for oral use may be obtained as solid excipients, optionally grinding a resulting mixture, and processing the mixture of granules, after adding suitable adjuvants, if desired, to obtain tablets or dragee cores. Suitable excipients may be, in particular, fillers such as sugars, including lactose, sucrose, mannitol, or sorbitol; cellulose formulation such as maize starch, wheat starch, rice starch, potato starch, gelatin, gum tragacanth, methyl cellulose, hydroxypropylmethyl-cellulose, sodium carboxymethylcellulose, and / or polyvinylpyrrolidone (PVP) formulation. Also, disintegrating agents may be employed, such as cross-linked polyvinylpyrrolidone, agar, or alginic acid or a salt thereof such as sodium alginate. Also, wetting agents, such as sodium dodecyl sulfate and the like, may be added. ActiveUS 205301239 38Attorney Docket No.: 0019240.01318WO1 Date of Electronic Filing: July 29, 2024 EXAMPLES

[0144] Examples are provided below to facilitate a more complete understanding of the invention. The following examples illustrate the exemplary modes of making and practicing the invention. However, the scope of the invention is not limited to specific embodiments disclosed in these Examples, which are for purposes of illustration only, since alternative methods can be utilized to obtain similar results. Synthesis of Compounds of Formula (I)

[0145] General schemes of synthesis for EZ115HAT are provided below.

[0146] FIG.2A provides a synthesis scheme in accordance with one embodiment. Reagents and conditions: CsOH (5.6M), THF, reflux 72h, 95% yielded; ii) DMAP, EDC HCl, CH2Cl2 in presence of 5-chloro-6-methoxypyridin-3-amine, overnight, RT, 76%yielded; iii) Cs2CO3, dioxane, in presence of 2-bromo-N,N-dimethylethan-1-amine, 70°C, 16 h, 57% yielded. i) ethyl 2-ethoxy-6-hydroxybenzoate (1) (10.00g, 0.045mol) was dissolved in THF (90ml), Cs(OH) 5.6 M solution (90.6ml) was poured into the solution. The mixture reaction was stirred at reflux for 72h. The reaction was cooled down and diluted with ethyl acetate (90ml) and then treated with HCl 1 N until pH=2. The white precipitate was filtrated, washed 3 times with H2O and dried under vacuum. The desired compound was used in the following step without any other purification.95 % yield. ii) 2-ethoxy-6-hydroxybenzoic acid (2) (10g, 0.05mol), DMAP (3.35g, 0.027mol) and the appropriate aromatic amine (0.06mol) were dissolved in CH2Cl2anhydrous (71.36ml) at room temperature under argon atmosphere. EDC HCl (13.42g, 0.07mol) was added portion-wise every 30 minutes at 0°C. The milky reaction mixture was stirred for 24h at room temperature under argon atmosphere. The solvent was evaporated off and the crude was treated with hot methanol (200ml) and a white precipitate was obtained. The precipitate was washed with hot methanol (100ml) and dried under vacuum to afford the desired product 3a (76% yielded). ActiveUS 205301239 39Attorney Docket No.: 0019240.01318WO1 Date of Electronic Filing: July 29, 2024 iii) 2-ethoxy-6-hydroxybenzoic acid (2) (10g, 0.05mol), DMAP (3.35g, 0.027mol) and the appropriate aromatic amine (0.06mol) were dissolved in CH2Cl2 anhydrous (71.36ml) at room temperature under argon atmosphere. EDC HCl (13.42g, 0.07mol) was added portion-wise every 30 minutes at 0°C. The milky reaction mixture was stirred for 24h at room temperature under argon atmosphere. The solvent was evaporated off and the crude was treated with hot methanol (200ml) and a white precipitate was obtained. The precipitate was washed with hot methanol (100ml) and dried under vacuum to afford the desired product 3a (76% yielded). iv) iii) N-(5-chloro-6-methoxypyridin-3-yl)-2-ethoxy-6-hydroxybenzamide (3a) (4.29g, 0.0133mol) was dissolved in 1-4 Dioxane anhydrous (30ml) , Cs2CO3 (0.0398mol) was added and the mixture was heated at 50°C for 1h.2-bromo-N,N- dimethylethan-1-amine (2.49g, 0.0173mol) was added and the reaction was stirred at 50°C for 16h. The crude was extracted with Ethyl-Acetate and water. The organic phase was collected by filtration and dried on Na2SO4. Flash chromatography, CH2Cl2 / MeOH 9:1 to give the desired product as free base EZ115HAT, 57% yielded (white solid).

[0147] The HCl salt form can be produced as follows in accordance with one embodiment: EZ115HAT free base (2g, 0.00507mol) was dissolved in 1:1 MTBE / Dichloromethane solution (50ml) and stirred a RT.4 N HCl in Dioxane anhydrous (0.1014mol, 3.69ml) solution was added dropwise. The reaction was stirred at RT overnight. The white precipitate was filtered under vacuum and washed with Dichloromethane (2 X 20ml). The white powder was dried under vacuum overnight to obtain EZ115HAT HCl salt, 97% yielded.

[0148] FIG.2B provides a synthesis scheme in accordance with another embodiment. This embodiment provides improved and optimized yield (>90%) and purity (>99%) of compound EZ115HAT, while also reducing the costs of the starting materials. i) Preparation of Phenoxyamine-209 on 140g scale. The intermediate Phenoxyamine-209 was prepared as a crude oil (143 gr) in 97% yield. The crude is ready to be used in the next carboxylation for process optimization. ActiveUS 205301239 40Attorney Docket No.: 0019240.01318WO1 Date of Electronic Filing: July 29, 2024 ii) Telescoping Carboxylation of Phenoxyamine-209 and Conversion to aminoacid- HCl. The telescoping process was developed for carboxylation of phenoxyamine- 209 to aminoacid-253 and conversion to aminoacid-HCl, using the optimized new process, the 40mol% of pentanoic-acid by-product in aminoacid-253-Li from carboxylation was completely purged after conversion of the Li-salt to aminoacid- 253-HCl and crystallization in MTBE / DCM, to provide aminoacid-253-HCl (2.4 gr assay wt, 97A% with 0 mol% pentanoic-acid) in 84% yield. iii) Amidation of aminoacid-253-HCl with pyridine-amine-158 to EZ115HAT using EDC and crystallization of EZ115HAT. The aminoacid-253-HCl (1.16 gr assay weight, 1 eq) in DCM (20V) was added with EDC-HCl (1.0eq) and then a solution of pyridinamine-158 (0.85eq) / DCM ( 10V) dropwise at room temperature and stirred for >1 hour at room temperature. 1h HPLC indicated 83% of EZ115HAT, 1.7% aminoacid-253 and 3.1% pyridine-amine and 9.6% of di-imine (MW= 313, the dimer of pyridine-amune-158). After work-up, crude EZ115HAT (1.2 gr assay wt. 83%) was obtained in 76% yield. The crude (1.2gr assay wt) was crystallized in 2-MeTHF (4V) and Heptane (8V) from 60 °C to 20 °C to give the purified final compound EZ115HAT.

[0149] A general scheme of synthesis for EZ199HAT in accordance with one embodiment is provided below.Reagents and conditions:

[0150] CsOH (5.6M), THF, reflux 72h, 95% yielded; ii) DMAP, EDC HCl, CH2Cl2 in presence of 5-chloro-6-methoxypyridin-3-amine, overnight, RT, 76%yielded; iii) Cs2CO3, dioxane, in presence of 2-bromo-N,N-dimethylethan-1-amine, 70oC, 16 h, 57% yielded. i) ethyl 2-ethoxy-6-hydroxybenzoate (1) (10.00g, 0.045mol) was dissolved in THF (90ml), Cs(OH) 5.6 M solution (90.6ml) was poured into the solution. The mixture reaction was stirred at reflux for 72h. The reaction was cooled down and diluted with ethyl acetate (90ml) ActiveUS 205301239 41Attorney Docket No.: 0019240.01318WO1 Date of Electronic Filing: July 29, 2024 and then treated with HC11 N until pH=2. The white precipitate was filtrated, washed 3 times with H2O and dried under vacuum. The desired compound was used in the following step without any other purification. 95% yield. ii) 2-ethoxy-6-hydroxybenzoic acid (2) (10g, 0.05mol), DMAP (3.35g, 0.027mol) and the appropriate aromatic amine (0.06mol) were dissolved in CH2Cl2 anhydrous (71.36ml) at room temperature under argon atmosphere. EDC HC1 (13.42g, 0.07mol) was added portion- wise every 30 minutes at 0°C. The milky reaction mixture was stirred for 24h at room temperature under argon atmosphere. The solvent was evaporated off and the crude was treated with hot methanol (200ml) and a white precipitate was obtained. The precipitate was washed with hot methanol (100ml) and dried under vacuum to afford the desired product 3b (65% yielded). iii) 2-ethoxy-6-hydroxy-N-(6-methoxy-5-(trifluoromethyl)pyridin-3-yl)benzamide (3b) (4.29g, 0.0133mol) was dissolved in 1-4Dioxane anhydrous (30ml) , Cs2CO3(0.0398mol) was added and the mixture was heated at 50°C for 1h. 2-bromo-N,N-dimethylethan-1-amine (2.49g, 0.0173mol) was added and the reaction and was stirred at 50°C for 16h. The crude was extracted with Ethyl-Acetate and water. The organic phase was collected by filtration and dried on Na2SO4. Flash chromatography, CH2Cl2 / MeOH 9:1 to give the desired product as free base EZ199HAT, 50% yielded (white solid).

[0151] A general scheme of synthesis for EZ202HAT in accordance with one embodiment is provided below.Reagents and conditions:

[0152] CsOH (5.6M), THF, reflux 72h, 95% yielded; ii) DMAP, EDC HCl, CH2Cl2in presence of 5-flouro-6-methoxypyridin-3-amine, overnight, RT, 76%yielded; iii) Cs2CO3, dioxane, in presence of 2-bromo-N,N-dimethylethan-1-amine, 70oC, 16 h, 50%yielded. ActiveUS 205301239 42Attorney Docket No.: 0019240.01318WO1 Date of Electronic Filing: July 29, 2024 i) ethyl 2-ethoxy-6-hydroxybenzoate (1) (10.00g, 0.045mol) was dissolved in THF (90ml), Cs(OH) 5.6 M solution (90.6ml) was poured into the solution. The mixture reaction was stirred at reflux for 72h. The reaction was cooled down and diluted with ethyl acetate (90ml) and then treated with HC11 N until pH=2. The white precipitate was filtrated, washed 3 times with H2O and dried under vacuum. The desired compound was used in the following step without any other purification.95% yield. ii) 2-ethoxy-6-hydroxybenzoic acid (2) (10g, 0.05mol), DMAP (3.35g, 0.027mol) and the appropriate aromatic amine (0.06mol) were dissolved in CH2Cl2 anhydrous (71.36ml) at room temperature under argon atmosphere. EDC HC1 (13.42g, 0.07mol) was added portion- wise every 30 minutes at 0°C. The milky reaction mixture was stirred for 24h at room temperature under argon atmosphere. The solvent was evaporated off and the crude was treated with hot methanol (200ml) and a white precipitate was obtained. The precipitate was washed with hot methanol (100ml) and dried under vacuum to afford the desired product 3c (73% yielded). iii) 2-ethoxy-N-(5-fluoro-6-methoxypyridin-3-yl)-6-hydroxybenzamide (3c) (4.29g, 0.0133mol) was dissolved in 1-4Dioxane anhydrous (30ml) , Cs2CO3(0.0398mol) was added and the mixture was heated at 50°C for 1h. 2-bromo-N,N-dimethylethan-1-amine (2.49g, 0.0173mol) was added and the reaction and was stirred at 50°C for 16h. The crude was extracted with Ethyl-Acetate and water. The organic phase was collected by filtration and dried on Na2SO4. Flash chromatography, CH2Cl2 / MeOH 9:1 to give the desired product as free base EZ101HAT, 57% yielded (white solid).

[0153] EZ115HAT compound showed improved physico-chemical properties compared to the other HAT activators, such as RA013915 HAT activators (disclosed in WO 2020 / 163731A1, hereby incorporated by reference), or YF2 HAT activator (disclosed in U.S. Pat. No.10,640,457 hereby incorporated by reference). For example, EZ115HAT provides a better partition coefficient (logD n-octanol / PBS, pH 7.4) 1.50 vs. >4.98 for RA013915, improved aqueous solubility in simulated gastric fluid 200μM vs.148.2μM for RA013915. Additionally, EZ115HAT provides a better human liver microsomal stability 39min at 1 μM (Clint0.0354 mL / min / mg protein) than YF2, 4.6min at 1 μM, (Clint0.299 mL / min / mg protein). Moreover, EZ115HAT shows improved hERG IC50and CAV 1.2 IC506.8 μM and >30 μM respectively, compared to YF2 hERG IC501.8 μM and CAV 1.2 IC503.24 μM. The IC50for compound EZ115HAT hERG is nearly 15-fold higher than RA013915. Moreover, ActiveUS 205301239 43Attorney Docket No.: 0019240.01318WO1 Date of Electronic Filing: July 29, 2024 the higher values for the Nav1.5 and Cav1.2 indicate that effect on those channels do not counteract the hERG inhibition.

[0154] Evaluating the Biological Activity of Compounds of Formula (I) Histone Acetyltransferase (HAT) Assay:

[0155] The aim of the in vitro acetylation assay is to measure the enzymatic activity of the various compounds towards p300.

[0156] First, the drug is prepared:

[0157] Second, dilute p300 in AM1 buffer to a concentration of 40 ng / µL (final concentration in the reaction 20 ng / µL). This is accomplished by diluting 1 µL of p300 (at 0.4 µg / µL) into 19 µL of AM1 buffer.

[0158] Third, prepare the Master Mix. Prepare the Master Mix in low protein binding tubes (ThermoFisher Cat. No.90410), 20 µL system.Incubate reactions at 30oC for 30 minutes. ActiveUS 205301239 44Attorney Docket No.: 0019240.01318WO1 Date of Electronic Filing: July 29, 2024Incubate reactions at 30oC for 1 hour.

[0159] Fourth, perform the western blot assay. • Add 6.7 µL of Laemmli Sample Buffer 4x (Bio-Rad Laboratories Cat. No.161-0737) for each reaction and boil samples at 95 °C for 5 min. • Charge 10 µL for each sample in 2 Tris Glycine 4-15 % gels (Bio-Rad Laboratories Cat. No.456-1086). Running Buffer: Tris-Glycine 1X (pour in the cell up to the writing “2 gels”). • Run at 90 V for 1 hour (until the gel front reaches the green line of cell). • Semi-dry transfer with Trans-Blot® Turbo™ Blotting System from Bio-Rad Laboratories (Trans-Blot® Turbo™ RTA midi PVDF transfer kit Cat. No1704275). • Activate PVDF membrane in methanol for 5 minutes and then wet in Trans- Blot® Turbo™ transfer buffer. • Wet stacks in transfer buffer. • Transfer at 23 V 1.3 A for 7 minutes. • Blocking buffer: 5% Non-Fat milk in TBST (tween 0.1%) 1 hour at room temperature. • Cut the membranes to get Ac-His 3 at 17 kDa and p300 at 300 kDa. • Incubate with primary antibodies overnight at 4°C. • The day after, make 3 washes in TBST (Tween 0.1%) of 10 minutes. • Incubate with secondary antibody at room temperature for 1 hour. • Make 3 washes in TBST (Tween 0.1%) of 5 minutes. • Incubate membranes with ECL (SuperSignal West Dura Extended Duration Substrate from ThermoFisher) for 5 minutes at room temperature. • Acquire the image with ChemiDoc Odyssey Fc 2 minutes.

[0160] To detect total H3 (loading control), strip the membranes with RestoreTMWestern Blot Stripping Buffer according to the manufacturer protocol. ActiveUS 205301239 45Attorney Docket No.: 0019240.01318WO1 Date of Electronic Filing: July 29, 2024Stability in Human Liver microsomes EXPERIMENTAL PROCEDURE STABILITY IN HUMAN LIVER MICROSOMES

[0161] Mixed-gender human liver microsomes (Lot# 1010420) were purchased from XenoTech. The reaction mixture, minus NADPH, was prepared as described below. The test article was added into the reaction mixture at a final concentration of 1 μM. The control compound, testosterone, was run simultaneously with the test article in a separate reaction. An aliquot of the reaction mixture (without cofactor) was equilibrated in a shaking water bath at 37°C for 5 minutes. The reaction was initiated by the addition of the cofactor, and the mixture was incubated in a shaking water bath at 37°C. Aliquots (100 μL) were withdrawn at 0, 10, 20, 30, and 60 minutes. Test article and testosterone samples were immediately combined with 400 μL of ice-cold 50 / 50 acetonitrile (ACN) / H2O containing 0.1% formic acid and internal standard to terminate the reaction. The samples were then mixed and centrifuged to precipitate proteins. All samples were assayed by LC-MS / MS using electrospray ionization. Analytical conditions are outlined in Appendix 1. The peak area response ratio (PARR) of analyte to internal standard at each time point was compared to the PARR at time 0 to determine the percent remaining at each time point. Half-lives and clearance were calculated using GraphPad software, fitting to a single-phase exponential decay equationaIntrinsic clearance (CLint) was calculated based on CLint= k / P, where k is the elimination rate constant and P is the protein concentration in the incubation. ActiveUS 205301239 46Attorney Docket No.: 0019240.01318WO1 Date of Electronic Filing: July 29, 2024

[0162] The samples were then mixed and centrifuged to precipitate proteins. All samples were assayed by LC-MS / MS using electrospray ionization. Analytical conditions are outlined in Appendix 1. The peak area response ratio (PARR) to internal standard was compared to the PARR at time 0 to determine the percent remaining at each time point. Half-lives and clearance were calculated using GraphPad software, fitting to a single-phase exponential decay equation. REACTION COMPOSITION Liver Microsomes 0.5 mg / mL NADPH (cofactor) 1 mM Potassium Phosphate, pH 7.4 100 mM Magnesium Chloride 5 mM Test Article 1 μM APPENDIX 1. ANALYTICAL METHOD Liquid Chromatography Column: Thermo BDS Hypersil C1830 × 2.1 mm, 3 μm, with guard column M.P. Buffer: 25 mM ammonium formate buffer, pH 3.5 Aqueous Reservoir (A): 90% water, 10% buffer Organic Reservoir (B): 90% acetonitrile, 10% buffer Flow Rate: 0.7 mL / minute Gradient Program:Total Run Time: 2.50 minute Autosampler: 1-4μL injection volume Strong Needle Wash: water / methanol / 2-propanol:1 / 1 / 1; with 0.2% formic acid Weak Needle Wash: 0.1% formic acid in water Seal Wash: 10% methanol, 90% water Mass Spectrometer Instrument: PE SCIEX API 4000 ActiveUS 205301239 47Attorney Docket No.: 0019240.01318WO1 Date of Electronic Filing: July 29, 2024 Interface: Turbo Ionspray Mode: Multiple reaction monitoring Method: 1.0 minute duration ENZYMATIC ACTIVITY

[0163] The activity of EZ115HAT was assessed through a cell-free assay in which the HAT enzyme, the acetyl acceptor histone 3, the acetyl donor acetyl-CoA, and the compound at the appropriate concentrations were mixed in vitro. Enzyme activity was assessed by measuring levels of acetylation of lysine residues through specific antibodies by immunoblot. Initially, acetylation of lysine 18 (K18) and 27 (K27) of histone 3 (H3) through p300 was measured (see, FIG. 2A and FIG. 2B). The values of EC50 for H3K18 were equal to 104.23 ± 0.38 nM, while EC50 for H3K27 was equal to 121.10 ± 0.51 nM. Assessment of activity was expaneded to other lysine residues of H3 and the other HAT, CREB-Binding-Protein (CBP). Table 1 shows the EC50 values of various lysine residues by p300. Interestingly, an increase in acetylation of H3K4 and H3K14 was observed, which have been shown to be decreased in specimens derived from the brain of Alzheimer’s disease patients. Table 1. Summary EZ115HAT biological activity of different lysine on p300 enzyme (EC50):EZ115HAT HDAC selectivity

[0164] Increase in histone acetylation might not only depend upon increase in activity of HATs, but also upon inhibition of histone deacetylase activity. The inhibitory activity of EZ115HAT on the enzymatic activities of human recombinant HDAC1,2,3,4, 5,6,7,8,9,11 and sirt1,2,3,5,6 was determined using in-vitro enzymatic assays. The enzymatic assays were performed by BPS Bioscience. The percent inhibition of EZ115HAT against the HDAC enzymes is summarized in Tables 2 and 3. As reported in the tables, EZ115HAT did not show significant inhibitory activity toward various HDAC isoforms. Table 2. Assessment of EZ115HAT activity versus HDAC1, 2, 3, 4, 5, 6, 7, 8, 9, 11. ActiveUS 205301239 48Attorney Docket No.: 0019240.01318WO1 Date of Electronic Filing: July 29, 2024Table 3. Assessment of EZ115HAT activity versus sirt1, 2, 3, 5, 6.EZ115HAT Geno-toxicity assessment (AMES TEST)

[0165] Compound EZ115HAT was evaluated for mutagenic activity in the in vitro bacterial reverse mutation assay. Four tester strains of Salmonella typhimurium (TA98, TA100, TA1535, and TA1537) and 1 Escherichia coli strain (WP2 uvrA) were used for mutagenicity testing. In the mutagenicity assay, EZ115HAT was tested at 25, 50, 100, 250, 500, 1000, 2500, and 5000 μg / plate using the plate incorporation method.

[0166] Precipitates were not observed in any strain with or without metabolic activation. Cytotoxicity (i.e., reduced background lawn and / or > 50% reduction in the mean number of revertant colonies compared to the vehicle control) was observed at ≥ 2500 μg / plate in strain TA1537 with metabolic activation; and at 5000 μg / plate in strains TA100 and TA1535 with and without metabolic activation, and strain TA1537 without metabolic activation (see, Table 4). ActiveUS 205301239 49Attorney Docket No.: 0019240.01318WO1 Date of Electronic Filing: July 29, 2024

[0167] In the mutagenicity assay, criteria for a negative response were met for all tester strains with and without metabolic activation. The data from the vehicle and positive controls demonstrated the validity and sensitivity of this test system for detecting chemical mutagens with and without metabolic activation.

[0168] Altogether these data demonstrate that EZ115HAT is negative for mutagenic activity in the Salmonella strains TA98, TA100, TA1535, and TA1537 and in the E. coli strain WP2 uvrA, with and without metabolic activation. Table 4. EZ115HAT (Test Article) summary table of Gen-toxicity assessment.ActiveUS 205301239 50Attorney Docket No.: 0019240.01318WO1 Date of Electronic Filing: July 29, 2024ActiveUS 205301239 51Attorney Docket No.: 0019240.01318WO1 Date of Electronic Filing: July 29, 20242NF 2-Nitrofluorene 2AA 2-Aminoanthracene ICR ICR-191 Acridine NQNO 4-nitroquinoline-N-oxide SA Sodium azide R Reduced

[0169] In conclusion, mean increases in the number of revertant colonies indicative of a positive response were not observed with EZ115HAT in the S. typhimurium strains TA98, TA100, TA1535, and TA1537 and in the E. coli strain WP2 uvrA, with and without metabolic activation. Therefore, EZ115HAT is considered to be negative for inducing mutagenicity. EZ115HAT Ion Channel Profilers and Cardiac Profiler Core panel.

[0170] Cardiotoxicity, specifically linked to the inhibition of the hERG (human ether-a- go-go related gene channel) responsible for potassium ion flux, is a particularly common concern. hERG channel inhibition can disrupt cardiac repolarization, leading to serious adverse cardiovascular effects such as arrhythmias. Identifying compounds that may exhibit h-ERG channel inhibition and subsequent cardiotoxicity is essential to ensure patient safety and the successful progression of drug candidates through clinical trials. ActiveUS 205301239 52Attorney Docket No.: 0019240.01318WO1 Date of Electronic Filing: July 29, 2024

[0171] Compound EZ115HAT was tested for electrophysiological assay for activities on the ion channel (voltage –gated Sodium HEK-Nav1.5; voltage-gated Calcium HEK-Cav 1.2; voltage-gated Potassium CHO-hERG) targets specified above using the Qube electrophysiological platform (Table 5). Table 5 EZ115HAT Cardiac Profiler Summary table.EZ115HAT Pharmacokinetic (PK) Properties

[0172] A PK study on Sprague Dawley Rats was conducted with EZ115HAT (FIGS.1A – 1D). The animals were treated with 10mg / kg dose via intravenous and 20mg / kg dose via oral gavage (PO) administration. In addition, brain and plasma concentrations were evaluated at various time points post dose up to 24h after dosing. Following IV administration, the average peak concentration of EZ115HAT was observed at the first time point, 0.083h, with a Cmaxof 2,590 ng / mL, while total exposure was observed to be 1,320 h*ng / mL for AUClast. The terminal phase half-life (T½ ) was 0.787 h. The clearance (Cl) after IV administration was 7,700 mL / h / kg and is approximately equal to hepatic blood flow, given body weight. The PK profile of EZ115HAT following PO administration had an average peak concentration at 1 h post-dose with a Cmax of 140 ng / mL and an average AUClast of 384 h*ng / mL. Following IV administration, the Cmax ratio between brain and plasma of EZ115HAT in the brain was 7.5-fold, and the AUClastratio of brain to plasma exposure of EZ115HAT was 7-fold. After PO administration, substantially good ratios were observed for EZ115HAT and the brain to ActiveUS 205301239 53Attorney Docket No.: 0019240.01318WO1 Date of Electronic Filing: July 29, 2024 plasma ratio was 2-fold based on Cmax and AUClast. These data indicate a significant drug penetration across the blood-brain barrier, following IV and PO dosing of EZ115HAT compound.

[0173] These studies demonstrated that EZ115HAT is rapidly absorbed into the brain with ample ability to cross the blood brain barrier (FIG. 4). Moreover, the concentration of EZ115HAT in the brain was higher than plasma (Table 6). In conclusion, following IV and PO administrations compound EZ115HAT showed a significant drug penetration across the blood-brain barrier and appeared to favor the CNS over peripheral circulation. Table 6. EZ115HAT PK data summaryIn vitro metabolite profiling of compound EZ115HAT.

[0174] The aim of this investigation is to identify the principal pathways of metabolism for compound EZ115HAT by profiling the metabolites produced in vitro in mouse, rat, rabbit dog and human hepatocytes. In this study, the metabolism of compound EZ115HAT was investigated in cryopreserved primary hepatocytes after 2h of incubation. The metabolites profiles were obtained by performing LC-MS / MS analysis of hepatocyte extracts of each species and subsequent analysis of the resulting mass spectra for each metabolite identified.

[0175] The data shown in table 7 indicate the major pathway of metabolism for compound EZ115HAT is the oxidative de-alkylation in all tested species. ActiveUS 205301239 54Attorney Docket No.: 0019240.01318WO1 Date of Electronic Filing: July 29, 2024 Table 7. LC-MS % Relative Abundance of EZ115HAT and its Metabolites in Mouse, Rat, Rabbit, Dog, Monkey, and Human Hepatocytes.aLC-MS % Relative Abundance= (LC-MS Peak Area of Analyte / Total LC-MS Peak Area of all Analytes in sample)x100%. Note: Conditional formatting applied to metabolite relative abundance only for clarity of major signals.bND = below limit of detection.

[0176] In conclusion, the most abundant metabolite varied across species, with M381 predominant in mice, rat, and rabbit, while M379-b was major in human, dog, and monkey. M379-b was also a major metabolite in mice. Glucuronidation of M365-b led to the formation of M541 in rabbit. The metabolite profiles of dog and human hepatocytes were similar, while extensive metabolism was observed in mouse, rat, monkey, and rabbit hepatocytes. No unique human metabolites were detected in vitro, and reactive metabolites M338 and M380 were mainly found in monkey, rabbit, and mouse hepatocytes. Prolonged administration of EZ115HAT rescues synaptic and memory defects in APP / PS1 and hTau / Mapt-KO mice. ActiveUS 205301239 55Attorney Docket No.: 0019240.01318WO1 Date of Electronic Filing: July 29, 2024

[0177] Experiments were performed in which APP / PS1 and hTau / Mapt-KO mice and non-transgenic littermates were treated with EZ115HAT (0.125 mg / kg, i.p., daily) or vehicle. APP / PS1 mice were chosen as models of Aβ elevation (Trinchese, F., et al., Progressive age- related development of Alzheimer-like pathology in APP / PS1 mice. Ann Neurol, 2004.55(6): p.801-14). These mice deposit Aβ after 6 weeks of age, and display impairments of long- term potentiation (LTP), a type of synaptic plasticity thought to underlie memory formation, and memory at 3 months of age. hTau / Mapt-KO mice were chosen as a model of tau elevation. (Wijesekara, N., et al., Tau ablation in mice leads to pancreatic beta cell dysfunction and glucose intolerance. FASEB J, 2018.32(6): p.3166-3173). They start showing tau oligomers at 8 months of age, display LTP impairment at 10 months and memory impairment at 15 months. (Puzzo, D., et al., Tau is not necessary for amyloid-beta- induced synaptic and memory impairments. J Clin Invest, 2020). The treatment of APP / PS1 mice started at 3.5 months of age and lasted until behavioral and electrophysiological assessments at 4.5-5 months of age, whereas hTau / Mapt-KO mice were treated from the age of 14-18 months after tau oligomer appearance, for 1.5 months. The behavior assessment included both fear conditioning and 2 day radial arm water maze (two types of tests assessing associative and spatial memory that are affected in AD patients), as well as control behavioral tasks (i) open field to exclude changes in mouse exploratory behavior complicating interpretation of memory tasks, ii) visible platform to exclude any visual, motor and motivational confounding effect, iii) cued memory to exclude any amygdala involvement, and iv) sensory threshold assessment to exclude a confounding effect onto fear conditioning of altered perception of the electric shock).

[0178] FIG.5A is a graph showing that EZ115HAT (0.125 mg / Kg, i.p., daily from the age of 3.5 months after amyloid plaque appearance, for 1.5 months) protected APP / PS1 mice against the impairment of LTP. EZ115HAT alone did not affect potentiation. LTP was induced through four pulses at 100 Hz, with the burst repeated at 5 Hz, and each tetanus including 3-ten burst trains separated by 15 sec, for this and the following LTP graphs. Overall Two-Way ANOVA: F(3,62)=5.861, p=0.0014, Two-Way ANOVA F(1,35)=8.367, p=0.0065 APP / PS1+vehicle vs. APP / PS1+EZ115HAT; F(1,30)=16.68, p=0.0003 APP / PS1+vehicle vs. WT+vehicle, F(1,27)=0.4683, p=0.4996 WT+vehicle vs. WT+ EZ115HAT: The number of slices is indicated on the graph in this and the following figures. ActiveUS 205301239 56Attorney Docket No.: 0019240.01318WO1 Date of Electronic Filing: July 29, 2024

[0179] FIG.5B is a graph showing that the treatment protected APP / PS1 mice against the impairment of contextual memory, while EZ115HAT alone did not affect freezing.1-Way ANOVA F(3,45)=2.983, p=0.0411; Bonferroni p=0.0453 WT+vehicle vs. APP / PS1+vehicle; p=0.0306 APP / PS1+vehicle vs. APP / PS1+EZ115HAT; p>0.05 WT+vehicle vs. WT+EZ115HAT. Baseline: 1-Way ANOVA F(3,45)=0.4177 p>0.05. The number of animals is indicated on the graph in this and the following figures.

[0180] FIG.5C is a graph showing that the treatment protected APP / PS1 mice against the impairment of spatial memory, while EZ115HAT alone did not affect performance. RAWM: 2-Way ANOVA (day 2): F (3,57) = 5.039 p = 0.0036. Bonferroni Block 10 p=0.0252 WT+vehicle vs. APP / PS1+vehicle; p=0.0096 APP / PS1+vehicle vs. APP / PS1+EZ115HAT; p>0.05 WT+vehicle vs. WT+EZ115HAT. Block 9 p>0.05 WT+vehicle vs. APP / PS1+vehicle; p>0.05 APP / PS1+vehicle vs. APP / PS1+EZ115HAT; p>0.05 WT+vehicle vs. WT+EZ115HAT. Block 8 p=0.0042 WT+vehicle vs. APP / PS1+vehicle; p=0.0412 APP / PS1+vehicle vs. APP / PS1+EZ115HAT; p>0.05 WT+vehicle vs. WT+EZ115HAT. Block 7 p=0.3233 WT+vehicle vs. APP / PS1+vehicle; p>0.05 APP / PS1+vehicle vs. APP / PS1+EZ115HAT; p>0.05 WT+vehicle vs. WT+EZ115HAT. Block 6 p>0.05 WT+vehicle vs. APP / PS1+vehicle; p>0.05 APP / PS1+vehicle vs. APP / PS1+EZ115HAT; p>0.05 WT+vehicle vs. WT+EZ115HAT. The number of animals is indicated on the graph in this and the following figures.

[0181] FIG.6A is a graph showing that EZ115HAT (0.125 mg / Kg, i.p., daily from the age of 14-18 months after tau oligomer appearance, for 1.5 months) protected hTau / Mapt-KO mice against the impairment of LTP. EZ115HAT alone did not affect potentiation. Overall Two-Way ANOVA: F(3,62)=4.707, p=0.0050, Two- Way ANOVA F(1,34)=10.63, p=0.0025 hTau / Mapt-KO +vehicle vs. hTau / Mapt-KO +EZ115HAT; F(1,33)=11.47, p=0.0018 hTau / Mapt- KO+vehicle vs. NonTg+vehicle, F(1,28)=0.05876, p=0.8102 NonTg+vehicle vs. NonTg+EZ115HAT.

[0182] FIG.6B is a graph showing that The treatment also protected hTau / Mapt-KO mice against the impairment of contextual memory, while EZ115HAT alone did not affect freezing.1-Way ANOVA F(3,54)=5.772, p=0.0017; Bonferroni p=0.0088 NonTg+vehicle vs. hTau / Mapt-KO+vehicle; p=0.0007 hTau / Mapt-KO+vehicle vs. hTau / Mapt- KO+EZ115HAT; p>0.05 NonTg+vehicle vs. NonTg+EZ115HAT. Baseline: 1-Way ANOVA F(3,54)=2.045 p>0.05. The number of animals is indicated on the graph. ActiveUS 205301239 57Attorney Docket No.: 0019240.01318WO1 Date of Electronic Filing: July 29, 2024

[0183] FIG.6C is a graph showing that the treatment protected hTau / Mapt-KO mice against the impairment of spatial memory, while EZ115HAT alone did not affect performance. RAWM: 2-Way ANOVA (day 2): F(3,54)=13.66, p<0.0001. Bonferroni’s Block 10 p<0.0001 NonTg+vehicle vs. hTau / Mapt-KO+vehicle; p=0.0002 hTau / Mapt- KO+vehicle vs. hTau / Mapt-KO+EZ115HAT; p>0.05 NonTg+vehicle vs. NonTg+EZ115HAT. Block 9 p<0.0001 NonTg+vehicle vs. hTau / Mapt-KO+vehicle; p=0.0002 hTau / Mapt-KO+vehicle vs. hTau / Mapt-KO+EZ115HAT; p>0.05 NonTg+vehicle vs. NonTg+EZ115HAT. Block 8 p=0.0008 NonTg+vehicle vs. hTau / Mapt-KO+vehicle; p=0.0010 hTau / Mapt-KO+vehicle vs. hTau / Mapt-KO+EZ115HAT; p>0.05 NonTg+vehicle vs. NonTg+EZ115HAT. Block 7 p=0.0025 NonTg+vehicle vs. hTau / Mapt-KO+vehicle; p=0.0484 hTau / Mapt-KO+vehicle vs. hTau / Mapt-KO+EZ115HAT; p>0.05 NonTg+vehicle vs. NonTg+EZ115HAT.

[0184] As shown above, chronic administration of EZ115HAT improved LTP and both contextual fear memory and spatial memory in both animal models. EZ115HAT alone did not affect them.

[0185] Finally, control behavioral tasks did not show any difference among different groups of mice indicating that the effect of EZ115HAT onto associative and spatial memory was a genuine effect against their impairment after Aβ and tau elevation as shown in FIGS. 7A – 7F. Thus, prolonged administration of EZ115HAT might effectively counteract AD progression.

[0186] FIGS.7A and 7B are graphs showing that EZ115HAT (0.125 mg / Kg, i.p., daily from the age of 14-18 months, for 1.5 months) has no effect on vision, motility, or motivation in hTau / Mapt-KO. Testing with the visible platform task does not reveal any difference in time to reach the visible platform (Two-way ANOVA: F(3,54)=2.501, p>0.05) (FIG.7A) and average speed (Two-way ANOVA: F(3,54)=0.9072, p>0.05) (FIG.7B).

[0187] FIG.7C is a chart showing that EZ115HAT has no effect on cued amygdala dependent memory in hTau / Mapt-KO. Freezing responses during the auditory cued conditioning are not significantly different among the groups (1-way ANOVA: F(3,54)=2.615 p>0.05).

[0188] FIGS.7D and 7E are charts showing that EZ115HAT has no effect on exploratory behavior in hTau / Mapt-KO. Open field test shows a similar percentage of time spent in the ActiveUS 205301239 58Attorney Docket No.: 0019240.01318WO1 Date of Electronic Filing: July 29, 2024 center (day 1: F(3,56)=0.2884, p>0.05; day 2: F(3,56)=0.3032, p>0.05) (D) and number of entries into the center among all conditions (day 1: F(3,56)=1.123, p>0.05; day 2: F(3,56)=0.5299, p>0.05) (E), indicating no differences in exploratory behavior.

[0189] FIG.7F is a chart showing that EZ115HAT has no effect on animal capability of perceiving the electric shock in hTau / Mapt-KO. No difference is detected among the groups during assessment of the sensory threshold. One-way ANOVA among all: for visible response F(3,55)=0.6861, p>0.05; for motor response F(3,55)=1.534, p>0.05 and for audible response F(3,55)=1.760, p>0.05. EZ115HAT rescues defects in synaptic plasticity following Aβ and tau oligomer elevation.

[0190] LTP was elicited in the presence of 200 nM Aβ42 (see Stine et al for their preparation (Stine, W.B., Jr., et al., In vitro characterization of conditions for amyloid-beta peptide oligomerization and fibrillogenesis. J Biol Chem, 2003.278(13): p.11612-22)), or 50 nM 4R / 2N tau oligomers (see Fa’ et al for their preparation (Fa, M., et al., Extracellular Tau Oligomers Produce An Immediate Impairment of LTP and Memory. Sci Rep, 2016.6: p. 19393)), or vehicle. Hippocampal slices were perfused with the oligomers through the bath solution for 20 min prior to the θ-burst. Aβ and tau reduced LTP. However, EZ115HAT (9.68 μM, for 20 min prior to the θ-burst) ameliorated LTP deficits. FIG.8 is a graph showing efficacy of EZ115HAT in AD relevant mouse models characterized by Aβ and tau oligomer elevation using LTP as a test of synaptic plasticity. Perfusion with EZ115HAT (9.68 μM) rescues LTP defect in hippocampal slices treated with 200 nM Aβ- or 50 nM tau-oligomers for 20 min prior to LTP induction through a theta-burst. EZ115HAT alone did not affect potentiation.2-Way ANOVA for repeated measures between groups: F(1,34)=6.612, p=0.0147 vehicle vs. Tau; F(1,28)=6.812, p=0.0144 vehicle vs. Aβ; F(1,23)=10.19, p=0.0040 Tau vs. Tau + EZ115HAT; F(1,22)=11.61, p=0.0025 Aβ vs. Aβ+EZ115HAT.). Thus, EZ115HAT is capable of rescuing the defects in synaptic plasticity following Tau and Aβ42oligomer elevation. EZ115HAT effect onto the LTP impairment induced by tau oligomers is dose- dependent.

[0191] The effect of EZ115HAT onto the oligomeric tau-induced defect in LTP was examined using different concentrations of the compound. The analysis revealed a dose- ActiveUS 205301239 59Attorney Docket No.: 0019240.01318WO1 Date of Electronic Filing: July 29, 2024 dependence with a plateau at 0.125 mg / Kg and an ED50 of 0.093 mg / kg dose. FIG.9 provides an EZ115HAT dose-response curve during assessment of LTP deficit due to oligomeric tau elevation. The measurement of ED50 revealed a low nanomolar efficacy of EZ1115HAT. BIOLOGICAL ACTIVITY (EFFICACY)

[0192] EZ115HAT was tested for efficacy in a test of long-term potentiation (LTP), a type of synaptic plasticity thought to underlie memory formation. For this test, we recorded extracellular field recordings on 400 μm transverse hippocampal slices. Hippocampal slices were cut through a tissue chopper, and transferred to a recording chamber and perfused (1–2 ml / min) with artificial cerebro-spinal fluid (ACSF) containing the following (in mM): NaCl (124), KCl (4.4), Na2HPO4 (1), NaHCO3 (25), CaCl2 (2), MgCl2 (2), glucose (10) kept at 29°C and continuously bubbled with an O2 / CO2 mixture at 95% and 5%. After 90 min recovery, field excitatory post-synaptic potential (fEPSPs) were recorded in CA1 stratum radiatum by a glass electrode filled with ACSF in response to Schaffer collateral stimulation by a bipolar tungsten electrode. Basal synaptic transmission was assayed by plotting the slope of the fEPSP at different intensities of stimulation to determine the intensity of stimulation equal to the 1 / 3 of the maximum response to be used for the baseline of the LTP experiment. Following recording for 30 min of a stable baseline LTP was evoked through a theta-burst stimulation (4 pulses at 100 Hz, with the bursts repeated at 5 Hz and each tetanus including 3 ten-burst trains separated by 15 sec) to evoke potentiation. LTP was followed for 120 min following its induction through the theta-burst. Oligomeric tau was applied for 20 min prior to the theta-burst either alone of in the presence of different concentration of EZ115HAT. The protocol to prepare human oligomeric tau has been described previously (Argyrousi, E.K. et al., Methods Mol Biol 1779, 85-9). Briefly, following synthesis of recombinant Tau 4R / 2N using Escherichia coli, tau was treated with TCEP-HCl and EDTA, and incubated at room temperature for 1 hr. Oligomerization was achieved via introduction of disulfide bonds through incubation with 1 mM H2O2 at room temperature for 20 hrs, followed by centrifugation in a PES at 4000 × g. The resulting material was stored ready to be used for the experiment, when tau was diluted to the final concentration of 50 nM in ACSF.

[0193] EZ115HAT has also demonstrated efficacy in an electrophysiological assessment of oligomeric tau-induced defect in long-term potentiation, a type of synaptic plasticity that is thought to underlie memory formation with dramatically improved potency at doses that are ActiveUS 205301239 60Attorney Docket No.: 0019240.01318WO1 Date of Electronic Filing: July 29, 2024 20 times lower than any other HAT activator so far tested in an ex-vivo model of Alzheimer’s disease and Alzheimer’s disease related dementia. In conclusion, EZ115HAT compound appeared to favor the CNS over peripheral circulation, showed a better drug availability via PO administration and better physical-chemical properties compared to RA013915, yet still showing improved excellent stability as RA013915 compared to YF2.

[0194] Chemical and biochemical properties of EZ115HAT are shown on Table 8 below. EZ115HAT Summary Table 8

[0195] Although the invention has been described and illustrated in the foregoing illustrative embodiments, it is understood that the present disclosure has been made only by ActiveUS 205301239 61Attorney Docket No.: 0019240.01318WO1 Date of Electronic Filing: July 29, 2024 way of example, and that numerous changes in the details of implementation of the invention can be made without departing from the spirit and scope of the invention, which is limited only by the claims that follow. Features of the disclosed embodiments can be combined and / or rearranged in various ways within the scope and spirit of the invention to produce further embodiments that are also within the scope of the invention. Those skilled in the art will recognize, or be able to ascertain, using no more than routine experimentation, numerous equivalents to the specific embodiments described specifically in this disclosure. Such equivalents are intended to be encompassed in the scope of the following claims. ActiveUS 205301239 62

Claims

Attorney Docket No.: 0019240.01318WO1 Date of Electronic Filing: July 29, 2024 CLAIMS What is claimed is:

1. A compound of Formula (I),wherein, Rais H, OH, CN, C1-C6-alkyl, O-(C3-C8-cycloalkyl), O-(C3-C8-heterocycloalkyl), O-(C2-C6- alkenyl), O-(C1-C6-alkyl), O-(C2-C6-alkyl)-N(R1)2, halogen, or haloalkyl; Rbis H, C1-C6-alkyl, C2-C6-alkenyl, C3-C8-cycloalkyl, C2-C6-heteroalkyl, C3-C8- heterocycloalkyl, aryl, heteroaryl, O-(C1-C6-alkyl), O-(C3-C8-cycloalkyl), O-(C2-C6-alkenyl), or O-(C3-C8-heterocycloalkyl); Rcis H, C1-C6-alkyl, C1-C6-haloalkyl, O-(C1-C6-alkyl), O-(C1-C6-haloalkyl), halogen, CN, or NO2; Rdis H, OH, C1-C6-alkyl, O-(C3-C8-cycloalkyl), O-(C3-C8-heterocycloalkyl), O-(C2-C6- alkenyl), O-(C1-C6-alkyl), or O-(C2-C6-alkyl)-N(R1)2; R1is independently H, -(C1-C4-alkyl), -(C1-C4-haloalkyl), -(C3-C8-cycloalkyl), -(C3-C8- heterocycloalkyl), aryl or heteroaryl; W is CH or N; X is -CO-, -CON(R10)-, -CON(R1)(CH2)n-, -(CH2)nN(R1)-, or -C=N-; Z is CH or N, and n is an integer from 1-3, or a pharmaceutically acceptable salt or hydrate thereof.

2. The compound of claim 1, wherein ActiveUS 205301239 63Attorney Docket No.: 0019240.01318WO1 Date of Electronic Filing: July 29, 2024 Rais OH, CN, C1-C6-alkyl, O-(C1-C6-alkyl), halogen, or haloalkyl; Rbis C1-C6-alkyl, C2-C6-heteroalkyl, aryl, heteroaryl, O-(C1-C6-alkyl); Rcis H, C1-C6-alkyl, C1-C6-haloalkyl, halogen, or CN; Rdis OH, C1-C6-alkyl, O-(C1-C6-alkyl), or O-(C2-C6-alkyl)-N(R1)2; R1is independently H, -(C1-C4-alkyl), or -(C1-C4-haloalkyl); W is CH; X is -CON(R1)-, -CON(R1)(CH2)n-; Z is CH, and n is an integer from 1-3, or a pharmaceutically acceptable salt or hydrate thereof.

3. The compound of claim 1or 2, wherein Rais OH, O-(C1-C6-alkyl), halogen, or haloalkyl; Rbis C1-C6-alkyl, O-(C1-C6-alkyl); Rcis C1-C6-alkyl, halogen, CF3or CN; Rdis OH, C1-C6-alkyl, O-(C1-C6-alkyl), or O-(C2-C6-alkyl)-N(R1)2; R1is independently H, -(C1-C4-alkyl), or -(C1-C4-haloalkyl); W is CH; X is -CON(R1)-; and Z is CH, or a pharmaceutically acceptable salt or hydrate thereof.

4. The compound of any one of claims 1-3, wherein X is –C(O)N(R1)-.

5. The compound of any one of claims 1-4, wherein Rais OH, O-(C1-C6-alkyl), halogen, or haloalkyl.

6. The compound of any one of claims 1-5, wherein Rbis C1-C6-alkyl, O-(C1-C6-alkyl). ActiveUS 205301239 64Attorney Docket No.: 0019240.01318WO1 Date of Electronic Filing: July 29, 2024 7. The compound of claim 1, wherein the compound of Formula (I) iswherein Rais OH, O-(C1-C6-alkyl), halogen, or haloalkyl; Rbis C1-C6-alkyl, O-(C1-C6-alkyl); Rcis C1-C6-alkyl, halogen, CF3 or CN; Rdis OH, C1-C6-alkyl, O-(C1-C6-alkyl), or O-(C2-C6-alkyl)-N(R1)2; R1is independently H, -(C1-C4-alkyl), or -(C1-C4-haloalkyl); W is CH; and Z is CH, or a pharmaceutically acceptable salt or hydrate thereof.

8. The compound of claim 1, wherein the compound of Formula (I) iswherein Rais OH, O-(C1-C3-alkyl), halogen, or haloalkyl; Rbis C1-C3-alkyl, O-(C1-C3-alkyl); Rcis C1-C6-alkyl, halogen, CF3or CN; Rdis O-(C1-C6-alkyl), or O-(C2-C6-alkyl)-N(R1)2; and ActiveUS 205301239 65Attorney Docket No.: 0019240.01318WO1 Date of Electronic Filing: July 29, 2024 R1is independently H, -(C1-C2-alkyl), or -(C1-C4-haloalkyl), or a pharmaceutically acceptable salt or hydrate thereof.

9. The compound of claim 8, wherein Rais O-(C1-C3-alkyl); Rbis O-(C1-C3-alkyl); Rcis C1-C6-alkyl, halogen, CF3or CN; Rdis O-(C2-C6-alkyl)-N(R1)2; and R1is independently H, -(C1-C2-alkyl), or a pharmaceutically acceptable salt or hydrate thereof.

10. The compound of claim 1 having the structure:or a pharmaceutically acceptable salt or solvate thereof.

11. The compound of any one of claims 1-10, wherein the compound is a HAT activator.

12. A pharmaceutical composition comprising a compound of any one of claims 1-11 and a pharmaceutically acceptable excipient.

13. A method of increasing histone acetylation in a subject, the method comprising administering to the subject a therapeutically effective amount of a compound of any one of claims 1-11, or the composition of claim 12. ActiveUS 205301239 66Attorney Docket No.: 0019240.01318WO1 Date of Electronic Filing: July 29, 2024 14. The method of claim 13, wherein histone acetylation occurs at K18 and / or K27 of histone H3.

15. A method of treating a neurodegenerative disease in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound of any one of claims 1-11, or the composition of claim 12.

16. A method of improving long term memory formation in a subject, the method comprising administering to the subject a therapeutically effective amount of a compound of any one of claims 1-11, or the composition of claim 12.

17. A method of enhancing memory retention in a subject comprising administering to the subject a therapeutically effective amount of a compound of any one of claims 1-11, or the composition of claim 12.

18. A method of enhancing learning or memory in a subject comprising administering to the subject a therapeutically effective amount of a compound of any one of claims 1-11, or the composition of claim 12.

19. The method of any one of claims 16-18, wherein the subject is afflicted with a neurodegenerative disease.

20. The method of claim 19, wherein the neurodegenerative disease is Adrenoleukodystrophy (ALD), Alcoholism, Alexander's disease, Alzheimer's disease Related Dementia (ADRD), Alper's disease, Alzheimer's disease, Amyotrophic lateral sclerosis (Lou Gehrig's Disease), Ataxia telangiectasia, Batten disease (also known as Spielmeyer-Vogt- Sjogren-Batten disease), Bovine spongiform encephalopathy (BSE), Canavan disease, Cockayne syndrome, Corticobasal degeneration, argyrophilic grain disease (AGD), and globular glial tauopathy (GGT), the neurofibrillary tangle-predominant senile dementia (now included also in the category of primary age-related tauopathy, PART), Behavioral variant frontotemporal dementia; Semantic variant primary progressive aphasia, non- fluent / agrammatic variant primary progressive aphasia, logopenic variant primary progressive aphasia, Creutzfeldt-Jakob disease, Familial fatal insomnia, Frontotemporal lobar degeneration, Huntington's disease, HIV -associated dementia, Kennedy's disease, Krabbe's disease, Lewy body dementia, Neuroborreliosis, Machado-Joseph disease (Spinocerebellar ataxia type 3), Multiple System Atrophy, Multiple sclerosis, Narcolepsy, ActiveUS 205301239 67Attorney Docket No.: 0019240.01318WO1 Date of Electronic Filing: July 29, 2024 Niemann Pick disease, Parkinson's disease, Pelizaeus-Merzbacher Disease, Pick's disease, Primary lateral sclerosis, Prion diseases, Progressive Supranuclear Palsy, Rett's syndrome, Tau-positive Pronto Temporal dementia, Tau-negative Frontotemporal dementia, Refsum's disease, tauopathy, Sandhoff disease, Schilder's disease, Subacute combined degeneration of spinal cord secondary to Pernicious Anaemia, Spielmeyer-Vogt-Sjogren-Batten disease, Batten disease, Spinocerebellar ataxia, Spinal muscular atrophy, Steele-Richardson- Olszewski disease, Tabes dorsalis, or Toxic encephalopathy.

21. The method of claim 20, wherein the neurodegenerative disease is Alzheimer’s Disease, ADRD, tauopathy, Parkinson’s Disease, ALS, or Huntington’s Disease.

22. A method of treating cancer in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a compound of any one of claims 1-11, or the composition of claim 12.

23. The method of claim 22, wherein the cancer is B cell lymphoma, colon cancer, lung cancer, renal cancer, bladder cancer, T cell lymphoma, myeloma, leukemia, chronic myeloid leukemia, acute myeloid leukemia, chronic lymphocytic leukemia, acute lymphocytic leukemia, hematopoietic neoplasias, thymoma, lymphoma, sarcoma, lung cancer, liver cancer, non-Hodgkin's lymphoma, Hodgkin's lymphoma, uterine cancer, renal cell carcinoma, hepatoma, adenocarcinoma, breast cancer, pancreatic cancer, liver cancer, prostate cancer, head and neck carcinoma, thyroid carcinoma, soft tissue sarcoma, ovarian cancer, primaiy or metastatic melanoma, squamous cell carcinoma, basal cell carcinoma, brain cancer, angiosarcoma, hemangiosarcoma, bone sarcoma, fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma, osteogenic sarcoma, chordoma, angiosarcoma, endotheliosarcoma, lymphangiosarcoma, lymphangioendotheliosarcoma, synovioma, testicular cancer, uterine cancer, cervical cancer, gastrointestinal cancer, mesothelioma, Ewing's tumor, leiomyosarcoma, rhabdomyosarcoma, colon carcinoma, pancreatic cancer, breast cancer, ovarian cancer, prostate cancer, squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, sweat gland carcinoma, sebaceous gland carcinoma, papillary carcinoma, Waldenstroom's macroglobulinemia, papillary adenocarcinomas, cystadenocarcinoma, bronchogenic carcinoma, bile duct carcinoma, choriocarcinoma, seminoma, embryonal carcinoma, Wilms' tumor, lung carcinoma, epithelial carcinoma, cervical cancer, testicular tumor, glioma, astrocytoma, medulloblastoma, craniopharyngioma, ependymoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma, ActiveUS 205301239 68Attorney Docket No.: 0019240.01318WO1 Date of Electronic Filing: July 29, 2024 meningioma, retinoblastoma, glioblastoma, diffuse midline glioma, leukemia, melanoma, neuroblastoma, small cell lung carcinoma, bladder carcinoma, multiple myeloma, follicular lymphoma or medullary carcinoma 24. The method of claim 22, wherein the cancer is Hodgkin's lymphoma, non-Hodgkin's lymphoma, B cell lymphoma, T cell lymphoma, follicular lymphoma, T cell leukemia, acute myeloid leukemia, acute lymphocytic leukemia, or myeloma.

25. The method of any one of claims 15-24, wherein the subject has at least one mutant HAT enzyme gene. ActiveUS 205301239 69

Images