BRM targeting compounds and associated methods of use

The present disclosure provides bifunctional compounds comprising a target protein binding moiety and a E3 ubiquitin ligase binding moiety, and associated methods of use.

SEQUENCE LISTING

The instant application contains a Sequence Listing which has been submitted electronically in ASCII format and is hereby incorporated by reference in its entirety. Said ASCII copy, created on Aug. 31, 2021, is named 105807_000171_SL.txt and is 4,020 bytes in size.

TECHNICAL FIELD

The description provides bifunctional compounds comprising a target protein binding moiety and a E3 ubiquitin ligase binding moiety, and associated methods of use. The bifunctional compounds are useful as modulators of targeted ubiquitination, especially with respect to Switch/Sucrose Non-Fermentable (SWI/SNF)-Related, Matrix-Associated, Actin-Dependent Regulator of Chromatin, Subfamily A, Member 2 (SMARCA2) (i.e. BRAHMA or BRM), which are degraded and/or otherwise inhibited by bifunctional compounds according to the present disclosure.

BACKGROUND

The human SWItch/Sucrose Non-Fermentable (SWI/SNF) complexes are ATP-dependent chromatin remodelers. These large complexes play important roles in essential cellular processes, such as transcription, DNA repair and replication by regulating DNA accessibility.

Mutations in the genes encoding up to 20 canonical SWI/SNF subunits are observed in nearly 20% of all human cancers with the highest frequency of mutations observed in rhabdoid tumors, female cancers (including ovarian, uterine, cervical and endometrial), lung adenocarcinoma, gastric adenocarcinoma, melanoma, esophageal, and renal clear cell carcinoma.

SMARCA2 (BRM) and SMARCA4 (BRG1) are the subunits containing catalytic ATPase domains and they are essential for the function of SWI/SNF in perturbation of histone-DNA contacts, thereby providing access points to transcription factors and cognate DNA elements that facilitate gene activation and repression.

SMARCA2 and SMARCA4 shares a high degree of homology (up to 75%). SMARCA4 is frequently mutated in primary tumors (i.e., deleted or inactivated), particularly in lung cancer (12%), melanoma, liver cancer and pancreatic cancer. SMARCA2 is one of the top essential genes in SMARCA4-mutant (deleted) cancer cell line. This is because SMARCA4 deleted cancer cells exclusively rely on SMARCA2 ATPase activity for their chromatin remodeling activity for cellular functions such as cell proliferation, survival and growth. Thus, targeting SMARCA2 may be promising therapeutic approach in SMARCA4-related or deficient cancers (genetic synthetic lethality).

Previous studies have demonstrated the strong synthetic lethality using gene expression manipulation such as RNAi; downregulating SMARCA2 gene expression in SMARCA4 mutated cancer cells results in suppression of cancer cell proliferation. However, SMARCA2/4 bromodomain inhibitors (e.g. PFI-3) exhibit none to minor effects on cell proliferation inhibition [Vangamudi et al. Cancer Res 2015]. This phenotypic discrepancy between gene expression downregulation and small molecule-based approach lead us to investigating protein degradation bispecific molecules in SMARCA4 deficient cancers.

SMARCA2 is also reported to play roles in multiple myeloma expressing t(4;14) chromosomal translocation [Chooi et al. Cancer Res abstract 2018]. SMARCA2 interacts with NSD2 and regulates gene expression such as PRL3 and CCND1. SMARCA2 gene expression downregulation with shRNA reduces cell cycle S phase and suppresses cell proliferation of t(4;14) MM cells.

SUMMARY

The present disclosure is directed to compounds of Formula (I):
PTM-ULM  (I)

or a pharmaceutically acceptable salt or solvate thereof,

wherein

PTM is a moiety of Formula IA:

whereinR1is a covalent bond, or chemical moiety that links PTM and ULM;* is a point of attachment to ULM;n=0-3;W is optionally substituted —CH2—, —C(O)—, —S(O)—, or —S(O)2—, wherein when n=2 or 3, only one W may be —C(O)—, —S(O)—, or —S(O)2—;Rc1and Rd1are independently H, D, Halo, C1-3alkyl, C1-3haloalkyl, or C1-4alkoxyl;Re3is H, —C(O)Rf, or —P(O)(ORg)2; wherein Rfand Rgare independently H, C1-4alkyl, C1-4substituted alkyl, C3-8cyclcoalkyl, C3-8substituted cyclcoalkyl, C3-8heterocyclcoalkyl, or C3-8substituted heterocyclcoalkyl;Z and Y are each independently N, CRhwherein Rh=H or absent; or, if R1is attached to Z, then Z is C and Y is N or CRhwherein Rhis H; or if R1is attached to Y, then Y is C and Z is N or CRhwherein Rhis H;B is an optionally substituted 5-7 membered cycloalkyl ring, an optionally substituted 5-7 membered heteroaryl ring, or an optionally substituted 5-7 membered heterocyclic ring, wherein ring B is fused to ring C through Y and Z; andULM is a small molecule E3 Ubiquitin Ligase binding moiety that binds a Von Hippel-Lindau E3 Ubiquitin Ligase.

DETAILED DESCRIPTION OF ILLUSTRATIVE EMBODIMENTS

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

The following terms are used to describe the present disclosure. In instances where a term is not specifically defined herein, that term is given an art-recognized meaning by those of ordinary skill applying that term in context to its use in describing the present disclosure.

The articles “a” and “an” as used herein and in the appended claims are used herein to refer to one or to more than one (e.g., to at least one) of the grammatical object of the article unless the context clearly indicates otherwise. By way of example, “an element” means one element or more than one element.

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

The term “compound”, as used herein, unless otherwise indicated, refers to any specific chemical compound disclosed herein and includes tautomers, regioisomers, geometric isomers, and where applicable, stereoisomers, including optical isomers (enantiomers) and other stereoisomers (diastereomers) thereof, as well as pharmaceutically acceptable salts and derivatives, including prodrug and/or deuterated forms thereof where applicable, in context. Deuterated small molecules contemplated are those in which one or more of the hydrogen atoms contained in the drug molecule have been replaced by deuterium.

Within its use in context, the term compound generally refers to a single compound, but also may include other compounds such as stereoisomers, regioisomers and/or optical isomers (including racemic mixtures) as well as specific enantiomers or enantiomerically enriched mixtures of disclosed compounds. The term also refers, in context to prodrug forms of compounds which have been modified to facilitate the administration and delivery of compounds to a site of activity. It is noted that in describing the present compounds, numerous substituents and variables associated with same, among others, are described. It is understood by those of ordinary skill that molecules which are described herein are stable compounds as generally described hereunder.

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

As used herein, the term “alkyl”, by itself or as part of another substituent, means, unless otherwise stated, a straight or branched chain hydrocarbon radical having up to twelve carbon atoms. In some embodiments, the number of carbon atoms is designated (i.e., C1-C8means one to eight carbons). Examples of alkyl groups include methyl, ethyl, n-propyl, iso-propyl, n-butyl, t-butyl, iso-butyl, sec-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, and the like. Alkyl groups may be optionally substituted as provided herein. In some embodiments, the alkyl group is a C1-C6alkyl; in some embodiments, it is a C1-C4alkyl.

When a range of carbon atoms is used herein, for example, C1-C6, all ranges, as well as individual numbers of carbon atoms are encompassed. For example, “C1-C3” includes C1-C3, C1-C2, C2-C3, C1, C2, and C3.

The term “optionally substituted”, as used in combination with a substituent defined herein, means that the substituent may, but is not required to be, substituted with one or more suitable functional groups or other substituents as provided herein. For example, a substituent may be optionally substituted with one or more of: halo, cyano, C1-6alkyl, C3-6cycloalkyl, C2-6alkenyl, C2-6alkynyl, halo(C1-6)alkyl, C1-6alkoxy, halo(C1-6alkoxy), C1-6alkylthio, C1-6alkylamino, NH2, NH(C1-6alkyl), N(C1-6alkyl)2, NH(C1-6alkoxy), N(C1-6alkoxy)2, —C(O)NHC1-6alkyl, —C(O)N(C1-6alkyl)2, —C(O)NH2, —C(O)C1-6alkyl, —C(O)2C1-6alkyl, —NHCO(C1-6alkyl), —N(C1-6alkyl)CO(C1-6alkyl), —S(O)C1-6alkyl, —S(O)2C1-6alkyl, oxo, 6-12 membered aryl, benzyl, pyridinyl, pyrazolyl, thiazolyl, isothiazolyl, or other 5 to 12 membered heteroaryl groups. In some embodiments, each of the above optional substituents are themselves optionally substituted by one or two groups.

The term “cycloalkyl” as used herein refers to a 3-12 membered cyclic alkyl group, and includes bridged and spirocycles (e.g., adamantine). Cycloalkyl groups may be fully saturated or partially unsaturated. The term “cycloalkyl” also includes multiple condensed ring systems (e.g., ring systems comprising 2, 3 or 4 rings) wherein a single cycloalkyl ring (as defined above) can be condensed with one or more groups selected from heterocycles, carbocycles, aryls, or heteroaryls to form the multiple condensed ring system. Such multiple condensed ring systems may be optionally substituted with one or more (e.g., 1, 2, 3 or 4) oxo groups on the carbocycle or heterocycle portions of the multiple condensed ring. The rings of the multiple condensed ring system can be connected to each other via fused, spiro and bridged bonds when allowed by valency requirements. It is to be understood that the individual rings of the multiple condensed ring system may be connected in any order relative to one another. It is also to be understood that the point of attachment of a multiple condensed ring system (as defined above for a cycloalkyl) can be at any position of the cycloalkylic ring. Examples of cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cycloheptyl, cyclohexyl, cycloheptyl, cyclooctyl, indenyl, bicyclo[2.2.1]heptanyl, bicyclo[3.1.1]heptanyl, bicyclo[4.1.0]heptanyl, spiro[3.3]heptanyl, and spiro[3.4]octanyl. In some embodiments, the cycloalkyl group is a 3-7 membered cycloalkyl.

The term “akenyl” as used herein refers to C2-C12alkyl group that contains at least one carbon-carbon double bond. In some embodiments, the alkenyl group is optionally substituted. In some embodiments, the alkenyl group is a C2-C6alkenyl.

The term “akynyl” as used herein refers to C2-C12alkyl group that contains at least one carbon-carbon triple bond. In some embodiments, the alkenyl group is optionally substituted. In some embodiments, the alkynyl group is a C2-C6alkynyl.

The terms “alkoxy,” “alkylamino” and “alkylthio”, are used in their conventional sense, and refer to those alkyl groups attached to the remainder of the molecule via an oxygen atom (“oxy”), an amino group (“amino”) or thio group. The term “alkylamino” includes mono-di-alkylamino groups, the alkyl portions can be the same or different.

The terms “halo” or “halogen”, by itself or as part of another substituent, means a fluorine, chlorine, bromine, or iodine atom.

The term “heteroalkyl” refers to an alkyl group in which one or more carbon atom has been replaced by a heteroatom selected from S, O, P and N. Exemplary heteroalkyls include alkyl ethers, secondary and tertiary alkyl amines, alkyl amides, alkyl sulfides, and the like. The group may be a terminal group or a bridging group. As used herein reference to the normal chain when used in the context of a bridging group refers to the direct chain of atoms linking the two terminal positions of the bridging group.

The term “aryl” as used herein refers to a single, all carbon aromatic ring or a multiple condensed all carbon ring system wherein at least one of the rings is aromatic. For example, in certain embodiments, an aryl group has 6 to 12 carbon atoms. Aryl includes a phenyl radical. Aryl also includes multiple condensed ring systems (e.g., ring systems comprising 2, 3 or 4 rings) having about 9 to 12 carbon atoms in which at least one ring is aromatic and wherein the other rings may be aromatic or not aromatic. Such multiple condensed ring systems are optionally substituted with one or more (e.g., 1, 2 or 3) oxo groups on any carbocycle portion of the multiple condensed ring system. The rings of the multiple condensed ring system can be connected to each other via fused, spiro and bridged bonds when allowed by valency requirements. It is to be understood that the point of attachment of a multiple condensed ring system, as defined above, can be at any position of the aromatic ring. Non-limiting examples of aryl groups include, but are not limited to, phenyl, indenyl, naphthyl, 1,2,3,4-tetrahydronaphthyl, and the like.

The term “heteroaryl” as used herein refers to a single aromatic ring that has at least one atom other than carbon in the ring, wherein the atoms are selected from the group consisting of oxygen, nitrogen and sulfur; “heteroaryl” also includes multiple condensed ring systems that have at least one such aromatic ring, which multiple condensed ring systems are further described below. Thus, “heteroaryl” includes single aromatic rings of from about 1 to 6 carbon atoms and about 1-4 heteroatoms selected from the group consisting of oxygen, nitrogen and sulfur. The sulfur and nitrogen atoms may also be present in an oxidized form provided the ring is aromatic. Exemplary heteroaryl ring systems include but are not limited to pyridyl, pyrimidinyl, oxazolyl or furyl. “Heteroaryl” also includes multiple condensed ring systems (e.g., ring systems comprising 2, 3 or 4 rings) wherein a heteroaryl group, as defined above, is condensed with one or more rings selected from heteroaryls (to form for example a naphthyridinyl such as 1,8-naphthyridinyl), heterocycles, (to form for example a 1, 2, 3, 4-tetrahydronaphthyridinyl such as 1,2,3,4-tetrahydro-1,8-naphthyridinyl), carbocycles (to form for example 5,6,7,8-tetrahydroquinolyl) and aryls (to form for example indazolyl) to form the multiple condensed ring system. Thus, a heteroaryl (a single aromatic ring or multiple condensed ring system) has about 1-20 carbon atoms and about 1-6 heteroatoms within the heteroaryl ring. A heteroaryl (a single aromatic ring or multiple condensed ring system) can also have about 5 to 12 or about 5 to 10 members within the heteroaryl ring. Multiple condensed ring systems may be optionally substituted with one or more (e.g., 1, 2, 3 or 4) oxo groups on the carbocycle or heterocycle portions of the condensed ring. The rings of a multiple condensed ring system can be connected to each other via fused, spiro and bridged bonds when allowed by valency requirements. It is to be understood that the individual rings of the multiple condensed ring system may be connected in any order relative to one another. It is also to be understood that the point of attachment of a multiple condensed ring system (as defined above for a heteroaryl) can be at any position of the heteroaryl ring. It is also to be understood that the point of attachment for a heteroaryl or heteroaryl multiple condensed ring system can be at any suitable atom of the heteroaryl ring including a carbon atom and a heteroatom (e.g., a nitrogen). Exemplary heteroaryls include but are not limited to pyridyl, pyrrolyl, pyrazinyl, pyrimidinyl, pyridazinyl, pyrazolyl, thienyl, indolyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, furyl, oxadiazolyl, thiadiazolyl, quinolyl, isoquinolyl, benzothiazolyl, benzoxazolyl, indazolyl, quinoxalyl, quinazolyl, 5,6,7,8-tetrahydroisoquinolinyl benzofuranyl, benzimidazolyl, thianaphthenyl, pyrrolo[2,3-b]pyridinyl, quinazolinyl-4(3H)-one, triazolyl, 4,5,6,7-tetrahydro-1H-indazole and 3b,4,4a,5-tetrahydro-1H-cyclopropa[3,4]cyclo-penta[1,2-c]pyrazole. In one embodiment the term “heteroaryl” refers to a single aromatic ring containing at least one heteroatom. For example, the term includes 5-membered and 6-membered monocyclic aromatic rings that include one or more heteroatoms. Non-limiting examples of heteroaryl include but are not limited to pyridyl, furyl, thiazole, pyrimidine, oxazole, and thiadiazole.

The term “heterocyclyl” or “heterocycle” as used herein refers to a single saturated or partially unsaturated ring that has at least one atom other than carbon in the ring, wherein the atom is selected from the group consisting of oxygen, nitrogen and sulfur; the term also includes multiple condensed ring systems that have at least one such saturated or partially unsaturated ring, which multiple condensed ring systems are further described below. Thus, the term includes single saturated or partially unsaturated rings (e.g., 3, 4, 5, 6 or 7-membered rings) from about 1 to 6 carbon atoms and from about 1 to 3 heteroatoms selected from the group consisting of oxygen, nitrogen and sulfur in the ring. The ring may be substituted with one or more (e.g., 1, 2 or 3) oxo groups and the sulfur and nitrogen atoms may also be present in their oxidized forms. Exemplary heterocycles include but are not limited to azetidinyl, tetrahydrofuranyl and piperidinyl. The term “heterocycle” also includes multiple condensed ring systems (e.g., ring systems comprising 2, 3 or 4 rings) wherein a single heterocycle ring (as defined above) can be condensed with one or more groups selected from heterocycles (to form for example a 1,8-decahydronapthyridinyl), carbocycles (to form for example a decahydroquinolyl) and aryls to form the multiple condensed ring system. Thus, a heterocycle (a single saturated or single partially unsaturated ring or multiple condensed ring system) has about 2-20 carbon atoms and 1-6 heteroatoms within the heterocycle ring. Such multiple condensed ring systems may be optionally substituted with one or more (e.g., 1, 2, 3 or 4) oxo groups on the carbocycle or heterocycle portions of the multiple condensed ring. The rings of the multiple condensed ring system can be connected to each other via fused, spiro and bridged bonds when allowed by valency requirements. It is to be understood that the individual rings of the multiple condensed ring system may be connected in any order relative to one another. Accordingly, a heterocycle (a single saturated or single partially unsaturated ring or multiple condensed ring system) has about 3-20 atoms including about 1-6 heteroatoms within the heterocycle ring system. It is also to be understood that the point of attachment of a multiple condensed ring system (as defined above for a heterocylyl) can be at any position of the heterocyclic ring. It is also to be understood that the point of attachment for a heterocycle or heterocycle multiple condensed ring system can be at any suitable atom of the heterocyclic ring including a carbon atom and a heteroatom (e.g., a nitrogen). In one embodiment the term heterocycle includes a C2-20heterocycle. In one embodiment the term heterocycle includes a C2-7heterocycle. In one embodiment the term heterocycle includes a C2-5heterocycle. In one embodiment the term heterocycle includes a C2-4heterocycle. Exemplary heterocycles include, but are not limited to aziridinyl, azetidinyl, pyrrolidinyl, piperidinyl, homopiperidinyl, morpholinyl, thiomorpholinyl, piperazinyl, tetrahydrofuranyl, dihydrooxazolyl, tetrahydropyranyl, tetrahydrothiopyranyl, 1,2,3,4-tetrahydroquinolyl, benzoxazinyl, dihydrooxazolyl, chromanyl, 1,2-dihydropyridinyl, 2,3-dihydrobenzofuranyl, 1,3-benzodioxolyl, 1,4-benzodioxanyl, spiro[cyclopropane-1,1′-isoindolinyl]-3′-one, isoindolinyl-1-one, 2-oxa-6-azaspiro[3.3]heptanyl, imidazolidin-2-one N-methylpiperidine, imidazolidine, pyrazolidine, butyrolactam, valerolactam, imidazolidinone, hydantoin, dioxolane, phthalimide, 1,4-dioxane, thiomorpholine, thiomorpholine-S-oxide, thiomorpholine-S,S-oxide, pyran, 3-pyrroline, thiopyran, pyrone, tetrhydrothiophene, quinuclidine, tropane, 2-azaspiro[3.3]heptane, (1R,5S)-3-azabicyclo[3.2.1]octane, (1s,4s)-2-azabicyclo[2.2.2]octane, (1R,4R)-2-oxa-5-azabicyclo[2.2.2]octane and pyrrolidin-2-one. In one embodiment the term “heterocycle” refers to a monocyclic, saturated or partially unsaturated, 3-8 membered ring having at least one heteroatom. For example, the term includes a monocyclic, saturated or partially unsaturated, 4, 5, 6, or 7 membered ring having at least one heteroatom. Non-limiting examples of heterocycle include aziridine, azetidine, pyrrolidine, piperidine, piperidine, piperazine, oxirane, morpholine, and thiomorpholine. The term “9- or 10-membered heterobicycle” as used herein refers to a partially unsaturated or aromatic fused bicyclic ring system having at least one heteroatom. For example, the term 9- or 10-membered heterobicycle includes a bicyclic ring system having a benzo ring fused to a 5-membered or 6-membered saturated, partially unsaturated, or aromatic ring that contains one or more heteroatoms.

As used herein, the term “heteroatom” is meant to include oxygen (O), nitrogen (N), sulfur (S) and silicon (Si). The nitrogen and sulfur can be in an oxidized form when feasible.

As used herein, the term “stereoisomers” refers to compounds which have identical chemical constitution, but differ with regard to the arrangement of the atoms or groups in space, e.g., enantiomers, diastereomers, tautomers.

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

The term “effective” is used to describe an amount of a compound, composition or component which, when used within the context of its intended use, effects an intended result. The term effective subsumes all other effective amount or effective concentration terms, which are otherwise described or used in the present application.

A “pharmaceutically acceptable excipient” refers to a substance that is non-toxic, biologically tolerable, and otherwise biologically suitable for administration to a subject, such as an inert substance, added to a pharmacological composition or otherwise used as a vehicle, carrier, or diluent to facilitate administration of an agent and that is compatible therewith. Examples of excipients include calcium carbonate, calcium phosphate, various sugars and types of starch, cellulose derivatives, gelatin, vegetable oils, and polyethylene glycols.

A “solvate” refers to a physical association of a compound of Formula I with one or more solvent molecules.

“Treating” or “treatment” of any disease or disorder refers, in one embodiment, to ameliorating the disease or disorder (e.g., arresting or reducing the development of the disease or at least one of the clinical symptoms thereof). In another embodiment “treating” or “treatment” refers to ameliorating at least one physical parameter, which may not be discernible by the subject. In yet another embodiment, “treating” or “treatment” refers to modulating the disease or disorder, either physically, (e.g., stabilization of a discernible symptom), physiologically, (e.g., stabilization of a physical parameter), or both. In yet another embodiment, “treating” or “treatment” refers to delaying the onset of the disease or disorder.

In one aspect, the disclosure is directed to a compound of Formula (I):
PTM-ULM  (I)
or a pharmaceutically acceptable salt or solvate thereof, wherein PTM is a moiety of Formula IA:

wherein

R1is a covalent bond, or chemical moiety that links PTM and ULM;

* is a point of attachment to ULM;

each W is independently optionally substituted —CH2—, —C(O)—, —S(O)—, or —S(O)2—, wherein when n=2 or 3, only one W may be —C(O)—, —S(O)—, or —S(O)2—;

Z and Y are each independently N, or CRhwherein Rh═H or absent or, if W is attached to Z, then Z is C and Y is N or CRhwherein Rhis H; or if W is attached to Y, then Y is C and Z is N or CRhwherein Rhis H;

B is an optionally substituted 5-7 membered cycloalkyl ring, an optionally substituted 5-7 membered heteroaryl ring, or an optionally substituted 5-7 membered heterocyclic ring, wherein ring B is fused to ring C through Y and Z; and ULM is a small molecule E3 Ubiquitin Ligase binding moiety that binds a Von Hippel-Lindau E3 Ubiquitin Ligase.

In some aspects, the compounds of Formula I include a protein targeting moiety (PTM). In some aspects, the PTM in the compounds of Formula I is a moiety of Formula IA

According to the disclosure, B is a ring fused to ring “C” via Y and Z.

In some aspects, B in Formula IA is an optionally substituted 5-7 membered cycloalkyl ring. an optionally substituted 5-7 membered heteroaryl ring, or an optionally substituted 5-7 membered heterocyclic ring.

In some embodiments, B in Formula IA is an optionally substituted 5-7 membered cycloalkyl ring.

In some embodiments, B in Formula IA is an optionally substituted 5-7 membered cycloalkyl ring wherein the optional substituents are hydroxy, halogen, alkoxy, alkyl, haloalkyl, amino, alkylamino, or cyano.

In some embodiments, B in Formula IA is an optionally substituted 5-7 membered heteroaryl ring.

In some embodiments, B in Formula IA is an optionally substituted 5-7 membered heteroaryl ring wherein the optional substituents are hydroxy, halogen, alkoxy, alkyl, haloalkyl, amino, alkylamino, or cyano.

In other embodiments, B in Formula IA is an optionally substituted 5-7 membered heterocyclic ring.

In some embodiments, B in Formula IA is an optionally substituted 5-7 membered heterocyclic ring wherein the optional substituents are hydroxy, halogen, alkoxy, alkyl, haloalkyl, amino, alkylamino, cyano.

In some aspects, n in Formula IA is 0-3. In some embodiments, n=0. In other embodiments, n=1. In other embodiments, n=2. In other embodiments, n=3.

In some aspects, each W in Formula IA is independently optionally substituted —CH2—, —C(O)—, —S(O)—, or —S(O)2—, wherein when n=2 or 3, only one W may be —C(O)—, —S(O)—, or —S(O)2—.

In some embodiments, W in Formula IA is optionally substituted —CH2—. In other embodiments, W in Formula IA is —CH2—.

In some embodiments, W in Formula IA is optionally substituted —CH2— wherein the optional substituent is an alkyl group, such as, for example methyl (—CH3), ethyl, propyl, and the like.

In other embodiments, W in Formula IA is —C(CH3)H—.

In some embodiments, W in Formula IA is —C(O)—.

In some embodiments, W in Formula IA is —S(O)—.

In some embodiments, W in Formula IA is optionally substituted —S(O)2—.

In embodiments of the disclosure wherein n is 2 or 3, then only one W may be —C(O)—, —S(O)—, or —S(O)2—.

In some aspects, Z and Y in Formula IA are each independently N, or CRhwherein Rh=H or may be absent when n=1-3 such that a double bond is formed between Z and Y, or, if R1is attached to Z, then Z is C and Y is N or CRhwherein Rhis H; or if R1is attached to Y, then Y is C and Z is N or CRhwherein Rhis H. Examples of these embodiments include:

In some embodiments, Z is N.

In other embodiments, Z is CRhwherein Rh=H.

In other embodiments, Z is CRhwherein Rh=absent, and Z is bonded to Y by a double bond.

In some embodiments, Z is C and is attached to R1.

In some embodiments, Y is N.

In other embodiments, Y is CRhwherein Rh=H.

In other embodiments, Y is CRhwherein Rh=absent, and Y is bonded to Z by a double bond.

In some embodiments, Y is C and is attached to R1.

In some embodiments, the PTM is a moiety of Formula IA wherein * is a point of attachment to ULM.

In some aspects, R1in Formula IA is a covalent bond, or chemical moiety that links PTM and ULM.

In some embodiments, R1in Formula IA is a covalent bond.

In other embodiments, R1in Formula IA is a chemical moiety that links PTM and ULM.

Chemical moieties that are used to link PTM and ULM moieties are known in the art. These moieties are sometimes referred to as “linkers” in the art. In some embodiments, R1in Formula IA is a chemical moiety that is used to link a PTM and ULM that is known in the art.

In some embodiments, R1in Formula IA is a chemical moiety that is used to link a PTM and ULM as described in U.S. Patent Application Publication No. 2019/0300521, the entirety of which is incorporated by reference herein.

In other embodiments, R1in Formula IA is a chemical moiety that is used to link a PTM and ULM as described in U.S. Patent Application Publication No. 2019/0255066, the entirety of which is incorporated by reference herein.

In other embodiments, R1in Formula IA is a chemical moiety that is used to link a PTM and ULM as described in WO 2019/084030, the entirety of which is incorporated by reference herein.

In other embodiments, R1in Formula IA is a chemical moiety that is used to link a PTM and ULM as described in WO 2019/084026, the entirety of which is incorporated by reference herein.

In some embodiments, R1in Formula IA is a chemical structural unit represented by the formula:
-(A)q-,
wherein:

q is an integer from 1 to 14;

In these embodiments, q represents the number of connected A groups. For example, when q=1, -(A)q- is -A1-; when q=2, -(A)q- is -A1-A2-; when q=3, -(A)q- is -A1-A2-A3-; when q=4, -(A)q- is -A1-A2-A3-A4-; when q=5, -(A)q- is -A1-A2-A3-A4-A5-; when q=6, -(A)q- is -A1-A2-A3-A4-A5-A6-; when q=7, -(A)q- is -A1-A2-A3-A4-A5-A6-A7-; when q=8, -(A)q- is -A1-A2-A3-A4-A5-A6-A7-A8-; when q=9, -(A)q- is -A1-A2-A3-A4-A5-A6-A7-A8-A9-; when q=10, -(A)q- is -A1-A2-A3-A4-A5-A6-A7-A8-A9-A10-; when q=11, -(A)q- is -A1-A2-A3-A4-A5-A6-A7-A8-A9-A10-A11-; when q=12, -(A)q- is -A1-A2-A3-A4-A5-A6-A7-A8-A9-A10-A11-A12-; when q=13, -(A)q- is -A1-A2-A3-A4-A5-A6-A7-A8-A9-A10-A11-A12-A13-; and when q=14, -(A)q- is -A1-A2-A3-A4-A5-A6-A7-A8-A9-A10-A11-A12-A13-A14-.

R1cand R1dare each independently selected from the group consisting of H, D, optionally substituted C1-4alkyl, C3-8cyclcoalkyl, C3-8heterocyclcoalkyl, aryl, or heteroaryl.

R1cand R1dare each independently selected from the group consisting of H, D, optionally substituted C1-4alkyl, C3-8cyclcoalkyl, C3-8heterocyclcoalkyl, aryl, or heteroaryl.

R1cand R1dare each independently selected from the group consisting of H, D, optionally substituted C1-4alkyl, C3-8cyclcoalkyl, C3-8heterocyclcoalkyl, aryl, or heteroaryl.

R1cand R1dare each independently selected from the group consisting of H, D, optionally substituted C1-4alkyl, C3-8cyclcoalkyl, C3-8heterocyclcoalkyl, aryl, or heteroaryl.

In some embodiments, R1is —(CR1aR1b)1-5, for example —(CH2)1-5—, —CH2—, —CH2CH2CH2— and the like.

In some embodiments, R1is —(CR1aR1b)1-5-A- wherein A is O, S, or NR1c, such as for example, —(CH2)1-5—O—, —(CH2)1-5—S—, —(CH2)1-5—NH—, or —(CH2)0-2—(C(CH3)2)—(CH2)0-2—O—.

In some embodiments, R1is —(C≡C)—(CR1aR1b)1-5, such as, for example, —(C≡C)—(CH2)2—, and the like.

In some aspects, the Y in the compound of Formula IA is CRhwherein Rhis H, and the compound of Formula IA has Formula IA-1:

In some embodiments, n in Formula IA-1 is 1.

In some embodiments of the compound of Formula IA-1, at least one W is optionally substituted —CH2—.

In some embodiments of the compound of Formula IA-1, at least one W is optionally substituted —CH2— wherein the optional substituents are alkyl, alkoxy, alkylamino.

In some embodiments of the compound of Formula IA-1, at least one W is —CH2—.

In some embodiments, W in Formula IA-1 is optionally substituted —CH2— wherein the optional substituent is an alkyl group, such as, for example methyl (—CH3), ethyl, propyl, and the like.

In other embodiments, W in Formula IA-1 is —CH(CH3)—.

In some embodiments of the compound of Formula IA-1, one W is —C(O)—.

In some embodiments of the compound of Formula IA-1, one W is —S(O)—.

In some embodiments of the compound of Formula IA-1, one W is —S(O)2—.

In some embodiments, B in Formula IA-1 is an optionally substituted 5-7 membered cycloalkyl ring.

In some embodiments, B in Formula IA-1 is an optionally substituted 5-7 membered cycloalkyl ring wherein the optional substituents are hydroxy, halogen, alkoxy, alkyl, haloalkyl, amino, alkylamino, or cyano.

In other embodiments, B in Formula IA-1 is an optionally substituted 5-7 membered heterocyclic ring.

In some embodiments, B in Formula IA-1 is an optionally substituted 5-7 membered heterocyclic ring wherein the optional substituents are hydroxy, halogen, alkoxy, alkyl, haloalkyl, amino, alkylamino, cyano.

In other aspects, the Y in the compound of Formula IA is N, and Z is CRhwherein Rhis H, and the compound of Formula IA has Formula IA-2.

In some embodiments, n in Formula IA-2 is 1.

In some embodiments of the compound of Formula IA-2, at least one W is optionally substituted —CH2—.

In some embodiments of the compound of Formula IA-2, at least one W is optionally substituted —CH2— wherein the optional substituents are alkyl, alkoxy, or alkylamino.

In some embodiments of the compound of Formula IA-2, at least one W is —CH2—.

In some embodiments, W in Formula IA-2 is optionally substituted —CH2— wherein the optional substituent is an alkyl group, such as, for example methyl (—CH3), ethyl, propyl, and the like.

In other embodiments, W in Formula IA-2 is —CH(CH3)—.

In some embodiments of the compound of Formula IA-2, one W is —C(O)—.

In some embodiments of the compound of Formula IA-2, one W is —S(O)—.

In some embodiments of the compound of Formula IA-2, one W is —S(O)2—.

In some embodiments, B in Formula IA-2 is an optionally substituted 5-7 membered heterocyclic ring.

In some embodiments, B in Formula IA-2 is an optionally substituted 5-7 membered heterocyclic ring wherein the optional substituents are hydroxy, halogen, alkoxy, alkyl, haloalkyl, amino, alkylamino, cyano.

In other embodiments, B in Formula IA-2 is an optionally substituted 5-7 membered heterocyclic ring.

In some embodiments, B in Formula IA-2 is an optionally substituted 5-7 membered heterocyclic ring wherein the optional substituents are hydroxy, halogen, alkoxy, alkyl, haloalkyl, amino, alkylamino, or cyano.

In some aspects, the compound of Formula IA is a compound of Formula IA-3:

X is optionally substituted —CH2—, or NH; or, if IV is attached to X, then X is —CH— or N; Q is optionally substituted —CH2—, optionally substituted —(CH2)2—, —C(O)—, optionally substituted —CH2C(O)—, —S(O)—, —S(O)2—, optionally substituted —CH2S(O)2—, or optionally substituted —CH2S(O)—; and wherein Rc1, Rd1, Re3, W, Z, B, n, and R1are as described above for Formula IA.

In some embodiments of the compound of Formula IA-3, n=1. In other embodiments of the compound of Formula IA-3, n=2. In other embodiments of the compound of Formula IA-3, n=3.

In some embodiments of the compound of Formula IA-3, X is —CH—.

In other embodiments of the compound of Formula IA-3, X is NH.

In some of those embodiments of the compound of Formula IA-3 wherein IV is attached to X, then X is CH.

In other of those embodiments of the compound of Formula IA-3 wherein IV is attached to X, then X is N.

In some embodiments of the compound of Formula IA-3, Q is optionally substituted —CH2—.

In some embodiments of the compound of Formula IA-3, Q is optionally substituted —CH2— wherein the optional substituents are alkyl, alkoxy, or alkylamino.

In some embodiments of the compound of Formula IA-3, Q is optionally substituted —(CH2)2—.

In some embodiments of the compound of Formula IA-3, Q is optionally substituted —(CH2)2— wherein the optional substituents are alkyl, alkoxy, or alkylamino.

In some embodiments of the compound of Formula IA-3, Q is —C(O)—.

In some embodiments of the compound of Formula IA-3, Q is optionally substituted —CH2C(O)—.

In some embodiments of the compound of Formula IA-3, Q is —S(O)—.

In some embodiments of the compound of Formula IA-3, Q is —S(O)2—.

In some embodiments of the compound of Formula IA-3, Q is optionally substituted —CH2S(O)2—.

In some embodiments of the compound of Formula IA-3, Q is optionally substituted —CH2S(O)—.

In some aspects, the compound of Formula IA is a compound of Formula IA-4

In some embodiments of the compound of Formula IA-4, n=1. In other embodiments of the compound of Formula IA-4, n=2. In other embodiments of the compound of Formula IA-4, n=3.

In some embodiments of the compound of Formula IA-4, m=1. In other embodiments of the compound of Formula IA-4, m=2. In other embodiments of the compound of Formula IA-4, m=3.

In some embodiments of the compound of Formula IA-4, s=0. In some embodiments of the compound of Formula IA-4, s=1. In other embodiments of the compound of Formula IA-4, s=2. In other embodiments of the compound of Formula IA-4, s=3.

In some embodiments of the compound of Formula IA-4, Rk=H.

In some embodiments of the compound of Formula IA-4, Rk=D.

In some embodiments of the compound of Formula IA-4, Rk=F.

In some embodiments of the compound of Formula IA-4, Rk=C1-3alkyl, for example, C1alkyl, C2alkyl, C3alkyl, —CH3, —CH2CH3, and the like.

In some embodiments of the compound of Formula IA-4, Rk=C1-3haloalkyl, for example, C1haloalkyl, C2haloalkyl, C3haloalkyl, —CF3, —CH2CF3, and the like.

In some embodiments of the compound of Formula IA-4, Rk=C1-4alkoxyl, for example, C1alkoxyl, C2alkoxyl, C3alkoxyl, —OCH3, —OCH2CH3, and the like.

In some embodiments of the compound of Formula IA-4, Rk=substituted C1-3alkyl, for example, substituted C1alkyl, substituted C2alkyl, substituted C3alkyl, and the like.

In some embodiments of the compound of Formula IA-4, Rk=substituted C1-3haloalkyl, for example, substituted C1haloalkyl, substituted C2haloalkyl, substituted C3haloalkyl, and the like.

In some embodiments of the compound of Formula IA-4, Rk=substituted C1-4alkoxyl, for example, substituted C1alkoxyl, substituted C2alkoxyl, substituted C3alkoxyl, and the like.

In some aspects, the compound of Formula IA is a compound of Formula IA-5:

In some embodiments of the compound of Formula IA-5, m=1. In other embodiments of the compound of Formula IA-5, m=2. In other embodiments of the compound of Formula IA-5, m=3.

In some embodiments of the compound of Formula IA-5, s=0. In some embodiments of the compound of Formula IA-5, s=1. In other embodiments of the compound of Formula IA-5, s=2. In other embodiments of the compound of Formula IA-5, s=3.

In some embodiments of the compound of Formula IA-5, Rk=H.

In some embodiments of the compound of Formula IA-5, Rk=D.

In some embodiments of the compound of Formula IA-5, Rk=F.

In some embodiments of the compound of Formula IA-5, Rk=C1-3alkyl, for example, C1alkyl, C2alkyl, C3alkyl, —CH3, —CH2CH3, and the like.

In some embodiments of the compound of Formula IA-5, Rk=C1-3haloalkyl, for example, C1haloalkyl, C2haloalkyl, C3haloalkyl, —CF3, —CH2CF3, and the like.

In some embodiments of the compound of Formula IA-5, Rk=H. or C1-4alkoxyl, for example, C1alkoxyl, C2alkoxyl, C3alkoxyl, —OCH3, —OCH2CH3, and the like.

In some aspects, the compound of Formula IA is a compound of Formula IA-6:

In some embodiments of the compound of Formula IA-6, s=0. In some embodiments of the compound of Formula IA-6, s=1. In other embodiments of the compound of Formula IA-6, s=2. In other embodiments of the compound of Formula IA-6, s=3.

In some embodiments of the compound of Formula IA-6, Rk=H.

In some embodiments of the compound of Formula IA-6, Rk=D.

In some embodiments of the compound of Formula IA-6, Rk=F.

In some embodiments of the compound of Formula IA-6, Rk=C1-3alkyl, for example, C1alkyl, C2alkyl, C3alkyl, —CH3, —CH2CH3, and the like.

In some embodiments of the compound of Formula IA-6, Rk=C1-3haloalkyl, for example, C1haloalkyl, C2haloalkyl, C3haloalkyl, —CF3, —CH2CF3, and the like.

In some embodiments of the compound of Formula IA-6, Rk=H. or C1-4alkoxyl, for example, C1alkoxyl, C2alkoxyl, C3alkoxyl, —OCH3, —OCH2CH3, and the like.

In some aspects, the ULM moiety in the compounds of the disclosure is a small molecule E3 Ubiquitin Ligase binding moiety that binds a Von Hippel-Lindau E3 Ubiquitin Ligase (VHL). Such ULM moieties that bind to VHL are known to those of skill in the art. Methods of determining whether a small molecule binds a Von Hippel-Lindau E3 Ubiguitin Ligase are known in the art.

In some embodiments, the ULM is a previously described ULM.

In some embodiments, the ULM is a ULM moiety described in U.S. Patent Application Publication No. 2019/0300521, the entirety of which is incorporated by reference herein.

In other embodiments, the ULM is a ULM moiety described in U.S. Patent Application Publication No. 2019/0255066, the entirety of which is incorporated by reference herein.

In other embodiments, the ULM is a ULM moiety described in WO 2019/084030, the entirety of which is incorporated by reference herein.

In other embodiments, the ULM is a ULM moiety described in WO 2019/084026, the entirety of which is incorporated by reference herein.

In some embodiments, the ULM is a moiety having the Formula ULM-I

wherein
------ indicates the position of attachment of the ULM to R1;
V is H or F;
R3is optionally substituted phenyl, optionally substituted napthyl, or an optionally substituted 5-10 membered heteroaryl;
one of R4or R5is H, D, haloalkyl, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, —CORd, CONRe1Re2;
the other of R4or R5is H or D;
or R4and R5, together with the carbon atom to which they are both attached, form an optionally substituted 3-5 membered cycloalkyl, or heterocyclyl;
W3is an optionally substituted aryl, optionally substituted heteroaryl, or

R6and R7are independently H, D, optionally substituted alkyl, optionally substituted cycloalkyl, or optionally substituted haloalkyl,
or R6, R7, and the carbon atom to which they are attached form an optionally substituted cycloalkyl or optionally substituted heterocyclyl;
R8is an optionally substituted heterocyclyl, optionally substituted heteroaryl, optionally substituted aryl, —C(O)NRaRb, —NRaRb,

Rais selected from H or optionally substituted alkyl;
Rbis selected from H, —C(O)—* wherein * is a point of attachment to R1, optionally substituted alkyl, optionally substituted alkylcarbonyl, optionally substituted (cycloalkyl)alkylcarbonyl, optionally substituted aralkylcarbonyl, optionally substituted arylcarbonyl, optionally substituted (cycloalkyl)carbonyl, optionally substituted (heterocyclyl)carbonyl, or optionally substituted aralkyl;
each Rcis independently H, halo, optionally substituted alkoxy, cyano, optionally substituted alkyl, haloalkyl, or haloalkoxy;
each Rdis independently selected from H, optionally substituted alkyl or NRe1Re2, each Re1and Re2is independently H, D, optionally substituted alkyl, or Re1and Re2together with the nitrogen atom to which they are attached form a 4-7 membered heterocyclyl; and
p is 0, 1, 2, 3, or 4.

In some embodiments of ULM-I, V is H.

In other embodiments of ULM-I, V is F.

In some embodiments of ULM-I, R3is optionally substituted phenyl having the formula:

In some embodiments wherein R3is optionally substituted phenyl, R9is an optionally substituted heteroaryl.

In some embodiments wherein R3is optionally substituted phenyl, R9is

each optionally substituted.

In other embodiments wherein R3is optionally substituted phenyl, R9is

In other embodiments wherein R3is optionally substituted phenyl, R9is

In other embodiments, R3is

In some embodiments of ULM-I, one of R4or R5is H, and the other of R4or R5is H or optionally substituted alkyl.

In other embodiments of ULM-I, one of R4or R5is H, and the other of R4or R5is optionally substituted C1-C6alkyl.

In other embodiments of ULM-I, one of R4or R5is H, and the other of R4or R5is C1-C6alkyl.

In other embodiments of ULM-I, one of R4or R5is H, and the other of R4or R5is —CH3.

In other embodiments of ULM-I, one of R4or R5is H, and the other of R4or R5is —CH2OH.

In other embodiments of ULM-I, both R4and R5are H.

In some embodiments of ULM-I, W3is

In some embodiments of ULM-I, R6is H.

In some embodiments of ULM-I, R7is H, or optionally substituted alkyl.

In some embodiments of ULM-I, R7is H.

In some embodiments of ULM-I, R7is optionally substituted alkyl.

In some embodiments of ULM-I, R7is optionally substituted C1-C6alkyl.

In some embodiments of ULM-I, R7is C1-C6alkyl.

In some embodiments of ULM-I, R7is C1-C6alk-OH, C1-C6alk-NH2, —C1-C6alk-CONH—*, or —C1-C6alk-NHCO—* wherein * is a point of attachment to R1.

In some embodiments of ULM-I, R7is -t-butyl or -isopropyl.

In some embodiments of ULM-I, R7is -t-butyl.

In some embodiments of ULM-I, R7is -isopropyl.

In some embodiments of ULM-I, R8is NRaRb.

In some embodiments, Rais H or optionally substituted alkyl.

In some embodiments, Rais H.

In some embodiments, Rbis H, optionally substituted alkyl, —C(O)—* wherein * is a point of attachment to R1, optionally substituted (cycloalkyl)carbonyl, or optionally substituted alkylcarbonyl.

In some embodiments, Rbis optionally substituted alkylcarbonyl.

In some embodiments, Rbis —C(O)—* wherein * is a point of attachment to R1.

In some embodiments of ULM-I, R8is CONRaRb.

In some embodiments of ULM-I, R8is

wherein * is a point of attachment to R1.

In some embodiments of ULM-I, R8is

wherein * is a point of attachment to R1.

In some embodiments of ULM-I, R8is

wherein * is a point of attachment to R1.

In some embodiments of ULM-I, R8is

In some embodiments, R8is —NH—* wherein * is a point of attachment to R1.

In some embodiments of ULM-I, R8is optionally substituted heteroaryl.

In some embodiments of ULM-I, R8is

In some embodiments, R8is

In some embodiments, R8is

wherein * is a point of attachment to R1.

In some embodiments, R8is

In some embodiments, R8is

In some embodiments, R8is

In some embodiments, R8is

In some embodiments, R8is

In some embodiments, R8is

In some embodiments, R8is

In some embodiments, R8is

In some embodiments, R8is

In some embodiments, ULM-I is a compound of formula:

* is a point of attachment of the ULM to R1.

In some embodiments of ULM-IA, ULM-IB, ULM-IC, or ULM-ID, R9is optionally substituted

In some embodiments, the ULM is a moiety having the Formula ULM-II

wherein
------ indicates the position of attachment of the ULM to R1;
R14is C1-C6alkyl, such as, for example, —CH3, —CH2CH3, —CH(CH3)2, and the like; and all of the other variables have the same scope as set forth above with respect to ULM-I.

In some aspects, the compounds of Formula I are those having the formula IA-7 or IA-8:

R7is optionally substituted alkyl, preferably optionally substituted C1-C6alkyl, and more preferably C1-C6alkyl; and

In some aspects, the compounds of Formula I are those having the formula IA-9 or IA-10:

R7is optionally substituted alkyl, preferably optionally substituted C1-C6alkyl, and more preferably C1-C6alkyl; and

each Re1and Re2is independently H, D, optionally substituted alkyl, or Re1and Re2together with the nitrogen atom to which they are attached form a 4-7 membered heterocyclyl.

In some embodiments of the compound of Formula IA-9 or IA-10, n=1. In other embodiments of the compound of Formula IA-9 or IA-10, n=2. In other embodiments of the compound of Formula IA-9 or IA-10, n=3.

In some embodiments of the compound of Formula IA-7 or IA-8, m=1. In other embodiments of the compound of Formula IA-7 or IA-8, m=2. In other embodiments of the compound of Formula IA-7 or IA-8, m=3.

In some embodiments of the compound of Formula IA-9 or IA-10, s=0. In some embodiments of the compound of Formula IA-9 or IA-10, s=1. In other embodiments of the compound of Formula IA-9 or IA-10, s=2. In other embodiments of the compound of Formula IA-9 or IA-10, p=3.

In some embodiments of the compound of Formula IA-9 or IA-10, Rk=H.

In some embodiments of the compound of Formula IA-9 or IA-10, Rk=D.

In some embodiments of the compound of Formula IA-9 or IA-10, Rk=F.

In some embodiments of the compound of Formula IA-9 or IA-10, Rk=C1-3alkyl, for example, C1alkyl, C2alkyl, C3alkyl, —CH3, —CH2CH3, and the like.

In some embodiments of the compound of Formula IA-9 or IA-10, Rk=C1-3haloalkyl, for example, C1haloalkyl, C2haloalkyl, C3haloalkyl, —CF3, —CH2CF3, and the like.

In some embodiments of the compound of Formula IA-9 or IA-10, Rk=H. or C1-4alkoxyl, for example, C1alkoxyl, C2alkoxyl, C3alkoxyl, —OCH3, —OCH2CH3, and the like.

In some embodiments of the compound of Formula IA-9 or IA-10, Rc1and Rd1are each H.

In some embodiments of the compound of Formula IA-9 or IA-10, Re3is H.

In some embodiments of the compound of Formula IA-9 or IA-10, Rc1, Rd1, and Re3and K are each H.

In some aspects, the compounds of Formula I are compounds of Formula IA-9a or IA-10a:

wherein X is N or CH, and the other variables are as set forth above with respect to Formula IA-9 and IA-10.

In some aspects, the compounds of Formula I are those having the formula IA-11 or IA-12:

In some embodiments, W is —CH2—.

In some embodiments, X is —N.

In other embodiments, X is —CH.

In some embodiments, s=0.

In some embodiments, R9is

In some embodiments of the compounds of formula IA-11 or formula IA-12, each R1aeach R1b, and each R1cis independently H or C1-C6alkyl.

In some embodiments of the compound of formula IA-11 or formula IA-12, R1is —(CR1aR1b)1-5, such as, for example, —CH2—, —CH2CH2—, —CH2CH2CH2—, —CH2CH2CH2CH2—, —CH2CH2CH2CH2CH2—, and the like.

In some embodiments of the compound of formula IA-11 or formula IA-12, R1is —(CR1aR1b)1-5-A- wherein A is O, S, or NR1c, such as, for example, —CH2—O—, —CH2CH2—O—, —CH2CH2CH2—O—, and the like.

In some embodiments of the compounds of formula I, R1is —(CR1aR1b)1-5-A-(CR1aR1b)1-5— wherein A is O, S, or NR1c, such as, for example, —CH2CH2CH2—N(CH3)—CH2CH2—, —CH2CH2—N(CH3)—CH2CH2—, —CH2CH2—O—CH2—, and the like.

In some embodiments of the compounds of formula I, R1is -(3-11 membered heterocyclyl optionally substituted with 0-6 R1aand/or R1bgroups)-(CO)—(CR1aR1b)1-5-A- wherein A is O, S, or NR1c, such as, for example, -piperidinyl-(CO)—CH(CH3)—O—, -pyrrolidinyl-(CO)—CH(CH3)—O—, -piperidinyl-(CO)—CH2—O—, -methylpiperidinyl-(CO)—CH2—O—, and the like.

In some embodiments of the compounds of formula I, R1is -(3-11 membered heterocyclyl optionally substituted with 0-6 R1aand/or R1bgroups)-(CO)-A-(CR1aR1b)1-5— wherein A is O, S, or NR1c, such as, for example, -piperidinyl-(CO)—O—CH2—, and the like.

In some embodiments of the compounds of formula I, R1is -(3-11 membered heterocyclyl optionally substituted with 0-6 R1aand/or R1bgroups)-(3-11 membered heterocyclyl optionally substituted with 0-6 R1aand/or R1bgroups)-(CR1aR1b)1-5-A- wherein A is O, S, or NR1c, such as, for example, -piperidinyl-pyrrolidinyl-CH2CH2—O—, -piperidinyl-piperidinyl-CH2CH2—O—, -pyrrolidinyl-piperidinyl-CH2CH2—O—, and the like.

In some embodiments of the compounds of formula I, R1is -(3-11 membered heterocyclyl optionally substituted with 0-6 R1aand/or R1bgroups)-(CR1aR1b)1-5(3-11 membered heterocyclyl optionally substituted with 0-6 R1aand/or R1bgroups)-(CR1aR1b)1-5-A- wherein A is O, S, or NR1c, such as, for example, -piperidinyl-CH2-piperidinyl-CH2CH2—O—, -piperidinyl-CH2CH2-piperidinyl-CH2CH2—O—, and the like.

In some embodiments of the compounds of formula I, R1is -(3-11 membered cycloalkyl optionally substituted with 0-6 R1aand/or R1bgroups)-A-(CR1aR1b)1-5— wherein each A is independently O, S, or NR1c, such as, for example, -cyclohexyl-N(CH3)—CH2CH2—O—, and the like.

In some embodiments of the compounds of formula I, R1is —(CO)-(3-11 membered heterocyclyl optionally substituted with 0-6 R1aand/or R1bgroups)-(CR1aR1b)1-5-A- wherein A is O, S, or NR1c, such as, for example, —(CO)-piperidinyl-CH2CH2—O—, and the like.

In some embodiments of the compounds of formula I, R1is —(CR1aR1b)1-5(3-11 membered heterocyclyl optionally substituted with 0-6 R1aand/or R1bgroups)-(CR1aR1b)1-5, such as, for example, —CH2-piperidinyl-CH2—, —CH2-piperidinyl-CH2CH2—, —CH2-piperidinyl-CH2CH2CH2—, and the like.

In some embodiments of the compounds of formula I, R1is —(CR1aR1b)1-5(3-11 membered heterocyclyl optionally substituted with 0-6 R1aand/or R1bgroups)-A- wherein A is O, S, or NR1c, such as, for example, —CH2CH2CH2-pyrrolidinyl-O—, and the like.

In some embodiments of the compounds of formula I, R1is -(heteroaryl optionally substituted with 0-4 R1aand/or R1bgroups)-A-(CR1aR1b)1-5— wherein A is O, S, or NR1c, such as, for example, -pyridinyl-O—CH2—, and the like.

In some embodiments of the compounds of formula I, R1is -A-(3-11 membered heterocyclyl optionally substituted with 0-6 R1aand/or R1bgroups)-(CR1aR1b)1-5-A- wherein each A is independently O, S, or NR1c, such as, for example, —N(CH3)-piperidinyl-CH2CH2—O—, and the like.

In some aspects, the compounds of Formula I are those having the formula IA-13a, IA-13b, IA-14a or IA-14b:

wherein X is N or CH.

In some embodiments of the compounds of formula IA-13a, IA-13b, IA-14a or IA-14b, R1is -(3-11 membered heterocyclyl optionally substituted with 0-6 R1aand/or R1bgroups)-(CO)—(CR1aR1b)1-5-A- wherein A is O, S, or NR1c, or -(3-11 membered heterocyclyl optionally substituted with 0-6 R1aand/or R1bgroups)-(CR1aR1b)1-5-A- wherein A is O, S, or NR1c,

In some embodiments of the compounds of formula IA-13a, IA-13b, IA-14a or IA-14b, R1is -(3-11 membered heterocyclyl)-(CO)—(CR1aR1b)1-3—O— wherein each R1ais H and each R1bis independently H or —C1-C8alkyl, preferably —CH3.

In other embodiments of the compounds of formula IA-13a, IA-13b, IA-14a or IA-14b, R1is -(3-11 membered heterocyclyl)-(CR1aR1b)1-3—O—, wherein each R1ais H and each R1bis independently H or —C1-C8alkyl, preferably —CH3.

In some aspects, the compounds of Formula I are those having the formula IA-15a, IA-15b, IA-16a, or IA-16b:

In some embodiments of the compound of formula IA-15a, IA-15b, IA-16a, or IA-16b, A is O and R1a1is —C1-C8alkyl, preferably —CH2CH3, or —CH3.

In some embodiments of the compound of formula IA-15a, IA-15b, IA-16a, or IA-16b, A1is a covalent bond.

In some embodiments of the compound of formula IA-15a, IA-15b, IA-16a, or IA-16b, A1is —(CR1aR1b)1-3.

In some embodiments of the compound of formula IA-15a, IA-15b, IA-16a, or IA-16b, A is O and R1a1is —CH3.

In some embodiments of the compound of formula IA-15a, IA-15b, IA-16a, or IA-16b, s=0.

In some embodiments of the compound of formula IA-15a, IA-15b, IA-16a, or IA-16b, s=1 and Rkis —CH3.

It will be apparent that the compounds of the invention, including all subgenera described herein, may have multiple stereogenic centers. As a result, there exist multiple stereoisomers (enantiomers and diastereomers) of the compounds (and subgenera described herein). The present disclosure contemplates and encompasses each stereoisomer of any compound encompassed by the disclosure as well as mixtures of said stereoisomers.

Pharmaceutically acceptable salts and solvates of the compounds of the disclosure (including all subgenera described herein) are also within the scope of the disclosure.

Isotopic variants of the compounds of the disclosure (including all subgenera described herein) are also contemplated by the present disclosure.

It is to be appreciated that certain features of the invention which are, for clarity, described herein in the context of separate embodiments, may also be provided in combination in a single embodiment. That is, unless obviously incompatible or specifically excluded, each individual embodiment is deemed to be combinable with any other embodiment(s) and such a combination is considered to be another embodiment. Conversely, various features of the invention that are, for brevity, described in the context of a single embodiment, may also be provided separately or in any sub-combination. While an embodiment may be described as part of a series of steps or part of a more general structure, each said step may also be considered an independent embodiment in itself, combinable with others.

Pharmaceutical Compositions and Methods of Administration

The subject pharmaceutical compositions are typically formulated to provide a therapeutically effective amount of a compound of the present disclosure as the active ingredient, or a pharmaceutically acceptable salt, ester, prodrug, solvate, hydrate or derivative thereof. Where desired, the pharmaceutical compositions contain pharmaceutically acceptable salt and/or coordination complex thereof, and one or more pharmaceutically acceptable excipients, carriers, including inert solid diluents and fillers, diluents, including sterile aqueous solution and various organic solvents, permeation enhancers, solubilizers and adjuvants.

The subject pharmaceutical compositions can be administered alone or in combination with one or more other agents, which are also typically administered in the form of pharmaceutical compositions. Where desired, the one or more compounds of the invention and other agent(s) may be mixed into a preparation or both components may be formulated into separate preparations to use them in combination separately or at the same time. In some embodiments, the concentration of one or more compounds provided in the pharmaceutical compositions of the present invention is less than 100%, 90%, 80%, 70%, 60%, 50%, 40%, 30%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.9%, 0.8%, 0.7%, 0.6%, 0.5%, 0.4%, 0.3%, 0.2%, 0.1%, 0.09%, 0.08%, 0.07%, 0.06%, 0.05%, 0.04%, 0.03%, 0.02%, 0.01%, 0.009%, 0.008%, 0.007%, 0.006%, 0.005%, 0.004%, 0.003%, 0.002%, 0.001%, 0.0009%, 0.0008%, 0.0007%, 0.0006%, 0.0005%, 0.0004%, 0.0003%, 0.0002%, or 0.0001% (or a number in the range defined by and including any two numbers above) w/w, w/v or v/v.

In some embodiments, the concentration of one or more compounds of the invention is in the range from approximately 0.0001% to approximately 50%, approximately 0.001% to approximately 40%, approximately 0.01% to approximately 30%, approximately 0.02% to approximately 29%, approximately 0.03% to approximately 28%, approximately 0.04% to approximately 27%, approximately 0.05% to approximately 26%, approximately 0.06% to approximately 25%, approximately 0.07% to approximately 24%, approximately 0.08% to approximately 23%, approximately 0.09% to approximately 22%, approximately 0.1% to approximately 21%, approximately 0.2% to approximately 20%, approximately 0.3% to approximately 19%, approximately 0.4% to approximately 18%, approximately 0.5% to approximately 17%, approximately 0.6% to approximately 16%, approximately 0.7% to approximately 15%, approximately 0.8% to approximately 14%, approximately 0.9% to approximately 12%, approximately 1% to approximately 10% w/w, w/v or v/v.

In some embodiments, the concentration of one or more compounds of the invention is in the range from approximately 0.001% to approximately 10%, approximately 0.01% to approximately 5%, approximately 0.02% to approximately 4.5%, approximately 0.03% to approximately 4%, approximately 0.04% to approximately 3.5%, approximately 0.05% to approximately 3%, approximately 0.06% to approximately 2.5%, approximately 0.07% to approximately 2%, approximately 0.08% to approximately 1.5%, approximately 0.09% to approximately 1%, approximately 0.1% to approximately 0.9% w/w, w/v or v/v.

In some embodiments, the amount of one or more compounds of the invention is in the range of 0.0001-10 g, 0.0005-9 g, 0.001-8 g, 0.005-7 g, 0.01-6 g, 0.05-5 g, 0.1-4 g, 0.5-4 g, or 1-3 g.

The compounds according to the invention are effective over a wide dosage range. For example, in the treatment of adult humans, dosages from 0.01 to 1000 mg, from 0.5 to 100 mg, from 1 to 50 mg per day, and from 5 to 40 mg per day are examples of dosages that may be used. An exemplary dosage is 10 to 30 mg per day. The exact dosage will depend upon the route of administration, the form in which the compound is administered, the subject to be treated, the body weight of the subject to be treated, and the preference and experience of the attending physician.

A pharmaceutical composition of the invention typically contains an active ingredient (e.g., a compound of the disclosure) of the present invention or a pharmaceutically acceptable salt and/or coordination complex thereof, and one or more pharmaceutically acceptable excipients, carriers, including but not limited to inert solid diluents and fillers, diluents, sterile aqueous solution and various organic solvents, permeation enhancers, solubilizers and adjuvants.

Described below are non-limiting exemplary pharmaceutical compositions and methods for preparing the same.

Pharmaceutical Compositions for Oral Administration.

In some embodiments, the invention provides a pharmaceutical composition for oral administration containing a compound of the invention, and a pharmaceutical excipient suitable for oral administration.

In some embodiments, the invention provides a solid pharmaceutical composition for oral administration containing: (i) an effective amount of a compound of the invention; optionally (ii) an effective amount of a second agent; and (iii) a pharmaceutical excipient suitable for oral administration. In some embodiments, the composition further contains: (iv) an effective amount of a third agent.

In some embodiments, the pharmaceutical composition may be a liquid pharmaceutical composition suitable for oral consumption. Pharmaceutical compositions of the invention suitable for oral administration can be presented as discrete dosage forms, such as capsules, cachets, or tablets, or liquids or aerosol sprays each containing a predetermined amount of an active ingredient as a powder or in granules, a solution, or a suspension in an aqueous or nonaqueous liquid, an oil-in-water emulsion, or a water-in-oil liquid emulsion. Such dosage forms can be prepared by any of the methods of pharmacy, but all methods include the step of bringing the active ingredient into association with the carrier, which constitutes one or more necessary ingredients. In general, the compositions are prepared by uniformly and intimately admixing the active ingredient with liquid carriers or finely divided solid carriers or both, and then, if necessary, shaping the product into the desired presentation. For example, a tablet can be prepared by compression or molding, optionally with one or more accessory ingredients. Compressed tablets can be prepared by compressing in a suitable machine the active ingredient in a free-flowing form such as powder or granules, optionally mixed with an excipient such as, but not limited to, a binder, a lubricant, an inert diluent, and/or a surface active or dispersing agent. Molded tablets can be made by molding in a suitable machine a mixture of the powdered compound moistened with an inert liquid diluent.

This invention further encompasses anhydrous pharmaceutical compositions and dosage forms comprising an active ingredient, since water can facilitate the degradation of some compounds. For example, water may be added (e.g., 5%) in the pharmaceutical arts as a means of simulating long-term storage in order to determine characteristics such as shelf-life or the stability of formulations over time. Anhydrous pharmaceutical compositions and dosage forms of the invention can be prepared using anhydrous or low moisture containing ingredients and low moisture or low humidity conditions. Pharmaceutical compositions and dosage forms of the invention which contain lactose can be made anhydrous if substantial contact with moisture and/or humidity during manufacturing, packaging, and/or storage is expected. An anhydrous pharmaceutical composition may be prepared and stored such that its anhydrous nature is maintained. Accordingly, anhydrous compositions may be packaged using materials known to prevent exposure to water such that they can be included in suitable formulary kits. Examples of suitable packaging include, but are not limited to, hermetically sealed foils, plastic or the like, unit dose containers, blister packs, and strip packs.

An active ingredient can be combined in an intimate admixture with a pharmaceutical carrier according to conventional pharmaceutical compounding techniques. The carrier can take a wide variety of forms depending on the form of preparation desired for administration. In preparing the compositions for an oral dosage form, any of the usual pharmaceutical media can be employed as carriers, such as, for example, water, glycols, oils, alcohols, flavoring agents, preservatives, coloring agents, and the like in the case of oral liquid preparations (such as suspensions, solutions, and elixirs) or aerosols; or carriers such as starches, sugars, microcrystalline cellulose, diluents, granulating agents, lubricants, binders, and disintegrating agents can be used in the case of oral solid preparations, in some embodiments without employing the use of lactose. For example, suitable carriers include powders, capsules, and tablets, with the solid oral preparations. If desired, tablets can be coated by standard aqueous or nonaqueous techniques.

Examples of suitable fillers for use in the pharmaceutical compositions and dosage forms disclosed herein include, but are not limited to, talc, calcium carbonate (e.g., granules or powder), microcrystalline cellulose, powdered cellulose, dextrates, kaolin, mannitol, silicic acid, sorbitol, starch, pre-gelatinized starch, and mixtures thereof.

Disintegrants may be used in the compositions of the invention to provide tablets that disintegrate when exposed to an aqueous environment. Too much of a disintegrant may produce tablets which may disintegrate in the bottle. Too little may be insufficient for disintegration to occur and may thus alter the rate and extent of release of the active ingredient(s) from the dosage form. Thus, a sufficient amount of disintegrant that is neither too little nor too much to detrimentally alter the release of the active ingredient(s) may be used to form the dosage forms of the compounds disclosed herein. The amount of disintegrant used may vary based upon the type of formulation and mode of administration, and may be readily discernible to those of ordinary skill in the art. About 0.5 to about 15 weight percent of disintegrant, or about 1 to about 5 weight percent of disintegrant, may be used in the pharmaceutical composition. Disintegrants that can be used to form pharmaceutical compositions and dosage forms of the invention include, but are not limited to, agar-agar, alginic acid, calcium carbonate, microcrystalline cellulose, croscarmellose sodium, crospovidone, polacrilin potassium, sodium starch glycolate, potato or tapioca starch, other starches, pre-gelatinized starch, other starches, clays, other algins, other celluloses, gums or mixtures thereof.

Lubricants which can be used to form pharmaceutical compositions and dosage forms of the invention include, but are not limited to, calcium stearate, magnesium stearate, mineral oil, light mineral oil, glycerin, sorbitol, mannitol, polyethylene glycol, other glycols, stearic acid, sodium lauryl sulfate, talc, hydrogenated vegetable oil (e.g., peanut oil, cottonseed oil, sunflower oil, sesame oil, olive oil, corn oil, and soybean oil), zinc stearate, ethyl oleate, ethyl laureate, agar, or mixtures thereof. Additional lubricants include, for example, a syloid silica gel, a coagulated aerosol of synthetic silica, or mixtures thereof. A lubricant can optionally be added, in an amount of less than about 1 weight percent of the pharmaceutical composition.

When aqueous suspensions and/or elixirs are desired for oral administration, the active ingredient therein may be combined with various sweetening or flavoring agents, coloring matter or dyes and, if so desired, emulsifying and/or suspending agents, together with such diluents as water, ethanol, propylene glycol, glycerin and various combinations thereof.

The tablets can be uncoated or coated by known techniques to delay disintegration and absorption in the gastrointestinal tract and thereby provide a sustained action over a longer period. For example, a time delay material such as glyceryl monostearate or glyceryl distearate can be employed. Formulations for oral use can also be presented as hard gelatin capsules wherein the active ingredient is mixed with an inert solid diluent, for example, calcium carbonate, calcium phosphate or kaolin, or as soft gelatin capsules wherein the active ingredient is mixed with water or an oil medium, for example, peanut oil, liquid paraffin or olive oil.

Surfactant which can be used to form pharmaceutical compositions and dosage forms of the invention include, but are not limited to, hydrophilic surfactants, lipophilic surfactants, and mixtures thereof. That is, a mixture of hydrophilic surfactants may be employed, a mixture of lipophilic surfactants may be employed, or a mixture of at least one hydrophilic surfactant and at least one lipophilic surfactant may be employed.

A suitable hydrophilic surfactant may generally have an HLB value of at least 10, while suitable lipophilic surfactants may generally have an HLB value of or less than about 10. An empirical parameter used to characterize the relative hydrophilicity and hydrophobicity of non-ionic amphiphilic compounds is the hydrophilic-lipophilic balance (“HLB” value). Surfactants with lower HLB values are more lipophilic or hydrophobic, and have greater solubility in oils, while surfactants with higher HLB values are more hydrophilic, and have greater solubility in aqueous solutions.

Hydrophilic surfactants are generally considered to be those compounds having an HLB value greater than about 10, as well as anionic, cationic, or zwitterionic compounds for which the HLB scale is not generally applicable. Similarly, lipophilic (e.g., hydrophobic) surfactants are compounds having an HLB value equal to or less than about 10. However, HLB value of a surfactant is merely a rough guide generally used to enable formulation of industrial, pharmaceutical and cosmetic emulsions.

Hydrophilic surfactants may be either ionic or non-ionic. Suitable ionic surfactants include, but are not limited to, alkylammonium salts; fusidic acid salts; fatty acid derivatives of amino acids, oligopeptides, and polypeptides; glyceride derivatives of amino acids, oligopeptides, and polypeptides; lecithins and hydrogenated lecithins; lysolecithins and hydrogenated lysolecithins; phospholipids and derivatives thereof lysophospholipids and derivatives thereof; carnitine fatty acid ester salts; salts of alkylsulfates; fatty acid salts; sodium docusate; acyl lactylates; mono- and di-acetylated tartaric acid esters of mono- and di-glycerides; succinylated mono- and di-glycerides; citric acid esters of mono- and di-glycerides; and mixtures thereof.

Within the aforementioned group, ionic surfactants include, by way of example: lecithins, lysolecithin, phospholipids, lysophospholipids and derivatives thereof; carnitine fatty acid ester salts; salts of alkylsulfates; fatty acid salts; sodium docusate; acylactylates; mono- and di-acetylated tartaric acid esters of mono- and di-glycerides; succinylated mono- and di-glycerides; citric acid esters of mono- and di-glycerides; and mixtures thereof.

Hydrophilic non-ionic surfactants may include, but are not limited to, alkylglucosides; alkylmaltosides; alkylthioglucosides; lauryl macrogolglycerides; polyoxyalkylene alkyl ethers such as polyethylene glycol alkyl ethers; polyoxyalkylene alkylphenols such as polyethylene glycol alkyl phenols; polyoxyalkylene alkyl phenol fatty acid esters such as polyethylene glycol fatty acids monoesters and polyethylene glycol fatty acids diesters; polyethylene glycol glycerol fatty acid esters; polyglycerol fatty acid esters; polyoxyalkylene sorbitan fatty acid esters such as polyethylene glycol sorbitan fatty acid esters; hydrophilic transesterification products of a polyol with at least one member of the group consisting of glycerides, vegetable oils, hydrogenated vegetable oils, fatty acids, and sterols; polyoxyethylene sterols, derivatives, and analogues thereof, polyoxyethylated vitamins and derivatives thereof, polyoxyethylene-polyoxypropylene block copolymers; and mixtures thereof; polyethylene glycol sorbitan fatty acid esters and hydrophilic transesterification products of a polyol with at least one member of the group consisting of triglycerides, vegetable oils, and hydrogenated vegetable oils. The polyol may be glycerol, ethylene glycol, polyethylene glycol, sorbitol, propylene glycol, pentaerythritol, or a saccharide.

Suitable lipophilic surfactants include, by way of example only: fatty alcohols; glycerol fatty acid esters; acetylated glycerol fatty acid esters; lower alcohol fatty acids esters; propylene glycol fatty acid esters; sorbitan fatty acid esters; polyethylene glycol sorbitan fatty acid esters; sterols and sterol derivatives; polyoxyethylated sterols and sterol derivatives; polyethylene glycol alkyl ethers; sugar esters; sugar ethers; lactic acid derivatives of mono- and di-glycerides; hydrophobic transesterification products of a polyol with at least one member of the group consisting of glycerides, vegetable oils, hydrogenated vegetable oils, fatty acids and sterols; oil-soluble vitamins/vitamin derivatives; and mixtures thereof. Within this group, preferred lipophilic surfactants include glycerol fatty acid esters, propylene glycol fatty acid esters, and mixtures thereof, or are hydrophobic transesterification products of a polyol with at least one member of the group consisting of vegetable oils, hydrogenated vegetable oils, and triglycerides.

In one embodiment, the composition may include a solubilizer to ensure good solubilization and/or dissolution of the compound of the present invention and to minimize precipitation of the compound of the present invention. This can be especially important for compositions for non-oral use, e.g., compositions for injection. A solubilizer may also be added to increase the solubility of the hydrophilic drug and/or other components, such as surfactants, or to maintain the composition as a stable or homogeneous solution or dispersion.

The amount of solubilizer that can be included is not particularly limited. The amount of a given solubilizer may be limited to a bioacceptable amount, which may be readily determined by one of skill in the art. In some circumstances, it may be advantageous to include amounts of solubilizers far in excess of bioacceptable amounts, for example to maximize the concentration of the drug, with excess solubilizer removed prior to providing the composition to a subject using conventional techniques, such as distillation or evaporation. Thus, if present, the solubilizer can be in a weight ratio of 10%, 25%, 50%), 100%, or up to about 200%>by weight, based on the combined weight of the drug, and other excipients. If desired, very small amounts of solubilizer may also be used, such as 5%>, 2%>, 1%) or even less. Typically, the solubilizer may be present in an amount of about 1%>to about 100%, more typically about 5%>to about 25%>by weight.

Pharmaceutical Compositions for Injection.

In some embodiments, the invention provides a pharmaceutical composition for injection containing a compound of the present invention and a pharmaceutical excipient suitable for injection. Components and amounts of agents in the compositions are as described herein.

The forms in which the novel compositions of the present invention may be incorporated for administration by injection include aqueous or oil suspensions, or emulsions, with sesame oil, corn oil, cottonseed oil, or peanut oil, as well as elixirs, mannitol, dextrose, or a sterile aqueous solution, and similar pharmaceutical vehicles.

Aqueous solutions in saline are also conventionally used for injection. Ethanol, glycerol, propylene glycol, liquid polyethylene glycol, and the like (and suitable mixtures thereof), cyclodextrin derivatives, and vegetable oils may also be employed. The proper fluidity can be maintained, for example, by the use of a coating, such as lecithin, for the maintenance of the required particle size in the case of dispersion and by the use of surfactants. The prevention of the action of microorganisms can be brought about by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like.

In some embodiments, the invention provides a pharmaceutical composition for transdermal delivery containing a compound of the present invention and a pharmaceutical excipient suitable for transdermal delivery.

Compositions of the present invention can be formulated into preparations in solid, semisolid, or liquid forms suitable for local or topical administration, such as gels, water soluble jellies, creams, lotions, suspensions, foams, powders, slurries, ointments, solutions, oils, pastes, suppositories, sprays, emulsions, saline solutions, dimethylsulfoxide (DMSO)-based solutions. In general, carriers with higher densities are capable of providing an area with a prolonged exposure to the active ingredients. In contrast, a solution formulation may provide more immediate exposure of the active ingredient to the chosen area.

The pharmaceutical compositions also may comprise suitable solid or gel phase carriers or excipients, which are compounds that allow increased penetration of, or assist in the delivery of, therapeutic molecules across the stratum corneum permeability barrier of the skin. There are many of these penetration-enhancing molecules known to those trained in the art of topical formulation.

Another exemplary formulation for use in the methods of the present invention employs transdermal delivery devices (“patches”). Such transdermal patches may be used to provide continuous or discontinuous infusion of a compound of the present invention in controlled amounts, either with or without another agent.

The construction and use of transdermal patches for the delivery of pharmaceutical agents is well known in the art. See, e.g., U.S. Pat. Nos. 5,023,252, 4,992,445 and 5,001,139. Such patches may be constructed for continuous, pulsatile, or on demand delivery of pharmaceutical agents.

Pharmaceutical Compositions for Inhalation.

Other Pharmaceutical Compositions.

Pharmaceutical compositions may also be prepared from compositions described herein and one or more pharmaceutically acceptable excipients suitable for sublingual, buccal, rectal, intraosseous, intraocular, intranasal, epidural, or intraspinal administration. Preparations for such pharmaceutical compositions are well-known in the art. See, e.g., Anderson, Philip O.; Knoben, James E.; Troutman, William G, eds., Handbook of Clinical Drug Data, Tenth Edition, McGraw-Hill, 2002; Pratt and Taylor, eds., Principles of Drug Action, Third Edition, Churchill Livingston, N.Y., 1990; Katzung, ed., Basic and Clinical Pharmacology, Ninth Edition, McGraw Hill, 20037ybg; Goodman and Gilman, eds., The Pharmacological Basis of Therapeutics, Tenth Edition, McGraw Hill, 2001; Remingtons Pharmaceutical Sciences, 20th Ed., Lippincott Williams & Wilkins, 2000; Martindale, The Extra Pharmacopoeia, Thirty-Second Edition (The Pharmaceutical Press, London, 1999); all of which are incorporated by reference herein in their entirety.

Administration of the compounds or pharmaceutical composition of the present invention can be effected by any method that enables delivery of the compounds to the site of action. These methods include oral routes, intraduodenal routes, parenteral injection (including intravenous, intraarterial, subcutaneous, intramuscular, intravascular, intraperitoneal or infusion), topical (e.g. transdermal application), rectal administration, via local delivery by catheter or stent or through inhalation. Compounds can also be administered intraadiposally or intrathecally.

In some embodiments, the compounds or pharmaceutical composition of the present invention are administered by intravenous injection.

The amount of the compound administered will be dependent on the subject being treated, the severity of the disorder or condition, the rate of administration, the disposition of the compound and the discretion of the prescribing physician. However, an effective dosage is in the range of about 0.001 to about 100 mg per kg body weight per day, preferably about 1 to about 35 mg/kg/day, in single or divided doses. For a 70 kg human, this would amount to about 0.05 to 7 g/day, preferably about 0.05 to about 2.5 g/day. In some instances, dosage levels below the lower limit of the aforesaid range may be more than adequate, while in other cases still larger doses may be employed without causing any harmful side effect, e.g. by dividing such larger doses into several small doses for administration throughout the day.

In some embodiments, a compound of the invention is administered in a single dose. Typically, such administration will be by injection, e.g., intravenous injection, in order to introduce the agent quickly. However, other routes may be used as appropriate. A single dose of a compound of the invention may also be used for treatment of an acute condition.

In some embodiments, a compound of the invention is administered in multiple doses. Dosing may be about once, twice, three times, four times, five times, six times, or more than six times per day. Dosing may be about once a month, once every two weeks, once a week, or once every other day. In another embodiment a compound of the invention and another agent are administered together about once per day to about 6 times per day. In another embodiment the administration of a compound of the invention and an agent continues for less than about 7 days. In yet another embodiment the administration continues for more than about 6, 10, 14, 28 days, two months, six months, or one year. In some cases, continuous dosing is achieved and maintained as long as necessary.

Administration of the compounds of the invention may continue as long as necessary. In some embodiments, a compound of the invention is administered for more than 1, 2, 3, 4, 5, 6, 7, 14, or 28 days. In some embodiments, a compound of the invention is administered for less than 28, 14, 7, 6, 5, 4, 3, 2, or 1 day. In some embodiments, a compound of the invention is administered chronically on an ongoing basis, e.g., for the treatment of chronic effects.

An effective amount of a compound of the invention may be administered in either single or multiple doses by any of the accepted modes of administration of agents having similar utilities, including rectal, buccal, intranasal and transdermal routes, by intra-arterial injection, intravenously, intraperitoneally, parenterally, intramuscularly, subcutaneously, orally, topically, or as an inhalant.

The compositions of the invention may also be delivered via an impregnated or coated device such as a stent, for example, or an artery-inserted cylindrical polymer. Such a method of administration may, for example, aid in the prevention or amelioration of restenosis following procedures such as balloon angioplasty. Without being bound by theory, compounds of the invention may slow or inhibit the migration and proliferation of smooth muscle cells in the arterial wall which contribute to restenosis. A compound of the invention may be administered, for example, by local delivery from the struts of a stent, from a stent graft, from grafts, or from the cover or sheath of a stent. In some embodiments, a compound of the invention is admixed with a matrix. Such a matrix may be a polymeric matrix, and may serve to bond the compound to the stent. Polymeric matrices suitable for such use, include, for example, lactone-based polyesters or copolyesters such as polylactide, polycaprolactonglycolide, polyorthoesters, polyanhydrides, polyaminoacids, polysaccharides, polyphosphazenes, poly (ether-ester) copolymers (e.g. PEO-PLLA); polydimethylsiloxane, poly(ethylene-vinylacetate), acrylate-based polymers or copolymers (e.g. polyhydroxyethyl methylmethacrylate, polyvinyl pyrrolidinone), fluorinated polymers such as polytetrafluoroethylene and cellulose esters. Suitable matrices may be nondegrading or may degrade with time, releasing the compound or compounds. Compounds of the invention may be applied to the surface of the stent by various methods such as dip/spin coating, spray coating, dip-coating, and/or brush-coating. The compounds may be applied in a solvent and the solvent may be allowed to evaporate, thus forming a layer of compound onto the stent. Alternatively, the compound may be located in the body of the stent or graft, for example in microchannels or micropores. When implanted, the compound diffuses out of the body of the stent to contact the arterial wall. Such stents may be prepared by dipping a stent manufactured to contain such micropores or microchannels into a solution of the compound of the invention in a suitable solvent, followed by evaporation of the solvent. Excess drug on the surface of the stent may be removed via an additional brief solvent wash. In yet other embodiments, compounds of the invention may be covalently linked to a stent or graft. A covalent linker may be used which degrades in vivo, leading to the release of the compound of the invention. Any bio-labile linkage may be used for such a purpose, such as ester, amide or anhydride linkages. Compounds of the invention may additionally be administered intravascularly from a balloon used during angioplasty. Extravascular administration of the compounds via the pericard or via advential application of formulations of the invention may also be performed to decrease restenosis.

A variety of stent devices which may be used as described are disclosed, for example, in the following references, all of which are hereby incorporated by reference: U.S. Pat. Nos. 5,451,233; 5,040,548; 5,061,273; 5,496,346; 5,292,331; 5,674,278; 3,657,744; 4,739,762; 5,195,984; 5,292,331; 5,674,278; 5,879,382; 6,344,053.

The compounds of the invention may be administered in dosages. It is known in the art that due to intersubject variability in compound pharmacokinetics, individualization of dosing regimen is necessary for optimal therapy. Dosing for a compound of the invention may be found by routine experimentation in light of the instant disclosure.

When a compound of the invention is administered in a composition that comprises one or more agents, and the agent has a shorter half-life than the compound of the invention unit dose forms of the agent and the compound of the invention may be adjusted accordingly. The subject pharmaceutical composition may, for example, be in a form suitable for oral administration as a tablet, capsule, pill, powder, sustained release formulations, solution, suspension, for parenteral injection as a sterile solution, suspension or emulsion, for topical administration as an ointment or cream or for rectal administration as a suppository. The pharmaceutical composition may be in unit dosage forms suitable for single administration of precise dosages. The pharmaceutical composition will include a conventional pharmaceutical carrier or excipient and a compound according to the invention as an active ingredient. In addition, it may include other medicinal or pharmaceutical agents, carriers, adjuvants, etc. Exemplary parenteral administration forms include solutions or suspensions of active compound in sterile aqueous solutions, for example, aqueous propylene glycol or dextrose solutions. Such dosage forms can be suitably buffered, if desired.

Methods of Use

The method typically comprises administering to a subject a therapeutically effective amount of a compound of the invention. The therapeutically effective amount of the subject combination of compounds may vary depending upon the intended application (in vitro or in vivo), or the subject and disease condition being treated, e.g., the weight and age of the subject, the severity of the disease condition, the manner of administration and the like, which can readily be determined by one of ordinary skill in the art. The term also applies to a dose that will induce a particular response in target cells, e.g., reduction of proliferation or downregulation of activity of a target protein. The specific dose will vary depending on the particular compounds chosen, the dosing regimen to be followed, whether it is administered in combination with other compounds, timing of administration, the tissue to which it is administered, and the physical delivery system in which it is carried.

In certain embodiment, the present invention provides a pharmaceutical composition comprising a compound of bispecific formula, or pharmaceutically acceptable salt thereof.

In certain embodiment, the present invention provides a pharmaceutical composition comprising a compound of bispecific formula for use in degrading a target protein in a cell.

In certain embodiment, a method of degrading a target protein comprising administering to a cell therapeutically effective amount of a bispecific compound, or pharmaceutically acceptable salt, wherein the compound is effective for degrading the target protein.

In certain embodiment, the present invention provides a pharmaceutical composition comprising a compound of bispecific formula, for use in treating or preventing of a disease or disorder in which SMARCA2 and/or SMARCA4 plays a role.

In certain embodiment, the present invention provides a pharmaceutical composition comprising a compound of bispecific formula, for use in treating or preventing of a disease or disorder in which SWI/SNF mutations plays a role.

In certain embodiment, target protein complex is SWI/SNF in a cell.

In certain embodiment, diseases or disorders dependent on SMARCA2 or SMARCA4 include cancers.

In certain embodiment, diseases or disorders dependent on SWI/SNF complex include cancers.

In certain further embodiment, the cancer is a SMARCA2 and/or SMARAC4-dependent cancer.

In some embodiments, the cancer harbors a SMARCA4 mutation.

In certain embodiment, the present invention provides a pharmaceutical composition comprising a compound of bispecific formula for use in the diseases or disorders dependent upon SMARCA2 and/or SMARCA4 is cancer.

Compounds of the disclosure, as well as pharmaceutical compositions comprising them, can be administered to treat any of the described diseases, alone or in combination with a medical therapy. Medical therapies include, for example, surgery and radiotherapy (e.g., gamma-radiation, neutron beam radiotherapy, electron beam radiotherapy, proton therapy, brachytherapy, systemic radioactive isotopes).

In other aspects, compounds of the disclosure, as well as pharmaceutical compositions comprising them, can be administered to treat any of the described diseases, alone or in combination with one or more other agents.

In other methods, the compounds of the disclosure, as well as pharmaceutical compositions comprising them, can be administered in combination with agonists of nuclear receptors agents.

In other methods, the compounds of the disclosure, as well as pharmaceutical compositions comprising them, can be administered in combination with antagonists of nuclear receptors agents.

In other methods, the compounds of the disclosure, as well as pharmaceutical compositions comprising them, can be administered in combination with an anti-proliferative agent.

Combination Therapies

In some embodiments, the compounds of the invention can be used in combination with a therapeutic agent that targets an epigenetic regulator. Examples of epigenetic regulators include bromodomain inhibitors, the histone lysine methyltransferase inhibitors, histone arginine methyl transferase inhibitors, histone demethylase inhibitors, histone deacetylase inhibitors, histone acetylase inhibitors, and DNA methyltransferase inhibitors. Histone deacetylase inhibitors include, e.g., vorinostat. Histone arginine methyl transferase inhibitors include inhibitors of protein arginine methyltransferases (PRMTs) such as PRMT5, PRMT1 and PRMT4. DNA methyltransferase inhibitors include inhibitors of DNMT1 and DNMT3.

In some embodiments, the inhibitor of an immune checkpoint molecule is an inhibitor of PD-1, e.g., an anti-PD-1 monoclonal antibody. In some embodiments, the anti-PD-1 monoclonal antibody is nivolumab, pembrolizumab (also known as MK-3475), or PDR001. In some embodiments, the anti-PD-1 monoclonal antibody is nivolumab or pembrolizumab. In some embodiments, the anti-PD1 antibody is pembrolizumab. In some embodiments, the inhibitor of an immune checkpoint molecule is an inhibitor of PD-L1, e.g., an anti-PD-L1 monoclonal antibody. In some embodiments, the anti-PD-L1 monoclonal antibody is atezolizumab, durvalumab, or BMS-935559. In some embodiments, the inhibitor of an immune checkpoint molecule is an inhibitor of CTLA-4, e.g., an anti-CTLA-4 antibody. In some embodiments, the anti-CTLA-4 antibody is ipilimumab.

In some embodiments, the agent is an alkylating agent, a proteasome inhibitor, a corticosteroid, or an immunomodulatory agent. Examples of an alkylating agent include cyclophosphamide (CY), melphalan (MEL), and bendamustine. In some embodiments, the proteasome inhibitor is carfilzomib. In some embodiments, the corticosteroid is dexamethasone (DEX). In some embodiments, the immunomodulatory agent is lenalidomide (LEN) or pomalidomide (POM).

Compounds of the present invention include, but are not limited to, those shown in Table 1.

EXAMPLES

The compounds of the Invention may be prepared using the general procedures described below.

Compounds of Formula (IA) can be synthesized using, for example, the sequences shown in Scheme A. The coupling between compounds A-1 and A-2 via a SNAr reaction or Pd-catalyzed cross coupling gives compound A-3. The following intramolecular SNAr reaction or amide formation can afford cyclized product A-4. Introduction of a protecting group to facilitate organometallic addition of Ar via, e.g., Suzuki conditions (e.g., in the presence of a palladium catalyst, such as but not limited to tetrakis(triphenylphosphine)palladium(0) or [1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium (II), complex with dichloromethane and a base (e.g., a carbonate base)) using the appropriate boronic acid or ester or other Pd catalyzed reactions gives A-6. Introduction of R1using appropriate synthetic methods (such as, but not limited to, SN2 reaction, SNAr reaction, reductive amination, Buchwald reaction, amide formation, Mitsunobu reaction, olefin metathesis, etc.) can give compounds A-7. The protecting groups on A-7 can be removed using standard conditions to give compounds IA.

Compounds of formula I-8 can be synthesized using, for example, the sequences shown in Scheme I. SNAr reaction between I-1 and compound I-2 in the presence of a base (e.g., Cs2CO3, NaHCO3, DIPEA) at elevated temperatures can give alcohol 1-3. Conversion of the hydroxyl group of 1-3 to a leaving group (LG) under appropriate conditions (such as, but not limited to, treatment with SOCl2, or CBr4/PPh3, or MsCl/Et3N) can afford compounds I-4. Treatment of 1-4 with NaN3gives compounds I-5. Reduction of the azido group of compounds I-5 using PPh3or Pd/H2to the corresponding amines, followed by intramolecular cyclization results in compounds I-6. Alternately compounds I-4 may be treated with ammonium hydroxide at elevated temperatures to give compounds I-6. Protection of the —NH group with an appropriate group (e.g., Boc, SEM, Bn, etc.) can afford compounds I-7, which can be converted to compounds I-8 under standard Suzuki conditions (e.g., in the presence of a palladium catalyst, such as but not limited to tetrakis(triphenylphosphine)palladium(0) or [1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium (II), complex with dichloromethane and a base (e.g., a carbonate base)) using the appropriate boronic acid or ester (e.g., 2-hydroxyphenylboronic acid).

Compounds of formula II-5 can be synthesized using, for example, the sequences shown in Scheme II. Coupling of compounds II-1 with R1using appropriate synthetic methods (such as but not limited to amide formation, SN2 reaction, reductive amination, etc.; e.g., RG1is a leaving group, such as a bromide and is displaced the amine of II-1) can afford compounds II-2. Compounds I-8 can be introduced using appropriate synthetic methods (such as, but not limited to, SN2 reaction, SNAr reaction, reductive amination, Buchwald reaction, amide formation, Mitsunobu reaction, olefin metathesis, etc.) to give compounds II-4. Alternatively, the synthesis of II-4 can be achieved by the coupling of I-8 with R1, followed by the introduction of II-1 using appropriate synthetic methods mentioned above. Removal of the protecting groups can afford compounds of formula II-5.

The compounds of formula III-5 can be synthesized using, for example, the sequences shown in Scheme III. Coupling of compounds III-1 with R1using appropriate synthetic methods (such as but not limited to SN2 reaction, SNAr reaction, Mitsunobu reaction, etc.) can afford compounds III-2. Compounds I-8 can be introduced using appropriate synthetic methods (such as, but not limited to, SN2 reaction, SNAr reaction, reductive amination, Buchwald reaction, amide formation, Mitsunobu reaction, olefin metathesis, etc.) to give compounds III-4. Alternatively, the synthesis of III-4 can be achieved by the coupling of 1-8 with R1, followed by the introduction of intermediates III-1 using appropriate synthetic methods mentioned above. Removal of the protecting groups can result in compounds of formula III-5.

The compounds of formula IV-4 can be synthesized using, for example, the sequences shown in Scheme IV. Coupling of compounds IV-1 with acids under standard amide coupling conditions (e.g., treatment with an appropriate base such as DIPEA or Et3N and in the presence of coupling agents such as HATU, HOBt, or PyBOP) gives amides IV-2. Nucleophilic addition of compounds IV-3 under basic conditions (e.g., in the presence of a carbonate base, DIPEA, Et3N, etc.) can afford compounds of formula IV-4.

An exemplary synthesis for preparing compound of V-4 is depicted in Scheme V. Nucleophilic substitution of haloalkylacids with compounds V-1 under basic conditions (e.g., in the presence of a carbonate base, DIPEA, Et3N, etc.) can give ethers V-2. Coupling of amines IV-3 with carboxylic acids V-2 under standard amide coupling conditions (e.g., treatment with an appropriate base such as DIPEA or trimethylamine and in the presence of coupling agents such as HATU, HOBt, or PyBOP) can give amides V-3. Removal of the protecting group using appropriate conditions can afford compounds of formula V-4.

An exemplary synthesis for preparing compound of VI-3 is depicted in Scheme VI. Nucleophilic substitution of alkyldihalide with alcohol V-1 under basic conditions (e.g., in the presence of a carbonate base, DIPEA, Et3N, etc.) can give ethers VI-1. Nucleophilic substitution of VI-1 where LG2is a leaving group, such as halo, with an amine IV-3 under basic conditions (e.g., in the presence of a carbonate base, DIPEA, Et3N, etc.) can give compounds VI-2. Removal of the protecting group appropriate conditions can afford compounds of formula VI-3.

Compounds of formula VII-9 can be synthesized using, for example, the sequences shown in Scheme VII. Coupling of amine VII-1 with carboxylic acid VII-2 under standard amide coupling conditions (treatment with an appropriate base such as, but not limited to, DIPEA or trimethylamine and in the presence of coupling agents such as HATU, HOBt, or PyBOP) gives amides VII-3. Nucleophilic substitution of VII-3 with an alkyl-dihalide under basic conditions (e.g., in the presence of a carbonate base, DIPEA, Et3N, etc.) can give compounds VII-4. A second nucleophilic substitution of VII-4 where LG2is halo with IV-3 under basic conditions (e.g., in the presence of a carbonate base, DIPEA, Et3N, etc.) can give compounds VII-5. Hydrolysis of t-butyl-ester using appropriate conditions can give compounds VII-6. Coupling of compounds VII-6 with compounds VII-7 under standard amide coupling conditions such as a base, (e.g., DIPEA or trimethylamino) and in the presence of coupling (e. HATU, HOBt or PyBOP) can afford amides VII-8. Deprotection of VII-8 can afford compounds of formula VII-9.

The compounds of formula VIII-5 can be synthesized using, for example, the sequences shown in Scheme VIII. The reductive amination between compounds IV-3 and VIII-1 can provide compounds VIII-2. Removal of the protecting groups, followed by a nucleophilic substitution of VIII-4 where LG is halo with VIII-3 under basic conditions (e.g., in the presence of a carbonate base, DIPEA, Et3N, etc.) can give compounds of formula VIII-5.

The compounds of formula IX-3 can be synthesized as shown in Scheme IX. A nucleophilic substitution of IX-1 where LG is halo with VIII-3 under basic conditions (e.g., in the presence of a carbonate base, DIPEA, Et3N, etc.) can give compounds IX-2. Removal of the protecting groups can afford compounds of formula IX-3.

Synthesis of Intermediates

To a solution of tert-butyl 3-(azidomethyl)-4-(3,6-dichloropyridazin-4-yl)piperazine-1-carboxylate (15) (2.0 g, 5.2 mmol) in DMF (10 mL) was added and triphenylphosphine (1.4 g, 5.2 mmol), the mixture was stirred at 45° C. for 18 h. The solution was added to a mixture of water (60 mL) and EtOAc (50 mL), extracted with EtOAc (60 mL×2), concentrated in vacuum and the residue was purified by gel silica chromatography (SiO2, 200-300 mesh, PE:EtOAc from 10:1 to 1:2) to give tert-butyl 2-chloro-5,6,6a,7,9,10-hexahydro-8H-pyrazino[1′,2′:4,5]pyrazino[2,3-c]pyridazine-8-carboxylate (5) (1500 mg, 4.6 mmol, 89% yield) as a yellow solid. LCMS calc'd for C14H20ClN5O2: 325.1; Found: LCMS [M+H]: 326.2.

Step b. Synthesis of 2-((5-(1-((2S,4R)-4-((tert-butyldimethylsilyl)oxy)-2-(((S)-1-(4-(4-methyl-1λ3,3λ2-thiazol-5-yl)phenyl)ethyl)carbamoyl)pyrrolidin-1-yl)-3-methyl-1-oxobutan-2-yl)isoxazol-3-yl)oxy)acetic acid (Int-2)

To a solution of (2S,4R)-4-((tert-butyldimethylsilyl)oxy)-1-(2-(3-hydroxyisoxazol-5-yl)-3-methylbutanoyl)-N—((S)-1-(4-(4-methyl-1λ3,3λ2-thiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide (9) (prepared using the procedure described in US2020/0038378, 100 mg, 0.16 mmol) and 1,2-dibromoethane (45 mg, 0.24 mmol) in DMF (3 mL) was added and NaHCO3(60 mg, 0.67 mmol) at 25° C., the mixture was stirred at 30° C. for 16 h. The reaction was taken up in EtOAc (20 mL) and the organics were washed with water (20 mL×3) and brine (20 mL). The organics were then separated and dried (MgSO4) before concentration to dryness. The crude product was then purified by prep-TLC (EtOAc:PE=2:1) to give (2S,4R)-1-(2-(3-(2-bromoethoxy)isoxazol-5-yl)-3-methylbutanoyl)-4-((tert-butyldimethylsilyl)oxy)-N—((S)-1-(4-(4-methyl-1λ3,3λ2-thiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide (Int-3) (50 mg, 0.07 mmol, 51% yield) as a sticky colorless solid. LCMS calc'd for C33H47BrN4O5SSi: 718.2; Found: LCMS [M+H]: 719.3.

To a solution of (2S,4R)-1-((S)-2-amino-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methyl-1λ3,3λ2-thiazol-5-yl)benzyl)pyrrolidine-2-carboxamide (prepared using the procedure described in WO2018/0140809, 500 mg, 1.2 mmol) in DCM (15 mL) and was added TEA (0.81 mL, 5.8 mmol), the reaction was cooled to 0° C., a solution of chloroacetyl chloride (262 mg, 2.3 mmol) in DCM (2 mL) was added, the reaction was stirred at 25° C. for 16 h under N2. The reaction was washed with water (10 mL×2), dried over Na2SO4, filtered and concentrated in vacuum, the residue was purified by silica gel column chromatography (100-200 mesh size, eluted with DCM:MeOH=25:1) to afford (2S,4R)-1-((S)-2-(2-chloroacetamido)-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methyl-1λ3,3λ2-thiazol-5-yl)benzyl)pyrrolidine-2-carboxamide (Int-5). (520 mg, 0.98 mmol, 84% yield) as a yellow solid. LCMS calc'd for C24H31ClN4O4S: 507.0; Found: LCMS [m/z]: 507.2.

The intermediates in the table below were prepared by the method used in preparing Int-5:

The intermediates in the table below were prepared by the method used in preparing Int-8 using appropriate starting materials:

To a solution of 3,4,6-trichloropyridazine (5.7 g, 31.1 mmol) in DMF (24 mL) was added N,N-diisopropylethylamine (5.9 mL, 34.2 mmol) and tert-butyl (R)-3-(hydroxymethyl) piperazine-1-carboxylate (7.1 g, 32.8 mmol). The reaction was stirred at 80° C. overnight. The reaction was cooled to 45° C. and water (17 mL) was added slowly. The resulted clear solution was stirred at 35° C. for 30 min until precipitate formed. Another portion of water (23 mL) was charged slowly and the mixture was stirred at 0° C. for an additional 1 h. The mixture was filtered and the resulting solid was washed with water and dried under vacuum to give tert-butyl (R)-4-(3,6-dichloropyridazin-4-yl)-3-(hydroxymethyl)piperazine-1-carboxylate (8.5 g, 75% yield) as an off-white solid. LCMS m/z calcd for C14H21Cl2N4O3[M+H]+: 363.1; found: 363.1.

To a solution of tert-butyl (R)-4-(3,6-dichloropyridazin-4-yl)-3-(hydroxymethyl) piperazine-1-carboxylate (5.45 g, 15 mmol) and triphenylphosphine (4.7 g, 18 mmol) in THF (150 mL) was added diisopropyl azodicarboxylate (3.5 mL, 18 mmol) and DPPA (3.9 mL, 18 mmol) at 0° C. The reaction was then stirred at RT overnight. The reaction mixture was cooled to 0° C., quenched with water and extracted with EtOAc. The combined organic layers were washed with brine and water, dried over Na2SO4and filtered. The filtrate was concentrated under reduced pressure to give crude tert-butyl (R)-3-(azidomethyl)-4-(3,6-dichloropyridazin-4-yl)piperazine-1-carboxylate (19.4 g, 100% yield), which was used without further purification. Assumed 100% yield, 30% purity. LCMS m/z calcd for C14H20Cl2N7O2[M+H]+: 388.1; found: 388.0.

To a stirred solution of crude tert-butyl (R)-3-(azidomethyl)-4-(3,6-dichloropyridazin-4-yl) piperazine-1-carboxylate (30% purity, 20.3 g, 15.7 mmol) in THF (200 mL), triphenylphosphine (4.9 g, 18.8 mmol) was added. The resulted solution was stirred at 60° C. for 3 h. Water (20 mL) and N,N-diisopropylethylamine (8.2 mL, 47.1 mmol) were added sequentially. After 20 h, the reaction mixture was diluted with EtOAc (100 mL) and water (100 mL). The aqueous layer was separated and extracted with EtOAc. The combined organic layers were washed with brine, dried over Na2SO4and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by silica gel chromatography, eluting with 0-100% EtOAc/hexanes to give tert-butyl (S)-2-chloro-5,6,6a,7,9,10-hexahydro-8H-pyrazino[1′,2′:4,5]pyrazino[2,3-c]pyridazine-8-carboxylate (3.1 g, 60% yield) as an off-white solid. LCMS m/z calcd for C14H21ClN5O2[M+H]+: 326.1; found: 326.2.

Step 6: Synthesis of (R)-2-(6,6a,7,8,9,10-hexahydro-5H-pyrazino[1′,2′:4,5]pyrazino[2,3-c]pyridazin-2-yl)phenol

The title compound was prepared using procedure analogous to those described for Int-9, using appropriate starting materials. LCMS m/z calcd for C15H18N5O [M+H]+: 284.2; found: 284.2.

To a solution of (2S,4R)-1-((R)-2-(3-(2-bromoethoxy)isoxazol-5-yl)-3-methylbutanoyl)-4-((tert-butyldimethylsilyl)oxy)-N—((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide (Int-4)(100 mg, 0.14 mmol) in THF (5 mL) were added TBAF in THF (0.1 mL, 7.3 mmol) the mixture solution was stirred at rt for 2 h. The reaction was diluted with EA (20 ml) and washed with brine (30 mL×2), the organic layer was concentrated to give the crude product, which was used in the next step without further purification. LCMS m/z calcd for C27H34BrN4O5S [M+H]+: 605.1; Found: 605.1.

The reaction mixture was diluted with EtOAc (20 mL) and the organic layer washed with water (2×10 mL) then saturated brine (1×10 mL). The organic layer was separated, dried over MgSO4, and filtered. The filtration was concentrated to dryness under reduced pressure. The residue was purified by prep-TLC (PE:EA=1:1) to get (3R,5S)-1-((R)-2-(3-(2-bromoethoxy)isoxazol-5-yl)-3-methylbutanoyl)-5-(((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)carbamoyl)pyrrolidin-3-yl acetate (40 mg, 0.06 mmol, 83% yield). LCMS m/z calcd for C29H36BrN4O6S [M+H]+: 647.1; Found: 647.1.

The title compound was prepared using procedure analogous to those described for Int-12, with isobutyric anhydride replacing acetic anhydride in step 2. LCMS m/z calcd for C31H40BrN4O6S [M+H]+: 675.2; Found: 675.2.

The title compound was prepared using procedure analogous to those described for Int-8, step a with appropriate starting materials. LCMS m/z calcd for C24H33N6O3[M+H]+: 453.3; Found: 453.3.

To a stirred solution of tert-butyl 3-((S)-2-(2-hydroxyphenyl)-5,6,6a,7,9,10-hexahydro-8H-pyrazino[1′,2′:4,5]pyrazino[2,3-c]pyridazin-8-yl)pyrrolidine-1-carboxylate (diastereomer 1, 2.0 g, 4.4 mmol) in DCM (10 mL) was added TFA (1.0 mL) at rt. After 16 h, the volatiles were removed under reduced pressure and the residue was used for next step without further purification. LCMS m/z calcd for C19H25N6O [M+H]+: 353.2; Found: 353.0.

To a stirred solution of tert-butyl 3-((S)-2-(2-hydroxyphenyl)-5,6,6a,7,9,10-hexahydro-8H-pyrazino[1′,2′:4,5]pyrazino[2,3-c]pyridazin-8-yl)pyrrolidine-1-carboxylate (diastereomer 2 from the synthesis of Int-19, step 2, 2.0 g, 4.42 mmol) in DCM (10 mL) was added TFA (1 mL) at rt. After 16 h, the volatiles were removed under reduced pressure and the residue was used for next step without further purification. LCMS m/z calcd for C19H25N6O [M+H]+: 353.2; Found: 353.1.

To a solution of tert-butyl (S)-4-(2-(4-(2-(2-hydroxyphenyl)-5,6,6a,7,9,10-hexahydro-8H-pyrazino[1′,2′:4,5]pyrazino[2,3-c]pyridazin-8-yl)piperidin-1-yl)ethyl)piperazine-1-carboxylate (40 mg, 0.07 mmol) in DCM (1 mL) was added TFA (0.03 mL, 0.16 mmol) at rt. The mixture was stirred at rt for 16 h. The volatiles were removed in vacuum and the residue was used for next step without further purification. LCMS m/z calcd for C26H39N8O [M+H]+: 479.3; Found: 479.2.

To a solution of tert-butyl (S)-4-(2-(4-(2-(2-hydroxyphenyl)-5,6,6a,7,9,10-hexahydro-8H-pyrazino[1′,2′:4,5]pyrazino[2,3-c]pyridazin-8-yl)piperidin-1-yl)ethyl)piperidine-1-carboxylate (40.0 mg, 0.07 mmol) in DCM (30 mL) was added TFA (0.3 mL) at rt. The mixture was stirred at rt for 16 h. LCMS showed the reaction was completed. The volatiles were removed in vacuum and the residue was used for next step without further purification. LCMS m/z calcd for C27H40N7O [M+H]+: 478.3; Found: 478.2.

To a solution of 1,4-bis(tert-butoxycarbonyl)piperazine-2-carboxylic acid (5.0 g, 15.1 mmol) and potassium carbonate (4.18 g, 30.3 mmol) in acetone (50 mL) was added iodomethane (2.17 g, 15.3 mmol) at rt. The mixture was stirred at rt for 16 h. The reaction mixture was filtered and the filtrate was concentrated in vacuum. The residue was dissolved in EA (100 ml) and washed with brine (100 mL×2). The organic layer was concentrated in vacuum to give 1,4-di-tert-butyl 2-methyl piperazine-1,2,4-tricarboxylate (5.2 g, 15.1 mmol, 99.7% yield) as a white solid. LCMS calc'd for C16H29N2O6[M+H]+: 345.2; Found: 345.2.

To a solution of 1,4-di-tert-butyl 2-methyl piperazine-1,2,4-tricarboxylate (5.2 g, 15.1 mmol) in THF (100 mL) was added LiHMDS (2.8 g, 16.6 mmol) at −78° C. The mixture was stirred at −78° C. for 2 h then iodomethane (6.4 g, 45.3 mmol) was added at −78° C. The resulted mixture was stirred at rt. for 16 h. The reaction was quenched with saturated aqueous NH4Cl (100 ml) at 0° C., diluted with EA (200 ml), and washed with water (2×100 mL) then brine (50 ml). The organic layer was dried (MgSO4), filtered, and the filtrate was concentrated to dryness. The crude was purified by silica gel column chromatography (100-200 mesh size), eluted with PE:EA=3:1 to 1:1 to give 1,4-di-tert-butyl 2-methyl 2-methylpiperazine-1,2,4-tricarboxylate (5.0 g, 13.9 mmol, 91.4% yield) as a yellow oil. LCMS calc'd for C17H31N2O6[M+H]+: 359.2; Found: 359.3.

To a solution of 1,4-di-tert-butyl 2-methyl 2-methylpiperazine-1,2,4-tricarboxylate (5.0 g, 13.9 mmol) in THF (12 mL)/methanol (2 mL)/water (2 mL) was added LiOH (713 mg, 17.0 mmol). The mixture was stirred at 50° C. for 16 h. TLC showed the reaction was complete. The reaction mixture was washed with PE (100 mL×2). The pH of the aqueous layer was adjusted to 3-4 with 1 N HCl, then extracted with EA (100 mL×3). The organic layers were combined, washed with brine (50 ml), and concentrated under reduced pressure to give the product 1,4-bis(tert-butoxycarbonyl)-2-methylpiperazine-2-carboxylic acid (4.5 g, 13.1 mmol, 93.7% yield) as a white solid. LCMS calc'd for C16H29N2O6[M+H]+: 345.2; Found: 345.2.

To a solution of 1,4-bis(tert-butoxycarbonyl)-2-methylpiperazine-2-carboxylic acid (4.2 g, 12.2 mmol) in DCM (25 mL) was added DMF (1 mL) and oxalyl chloride (4.6 g, 36.6 mmol). The mixture was stirred at rt for 30 min. The volatiles were removed under reduced pressure and DMF (25 mL), DIEA (10.1 mL, 61.0 mmol) and 5-bromo-6-chloropyridazin-3-amine (5.1 g, 24.4 mmol) were added sequentially. The resulted mixture was stirred at 120° C. for 16 h. The reaction mixture was diluted with EA (100 ml) and washed with brine (30 mL×2). The organic layer was concentrated in vacuum and purified by prep-TLC, eluting with PE:EA=1:1 to give tert-butyl 2-chloro-6a-methyl-6-oxo-5,6,6a,7,9,10-hexahydro-8H-pyrazino[1′,2′:4,5]pyrazino[2,3-c]pyridazine-8-carboxylate (1.5 g, 4.2 mmol, 34.7% yield) as a yellow solid. LCMS calc'd for C15H21ClN5O3[M+H]+: 354.1; Found: 354.1.

To a solution of tert-butyl 2-chloro-6a-methyl-6-oxo-5,6,6a,7,9,10-hexahydro-8H-pyrazino[1′,2′:4,5]pyrazino[2,3-c]pyridazine-8-carboxylate (87.3 mg, 0.25 mmol) in THF (8 mL) was added BH3in THF (1 M, 0.74 mL, 0.74 mmol). The resulted mixture was stirred at 80° C. for 16 h. The reaction was diluted with MeOH (20 ml) and was stirred at 80° C. for additional 16 h. The volatiles were removed under reduce pressure and the residue was purified by prep-TLC (DCM:MeOH=10:1) to give tert-butyl 2-chloro-6a-methyl-5,6,6a,7,9,10-hexahydro-8H-pyrazino[1′,2′:4,5]pyrazino[2,3-c]pyridazine-8-carboxylate (40.0 mg, 0.12 mmol, 47.7% yield) as a yellow solid. LCMS calc'd for C15H23ClN5O2[M+H]+: 340.2; Found: 340.1.

To a solution of 2-hydroxyphenylboronic acid (731 mg, 5.3 mmol), potassium carbonate (1.1 g, 7.95 mmol) and tert-butyl 2-chloro-6a-methyl-5,6,6a,7,9,10-hexahydro-8H-pyrazino[1′,2′:4,5]pyrazino[2,3-c]pyridazine-8-carboxylate (900 mg, 2.65 mmol) in 1,4-dioxane (10 mL) and water (1 mL) was added Pd(dppf)2Cl2(216 mg, 0.26 mmol). The mixture was stirred at 105° C. for 16 h under N2. The reaction was diluted with EA (200 ml) and washed with brine (100 mL×2). The organic layer was concentrated and the residue was purified by silica gel column chromatography (100-200 mesh size), eluting with PE:EA=3:1 to 1:1 to give tert-butyl 2-(2-hydroxyphenyl)-6a-methyl-5,6,6a,7,9,10-hexahydro-8H-pyrazino[1′,2′:4,5]pyrazino[2,3-c]pyridazine-8-carboxylate (1.0 g, 2.51 mmol, 95.0% yield) as a yellow solid. LCMS calc'd for C21H28N5O3[M+H]+: 398.2; Found: 398.2.

To a solution of 2-(benzyloxy)ethan-1-ol (1.0 g, 6.6 mmol) in MeCN (30 mL) was added IBX (8.4 g, 19.7 mmol). The mixture was stirred at 90° C. for 10 min. The reaction was diluted with EtOAc (30 mL) and was washed with water (2×10 mL) then saturated brine (1×10 mL). The organic layer was dried over MgSO4, filtered and concentrated to dryness. The crude was then purified by silica gel chromatography (EA:PE=1:5) to give 2-(benzyloxy)acetaldehyde (150 mg, 1.00 mmol). LCMS calc'd for C9H11O2[M+H]+: 151.1; Found: 151.1.

To a solution of 2-(benzyloxy)acetaldehyde (1.0 g, 6.7 mmol) and hydroxylamine hydrochloride (508 mg, 7.33 mmol) in ethanol (10 mL) and water (30 mL) was added NaOH (666 mg, 16.6 mmol) at 0° C. The reaction was stirred at 0° C. for 2 h. The resulting mixture was acidified with HCl (5 N) to pH 2. The mixture was extracted with EA (30 mL×2). The combined organic layers were washed with brine (8.0 mL), dried over MgSO4, and concentrated to give (E)-2-(benzyloxy)acetaldehyde oxime (500 mg, 3.0 mmol, 44.5% yield) as a colorless oil. LCMS calc'd for C9H12NO2[M+H]+: 166.1; Found: 166.1.

To a solution of (Z)-2-(benzyloxy)-N-hydroxyacetimidoyl chloride (6.4 g, 32.1 mmol) and NaHCO3(3.4 g, 40.1 mmol) in EA (20 mL) and water (20 mL) was added 3-butyn-1-ol (2.76 g, 39.4 mmol). The reaction was stirred at 25° C. for 2 h. The resulted mixture was diluted with EA (50 mL) then washed with water (2×30 mL) and brine (1×10 mL). The organic layer was dried over MgSO4and concentrated to dryness. The residue was purified by silica gel chromatography (PE:EA=1:1) to afford 2-(3-((benzyloxy)methyl)isoxazol-5-yl)ethan-1-ol (2.7 g, 11.6 mmol, 36.1% yield) as an oil. LCMS calc'd for C13H16NO3[M+H]+: 234.1; Found: 234.0.

To a solution of 2-(3-((benzyloxy)methyl)isoxazol-5-yl)acetic acid (80.0 mg, 0.32 mmol) in ethanol (5 mL) was added sulfuric acid (0.1 mL, 0.32 mmol). The reaction was stirred at 70° C. for 2 h. The reaction mixture was diluted with water (20 mL) and extracted with EA (2×30 mL). The combined organic layers were concentrated to afford ethyl 2-(3-((benzyloxy)methyl)isoxazol-5-yl)acetate (60 mg, 0.21 mmol, 66.0% yield) as a colorless oil. LCMS calc'd for C15H18NO4[M+H]+: 276.1; Found: 276.1.

To a solution of ethyl 2-[3-(phenylmethoxymethyl)-1,2-oxazol-5-yl]acetate (1.0 g, 3.63 mmol) and potassium tert-butoxide (815 mg, 7.26 mmol) in THF (15 mL) was added 2-iodopropane (926 mg, 5.45 mmol) at 0° C. The resulted solution was stirred at rt for 3 h. The mixture was diluted with EA (20 mL) and was washed with water (2×10 mL) and brine (1×10 mL). The organic layer was separated, dried over MgSO4, and concentrated to dryness to give ethyl 2-(3-((benzyloxy)methyl)isoxazol-5-yl)-3-methylbutanoate (800 mg, 2.52 mmol, 69.4% yield) as an oil.

To a solution of 2-(3-(hydroxymethyl)isoxazol-5-yl)-3-methylbutanoic acid (300.0 mg, 1.51 mmol) in DMF (2 mL) was added (2S,4R)-4-((tert-butyldimethylsilyl)oxy)-N—((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide (1.01 g, 2.26 mmol), HATU (859 mg, 2.26 mmol), DIEA (0.75 mL, 4.52 mmol). The mixture was stirred at 25° C. for 16 h. The reaction mixture was diluted with H2O and extracted with EA (20 mL×3). The organic layers were combined and the volatiles were removed under reduced pressure. The residue was purified by silica gel chromatography to give (2S,4R)-4-((tert-butyldimethylsilyl)oxy)-1-(2-(3-(hydroxymethyl)isoxazol-5-yl)-3-methylbutanoyl)-N—((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide (550 mg, 0.88 mmol, 58.3% yield) as a yellow oil. LCMS calc'd for C32H47N4O5SSi [M+H]+: 627.3; Found: 627.4.

To a solution of (2S,4R)-4-((tert-butyldimethylsilyl)oxy)-1-(2-(3-(hydroxymethyl)isoxazol-5-yl)-3-methylbutanoyl)-N—((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide (60 mg, 0.10 mmol) in DCM (2 mL) was added MnO2(167 mg, 1.91 mmol). The mixture was stirred at 45° C. for 1 h. The reaction mixture was diluted with H2O and extracted with EA (10 mL×3). The organic layers were combined and the volatiles were removed under reduced pressure. The residue was purified by silica gel chromatography to give (2S,4R)-4-((tert-butyldimethylsilyl)oxy)-1-(2-(3-formylisoxazol-5-yl)-3-methylbutanoyl)-N—((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide (40 mg, 0.06 mmol, 66.9% yield) as a yellow oil. LCMS calc'd for C32H45N4O5SSi [M+H]+: 625.3; Found: 625.2.

The title compound was prepared using procedures analogous to those described for Int-32, from step 1 to step 10, with appropriate starting materials. LCMS m/z calcd for C34H51N4O5SSi [M+H]+: 655.3; Found: 655.3.

The title compound was prepared using procedures analogous to those described for Int-33 with appropriate starting materials. LCMS m/z calcd for C33H47N4O5SSi [M+H]+: 639.3; Found: 639.3.

To a solution of 1,2-dibromoethane (15.3 g, 81.6 mmol) and methyl (2S,4R)-4-((tert-butyldimethylsilyl)oxy)-1-((R)-2-(3-hydroxyisoxazol-5-yl)-3-methylbutanoyl)pyrrolidine-2-carboxylate (3.5 g, 8.16 mmol) in DMF (5 mL) was added potassium carbonate (3.4 g, 24.5 mmol) at rt. The mixture was stirred at rt for 16 h. The reaction mixture was diluted with EA (50 mL) and washed with water (2×20 mL) and saturated brine (1×10 mL). The organic layer was separated, dried over MgSO4, and concentrated to dryness. The residue was purified by prep-HPLC, eluted with CH3CN in H2O (0.1% NH3·H2O) from 5.0% to 95% to give methyl (2S,4R)-1-((R)-2-(3-(2-bromoethoxy)isoxazol-5-yl)-3-methylbutanoyl)-4-((tert-butyldimethylsilyl)oxy)pyrrolidine-2-carboxylate (1.0 g, 1.87 mmol, 22.9% yield). LCMS m/z calcd for C22H38BrN2O6Si [M+H]+: 533.2; Found: 533.3.

To a solution of (S)-2-(8-(piperidin-4-ylmethyl)-6,6a,7,8,9,10-hexahydro-5H-pyrazino[1′,2′:4,5]pyrazino[2,3-c]pyridazin-2-yl)phenol (60 mg, 0.16 mmol) and methyl (2S,4R)-1-((R)-2-(3-(2-bromoethoxy)isoxazol-5-yl)-3-methylbutanoyl)-4-((tert-butyldimethylsilyl)oxy)pyrrolidine-2-carboxylate (92.5 mg, 0.17 mmol) in DMF (5 mL) was added NaHCO3(132 mg, 1.58 mmol) at rt. The mixture was stirred at rt for 16 h. The mixture was diluted with EA (50 mL) and washed with water (2×20 mL) and brine (1×10 mL). The organic layer was dried over MgSO4, and concentrated to dryness. The residue was purified by silica gel chromatography to afford methyl (2S,4R)-4-((tert-butyldimethylsilyl)oxy)-1-((R)-2-(3-(2-(4-(((S)-2-(2-hydroxyphenyl)-5,6,6a,7,9,10-hexahydro-8H-pyrazino[1′,2′:4,5]pyrazino[2,3-c]pyridazin-8-yl)methyl)piperidin-1-yl)ethoxy)isoxazol-5-yl)-3-methylbutanoyl)pyrrolidine-2-carboxylate (18.0 mg, 0.021 mmol, 13.7% yield). LCMS m/z calcd for C43H65N8O7Si [M+H]+: 833.5; Found: 833.3.

To a solution of methyl (2S,4R)-4-((tert-butyldimethylsilyl)oxy)-1-((R)-2-(3-(2-(4-(((S)-2-(2-hydroxyphenyl)-5,6,6a,7,9,10-hexahydro-8H-pyrazino[1′,2′:4,5]pyrazino[2,3-c]pyridazin-8-yl)methyl)piperidin-1-yl)ethoxy)isoxazol-5-yl)-3-methylbutanoyl)pyrrolidine-2-carboxylate (30.0 mg, 0.04 mmol) in THF (2 mL) and water (2 mL) was added LiOH (15.1 mg, 0.36 mmol) at rt. The reaction was stirred at rt for 16 h. The mixture was diluted with EA (30 mL) and washed with water (2×10 mL) and brine (1×10 mL). The organic layer was dried over MgSO4, and concentrated to dryness. The residue was purified by prep-HPLC (eluting with MeCN in H2O, 0.1% HCl) to get the des-TBS product (2S,4R)-4-hydroxy-1-((R)-2-(3-(2-(4-(((S)-2-(2-hydroxyphenyl)-5,6,6a,7,9,10-hexahydro-8H-pyrazino[1′,2′:4,5]pyrazino[2,3-c]pyridazin-8-yl)methyl)piperidin-1-yl)ethoxy)isoxazol-5-yl)-3-methylbutanoyl)pyrrolidine-2-carboxylic acid (18.0 mg, 0.025 mmol, 70.9% yield). LCMS m/z calcd for C36H49H8O7[M+H]+: 705.4; Found: 705.5.

To a solution of tert-butyl (S)-(2-(2-(2-hydroxyphenyl)-5,6,6a,7,9,10-hexahydro-8H-pyrazino[1′,2′:4,5]pyrazino[2,3-c]pyridazin-8-yl)ethyl)carbamate (40 mg, 0.094 mmol) in DCM (10 mL) was added TFA (1.0 mL). The mixture was stirred at 25° C. for 10 h. The volatiles were removed under reduced pressure to afford crude (S)-2-(8-(2-aminoethyl)-6,6a,7,8,9,10-hexahydro-5H-pyrazino[1′,2′:4,5]pyrazino[2,3-c]pyridazin-2-yl)phenol as its TFA salt (40 mg). LCMS m/z calcd for C17H23N6O [M+H]+: 327.2; Found: 327.2.

Palladium acetate (5.71 mg, 0.03 mmol) was added to the solution of tert-butyl (S)-(1-(4-bromo-2-fluorophenyl)ethyl)carbamate (90.0 mg, 0.25 mmol), potassium acetate (50 mg, 0.51 mmol) and 4-methyl-1,3-thiazole (50.5 mg, 0.51 mmol) in DMF (2 mL). The resulting mixture was purged with nitrogen for 3 times. The reaction mixture was heated to 90° C. and stirred for 16 h. The combined reaction mixture was diluted with EtOAc (25 mL) and washed with water (4×10 mL). The organic layer was dried over sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by prep-TLC with PE/EA (6/1) to afford tert-butyl (S)-(1-(2-fluoro-4-(4-methylthiazol-5-yl)phenyl)ethyl)carbamate (36 mg, 0.093 mmol, 36.5% yield). LCMS m/z calcd for C17H22FN2O2S [M+H]+: 337.1; Found: 337.3.

The title compound was prepared using procedure analogous to those described for Int-39, step 1 with appropriate starting materials. LCMS m/z calcd for C9H11BrNO2[M+H-tBu]+: 244.0; Found: 244.2.

To a solution of 1-(4-bromo-2-methoxyphenyl)ethanone (1.0 g, 4.4 mmol) in THF (10 mL) was added titanium ethoxide (2.0 g, 8.7 mmol) and (R)-tert-butanesulfinamide (635 mg, 5.24 mmol). After being purged with N23 times, the mixture was stirred at 70° C. for 12 h. The reaction mixture was diluted with water (20 mL) and extracted with EA (3×30 mL). The organic layers were combined, washed with brine, concentrated in vacuo and the residue was purified by silica gel chromatography (PE:EA=20:1 to 3:1) to give (S,E)-N-(1-(4-bromo-2-methoxyphenyl)ethylidene)-2-methylpropane-2-sulfinamide (400 mg, 1.2 mmol, 27% yield) as a yellow oil. LCMS m/z calcd for C13H19BrNO2S [M+1-1]+: 332.0; Found: 332.0.

To a solution of (SE)-N-(1-(4-bromo-2-methoxyphenyl)ethylidene)-2-methylpropane-2-sulfinamide (400 mg, 1.2 mmol) in THF (20 mL) was added and L-selectride (1 M, 8.73 mL, 8.73 mmol) at 0° C. After being purged with N23 times, the mixture was stirred at 70° C. for 12 h. The reaction mixture was diluted with water (20 mL) and extracted with EA (3×30 mL). The organic layers were combined, washed with brine, concentrated in vacuo and the residue was purified by silica gel chromatography (PE:EA=20:1 to 1:1) to give N—((S)-1-(4-bromo-2-methoxyphenyl)ethyl)-2-methylpropane-2-sulfinamide (206 mg, 0.59 mmol, 51.2% yield) as a yellow oil. LCMS m/z calcd for C13H21BrNO2S [M+1-1]+: 334.0; Found: 334.1.

A solution of N—((S)-1-(2-methoxy-4-(4-methylthiazol-5-yl)phenyl)ethyl)-2-methylpropane-2-sulfinamide (100 mg, 0.28 mmol) in HCl in dioxane (1 M, 1.42 mL, 1.42 mmol) was stirred at rt for 2 h. The volatiles were removed under reduced pressure to give (S)-1-(2-methoxy-4-(4-methylthiazol-5-yl)phenyl)ethan-1-amine (60 mg, 0.24 mmol, 85% yield) as a yellow solid which was used for the next step directly. LCMS m/z calcd for C13H17N2OS [M+H]+: 249.1; Found: 249.2.

The title compound was prepared using procedure analogous to those described for Int-39, step 2 with appropriate starting materials. LCMS m/z calcd for C22H31N2O5S [M+H]+: 435.2; Found: 435.1.

To a solution of (R)-2-amino-2-(4-(4-methylthiazol-5-yl)phenyl)ethan-1-ol (850 mg, 3.63 mmol) in DCM (10 mL) was added imidazole (741 mg, 10.9 mmol) and tert-butyl dimethylchlorosilane (820 mg, 5.44 mmol). The mixture was stirred at rt for 16 h. The reaction mixture was diluted with water and extracted with DCM. The organic layers were combined, dried over MgSO4, filtered and concentrated to dryness. The residue was purified by silica gel chromatography (DCM:MeOH=20:1) to get (R)-2-((tert-butyldimethylsilyl)oxy)-1-(4-(4-methylthiazol-5-yl)phenyl)ethan-1-amine (500 mg, 1.24 mmol, 34.3% yield) as a yellow oil. LCMS m/z calcd for C18H29N2OSSi [M+H]+: 349.2; Found: 349.2.

To a stirred solution of (2S,4R)-4-((tert-butyldimethylsilyl)oxy)-N—((R)-2-((tert-butyldimethylsilyl)oxy)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)-1-(2-(3-hydroxyisoxazol-5-yl)-3-methylbutanoyl)pyrrolidine-2-carboxamide (40.0 mg, 0.05 mmol) and 1,2-dibromoethane (15.2 mg, 0.08 mmol) in DMF (10 mL) was added potassium carbonate (0.06 mL, 0.11 mmol) at rt. After 16 h, water (20 ml) was added and the resulted mixture was extracted with EA (20 ml×3). The organic layers were combined, dried over MgSO4, filtered, and concentrated to dryness. The residue was purified by silica gel chromatography column (DCM:MeOH=20:1) to give (2S,4R)-1-(2-(3-(2-bromoethoxy)isoxazol-5-yl)-3-methylbutanoyl)-4-((tert-butyldimethylsilyl)oxy)-N—((R)-2-((tert-butyldimethylsilyl)oxy)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide (40.6 mg, 0.048 mmol, 89.2% yield) as a yellow solid. LCMS m/z calcd for C39H62BrN4O6SSi2[M+H]+: 849.3; Found: 849.5.

The title compound was prepared using procedure analogous to those described for Int-37 with appropriate starting materials. LCMS m/z calcd for C35H52BrN4O5SSi [M+H]+: 747.3; Found: 747.3.

To a solution of (2S,4R)-1-((S)-2-amino-3,3-dimethylbutanoyl)-4-hydroxy-N—((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide (50 mg, 0.11 mmol) in DMF (2 mL) was added (2-chloroethoxy)acetic acid (18.7 mg, 0.13 mmol), 2-(7-Azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (64 mg, 0.17 mmol) and TEA (0.08 mL, 0.45 mmol). The mixture was stirred at 110° C. for 4 h under the atmosphere of N2. The reaction was quenched with H2O and extracted with EA (10.0 mL×3). The organic layers were combined, dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by prep-TLC (DCM:MeOH=10:1) to give (2S,4R)-1-((S)-2-(2-(2-chloroethoxy)acetamido)-3,3-dimethylbutanoyl)-4-hydroxy-N—((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide (57 mg, 0.10 mmol, 89.7% yield) as a white solid. LCMS m/z calcd for C27H38ClN4O5S [M+H]+: 565.2; Found: 565.0.

To a solution of (R)-2-(6,6a,7,8,9,10-hexahydro-5H-pyrazino[1′,2′:4,5]pyrazino[2,3-c]pyridazin-2-yl)phenol (50 mg, 0.18 mmol) in ethanol (5 mL) was added tert-butyl 6-oxa-3-azabicyclo[3.1.0]hexane-3-carboxylate (327 mg, 1.76 mmol) and the solution was stirred at 80° C. for 6 h. The volatiles were removed and the residue was purified by prep-TLC to get the desired product (60 mg, 0.12 mmol, 72.5% yield). LCMS m/z calcd for C24H33N6O4[M+H]+: 469.2; Found: 469.2.

The title compound was prepared using a procedure analogous to those described for Int-64 with appropriate starting materials. LCMS m/z calcd for C20H276N6O2[M+H]+: 383.2; Found: 383.2.

To a stirred solution of tert-butyl (S)-4-(2-(2-hydroxyphenyl)-6,6a,7,8,9,10-hexahydro-5H-pyrazino[1′,2′:4,5]pyrazino[2,3-c]pyridazine-8-carbonyl)piperidine-1-carboxylate (170 mg, 0.34 mmol) in DCM (2 mL) was added trifluoroacetic acid (0.5 mL) at rt. After 16 h, the volatiles were removed under reduced pressure and the residue was purified by prep-HPLC (eluting with CH3CN in H2O: (0.1% NH3·H2O) from 10% to 95%) to give the desired product (150 mg, 0.38 mmol, 99% yield) as a white solid. LCMS m/z calcd for C21H27N6O2[M+H]+: 395.2; Found: 395.2.

The title compound was prepared using procedure analogous to those described for Int-9, step 1 to step 4 with appropriate starting materials. LCMS m/z calcd for CIII-136ClN4O3Si [M+H]+: 455.2; Found: 455.3.

To a stirred solution of tert-butyl 8-((tert-butyldimethylsilyl)oxy)-2-chloro-6,6a,7,8,9,10-hexahydro-5H-pyrido[1′,2′:4,5]pyrazino[2,3-c]pyridazine-5-carboxylate (170 mg, 0.37 mmol) in THF (5 mL) was added TBAF (0.75 mL, 0.75 mmol) at 0° C. After 1 h, the reaction was diluted with EA (30 mL) and washed with H2O (20 mL×3) and brine (20 mL). The organic layer was separated, dried over Na2SO4, filtered, and concentrated to dryness. The residue was purified by prep-TLC (DCM:MeOH=20:1) to give the desired product (120 mg, 0.35 mmol, 94.2% yield) as a white solid. LCMS m/z calcd for C15H22ClN4O3[M+H]+: 341.1; Found: 341.1.

To a stirred solution of tert-butyl 2-chloro-8-hydroxy-6,6a,7,8,9,10-hexahydro-5H-pyrido[1′,2′:4,5]pyrazino[2,3-c]pyridazine-5-carboxylate (110 mg, 0.32 mmol) in DCM (5 mL) was added Dess-Martin periodinane (274 mg, 0.65 mmol) at 0° C. The resulted mixture was stirred at 25° C. for 2 h. The reaction was diluted with DCM (30 mL), washed with aqueous NaHCO3(20 mL) and brine (20 mL). The organic layer was separated, dried over Na2SO4, filtered, and concentrated to dryness. The residue was purified by prep-TLC (DCM:MeOH=20:1) to give the desired product (100 mg, 0.29 mmol, 91.4% yield) as a white solid. LCMS m/z calcd for C15H20ClN4O3[M+H]+: 339.1; Found: 339.2.

To a stirred suspension solution of O-benzyl-N-(tert-butoxycarbonyl)-L-serine (20.0 g, 67.7 mmol) and 1-hydroxybenzotriazole hydrate (11.0 g, 81.3 mmol) in CH2Cl2(451 mL) was added DIPEA (14.2 mL, 81.3 mmol) at 0° C. The reaction mixture was added EDCI (15.6 g, 81.3 mmol) and stirred at 0° C. for 15 minutes. Then, the reaction mixture was added to a mixture of L-serine methyl ester hydrochloride (11.3 g, 81.3 mmol) in DIPEA (14.2 mL, 81.3 mmol) and DMF (30 mL) dropwise at 0° C. over 5 minutes. The reaction was warmed up to room temperature and stirred for 3 hours. The reaction was added water (500 mL), and extracted with DCM (300 mL×3). The organic phases were dried over Na2SO4, filtered and concentrated. The residue was purified by flash silica gel column chromatography (ethyl acetate and heptane, 0% to 100%) to give methyl O-benzyl-N-(tert-butoxycarbonyl)-L-seryl-L-alaninate (26.1 g, yield: 99%). LCMS calculated for C19H28N2O6(M+H)+: m/z=381.2; found: 381.0.

To a solution of methyl O-benzyl-N-(tert-butoxycarbonyl)-L-seryl-L-alaninate (26.1 g, 68.6 mmol) in DCM (260 mL) was added TFA (51.4 mL, 672.3 mmol) at room temperature. The reaction was stirred at room temperature for 3 hours. The reaction mixture was basified to between pH 7 and pH 8 via saturated aqueous NaHCO3solution, extracted with DCM (100 mL×3), and washed with brine (100 mL×1). The combined organic phases were dried over Na2SO4, filtered and concentrated. The residue was directly used for the next step without purification (16.9 g crude). LCMS calculated for C14H20N2O4(M+H)+: m/z=281.1; found: 281.0.

To a solution of methyl O-benzyl-L-seryl-L-alaninate (16.9 g, 60.3 mmol) in dioxane (169 mL) was stirred at 100° C. for overnight. The reaction was cooled to room temperature (white solid was precipitated out). The white precipitate was filtered, collected, and washed with cold MTBE (100 mL) to give (3S,6S)-3-((benzyloxy)methyl)-6-methylpiperazine-2,5-dione (11 g, yield: 73%).

To a solution of (3S,6S)-3-((benzyloxy)methyl)-6-methylpiperazine-2,5-dione (9.0 g, 36.3 mmol) in THF (201 mL) was added borane dimethyl sulfide complex (27.5 mL, 290.0 mmol) under ice-water bath. The reaction was stirred at 60° C. for overnight. The reaction was cooled under ice-water bath, and slowly added MeOH (200 mL). The reaction mixture was warmed up to room temperature, added 1 N HCl aqueous solution to pH 3, and then stirred at 50° C. for 3 hours. The reaction mixture was basified to pH 12 via 1 N NaOH aqueous solution and extracted with CHCl3(200 mL×3). The combined organic phases were dried over Na2SO4, filtered and concentrated. The residue was directly used for the next step without purification (9.8 g crude). LCMS calculated for C13H20N2O (M+H)+: m/z=221.2; found: 221.2.

To a solution of (2R,5S)-2-((benzyloxy)methyl)-5-methylpiperazine (0.29 g, 1.3 mmol) in DCM (13 mL) was added 1 M BCl3in DCM solution (5.2 mL, 5.2 mmol) at 78° C. The reaction was slowly warmed up to room temperature and stirred for overnight. The reaction was cooled under ice-water bath, and slowly added MeOH (10 mL). The reaction mixture was concentrated to dryness. The residue was directly used for the next step without purification (0.23 g crude). LCMS calculated for C6H14N2O (M+H)+: m/z=131.1; found: 131.0.

To a solution of ((2R,5S)-5-methylpiperazin-2-yl)methanol (9.0 g, 69.1 mmol) in DCM (376 mL) was added TEA (120.0 mL, 864.0 mmol), and di-tert-butyl dicarbonate (45.3 g, 207.0 mmol) at 0° C. The reaction was stirred at room temperature for overnight, and then concentrated to dryness. The residue was directly used for the next step without purification (24.0 g crude). LCMS calculated for C16H30N2O5(M+H)+: m/z=331.2; found: 331.0.

To a solution of di-tert-butyl (2R,5S)-2-(hydroxymethyl)-5-methylpiperazine-1,4-dicarboxylate (14.0 g, 42.4 mmol) in EtOH (78.5 mL) was added a solution of NaOH (8.5 g, 211.9 mmol) in water (78.5 mL). The reaction mixture was stirred at 80° C. for overnight. The reaction was cooled to room temperature, added 1 N HCl aqueous solution to pH 9, and extracted with CHCl3(100 mL×3). The combined organic phases were dried over Na2SO4, filtered and concentrated. The residue was purified by flash silica gel column chromatography (DCM and MeOH with 0.1% TEA, 0% to 10%) to give tert-butyl (2S,5R)-5-(hydroxymethyl)-2-methylpiperazine-1-carboxylate (2.7 g, yield: 28%). LCMS calculated for C11H22N2O3(M+H)+: m/z=231.2; found: 231.0.

The title compound was prepared using procedure analogous to those described for Int-9, step 1 to step 5, using appropriate starting materials. LCMS calc. for C26H35N5O5[M+H]+: m/z=498.3; Found: 498.5.

SYNTHESIS OF EXAMPLES

The Examples disclosed herein are embodiments of the invention.

The examples in the table below were prepared according to the same method as example 1.

The examples in the table below were prepared according to the same method as example 3 using appropriate starting materials.

To a solution of 2-(6,6a,7,8,9,10-hexahydro-5H-pyrazino[1′,2′:4,5]pyrazino[2,3-c]pyridazin-2-yl)phenol (Int-1) (22.0 mg, 0.06 mmol) and (2S,4R)-1-((R)-2-(3-(2-bromoethoxy)isoxazol-5-yl)-3-methylbutanoyl)-4-((tert-butyldimethylsilyl)oxy)-N—((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide (Int-4) (30 mg, 0.04 mmol) in DMF (3 mL) was added NaHCO3(36.0 mg, 0.4 mmol) at rt, the mixture was stirred at 60° C. for 16 h. The reaction was taken up in EtOAc (20 ml) and the organics were washed with water (20 mL×3) and brine (20 mL). The organics were then separated and dried (MgSO4) before concentration in vacuum to dryness. The crude product was then purified by prep-HPLC (0.1% HCl, MeCN in water from 10%-90%) to give (2S,4R)-4-((tert-butyldimethylsilyl)oxy)-1-((2R)-2-(3-(2-(2-(2-hydroxyphenyl)-5,6,6a,7,9,10-hexahydro-8H-pyrazino[1′,2′:4,5]pyrazino[2,3-c]pyridazin-8-yl)ethoxy)isoxazol-5-yl)-3-methylbutanoyl)-N—((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide (17) (15 mg, 0.02 mmol, 31% yield) as a sticky colorless solid. LCMS calc'd for C48H63N9O6SSi: 921.4; Found: LCMS [M+H]: 922.5.

The title compound was prepared using procedure analogous to those described for example 4 with appropriate starting materials. LCMS m/z calcd for C42H50N9O6S [M+H]+: 808.4; Found: 808.0.

To a solution of (2S,4R)-1-((R)-2-(3-(2-bromoethoxy)isoxazol-5-yl)-3-methylbutanoyl)-4-((tert-butyldimethylsilyl)oxy)-N—((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide (Int-4) (55.8 mg, 0.08 mmol) and 2-(8-(piperidin-4-ylmethyl)-6,6a,7,8,9,10-hexahydro-5H-pyrazino[1′,2′:4,5]pyrazino[2,3-c]pyridazin-2-yl)phenol (Int-8) (25 mg, 0.07 mmol) in DMF (3 mL) were added NaHCO3(55.2 mg, 0.66 mmol) and sodium iodide (0.48 mg, 0.003 mmol) at rt, the mixture solution was stirred at 60° C. for 16 h. The residue was extracted with EtOAc (10 mL) and the organics were washed with water (30 mL×2) and saturated brine (30 ml). The organics were then separated and dried over MgSO4before concentration to dryness. The crude product was then purified by prep-TLC (PE:EtOAc=1:2) to give (2S,4R)-4-[tert-butyl(dimethyl)silyl]oxy-1-[(2R)-2-[3-[2-[4-[[4-(2-hydroxyphenyl)-1,5,6,8,12-pentazatricyclo[8.4.0.02,7]tetradeca-2(7),3,5-trien-12-yl]methyl]piperidin-1-yl]ethoxy]-1,2-oxazol-5-yl]-3-methylbutanoyl]-N-[(1S)-1-[4-(4-methyl-1,3-thiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (18) (50 mg, 0.05 mmol, 75% yield) as a solid. LCMS calc'd for C54H74N10O6SSi: 1018.4; Found: LCMS [M+H]: 1019.6.

The title compound was prepared using procedure analogous to those described for Example 3, with tert-butyl 3-bromopropanoate replacing Int-6. LCMS m/z calcd for C28H41N6O3[M+H]+: 509.3; Found: 509.1.

To a solution of tert-butyl 3-(4-((2-(2-hydroxyphenyl)-5,6,6a,7,9,10-hexahydro-8H-pyrazino[1′,2′:4,5]pyrazino[2,3-c]pyridazin-8-yl)methyl)piperidin-1-yl)propanoate (27.0 mg, 0.05 mmol) in DCM (3 mL) and TFA (0.5 mL, 6.53 mmol), the mixture solution was stirred at 25° C. for 1 h. The residue was diluted with EtOAc (10 mL) and the organic layer washed with water (2×10 mL) then saturated brine (1×10 mL). The organic layer was separated, dried over MgSO4, and filtered. The filtration was concentrated to dryness under reduced pressure. The residue was purified by prep-TLC (PE:EA=1:2) to give 3-(4-((2-(2-hydroxyphenyl)-5,6,6a,7,9,10-hexahydro-8H-pyrazino[1′,2′:4,5]pyrazino[2,3-c]pyridazin-8-yl)methyl)piperidin-1-yl)propanoic acid (20 mg, 0.04 mmol, 83.3% yield) as a solid. LCMS m/z calcd for C24H33N6O3[M+H]+: 453.2; Found: 453.2.

The examples in the table below were prepared according to the same method as example 27 using appropriate starting materials.

The examples in the table below were prepared according to the same method as example 32 using appropriate starting materials.

To a solution of (S)-2-(8-(2-aminoethyl)-6,6a,7,8,9,10-hexahydro-5H-pyrazino[1′,2′:4,5]pyrazino[2,3-c]pyridazin-2-yl)phenol (20 mg, 0.06 mmol) and (2S,4R)-4-((tert-butyldimethylsilyl)oxy)-1-(3-methyl-2-(3-(2-oxoethyl)isoxazol-5-yl)butanoyl)-N—((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide (58.7 mg, 0.09 mmol) in DCM (30 mL) was added NaBH(OAc)3(25.4 mg, 0.12 mmol) at rt. The mixture was stirred at rt for 16 h. The volatiles were removed under reduced pressure and the residue was purified by prep-TLC (MeOH:DCM=1:10). TBS group removal was achieved upon work-up and purification to give (2S,4R)-4-hydroxy-1-(2-(3-(2-((2-((S)-2-(2-hydroxyphenyl)-5,6,6a,7,9,10-hexahydro-8H-pyrazino[1′,2′:4,5]pyrazino[2,3-c]pyridazin-8-yl)ethyl)amino)ethyl)isoxazol-5-yl)-3-methylbutanoyl)-N—((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide (20 mg, 0.024 mmol, 39.1% yield) as a yellow solid). LCMS m/z calcd for C44H55N10O5S [M+H]+: 835.4; Found: 835.5.

The examples in the table below were prepared according to the same method as example 34 using appropriate starting materials.

The examples in the table below were prepared according to the same method as example 35 using appropriate starting materials.

A solution of methyl (S)-2-((5-(((S)-1-((2S,4R)-4-((tert-butyldimethylsilyl)oxy)-2-(((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)carbamoyl)pyrrolidin-1-yl)-3-methyl-1-oxobutan-2-yl)isoxazol-3-yl)oxy)propanoate (6.0 g, 8.6 mmol) in MeOH (75 mL) was added NaBH4(1.3 g, 4.8 mmol). The resulting mixture was stirred at rt for 3 h. The reaction was quenched with water (50 mL) and diluted with DCM (100 mL). The layers were separated, and the aqueous phase was extracted with DCM (1×100 mL). The combined organic layers were washed with brine (50 mL), dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to afford crude product as colorless oil, which was used directly in the next step. LCMS m/z calcd for C34H51N4O6SSi [M+H]+: 671.3; Found: 671.2.

A solution of crude (2S,4R)-4-((tert-butyldimethylsilyl)oxy)-1-((R)-2-(3-(((S)-1-hydroxypropan-2-yl)oxy)isoxazol-5-yl)-3-methylbutanoyl)-N—((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide (2.6 g, 3.9 mmol) in ACN (30 mL) was added IBX (45 wt %, 2.7 g, 9.7 mmol). The resulting mixture was heated at 80° C. for 3 h. The reaction crude was cooled to rt and filtered under reduced pressure. The filter cake was washed by ACN (50 mL). The filtrate was concentrated in vacuum to give the crude product, which was purified by silica gel chromatography column, using EA/Heptane as eluents (20-80% EA in heptane) to afford the desired product (2.1 g, 72.9% yield over two steps) as a colorless oil. LCMS m/z calcd for C34H49N4O6SSi [M+H]+: 669.3; Found: 669.2.

A suspension of (2S,4R)-4-((tert-butyldimethylsilyl)oxy)-1-((R)-3-methyl-2-(3-(((S)-1-oxopropan-2-yl)oxy)isoxazol-5-yl)butanoyl)-N—((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide (2.1 g, 3.14 mmol) in DCM (20 mL) was added STAB (2000 mg, 9.4 mmol, 3.0 eq) and NaHCO3(1350 mg, 16 mmol, 5.0 eq). Then a solution of 2-((6aS)-8-(pyrrolidin-3-yl)-6,6a,7,8,9,10-hexahydro-5H-pyrazino[1′,2′:4,5]pyrazino[2,3-c]pyridazin-2-yl)phenol (Int-20, 1.7 g, 4.8 mmol) in DCM (20 mL) was added dropwise at rt. The resulting suspension was stirred at rt for 18 h. The reaction was quenched with water (50 mL). The organic layer was separated, washed with brine (50 mL), dried over Na2SO4, filtered, and the filtrate was concentrated in vacuo. The residue was purified by silica gel chromatography column using MeOH/DCM as eluents (0-10% MeOH in DCM) to afford the desired product (2.3 g, 72.9% yield) as a white solid. LCMS calc'd for C53H73N10O6SSi [M+1-1]+: 1005.5; Found: 1005.6.

The examples in the table below were prepared according to the same method as example 48 using appropriate starting materials.

Biological Assays

SMARCA2 Bromodomain Binary and Ternary Binding Assay

The inhibitory activity of compounds was evaluated in vitro using TR-FRET assay with white 384-well low volume microplate (PerkinElmer ProxiPlate Plus), in which the compound competes the same binding site with the ligand, and thus lead to dose-dependent TR-FRET signal reduction. The cooperativity of the compounds for ternary complex formation with E3 ligase was evaluated in the absence or presence of saturating concentrations of VCB. Testing compounds were dissolved in DMSO at 10 μM and tested in 9-dose IC50. The assay mixture was prepared by mixing SMARCA2 (10 nM final), biotinylated probe (25 nM final), and assay buffer or VCB (5 μM) in 1× AlphaLISA Epigenetics Buffer (PerkinElmer AL008F) with 1 mM TCEP. The compounds in DMSO were added to each well in 3-fold serial dilution by dispenser (TECAN D300E) and incubate for 20 minutes at room temperature before addition of detection reagents, Lance Eu W1024 anti-6×His (0.6 nM final, PerkinElmer AD0110) and Streptavidin Surelight APC (6 nM final, PerkinElmer CR130-100). The plate was then sealed and further incubated at 4° C. overnight in dark, and then was read by Envision multimode plate reader (PerkinElmer, 2102-0010). The ratio of florescence signal at 665/620 was used in data analysis. Percentage inhibition was calculated by % inhibition=100×(FDMSO−F)/(FDMSO−FPC), in which FDMSOis DMSO control, and FPCis positive control. IC50values were determined from dose response curve by fitting the percent inhibition against compound concentration using GraphPad Prism software.

SMARCA4 Bromodomain Binary and Ternary Binding Assay

Recombinant bromo domain of SMARCA4 protein, was purchased from Active Motif (31401). The inhibitory activity of compounds was evaluated in vitro using TR-FRET assay with white 384-well low volume microplate (PerkinElmer ProxiPlate Plus), in which the compound competes the same binding site with the ligand, and thus lead to dose-dependent TR-FRET signal reduction. The cooperativity of the compounds for ternary complex formation with E3 ligase was evaluated in the absence or presence of saturating concentrations of VCB. Testing compounds were dissolved in DMSO at 10 mM and tested in 9-point IC50mode. The assay mixture was prepared by mixing SMARCA4 (20 nM final), biotinylated probe (25 nM final), and assay buffer or VCB (5 μM) in 1× AlphaLISA Epigenetics Buffer (PerkinElmer AL008F) with 1 mM TCEP. The compounds of interest in DMSO were added to each well in 3-fold serial dilution by dispenser (TECAN D300E). and incubate for 20 minutes at room temperature before addition of detection reagents, Lance Eu W1024 anti-6×His (0.6 nM final, PerkinElmer AD0110) and Streptavidin Surelight APC (6 nM final, PerkinElmer CR130-100). The plate was then sealed and further incubated at 4° C. overnight in dark, and then was read by Envision multimode plate reader (PerkinElmer, 2102-0010). The ratio of florescence signal at 665/620 was used in data analysis. Percentage inhibition was calculated by % inhibition=100×(FDMSO−F)/(FDMSO−FPC), in which FDMSOis DMSO control, and FPCis positive control. IC50values were determined from dose response curve by fitting the percent inhibition against compound concentration using GraphPad Prism software.

Cell Treatment and in Cell Western (ICW) for Detecting SMARCA Proteins

Compound titration and cell culture: Compounds were dissolved in DMSO to make 10 mM stock and 3-fold series dilutions were further conducted keeping the highest concentration 10 μM. NCIH1693 and NCIH520 cells were maintained in PRMI 1640 medium (Corning Cellgro, Catalog #:10-040-CV) supplemented with 10% v/v FBS (GE Healthcare, Catalog #: SH30910.03) by splitting 1:3 twice a week.

To determine SMACRA2 and SMARCA4 protein degradation DC50values in NCIH1693 and NCIH520 cells by In Cell Western (ICW) analysis. Cells were trypsinized and 30 thousand cells/well were seeded into 384-well plates and were allowed to grow for 5 hours at 37° C. Eight-point, 3-fold serial dilutions of compounds from 10 mM stocks were added to the cells (using digital Dispenser D300-Tecan, keeping highest concentration 10 μM and normalizing with DMSO at the highest dispensed volume). Plates were incubated at 37° C. for overnight (maximum 18 hours). Cells incubated with DMSO was used as a vehicle control.

To perform In Cell Western, medium was removed from all the wells leaving cells attached to the surface. After removing the medium, cells were fixed within the plates with 40 μL of 4% formaldehyde by incubating at room temperature for 30 minutes, and then permeabilized with wash buffer (1×PBS with 0.1% Triton X-100) by washing the plate 5 times with 50 μL/well. Before labeling with primary antibodies, cells were blocked with 30 μL/well of blocking buffer (Licor Odyssey blocking buffer PBS #927-40000) for 30 minutes at room temperature. To measure SMARCA2 or SMARCA4 proteins, cells were labeled with 20 μL/well of anti SMARCA2 or SMACRA4 antibodies (Cell Signaling BRM #11966S 1:1000, Cell Signaling BRG #49360S 1:1000) diluted in Li-Cor Odyssey blocking buffer-PBS #927-40000, followed by overnight incubation at 4° C.

The next day, plates were washed 5×5 minutes with 50 μL/well of washing buffer to remove all the excess primary antibody and then 20 μL from a mixture of secondary antibody and fluorescent DNA specific dye (Goat anti rabbit 1:500 IRDye-800CW #92632211, and DRAQ5™ 1:2000-#ab108410) was added to each well. Plates were incubated for 1 hour at room temperature with gentle rocking. Cells were washed 5 times with 50 μL/well wash buffer, followed by one last wash with DI water, followed by 10 mins drying at 37° C. oven before scanning. Plates were scanned using Li-Cor Odyssey CLx imaging system to acquire integrated intensities at 700 nm and 800 nm. SMARCA signals were normalized to total cell count and then these normalized values were used to calculate the percent degradation relative to DMSO control and maximum inhibition. DC50s were calculated by using the GraphPad Prism4 program based on sigmoidal dose response equation ([Inhibitor] vs. normalized response—Variable slope).

Cell Proliferation Assay to Determine IC50in NCIH1693 and NCIH1708 Cells.

One thousand NCIH1693 or NCIH1703 cells per well were seeded in 96 well plate and incubated at 37° C. for 5 hrs. A series of three-fold dilution of compounds from 10 mM stock concentration were added to the cells by using digital Dispenser D300-Tecan, keeping highest concentration 10 μM and normalizing with DMSO at the highest dispensed volume). Cells incubated with DMSO was used as a vehicle control. After compound treatment plates were incubated at 37° C. for 4 days and cell viability was measured by measuring ATP content from the cell lysate using ATPlite Luminescence assay system (PerkinElmer Cat. no #6016941). Percentage of viable cells, relative to DMSO vehicle control and maximum inhibition, was calculated and plotted in Graphpad Prism ([Inhibitor] vs. normalized response Variable slope) to determine cell proliferation IC50values on day 4.

HiBiT peptide knock-in of SMARCA2 in LgBiT expressing HEK293T cells was performed by CRISPR-mediated tagging system as described Promega. The homozygous HiBiT knock-in on c-terminus SMARCA2 was confirmed by sanger sequence. SMARCA2-HiBiT knock-in Hela monoclonal cell (CS302366) and SMARCA4-HiBiT knock-in Hela monoclonal cell (CS3023226) were purchased from Promega. The heterozygous HiBiT-knock-in was confirmed by sanger sequence in both SMARCA2-HiBiT and SMARCA4-HiBiT monoclonal cells.

SMARCA2 HiBiT Degradation Assay in HEK293T Cells

Compounds were prepared in a low dead volume plate at 3-fold serial dilution, and then transferred 25 nL/well into a 384 well plate (Corning: #356661) by Agilent Bravo liquid dispenser. SMARCA2-HiBiT monoclonal HEK293T cells were added at 5,000/well/25 uL into the compound containing 384 well plate. After 24 h incubation, 25 uL/well of Nano-Glo HiBiT Lytic detection buffer (Promega: N3050) was added to the wells and incubate for 10 min on a shaker, centrifuged for 5 min, and then RLU was detected on a microplate reader (Envision, PerkinElmer). The RLU ratio values are normalized to percent inhibition as follows: % inhibition=((HC−LC)−(compound−LC))/(HC−LC))*100, where HC=high control=mean signal of DMSO only; LC=low control=mean signal of 100% inhibition of RLU by 1 μM PRT1001728. An 11-point dose response curve for each compound will be generated using normalized % inhibition to determine IC50 values based on the equation: Y=Bottom+((Top−Bottom)/(1+((IC50/X){circumflex over ( )}Slope))), where Y is the % inhibition in the presence of X inhibitor concentration; Top=the top plateau of the curve; Bottom=the bottom plateau of the curve; Slope=Hill coefficient; DC50=concentration of compound with 50% degradation in relation to top/high control. DC50 values were determined by using XLfit Model 205.

SMARCA2 HiBiT and SMARCA4 HiBiT Degradation Assay in HeLa Cells

Dispense 10 μL aliquot of prepared Hela-SMARCA2-HiBiT or Hela-SMARCA4-HiBiT cells (1:1 ratio of cells:Trypan Blue (#1450013, Bio-Rad)) onto cell counting slide (#145-0011, Bio-Rad) and obtain cell density and cell viability using cell counter (TC20, Bio-Rad). Remove appropriate volume of resuspended cells from culture flask to accommodate 2500 cells/well@20 μL/well. Transfer Hela-HiBiT cells to 50 ml conical (#430290, Corning). Spin down at 1000 rpm for 5 min using tabletop centrifuge (SPINCHRON 15, Beckman). Discard supernatant and resuspend cell pellet in modified EMEM (#30-2003, ATCC) cell culture media containing 10% FBS (F2422-500ML, Sigma), and 1× Penicillin/Streptomycin (200 g/1) (30-002-CI, Corning) to a cell density of 125,000 cells/ml. Dispense 20 μL of resuspended Hela-HiBit cells per well in 384-well TC treated plate (#12-565-343, Thermo Scientific) using standard cassette (#50950372, Thermo Scientific) on Multidrop Combi (#5840310, Thermo Scientific) inside laminar flow cabinet. Dispense test compounds onto plates using digital liquid dispenser (D300E, Tecan). Incubate plates in humidified tissue culture incubator @37° C. for 18 hours. Add 20 ul of prepared Nano-Glo® HiBiT Lytic detection buffer (N3050, Promega) to each well of 384-well plate using small tube cassette (#24073295, Thermo Scientific) on Multidrop Combi, incubate @RT for 30-60 min. Read plates on microplate reader (Envision 2105, PerkinElmer) using 384 well Ultra-Sensitive luminescence mode. Raw data files and compound information reports are swept into centralized data lake and deconvoluted using automated scripts designed by TetraScience, Inc. Data analysis, curve-fitting and reporting done in Dotmatics Informatics Suite using Screening Ultra module.

In some embodiments, the disclosure is directed to the following aspects:Aspect 1. A compounds of Formula (I):
PTM-ULM  (I)or a pharmaceutically acceptable salt or solvate thereof,whereinPTM is a moiety of Formula IA:

Aspect 7. The compound according to any one of aspects 1-5, wherein the compound of Formula IA is a compound of Formula IA-2:

Aspect 8. The compound according to any one of aspects 1-5 or 7, wherein the compound of Formula IA is a compound of Formula IA-3:

wherein m=1 to 3;X is optionally substituted —CH2—, or NH; or, if R1is attached to X, then X is —CH— or N;and Q is optionally substituted —CH2—, optionally substituted —(CH2)2—, —C(O)—, optionally substituted —CH2C(O)—, —S(O)—, —S(O)2—, optionally substituted —CH2S(O)2—, or optionally substituted —CH2S(O)—.Aspect 9. The compound according to any one of aspects 1-5 or 7-8, wherein the compound of Formula IA is a compound of Formula IA-4:

wherein m=1 to 3; each Rkis independently H, D, F, C1-3alkyl, C1-3haloalkyl, C1-4alkoxyl, substituted C1-3alkyl, substituted C1-3haloalkyl, or substituted C1-4alkoxyl; and s=0-7.Aspect 10. The compound according to aspect 9, wherein the compound of Formula IA-4 is a compound of Formula IA-5:

Aspect 11. The compound according to any one of aspects 8-10, wherein m=2.Aspect 12. The compound according to any one of aspects 8-11, wherein at least one W is optionally substituted —CH2; and wherein when n=2 or 3, only one W may be —C(O)—, —S(O)—, or —S(O)2—Aspect 13. The compound according to any one of aspects 8-11, wherein at least one W is —C(O)—.Aspect 14. The compound according to aspect 11, wherein the compound of Formula IA-5 is a compound of Formula IA-6:

Aspect 15. The compound according to any one of the preceding aspects, wherein Re3is H.Aspect 16. The compound according to any one of the preceding aspects, wherein Rd1is H.Aspect 17. The compound according to any one of aspect 1-13, wherein Rc1is H.Aspect 18. The compound according to any one of the preceding aspects, wherein ULM is a moiety having the Formula ULM-I

whereinthe dashed line (----) indicates the position of attachment of ULM-I to R1;V is H or F;R3is optionally substituted phenyl, optionally substituted napthyl, or an optionally substituted 5-10 membered heteroaryl;one of R4or R5is H, D, haloalkyl, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, —CORd, CONRe1Re2;the other of R4or R5is H or D;or R4and R5, together with the carbon atom to which they are both attached, form an optionally substituted 3-5 membered cycloalkyl, heterocyclyl;W3is an optionally substituted aryl, optionally substituted heteroaryl, or

R6and R7are independently H, D, optionally substituted alkyl, optionally substituted cycloalkyl, or optionally substituted haloalkyl,or R6, R7, and the carbon atom to which they are attached form an optionally substituted cycloalkyl or optionally substituted heterocyclyl;R8is an optionally substituted heterocyclyl, optionally substituted heteroaryl, optionally substituted aryl, CONRaRb, NRaRb,

Rais selected from H or optionally substituted alkyl;Rbis selected from H, —C(O)—* wherein * is a point of attachment to R1, optionally substituted alkyl, optionally substituted alkylcarbonyl, optionally substituted (cycloalkyl)alkylcarbonyl, optionally substituted aralkylcarbonyl, optionally substituted arylcarbonyl, optionally substituted (cycloalkyl)carbonyl, optionally substituted (heterocyclyl) carbonyl, or optionally substituted aralkyl;each Rcis independently H, halo, optionally substituted alkoxy, cyano, optionally substituted alkyl, haloalkyl, or haloalkoxy;each Rdis independently selected from H, optionally substituted alkyl or NRe1Re2;each Re1and Re2is independently H, D, optionally substituted alkyl,or Re1and Re2together with the nitrogen atom to which they are attached form a 4-7 membered heterocyclyl; andp is 0, 1, 2, 3, or 4.Aspect 19. The compound according to aspect 18, wherein R8is optionally substituted heterocyclyl.Aspect 20. The compound according to aspect 18, wherein R8is optionally substituted heteroaryl.Aspect 21. The compound according to aspect 18, wherein R8is optionally substituted aryl.Aspect 22. The compound according to aspect 18, wherein R8is CONRaRb.Aspect 23. The compound according to aspect 18, wherein R8is NRaRb.Aspect 24. The compound according to aspect 18, wherein R8is

Aspect 25. The compound according to aspect 18, wherein R8is

Aspect 26. The compound according to aspect 18, wherein R8is

Aspect 27. The compound according to aspect 18, wherein R8is

Aspect 28. The compound according to aspect 18, wherein R8is

Aspect 29. The compound according to aspect 18, wherein R8is

Aspect 30. The compound according to aspect 18, wherein R8is

Aspect 31. The compound according to aspect 18, wherein R8is

Aspect 32. The compound according to aspect 18, wherein R8is

Aspect 33. The compound according to any one of aspects 18-32, wherein R3is optionally substituted phenyl having the formula:

each optionally substituted.Aspect 35. The compound according to aspect 34, wherein R9is

Aspect 36. The compound according to any one of aspects 33-35, wherein R10is H, D, hydroxy, halogen, —NH(C1-C4alkyl), or C1-C6alkoxy, and z is 0, 1, 2, 3, or 4.Aspect 37. The compound according to any one of aspects 18-36, whereinW3is

Rais H or optionally substituted alkyl;Rbis H, —C(O)—* wherein * is a point of attachment to R1, optionally substituted alkyl, optionally substituted alkylcarbonyl, or optionally substituted (cycloalkyl)carbonyl.Aspect 38. The compound according to aspect 37, whereinR7is H, C1-C6alkyl, C1-C6alk-OH, C1-C6alk-NH2, —C1-C6alk-CONH—*, or —C1-C6alk-NHCO—*;R8is —NH—*, or —NHCOR11;* is a point of attachment of the ULM to R1; andR11is

Aspect 39. The compound according to any one of aspects 18-36, whereinW3is

Rcis H or optionally substituted alkyl; and p=1.Aspect 40. The compound according to any one of aspects 18-39, wherein ULM-I is a compound of formula:

* is a point of attachment of the ULM to R1.Aspect 41. The compound of aspect 40, whereinR9is optionally substituted

and R10is H, D, hydroxy, halogen, —NH(C1-C6alkyl), or —OC1-C6alkyl.Aspect 42. The compound according to aspect 40 or 41, wherein the compound of Formula I is a compound of Formula IA-7 or IA-8:

Aspect 43. The compound according to aspect 42, wherein the compound of Formula I is a compound of Formula IA-9 or IA-10:

Aspect 44. The compound according to any one of aspects 1-43, wherein R1is —CR1a═CR1b—.Aspect 45. The compound according to any one of aspects 1-43, wherein R1is —(CR1aR1b)1-5.Aspect 46. The compound according to any one of aspects 1-43, wherein R1is —(CR1aR1b)1-5-A- wherein A is O, S, or NR1c.Aspect 47. The compound according to any one of aspects 1-43, wherein R1is —(CR1aR1b)1-5-A-(CR1aR1b)1-5— wherein A is O, S, or NR1c.Aspect 48. The compound according to any one of aspects 1-43, wherein R1is —(C≡C)—(CR1aR1b)1-5.Aspect 49. The compound according to any one of aspects 1-43, wherein R1is —(CR1aR1b)1-5-(3-11 membered cycloalkyl optionally substituted with 0-6 R1aand/or R1bgroups)-.Aspect 50. The compound according to any one of aspects 1-43, wherein R1is —(CR1aR1b)1-5-(3-11 membered cycloalkyl optionally substituted with 0-6 R1aand/or R1bgroups)-(CR1aR1b)1-5.Aspect 51. The compound according to any one of aspects 1-43, wherein R1is —(CR1aR1b)1-5-(3-11 membered heterocyclyl optionally substituted with 0-6 R1aand/or R1bgroups)-(CR1aR1b)1-5.Aspect 52. The compound according to any one of aspects 1-43, wherein R1is —(CR1aR1b)1-5-(3-11 membered heterocyclyl optionally substituted with 0-6 R1aand/or R1bgroups)-.Aspect 53. The compound according to any one of aspects 1-43, wherein R1is -(3-11 membered heterocyclyl optionally substituted with 0-6 R1aand/or R1bgroups) —(CR1aR1b)1-5—.Aspect 54. The compound according to any one of aspects 1-43, wherein R1is —(CR1aR1b)1-5-(3-11 membered cycloalkyl optionally substituted with 0-6 R1aand/or R1bgroups)-(CR1aR1b)1-5-A- wherein A is O, S, or NR1c.Aspect 55. The compound according to any one of aspects 1-43, wherein R1is —(CR1aR1b)1-5-(3-11 membered heterocyclyl optionally substituted with 0-6 R1aand/or R1bgroups)-(CR1aR1b)1-5-A- wherein A is O, S, or NR1c.Aspect 56. The compound according to any one of aspects 1-43, wherein R1is —(CR1aR1b)1-5-(3-11 membered heterocyclyl optionally substituted with 0-6 R1aand/or R1bgroups)-A- wherein A is O, S, or NR1c.Aspect 57. The compound according to any one of aspects 1-43, wherein R1is —(CR1aR1b)1-5-A-(3-11 membered heterocyclyl optionally substituted with 0-6 R1aand/or R1bgroups)- wherein A is O, S, or NR1c.Aspect 58. The compound according to any one of aspects 1-43, wherein R1is —(CR1aR1b)1-5-A-(3-11 membered cycloalkyl optionally substituted with 0-6 R1aand/or R1bgroups)- wherein A is O, S, or NR1c.Aspect 59. The compound according to any one of aspects 1-43, wherein R1is —(CR1aR1b)1-5-A-(CR1aR1b)1-5-A- wherein A is O, S, or NR1c.Aspect 60. A pharmaceutical composition comprising a compound according to any one of aspects 1 to 59 and a pharmaceutically acceptable excipient.Aspect 61. A method of treating cancer in a subject in need thereof comprising administering to the subject a compound of any one of aspects 1 to 59.Aspect 62. The method of aspect 61, wherein the cancer is SMARCA4 deleted cancer.Aspect 63. The method of either one of aspect 61 or 62, wherein the cancer is squamous-cell carcinoma, basal cell carcinoma, adenocarcinoma, hepatocellular carcinomas, and renal cell carcinomas, cancer of the bladder, bowel, breast, cervix, colon, esophagus, head, kidney, liver, lung, neck, ovary, pancreas, prostate, and stomach; leukemias; benign and malignant lymphomas, particularly Burkitt's lymphoma and Non-Hodgkin's lymphoma; benign and malignant melanomas; myeloproliferative diseases; sarcomas, including Ewing's sarcoma, hemangiosarcoma, Kaposi's sarcoma, liposarcoma, myosarcomas, peripheral neuroepithelioma, synovial sarcoma, gliomas, astrocytomas, oligodendrogliomas, ependymomas, glioblastomas, neuroblastomas, ganglioneuromas, gangliogliomas, medulloblastomas, pineal cell tumors, meningiomas, meningeal sarcomas, neurofibromas, and Schwannomas; bowel cancer, breast cancer, prostate cancer, cervical cancer, uterine cancer, lung cancer, ovarian cancer, testicular cancer, thyroid cancer, astrocytoma, esophageal cancer, pancreatic cancer, stomach cancer, liver cancer, colon cancer, melanoma; carcinosarcoma, Hodgkin's disease, Wilms' tumor and teratocarcinomas. Additional cancers which may be treated using compounds according to the present disclosure include, for example, T-lineage Acute lymphoblastic Leukemia (T-ALL), T-lineage lymphoblastic Lymphoma (T-LL), Peripheral T-cell lymphoma, Adult T-cell Leukemia, Pre-B ALL, Pre-B Lymphomas, Large B-cell Lymphoma, Burkitts Lymphoma, B-cell ALL, Philadelphia chromosome positive ALL and Philadelphia chromosome positive CML.