Oxysterols and methods of use thereof

Compounds are provided according to Formula (I) and pharmaceutically acceptable salts thereof, and pharmaceutical compositions thereof; wherein R1, R2, and R3 are as defined herein. Compounds of the present invention are contemplated useful for the prevention and treatment of a variety of conditions.

BACKGROUND OF THE INVENTION

NMDA receptors are heteromeric complexes comprised of NR1, NR2, and/or NR3 subunits and possess distinct recognition sites for exogenous and endogenous ligands. These recognition sites include binding sites for glycine, and glutamate agonists and modulators. NMDA receptors are expressed in the peripheral tissues and the CNS, where they are involved in excitatory synaptic transmission. Activating these receptors contributes to synaptic plasticity in some circumstances and excitotoxicity in others. These receptors are ligand-gated ion channels that admit Ca2+ after binding of the glutamate and glycine, and are fundamental to excitatory neurotransmission and normal CNS function. Positive modulators may be useful as therapeutic agents with potential clinical uses as cognitive enhancers and in the treatment of psychiatric disorders in which glutamatergic transmission is reduced or defective (see, e.g., Horak et al., J. of Neuroscience, 2004, 24(46), 10318-10325). In contrast, negative modulators may be useful as therapeutic agents with potential clinical uses in the treatment of psychiatric disorders in which glutamatergic transmission is pathologically increased (e.g., treatment resistant depression).

Oxysterols are derived from cholesterol and have been shown to potently and selectively modulate NMDA receptor function. New and improved oxysterols are needed that modulate the NMDA receptor for the prevention and treatment of conditions associated with NMDA expression and function. Compounds, compositions, and methods described herein are directed toward this end.

SUMMARY OF THE INVENTION

Provided herein are substituted oxysterols useful for preventing and/or treating a broad range of disorders, including, but not limited to, NMDA-mediated disorders. These compounds are expected to show improved in vivo potency, pharmacokinetic (PK) properties, oral bioavailability, formulatability, stability, and/or safety as compared to other oxysterols. Further provided are pharmaceutical compositions comprising the compounds of the present invention, and methods of their use and treatment.

In one aspect, provided herein are compounds according to Formula (I):

or a pharmaceutically acceptable salt thereof, wherein: R1is hydrogen or C1-6alkyl; R2is C1-6alkyl, carbocyclyl, or heterocyclyl; R5is absent or hydrogen; andrepresents a single or double bond, wherein when oneis a double bond, the otheris a single bond and R5is absent.

In some embodiments,represents a single bond.

In some embodiments, the compound of Formula (I) is a compound of Formula (I-A) or Formula (I-B):

In some embodiments, the compound of Formula (I) is a compound of Formula (I-B-i) or Formula (I-B-ii):

In some embodiments, the compound of Formula (I-C) is a compound of Formula (I-C-i) or (I-C-ii):

In some embodiments, the compound of Formula (I-C-i) is a compound of Formula (I-C-i-a) or (I-C-i-b):

In some embodiments, the compound of Formula (I-C-ii) is a compound of Formula (I-C-ii-a) or (I-C-ii-b):

or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound of Formula (II) is a compound of Formula (II-A) or Formula (II-B):

In some embodiments, the compound of Formula (II) is a compound of Formula (II-C) or Formula (II-D):

In some embodiments, the compound of Formula (II) is a compound of Formula (II-E):

In some embodiments, the compound of Formula (II-E) is a compound of Formula (II-E-i) or Formula (II-E-ii):

In some embodiments, the compound of Formula (II-E-i) is a compound of Formula (II-E-i-a) or Formula (II-E-i-b):

In some embodiments, the compound of Formula (II-E-ii) is a compound of Formula (II-E-ii-a) or Formula (II-E-ii-b):

or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound of Formula (VII) is a compound of Formula (VII-A) or Formula (VII-B):

In some embodiments, the compound of Formula (VII) is a compound of Formula (VII-C) or Formula (VII-D):

In some embodiments, the compound of Formula (VII) is a compound of Formula (VII-E):

In some embodiments, the compound of Formula (VII-E) is a compound of Formula (VII-E-i) or Formula (VII-E-ii):

In some embodiments, the compound of Formula (VII-E) is a compound of Formula (VII-E-i-a) or Formula (VII-E-i-b):

In some embodiments, the compound of Formula (VII-E-ii) is a compound of Formula (VII-E-ii-a) or Formula (VII-E-ii-b):

or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound of Formula (III) is a compound of Formula (III-A) or Formula (III-B):

or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound of Formula (IV) is a compound of Formula (IV-A) or Formula (IV-B):

or a pharmaceutically acceptable salt thereof.

or a pharmaceutically acceptable salt thereof, wherein X is —CH2—, —O—, —S—, or —NRA—, and m is an integer selected from 0, 1, 2, 3, 4, or 5; wherein RAis hydrogen, alkyl, —C(O)RC, —C(O)N(RC)2, or —SO2N(RC)2; and each RCis independently hydrogen, alkyl, aryl, or heteroaryl. In some embodiments, X is —CH2—, —O—, —S—, or —NH—.

In some embodiments, the compound of Formula (V) is a compound of Formula (V-A-i) or Formula (V-A-ii):

In some embodiments, the compound of Formula (V) is a compound of Formula (V-B):

In some embodiments, X is —CH2—.

In some embodiments, X is —O—.

In some embodiments, m is 0, 1, 2, or 3.

In some embodiments, the compound of Formula (V) is a compound of Formula (V-B-i):

or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound of Formula (V) is a compound of Formula (V-C):

or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound of Formula (I) is a compound of Formula (VI-A) or Formula (VI-B):

or a pharmaceutically acceptable salt thereof.

or a pharmaceutically acceptable salt thereof.

In an aspect, provided herein is a pharmaceutical composition comprising a compound described herein (e.g., a compound of Formula (I), or pharmaceutically acceptable salt thereof), and a pharmaceutically acceptable carrier.

In an aspect, provided herein is a method of inducing sedation or anesthesia comprising administering to a subject an effective amount of a compound described herein (e.g., a compound of Formula (I), or pharmaceutically acceptable salt thereof), or pharmaceutical composition thereof.

In an aspect, provided herein is a method for treating or preventing a disorder described herein, comprising administering to a subject in need thereof an effective amount of a compound described herein (e.g., a compound of Formula (I), or pharmaceutically acceptable salt thereof), or pharmaceutical composition thereof.

In some embodiments, the disorder is inflammatory bowel disease.

In some embodiments, the disorder is cancer, diabetes, or a sterol synthesis disorder.

In an aspect, provided herein is a method for treating or preventing a CNS-related condition comprising administering to a subject in need thereof an effective amount of a compound described herein (e.g., a compound of Formula (I), or pharmaceutically acceptable salt thereof), or pharmaceutical composition thereof. In some embodiments, the CNS-related condition is an adjustment disorder, anxiety disorder (including obsessive-compulsive disorder, posttraumatic stress disorder, and social phobia), cognitive disorder (including Alzheimer's disease and other forms of dementia), dissociative disorder, eating disorder, mood disorder (including depression (e.g., postpartum depression), bipolar disorder, dysthymic disorder, suicidality), schizophrenia or other psychotic disorder (including schizoaffective disorder), sleep disorder (including insomnia), substance-related disorder, personality disorder (including obsessive-compulsive personality disorder), autism spectrum disorders (including those involving mutations to the Shank group of proteins (e.g., Shank3)), neurodevelopmental disorder (including Rett syndrome, Tuberous Sclerosis complex), multiple sclerosis, sterol synthesis disorders, pain (including acute and chronic pain), encephalopathy secondary to a medical condition (including hepatic encephalopathy and anti-NMDA receptor encephalitis), seizure disorder (including status epilepticus and monogenic forms of epilepsy such as Dravet's disease), stroke, traumatic brain injury, movement disorder (including Huntington's disease and Parkinson's disease), vision impairment, hearing loss, and tinnitus.

In some embodiments, the disorder is sterol synthesis disorder.

Other objects and advantages will become apparent to those skilled in the art from a consideration of the ensuing Detailed Description, Examples, and Claims.

DEFINITIONS

Chemical Definitions

Compound described herein may also comprise one or more isotopic substitutions. For example, H may be in any isotopic form, including1H,2H (D or deuterium), and3H (T or tritium); C may be in any isotopic form, including12C,13C, and14C; O may be in any isotopic form, including16O and18O; and the like.

The following terms are intended to have the meanings presented therewith below and are useful in understanding the description and intended scope of the present invention. When describing the invention, which may include compounds, pharmaceutical compositions containing such compounds and methods of using such compounds and compositions, the following terms, if present, have the following meanings unless otherwise indicated. It should also be understood that when described herein any of the moieties defined forth below may be substituted with a variety of substituents, and that the respective definitions are intended to include such substituted moieties within their scope as set out below. Unless otherwise stated, the term “substituted” is to be defined as set out below. It should be further understood that the terms “groups” and “radicals” can be considered interchangeable when used herein. The articles “a” and “an” may be used herein to refer to one or to more than one (i.e. at least one) of the grammatical objects of the article. By way of example “an analogue” means one analogue or more than one analogue.

“Alkyl” refers to a radical of a straight-chain or branched saturated hydrocarbon group having from 1 to 20 carbon atoms (“C1-20alkyl”). In some embodiments, an alkyl group has 1 to 12 carbon atoms (“C1-12alkyl”). In some embodiments, an alkyl group has 1 to 10 carbon atoms (“C1-10alkyl”). In some embodiments, an alkyl group has 1 to 9 carbon atoms (“C1-9alkyl”). In some embodiments, an alkyl group has 1 to 8 carbon atoms (“C1-8alkyl”). In some embodiments, an alkyl group has 1 to 7 carbon atoms (“C1-7alkyl”). In some embodiments, an alkyl group has 1 to 6 carbon atoms (“C1-6alkyl”, also referred to herein as “lower alkyl”). In some embodiments, an alkyl group has 1 to 5 carbon atoms (“C1-5alkyl”). In some embodiments, an alkyl group has 1 to 4 carbon atoms (“C1-4alkyl”). In some embodiments, an alkyl group has 1 to 3 carbon atoms (“C1-3alkyl”). In some embodiments, an alkyl group has 1 to 2 carbon atoms (“C1-2alkyl”). In some embodiments, an alkyl group has 1 carbon atom (“C1alkyl”). In some embodiments, an alkyl group has 2 to 6 carbon atoms (“C2-6alkyl”). Examples of C1-6alkyl groups include methyl (C1), ethyl (C2), n-propyl (C3), isopropyl (C3), n-butyl (C4), tert-butyl (C4), sec-butyl (C4), iso-butyl (C4), n-pentyl (C5), 3-pentanyl (C5), amyl (C5), neopentyl (C5), 3-methyl-2-butanyl (C5), tertiary amyl (C5), and n-hexyl (C6). Additional examples of alkyl groups include n-heptyl (C7), n-octyl (C8) and the like. Unless otherwise specified, each instance of an alkyl group is independently optionally substituted, i.e., unsubstituted (an “unsubstituted alkyl”) or substituted (a “substituted alkyl”) with one or more substituents; e.g., for instance from 1 to 5 substituents, 1 to 3 substituents, or 1 substituent. In certain embodiments, the alkyl group is unsubstituted C1-10alkyl (e.g., —CH3). In certain embodiments, the alkyl group is substituted C1-10alkyl. Common alkyl abbreviations include Me (—CH3), Et (—CH2CH3), iPr (—CH(CH3)2), nPr (—CH2CH2CH3), n-Bu (—CH2CH2CH2CH3), or i-Bu (—CH2CH(CH3)2).

As used herein, “alkylene,” “alkenylene,” and “alkynylene,” refer to a divalent radical of an alkyl, alkenyl, and alkynyl group, respectively. When a range or number of carbons is provided for a particular “alkylene,” “alkenylene,” and “alkynylene” group, it is understood that the range or number refers to the range or number of carbons in the linear carbon divalent chain. “Alkylene,” “alkenylene,” and “alkynylene” groups may be substituted or unsubstituted with one or more substituents as described herein.

“Alkylene” refers to an alkyl group wherein two hydrogens are removed to provide a divalent radical, and which may be substituted or unsubstituted. Unsubstituted alkylene groups include, but are not limited to, methylene (—CH2—), ethylene (—CH2CH2—), propylene (—CH2CH2CH2—), butylene (—CH2CH2CH2CH2—), pentylene (—CH2CH2CH2CH2CH2—), hexylene (—CH2CH2CH2CH2CH2CH2—), and the like. Exemplary substituted alkylene groups, e.g., substituted with one or more alkyl (methyl) groups, include but are not limited to, substituted methylene (—CH(CH3)—, (—C(CH3)2—), substituted ethylene (—CH(CH3)CH2—, —CH2CH(CH3)—, —C(CH3)2CH2—, —CH2C(CH3)2—), substituted propylene (—CH(CH3)CH2CH2—, —CH2CH(CH3)CH2—, —CH2CH2CH(CH3)—, —C(CH3)2CH2CH2—, —CH2C(CH3)2CH2—, —CH2CH2C(CH3)2—), and the like.

“Alkenyl” refers to a radical of a straight-chain or branched hydrocarbon group having from 2 to 20 carbon atoms, one or more carbon-carbon double bonds (e.g., 1, 2, 3, or 4 carbon-carbon double bonds), and optionally one or more carbon-carbon triple bonds (e.g., 1, 2, 3, or 4 carbon-carbon triple bonds) (“C2-20alkenyl”). In certain embodiments, alkenyl does not contain any triple bonds. In some embodiments, an alkenyl group has 2 to 10 carbon atoms (“C2-10alkenyl”). In some embodiments, an alkenyl group has 2 to 9 carbon atoms (“C2-9alkenyl”). In some embodiments, an alkenyl group has 2 to 8 carbon atoms (“C2-8alkenyl”). In some embodiments, an alkenyl group has 2 to 7 carbon atoms (“C2-7alkenyl”). In some embodiments, an alkenyl group has 2 to 6 carbon atoms (“C2-6alkenyl”). In some embodiments, an alkenyl group has 2 to 5 carbon atoms (“C2-5alkenyl”). In some embodiments, an alkenyl group has 2 to 4 carbon atoms (“C2-4alkenyl”). In some embodiments, an alkenyl group has 2 to 3 carbon atoms (“C2-3alkenyl”). In some embodiments, an alkenyl group has 2 carbon atoms (“C2alkenyl”). The one or more carbon-carbon double bonds can be internal (such as in 2-butenyl) or terminal (such as in 1-butenyl). Examples of C2-4alkenyl groups include ethenyl (C2), 1-propenyl (C3), 2-propenyl (C3), 1-butenyl (C4), 2-butenyl (C4), butadienyl (C4), and the like. Examples of C2-6alkenyl groups include the aforementioned C2-4alkenyl groups as well as pentenyl (C5), pentadienyl (C5), hexenyl (C6), and the like. Additional examples of alkenyl include heptenyl (C7), octenyl (C8), octatrienyl (C8), and the like. Unless otherwise specified, each instance of an alkenyl group is independently optionally substituted, i.e., unsubstituted (an “unsubstituted alkenyl”) or substituted (a “substituted alkenyl”) with one or more substituents e.g., for instance from 1 to 5 substituents, 1 to 3 substituents, or 1 substituent. In certain embodiments, the alkenyl group is unsubstituted C2-10alkenyl. In certain embodiments, the alkenyl group is substituted C2-10alkenyl.

“Alkenylene” refers to an alkenyl group wherein two hydrogens are removed to provide a divalent radical, and which may be substituted or unsubstituted. Exemplary unsubstituted divalent alkenylene groups include, but are not limited to, ethenylene (—CH═CH—) and propenylene (e.g., —CH═CHCH2—, —CH2—CH═CH—). Exemplary substituted alkenylene groups, e.g., substituted with one or more alkyl (methyl) groups, include but are not limited to, substituted ethylene (—C(CH3)═CH—, —CH═C(CH3)—), substituted propylene (e.g., —C(CH3)═CHCH2—, —CH═C(CH3)CH2—, —CH═CHCH(CH3)—, —CH═CHC(CH3)2—, —CH(CH3)—CH═CH—, —C(CH3)2—CH═CH—, —CH2—C(CH3)═CH—, —CH2—CH═C(CH3)—), and the like.

“Alkynylene” refers to a linear alkynyl group wherein two hydrogens are removed to provide a divalent radical, and which may be substituted or unsubstituted. Exemplary divalent alkynylene groups include, but are not limited to, substituted or unsubstituted ethynylene, substituted or unsubstituted propynylene, and the like.

The term “heteroalkyl,” as used herein, refers to an alkyl group, as defined herein, which further comprises 1 or more (e.g., 1, 2, 3, or 4) heteroatoms (e.g., oxygen, sulfur, nitrogen, boron, silicon, phosphorus) within the parent chain, wherein the one or more heteroatoms is inserted between adjacent carbon atoms within the parent carbon chain and/or one or more heteroatoms is inserted between a carbon atom and the parent molecule, i.e., between the point of attachment. In certain embodiments, a heteroalkyl group refers to a saturated group having from 1 to 10 carbon atoms and 1, 2, 3, or 4 heteroatoms (“heteroC1-10alkyl”). In some embodiments, a heteroalkyl group is a saturated group having 1 to 9 carbon atoms and 1, 2, 3, or 4 heteroatoms (“heteroC1-9alkyl”). In some embodiments, a heteroalkyl group is a saturated group having 1 to 8 carbon atoms and 1, 2, 3, or 4 heteroatoms (“heteroC1-8alkyl”). In some embodiments, a heteroalkyl group is a saturated group having 1 to 7 carbon atoms and 1, 2, 3, or 4 heteroatoms (“heteroC1-7alkyl”). In some embodiments, a heteroalkyl group is a group having 1 to 6 carbon atoms and 1, 2, or 3 heteroatoms (“heteroC1-6alkyl”). In some embodiments, a heteroalkyl group is a saturated group having 1 to 5 carbon atoms and 1 or 2 heteroatoms (“heteroC1-5alkyl”). In some embodiments, a heteroalkyl group is a saturated group having 1 to 4 carbon atoms and 1 or 2 heteroatoms (“heteroC1-4alkyl”). In some embodiments, a heteroalkyl group is a saturated group having 1 to 3 carbon atoms and 1 heteroatom (“heteroC1-3alkyl”). In some embodiments, a heteroalkyl group is a saturated group having 1 to 2 carbon atoms and 1 heteroatom (“heteroC1-2alkyl”). In some embodiments, a heteroalkyl group is a saturated group having 1 carbon atom and 1 heteroatom (“heteroC1alkyl”). In some embodiments, a heteroalkyl group is a saturated group having 2 to 6 carbon atoms and 1 or 2 heteroatoms (“heteroC2-6alkyl”). Unless otherwise specified, each instance of a heteroalkyl group is independently unsubstituted (an “unsubstituted heteroalkyl”) or substituted (a “substituted heteroalkyl”) with one or more substituents. In certain embodiments, the heteroalkyl group is an unsubstituted heteroC1-10alkyl. In certain embodiments, the heteroalkyl group is a substituted heteroC1-10alkyl.

The term “heteroalkenyl,” as used herein, refers to an alkenyl group, as defined herein, which further comprises one or more (e.g., 1, 2, 3, or 4) heteroatoms (e.g., oxygen, sulfur, nitrogen, boron, silicon, phosphorus) wherein the one or more heteroatoms is inserted between adjacent carbon atoms within the parent carbon chain and/or one or more heteroatoms is inserted between a carbon atom and the parent molecule, i.e., between the point of attachment. In certain embodiments, a heteroalkenyl group refers to a group having from 2 to 10 carbon atoms, at least one double bond, and 1, 2, 3, or 4 heteroatoms (“heteroC2-10alkenyl”). In some embodiments, a heteroalkenyl group has 2 to 9 carbon atoms at least one double bond, and 1, 2, 3, or 4 heteroatoms (“heteroC2-9alkenyl”). In some embodiments, a heteroalkenyl group has 2 to 8 carbon atoms, at least one double bond, and 1, 2, 3, or 4 heteroatoms (“heteroC2-8alkenyl”). In some embodiments, a heteroalkenyl group has 2 to 7 carbon atoms, at least one double bond, and 1, 2, 3, or 4 heteroatoms (“heteroC2-7alkenyl”). In some embodiments, a heteroalkenyl group has 2 to 6 carbon atoms, at least one double bond, and 1, 2, or 3 heteroatoms (“heteroC2-6alkenyl”). In some embodiments, a heteroalkenyl group has 2 to 5 carbon atoms, at least one double bond, and 1 or 2 heteroatoms (“heteroC2-5alkenyl”). In some embodiments, a heteroalkenyl group has 2 to 4 carbon atoms, at least one double bond, and 1 or 2 heteroatoms (“heteroC2-4alkenyl”). In some embodiments, a heteroalkenyl group has 2 to 3 carbon atoms, at least one double bond, and 1 heteroatom (“heteroC2-3alkenyl”). In some embodiments, a heteroalkenyl group has 2 to 6 carbon atoms, at least one double bond, and 1 or 2 heteroatoms (“heteroC2-6alkenyl”). Unless otherwise specified, each instance of a heteroalkenyl group is independently unsubstituted (an “unsubstituted heteroalkenyl”) or substituted (a “substituted heteroalkenyl”) with one or more substituents. In certain embodiments, the heteroalkenyl group is an unsubstituted heteroC2-10alkenyl. In certain embodiments, the heteroalkenyl group is a substituted heteroC2-10alkenyl.

The term “heteroalkynyl,” as used herein, refers to an alkynyl group, as defined herein, which further comprises one or more (e.g., 1, 2, 3, or 4) heteroatoms (e.g., oxygen, sulfur, nitrogen, boron, silicon, phosphorus) wherein the one or more heteroatoms is inserted between adjacent carbon atoms within the parent carbon chain and/or one or more heteroatoms is inserted between a carbon atom and the parent molecule, i.e., between the point of attachment. In certain embodiments, a heteroalkynyl group refers to a group having from 2 to 10 carbon atoms, at least one triple bond, and 1, 2, 3, or 4 heteroatoms (“heteroC2-10alkynyl”). In some embodiments, a heteroalkynyl group has 2 to 9 carbon atoms, at least one triple bond, and 1, 2, 3, or 4 heteroatoms (“heteroC2-9alkynyl”). In some embodiments, a heteroalkynyl group has 2 to 8 carbon atoms, at least one triple bond, and 1, 2, 3, or 4 heteroatoms (“heteroC2-8alkynyl”). In some embodiments, a heteroalkynyl group has 2 to 7 carbon atoms, at least one triple bond, and 1, 2, 3, or 4 heteroatoms (“heteroC2-7alkynyl”). In some embodiments, a heteroalkynyl group has 2 to 6 carbon atoms, at least one triple bond, and 1, 2, or 3 heteroatoms (“heteroC2-6alkynyl”). In some embodiments, a heteroalkynyl group has 2 to 5 carbon atoms, at least one triple bond, and 1 or 2 heteroatoms (“heteroC2-5alkynyl”). In some embodiments, a heteroalkynyl group has 2 to 4 carbon atoms, at least one triple bond, and 1 or 2 heteroatoms (“heteroC2-4alkynyl”). In some embodiments, a heteroalkynyl group has 2 to 3 carbon atoms, at least one triple bond, and 1 heteroatom (“heteroC2-3alkynyl”). In some embodiments, a heteroalkynyl group has 2 to 6 carbon atoms, at least one triple bond, and 1 or 2 heteroatoms (“heteroC2-6alkynyl”). Unless otherwise specified, each instance of a heteroalkynyl group is independently unsubstituted (an “unsubstituted heteroalkynyl”) or substituted (a “substituted heteroalkynyl”) with one or more substituents. In certain embodiments, the heteroalkynyl group is an unsubstituted heteroC2-10alkynyl. In certain embodiments, the heteroalkynyl group is a substituted heteroC2-10alkynyl.

As used herein, “alkylene,” “alkenylene,” “alkynylene,” “heteroalkylene,” “heteroalkenylene,” and “heteroalkynylene,” refer to a divalent radical of an alkyl, alkenyl, alkynyl group, heteroalkyl, heteroalkenyl, and heteroalkynyl group respectively. When a range or number of carbons is provided for a particular “alkylene,” “alkenylene,” “alkynylene,” “heteroalkylene,” “heteroalkenylene,” or “heteroalkynylene,” group, it is understood that the range or number refers to the range or number of carbons in the linear carbon divalent chain. “Alkylene,” “alkenylene,” “alkynylene,” “heteroalkylene,” “heteroalkenylene,” and “heteroalkynylene” groups may be substituted or unsubstituted with one or more substituents as described herein.

“Aryl” refers to a radical of a monocyclic or polycyclic (e.g., bicyclic or tricyclic) 4n+2 aromatic ring system (e.g., having 6, 10, or 14 π electrons shared in a cyclic array) having 6-14 ring carbon atoms and zero heteroatoms provided in the aromatic ring system (“C6-14aryl”). In some embodiments, an aryl group has six ring carbon atoms (“C6aryl”; e.g., phenyl). In some embodiments, an aryl group has ten ring carbon atoms (“C10aryl”; e.g., naphthyl such as 1-naphthyl and 2-naphthyl). In some embodiments, an aryl group has fourteen ring carbon atoms (“C14aryl”; e.g., anthracyl). “Aryl” also includes ring systems wherein the aryl ring, as defined above, is fused with one or more carbocyclyl or heterocyclyl groups wherein the radical or point of attachment is on the aryl ring, and in such instances, the number of carbon atoms continue to designate the number of carbon atoms in the aryl ring system. Typical aryl groups include, but are not limited to, groups derived from aceanthrylene, acenaphthylene, acephenanthrylene, anthracene, azulene, benzene, chrysene, coronene, fluoranthene, fluorene, hexacene, hexaphene, hexalene, as-indacene, s-indacene, indane, indene, naphthalene, octacene, octaphene, octalene, ovalene, penta-2,4-diene, pentacene, pentalene, pentaphene, perylene, phenalene, phenanthrene, picene, pleiadene, pyrene, pyranthrene, rubicene, triphenylene, and trinaphthalene. Particularly aryl groups include phenyl, naphthyl, indenyl, and tetrahydronaphthyl. Unless otherwise specified, each instance of an aryl group is independently optionally substituted, i.e., unsubstituted (an “unsubstituted aryl”) or substituted (a “substituted aryl”) with one or more substituents. In certain embodiments, the aryl group is unsubstituted C6-14aryl. In certain embodiments, the aryl group is substituted C6-14aryl.

In certain embodiments, an aryl group substituted with one or more of groups selected from halo, C1-C8alkyl, C1-C8haloalkyl, cyano, hydroxy, C1-C8alkoxy, and amino.

Examples of representative substituted aryls include the following

“Fused aryl” refers to an aryl having two of its ring carbon in common with a second aryl or heteroaryl ring or with a carbocyclyl or heterocyclyl ring.

“Aralkyl” is a subset of alkyl and aryl, as defined herein, and refers to an optionally substituted alkyl group substituted by an optionally substituted aryl group.

Examples of representative heteroaryls include the following:

“Heteroaralkyl” is a subset of alkyl and heteroaryl, as defined herein, and refers to an optionally substituted alkyl group substituted by an optionally substituted heteroaryl group.

“Heterocyclyl” or “heterocyclic” refers to a radical of a 3- to 10-membered non-aromatic ring system having ring carbon atoms and 1 to 4 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, sulfur, boron, phosphorus, and silicon (“3-10 membered heterocyclyl”). In heterocyclyl groups that contain one or more nitrogen atoms, the point of attachment can be a carbon or nitrogen atom, as valency permits. A heterocyclyl group can either be monocyclic (“monocyclic heterocyclyl”) or a fused, bridged or spiro ring system such as a bicyclic system (“bicyclic heterocyclyl”), and can be saturated or can be partially unsaturated. Heterocyclyl bicyclic ring systems can include one or more heteroatoms in one or both rings. “Heterocyclyl” also includes ring systems wherein the heterocyclyl ring, as defined above, is fused with one or more carbocyclyl groups wherein the point of attachment is either on the carbocyclyl or heterocyclyl ring, or ring systems wherein the heterocyclyl ring, as defined above, is fused with one or more aryl or heteroaryl groups, wherein the point of attachment is on the heterocyclyl ring, and in such instances, the number of ring members continue to designate the number of ring members in the heterocyclyl ring system. Unless otherwise specified, each instance of heterocyclyl is independently optionally substituted, i.e., unsubstituted (an “unsubstituted heterocyclyl”) or substituted (a “substituted heterocyclyl”) with one or more substituents. In certain embodiments, the heterocyclyl group is unsubstituted 3-10 membered heterocyclyl. In certain embodiments, the heterocyclyl group is substituted 3-10 membered heterocyclyl.

“Hetero” when used to describe a compound or a group present on a compound means that one or more carbon atoms in the compound or group have been replaced by a nitrogen, oxygen, or sulfur heteroatom. Hetero may be applied to any of the hydrocarbyl groups described above such as alkyl, e.g., heteroalkyl, cycloalkyl, e.g., heterocyclyl, aryl, e.g., heteroaryl, cycloalkenyl, e.g., cycloheteroalkenyl, and the like having from 1 to 5, and particularly from 1 to 3 heteroatoms.

“Amino” refers to the radical —NH2.

“Substituted amino” refers to an amino group of the formula —N(R38)2wherein R38is hydrogen, substituted or unsubstituted alkyl, substituted or unsubstitued alkenyl, substituted or unsubstitued alkynyl, substituted or unsubstitued carbocyclyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted aryl, substituted or unsubstitued heteroaryl, or an amino protecting group, wherein at least one of R38is not a hydrogen. In certain embodiments, each R38is independently selected from hydrogen, C1-C8alkyl, C3-C8alkenyl, C3-C8alkynyl, C6-C10aryl, 5-10 membered heteroaryl, 4-10 membered heterocyclyl, or C3-C10cycloalkyl; or C1-C8alkyl, substituted with halo or hydroxy; C3-C8alkenyl, substituted with halo or hydroxy; C3-C8alkynyl, substituted with halo or hydroxy, or —(CH2)t(C6-C10aryl), —(CH2)t(5-10 membered heteroaryl), —(CH2)t(C3-C10cycloalkyl), or —(CH2)t(4-10 membered heterocyclyl), wherein t is an integer between 0 and 8, each of which is substituted by unsubstituted C1-C4alkyl, halo, unsubstituted C1-C4alkoxy, unsubstituted C1-C4haloalkyl, unsubstituted C1-C4hydroxyalkyl, or unsubstituted C1-C4haloalkoxy or hydroxy; or both R38groups are joined to form an alkylene group.

Exemplary “substituted amino” groups include, but are not limited to, —NR39—C1-C8alkyl, —NR39—(CH2)t(C6-C10aryl), —NR39—(CH2)t(5-10 membered heteroaryl), —NR39—(CH2)t(C3-C10cycloalkyl), and —NR39—(CH2)t(4-10 membered heterocyclyl), wherein t is an integer from 0 to 4, for instance 1 or 2, each R39independently represents H or C1-C8alkyl; and any alkyl groups present, may themselves be substituted by halo, substituted or unsubstituted amino, or hydroxy; and any aryl, heteroaryl, cycloalkyl, or heterocyclyl groups present, may themselves be substituted by unsubstituted C1-C4alkyl, halo, unsubstituted C1-C4alkoxy, unsubstituted C1-C4haloalkyl, unsubstituted C1-C4hydroxyalkyl, or unsubstituted C1-C4haloalkoxy or hydroxy. For the avoidance of doubt the term ‘substituted amino’ includes the groups alkylamino, substituted alkylamino, alkylarylamino, substituted alkylarylamino, arylamino, substituted arylamino, dialkylamino, and substituted dialkylamino as defined below. Substituted amino encompasses both monosubstituted amino and disubstituted amino groups.

“Carboxy” refers to the radical —C(O)OH.

“Cyano” refers to the radical —CN.

“Halo” or “halogen” refers to fluoro (F), chloro (Cl), bromo (Br), and iodo (I). In certain embodiments, the halo group is either fluoro or chloro.

“Hydroxy” refers to the radical —OH.

“Nitro” refers to the radical —NO2.

“Cycloalkylalkyl” refers to an alkyl radical in which the alkyl group is substituted with a cycloalkyl group. Typical cycloalkylalkyl groups include, but are not limited to, cyclopropylmethyl, cyclobutylmethyl, cyclopentylmethyl, cyclohexylmethyl, cycloheptylmethyl, cyclooctylmethyl, cyclopropylethyl, cyclobutylethyl, cyclopentylethyl, cyclohexylethyl, cycloheptylethyl, and cyclooctylethyl, and the like.

“Heterocyclylalkyl” refers to an alkyl radical in which the alkyl group is substituted with a heterocyclyl group. Typical heterocyclylalkyl groups include, but are not limited to, pyrrolidinylmethyl, piperidinylmethyl, piperazinylmethyl, morpholinylmethyl, pyrrolidinylethyl, piperidinylethyl, piperazinylethyl, morpholinylethyl, and the like.

“Thioketo” refers to the group ═S.

or two geminal hydrogens on a carbon atom are replaced with the group ═O, ═S, ═NN(Rbb)2, ═NNRbbC(═O)Raa, ═NNRbbC(═O)ORaa, ═NNRbbS(═O)2Raa, ═NRbb, or ═NORcc;

These and other exemplary substituents are described in more detail in the Detailed Description, Examples, and claims. The invention is not intended to be limited in any manner by the above exemplary listing of substituents.

Other Definitions

A “subject” to which administration is contemplated includes, but is not limited to, humans (i.e., a male or female of any age group, e.g., a pediatric subject (e.g, infant, child, adolescent) or adult subject (e.g., young adult, middle-aged adult or senior adult)) and/or a non-human animal, e.g., a mammal such as primates (e.g., cynomolgus monkeys, rhesus monkeys), cattle, pigs, horses, sheep, goats, rodents, cats, and/or dogs. In certain embodiments, the subject is a human. In certain embodiments, the subject is a non-human animal. The terms “human,” “patient,” and “subject” are used interchangeably herein.

Disease, disorder, and condition are used interchangeably herein.

As used herein, and unless otherwise specified, the terms “treat,” “treating” and “treatment” contemplate an action that occurs while a subject is suffering from the specified disease, disorder or condition, which reduces the severity of the disease, disorder or condition, or retards or slows the progression of the disease, disorder or condition (“therapeutic treatment”), and also contemplates an action that occurs before a subject begins to suffer from the specified disease, disorder or condition (“prophylactic treatment”).

In general, the “effective amount” of a compound refers to an amount sufficient to elicit the desired biological response. As will be appreciated by those of ordinary skill in this art, the effective amount of a compound of the invention may vary depending on such factors as the desired biological endpoint, the pharmacokinetics of the compound, the disease being treated, the mode of administration, and the age, health, and condition of the subject. An effective amount encompasses therapeutic and prophylactic treatment.

As used herein, and unless otherwise specified, a “therapeutically effective amount” of a compound is an amount sufficient to provide a therapeutic benefit in the treatment of a disease, disorder or condition, or to delay or minimize one or more symptoms associated with the disease, disorder or condition. A therapeutically effective amount of a compound means an amount of therapeutic agent, alone or in combination with other therapies, which provides a therapeutic benefit in the treatment of the disease, disorder or condition. The term “therapeutically effective amount” can encompass an amount that improves overall therapy, reduces or avoids symptoms or causes of disease or condition, or enhances the therapeutic efficacy of another therapeutic agent.

As generally described herein, the present invention provides oxysterols useful for preventing and/or treating a broad range of disorders, including, but not limited to, NMDA-mediated disorders. These compounds are expected to show improved in vivo potency, pharmacokinetic (PK) properties, oral bioavailability, formulatability, stability, and/or safety as compared to other oxysterols.

Compounds

In one aspect, the present invention features a compound of Formula (I):

or a pharmaceutically acceptable salt thereof, wherein: R1is hydrogen or C1-6alkyl; R2is C1-6alkyl, carbocyclyl, or heterocyclyl; R5is absent or hydrogen; andrepresents a single or double bond, wherein when oneis a double bond, the otheris a single bond and R5is absent.

In some embodiments, R1is hydrogen. In some embodiments, the compound of Formula (I) is a compound of Formula (X):

In some embodiments,represents a single bond.

In some embodiments, the compound of Formula (I) is a compound of Formula (I-A) or Formula (I-B):

In some embodiments, the compound of Formula (I) is a compound of Formula (I-B-i) or Formula (I-B-ii):

In some embodiments, the compound of Formula (I-C) is a compound of Formula (I-C-i) or (I-C-ii):

In some embodiments, the compound of Formula (I-C-i) is a compound of Formula (I-C-i-a) or (I-C-i-b):

In some embodiments, the compound of Formula (I-C-ii) is a compound of Formula (I-C-ii-a) or (I-C-ii-b):

In some embodiments, R2is C1-6alkyl. In some embodiments, R2is substituted C1-6alkyl. In some embodiments, the compound of Formula (I) is a compound of Formula (II):

or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound of Formula (II) is a compound of Formula (II-A) or Formula (II-B):

In some embodiments, the compound of Formula (II) is a compound of Formula (II-C) or Formula (II-D):

In some embodiments, the compound of Formula (II) is a compound of Formula (II-E):

In some embodiments, the compound of Formula (II-E) is a compound of Formula (II-E-i) or Formula (II-E-ii):

In some embodiments, the compound of Formula (II-E-i) is a compound of Formula (II-E-i-a) or Formula (II-E-i-b):

In some embodiments, the compound of Formula (II-E-ii) is a compound of Formula (II-E-ii-a) or Formula (II-E-ii-b):

or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound of Formula (VII) is a compound of Formula (VII-A) or Formula (VII-B):

In some embodiments, the compound of Formula (VII) is a compound of Formula (VII-C) or Formula (VII-D):

In some embodiments, the compound of Formula (VII) is a compound of Formula (VII-E):

In some embodiments, the compound of Formula (VII-E) is a compound of Formula (VII-E-i) or Formula (VII-E-ii):

In some embodiments, the compound of Formula (VII-E-i) is a compound of Formula (VII-E-i-a) or Formula (VII-E-i-b):

In some embodiments, the compound of Formula (VII-E-ii) is a compound of Formula (VII-E-ii-a) or Formula (VII-E-ii-b):

or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound of Formula (III) is a compound of Formula (III-A) or Formula (III-B):

or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound of Formula (IV) is a compound of Formula (IV-A) or Formula (IV-B):

or a pharmaceutically acceptable salt thereof.

or a pharmaceutically acceptable salt thereof, wherein X is —CH2—, —O—, —S—, or —NRA—, and m is an integer selected from 0, 1, 2, 3, 4, or 5; wherein RAis hydrogen, alkyl, —C(O)RC, —C(O)N(RC)2, or —SO2N(RC)2; and each RCis independently hydrogen, alkyl, aryl, or heteroaryl. In some embodiments, X is —CH2—, —O—, —S—, or —NH—.

In some embodiments, the compound of Formula (V) is a compound of Formula (V-A-i) or Formula (V-A-ii):

In some embodiments, the compound of Formula (V) is a compound of Formula (V-B):

In some embodiments, X is —CH2—.

In some embodiments, X is —O—.

In some embodiments, m is 0, 1, 2, or 3.

In some embodiments, the compound of Formula (V) is a compound of Formula (V-B-i):

or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound of Formula (V) is a compound of Formula (V-C):

or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound of Formula (VI) is a compound of Formula (VI-A) or Formula (VI-B):

or a pharmaceutically acceptable salt thereof.

or a pharmaceutically acceptable salt thereof.
Pharmaceutical Compositions

In another aspect, the invention provides a pharmaceutical composition comprising a pharmaceutically acceptable carrier and a effective amount of a compound described herein (e.g., a compound of Formula (I).

When employed as pharmaceuticals, the compounds provided herein are typically administered in the form of a pharmaceutical composition. Such compositions can be prepared in a manner well known in the pharmaceutical art and comprise at least one active compound.

In one embodiment, with respect to the pharmaceutical composition, the carrier is a parenteral carrier, oral or topical carrier.

The present invention also relates to a compound described herein (e.g., a compound of Formula (I), or pharmaceutical composition thereof) for use as a pharmaceutical or a medicament.

Generally, the compounds provided herein are administered in a therapeutically effective amount. The amount of the compound actually administered will typically be determined by a physician, in the light of the relevant circumstances, including the condition to be treated, the chosen route of administration, the actual compound administered, the age, weight, and response of the individual patient, the severity of the patient's symptoms, and the like.

The pharmaceutical compositions provided herein can be administered by a variety of routes including oral, rectal, transdermal, subcutaneous, intravenous, intramuscular, and intranasal. Depending on the intended route of delivery, the compounds provided herein are preferably formulated as either injectable or oral compositions or as salves, as lotions or as patches all for transdermal administration.

Injectable compositions are typically based upon injectable sterile saline or phosphate-buffered saline or other injectable carriers known in the art. As before, the active compound in such compositions is typically a minor component, often being from about 0.05 to 10% by weight with the remainder being the injectable carrier and the like.

Transdermal compositions are typically formulated as a topical ointment or cream containing the active ingredient(s), generally in an amount ranging from about 0.01 to about 20% by weight, preferably from about 0.1 to about 20% by weight, preferably from about 0.1 to about 10% by weight, and more preferably from about 0.5 to about 15% by weight. When formulated as a ointment, the active ingredients will typically be combined with either a paraffinic or a water-miscible ointment base. Alternatively, the active ingredients may be formulated in a cream with, for example an oil-in-water cream base. Such transdermal formulations are well-known in the art and generally include additional ingredients to enhance the dermal penetration of stability of the active ingredients or the formulation. All such known transdermal formulations and ingredients are included within the scope provided herein.

The compounds provided herein can also be administered by a transdermal device. Accordingly, transdermal administration can be accomplished using a patch either of the reservoir or porous membrane type, or of a solid matrix variety.

The above-described components for orally administrable, injectable or topically administrable compositions are merely representative. Other materials as well as processing techniques and the like are set forth in Part 8 ofRemington's Pharmaceutical Sciences,17th edition, 1985, Mack Publishing Company, Easton, Pa., which is incorporated herein by reference.

The above-described components for orally administrable, injectable, or topically administrable compositions are merely representative. Other materials as well as processing techniques and the like are set forth in Part 8 ofRemington's The Science and Practice of Pharmacy,21st edition, 2005, Publisher: Lippincott Williams & Wilkins, which is incorporated herein by reference.

The compounds of this invention can also be administered in sustained release forms or from sustained release drug delivery systems. A description of representative sustained release materials can be found inRemington's Pharmaceutical Sciences.

The present invention also relates to the pharmaceutically acceptable formulations of a compound described herein (e.g., a compound of Formula (I)). In one embodiment, the formulation comprises water. In another embodiment, the formulation comprises a cyclodextrin derivative. The most common cyclodextrins are α-, β- and γ-cyclodextrins consisting of 6, 7 and 8 α-1,4-linked glucose units, respectively, optionally comprising one or more substituents on the linked sugar moieties, which include, but are not limited to, methylated, hydroxyalkylated, acylated, and sulfoalkylether substitution. In certain embodiments, the cyclodextrin is a sulfoalkyl ether β-cyclodextrin, e.g., for example, sulfobutyl ether β-cyclodextrin, also known as CAPTISOL (sulfobutyl ether β-cyclodextrin) See, e.g., U.S. Pat. No. 5,376,645. In certain embodiments, the formulation comprises hexapropyl-β-cyclodextrin. In a more particular embodiment, the formulation comprises hexapropyl-β-cyclodextrin (10-50% in water).

The present invention also relates to the pharmaceutically acceptable acid addition salt of a compound described herein (e.g., a compound of Formula (I)). The acid which may be used to prepare the pharmaceutically acceptable salt is that which forms a non-toxic acid addition salt, i.e., a salt containing pharmacologically acceptable anions such as the hydrochloride, hydroiodide, hydrobromide, nitrate, sulfate, bisulfate, phosphate, acetate, lactate, citrate, tartrate, succinate, maleate, fumarate, benzoate, para-toluenesulfonate, and the like.

The following formulation examples illustrate representative pharmaceutical compositions that may be prepared in accordance with this invention. The present invention, however, is not limited to the following pharmaceutical compositions.

A compound described herein (e.g., a compound of Formula (I), or pharmaceutically acceptable salt thereof), may be admixed as a dry powder with a dry gelatin binder in an approximate 1:2 weight ratio. A minor amount of magnesium stearate is added as a lubricant. The mixture is formed into 240-270 mg tablets (80-90 mg of active compound per tablet) in a tablet press.

A compound described herein (e.g., a compound of Formula (I), or pharmaceutically acceptable salt thereof), may be admixed as a dry powder with a starch diluent in an approximate 1:1 weight ratio. The mixture is filled into 250 mg capsules (125 mg of active compound per capsule).

A compound described herein (e.g., a compound of Formula (I), or pharmaceutically acceptable salt thereof), (125 mg) may be admixed with sucrose (1.75 g) and xanthan gum (4 mg) and the resultant mixture may be blended, passed through a No. 10 mesh U.S. sieve, and then mixed with a previously made solution of microcrystalline cellulose and sodium carboxymethyl cellulose (11:89, 50 mg) in water. Sodium benzoate (10 mg), flavor, and color are diluted with water and added with stirring. Sufficient water may then be added to produce a total volume of 5 mL.

A compound described herein (e.g., a compound of Formula (I) or pharmaceutically acceptable salt thereof), may be admixed as a dry powder with a dry gelatin binder in an approximate 1:2 weight ratio. A minor amount of magnesium stearate is added as a lubricant. The mixture is formed into 450-900 mg tablets (150-300 mg of active compound) in a tablet press.

A compound described herein (e.g., a compound of Formula (I), or pharmaceutically acceptable salt thereof), may be dissolved or suspended in a buffered sterile saline injectable aqueous medium to a concentration of approximately 5 mg/mL.

A compound described herein (e.g., a compound of Formula (I) or pharmaceutically acceptable salt thereof), may be admixed as a dry powder with a dry gelatin binder in an approximate 1:2 weight ratio. A minor amount of magnesium stearate is added as a lubricant. The mixture is formed into 90-150 mg tablets (30-50 mg of active compound per tablet) in a tablet press.

A compound described herein (e.g., a compound of Formula (I) or pharmaceutically acceptable salt thereof), may be may be admixed as a dry powder with a dry gelatin binder in an approximate 1:2 weight ratio. A minor amount of magnesium stearate is added as a lubricant. The mixture is formed into 30-90 mg tablets (10-30 mg of active compound per tablet) in a tablet press.

A compound described herein (e.g., a compound of Formula (I), or pharmaceutically acceptable salt thereof), may be admixed as a dry powder with a dry gelatin binder in an approximate 1:2 weight ratio. A minor amount of magnesium stearate is added as a lubricant. The mixture is formed into 0.3-30 mg tablets (0.1-10 mg of active compound per tablet) in a tablet press.

A compound described herein (e.g., a compound of Formula (I) or pharmaceutically acceptable salt thereof), may be admixed as a dry powder with a dry gelatin binder in an approximate 1:2 weight ratio. A minor amount of magnesium stearate is added as a lubricant. The mixture is formed into 150-240 mg tablets (50-80 mg of active compound per tablet) in a tablet press.

A compound described herein (e.g., a compound of Formula (I), or pharmaceutically acceptable salt thereof), may be admixed as a dry powder with a dry gelatin binder in an approximate 1:2 weight ratio. A minor amount of magnesium stearate is added as a lubricant. The mixture is formed into 270-450 mg tablets (90-150 mg of active compound per tablet) in a tablet press.

Injection dose levels range from about 0.1 mg/kg/hour to at least 10 mg/kg/hour, all for from about 1 to about 120 hours and especially 24 to 96 hours. A preloading bolus of from about 0.1 mg/kg to about 10 mg/kg or more may also be administered to achieve adequate steady state levels. The maximum total dose is not expected to exceed about 2 g/day for a 40 to 80 kg human patient.

For the prevention and/or treatment of long-term conditions the regimen for treatment usually stretches over many months or years so oral dosing is preferred for patient convenience and tolerance. With oral dosing, one to five and especially two to four and typically three oral doses per day are representative regimens. Using these dosing patterns, each dose provides from about 0.01 to about 20 mg/kg of the compound provided herein, with preferred doses each providing from about 0.1 to about 10 mg/kg, and especially about 1 to about 5 mg/kg.

Transdermal doses are generally selected to provide similar or lower blood levels than are achieved using injection doses.

When used to prevent the onset of a CNS-disorder, the compounds provided herein will be administered to a subject at risk for developing the condition, typically on the advice and under the supervision of a physician, at the dosage levels described above. Subjects at risk for developing a particular condition generally include those that have a family history of the condition, or those who have been identified by genetic testing or screening to be particularly susceptible to developing the condition.

Methods of Treatment and Use

Compounds of the present invention (e.g., a compound of Formula (I), and pharmaceutically acceptable salts thereof), as described herein, are generally designed to modulate NMDA function, and therefore to act as oxysterols for the treatment and prevention of, e.g., CNS-related conditions in a subject. In some embodiments, the compounds described herein (e.g., a compound of Formula (I), and pharmaceutically acceptable salts thereof), as described herein, are generally designed to penetrate the blood brain barrier (e.g., designed to be transported across the blood brain barrier). Modulation, as used herein, refers to, for example, the inhibition or potentiation of NMDA receptor function. In certain embodiments, the compound described herein (e.g., a compound of Formula (I), and pharmaceutically acceptable salts thereof), may act as a negative allosteric modulator (NAM) of NMDA, and inhibit NMDA receptor function. In certain embodiments, the present invention, e.g., a compound of Formula (I), or pharmaceutically acceptable salt thereof, may act as positive allosteric modulators (PAM) of NMDA, and potentiate NMDA receptor function. In certain embodiments, the compound described herein (e.g., a compound of Formula (I), and pharmaceutically acceptable salts thereof), modulates NMDA function, but does not act as a negative allosteric modulator (NAM) or positive allosteric modulator (PAM) of NMDA.

In some embodiments, the disorder is cancer. In some embodiments, the disorder is diabetes. In some embodiments, the disorder is a sterol synthesis disorder. In some embodiments, the disorder is a gastrointestinal (GI) disorder, e.g., constipation, irritable bowel syndrome (IBS), inflammatory bowel disease (IBD) (e.g., ulcerative colitis, Crohn's disease), structural disorders affecting the GI, anal disorders (e.g., hemorrhoids, internal hemorrhoids, external hemorrhoids, anal fissures, perianal abscesses, anal fistula), colon polyps, cancer, colitis. In some embodiments, the disorder is inflammatory bowel disease.

In some embodiments, the disorder is Smith-Lemli-Opitz Syndrome (SLOS). In some embodiments, the disorder is desmosterolosis. In some embodiments, the disorder is sitosterolemia. In some embodiments, the disorder is cerebrotendinous xanthomatosis (CTX). In some embodiments, the disorder is Mevalonate Kinase Deficiency (MKD). In some embodiments, the disorder is SC4MOL gene mutation (SMO Deficiency). In some embodiments, the disorder is Niemann-Pick disease. In some embodiments, the disorder is autism spectrum disorder (ASD). In some embodiments, the disorder is associated with phenylketomuria.

Exemplary CNS conditions related to NMDA-modulation include, but are not limited to, adjustment disorders, anxiety disorders (including obsessive-compulsive disorder, posttraumatic stress disorder, social phobia, generalized anxiety disorder), cognitive disorders (including Alzheimer's disease and other forms of dementia (e.g., frontotemporal dementia)), dissociative disorders, eating disorders, mood disorders (including depression (e.g., postpartum depression), bipolar disorder, dysthymic disorder, suicidality), schizophrenia or other psychotic disorders (including schizoaffective disorder), sleep disorders (including insomnia), substance abuse-related disorders, personality disorders (including obsessive-compulsive personality disorder), autism spectrum disorders (including those involving mutations to the Shank group of proteins (e.g., Shank3)), neurodevelopmental disorders (including Rett syndrome), multiple sclerosis, sterol synthesis disorders, pain (including acute and chronic pain; headaches, e.g., migraine headaches), seizure disorders (including status epilepticus and monogenic forms of epilepsy such as Dravet's disease, and Tuberous Sclerosis Complex (TSC)), stroke, traumatic brain injury, movement disorders (including Huntington's disease and Parkinson's disease) and tinnitus. In certain embodiments, the compound of the present invention, e.g., a compound of Formula (I), and pharmaceutically acceptable salts thereof, can be used to induce sedation or anesthesia. In certain embodiments, the compound described herein (e.g., a compound of Formula (I), and pharmaceutically acceptable salts thereof), is useful in the treatment or prevention of adjustment disorders, anxiety disorders, cognitive disorders, dissociative disorders, eating disorders, mood disorders, schizophrenia or other psychotic disorders, sleep disorders, substance-related disorders, personality disorders, autism spectrum disorders, neurodevelopmental disorders, sterol synthesis disorders, pain, seizure disorders, stroke, traumatic brain injury, movement disorders and vision impairment, hearing loss, and tinnitus. In some embodiments, the disorder is Huntington's disease. In some embodiments, the disorder is Parkinson's disease. In some embodiments, the disorder is an inflammatory disease (e.g., lupus).

In another aspect, provided is a method of treating or preventing brain excitability in a subject susceptible to or afflicted with a condition associated with brain excitability, comprising administering to the subject an effective amount of a compound of the present invention, e.g., a compound of Formula (I) or a pharmaceutically acceptable salt thereof.

In yet another aspect, the present invention provides a combination of a compound of the present invention, e.g., a compound of Formula (I), or pharmaceutically acceptable salt thereof, and another pharmacologically active agent. The compounds provided herein can be administered as the sole active agent or they can be administered in combination with other agents. Administration in combination can proceed by any technique apparent to those of skill in the art including, for example, separate, sequential, concurrent and alternating administration.

Diseases and Disorders

Described herein are methods of treating a sterol synthesis disorder. Exemplary disorders are described herein. The methods include administering to a subject, e.g., a subject suffering from a sterol synthesis disorder such as SLOS, a NMDA receptor modulating compound. Exemplary compounds are described herein.

Sterol Synthesis Disorders

In one aspect, described herein are methods for treating a sterol synthesis disorder. Cholesterol has an essential rule in growth and development. It is a membrane lipid and a precursor to many molecules that play important roles in cellular growth and differentiation, protein glycosylation, and signaling pathways. Biosynthesis of cholesterol involves a number of enzymes and intermediates. Disorders resulting from a deficiency in any of the enzymes involved in cholesterol biosynthesis lead to the accumulation of intermediates and imbalance in biomolecules, resulting in disorders including congenital skeletal malformations, dysmorphic facial features, psychomotor retardation, and failure to thrive. In an embodiment, a sterol synthesis disorder or symptom of a sterol synthesis disorder can be treated by administering to a subject suffering from a sterol synthesis disorder a compound described herein, such as a NMDA receptor modulating compound as described herein. Additional disorders are described below.

In one aspect, described herein are methods for treating Smith-Lemli-Opitz Syndrome (or SLOS, or 7-dehydrocholesterol reductase deficiency). SLOS is an inborn error of cholesterol synthesis. In addition to microcephaly, moderate to severe intellectual disability, sensory hypersensitivity, stereotyped behaviors, dysmorphic facial features, and syndactyly of the second/third toes, a feature of the disease is reduced cerebrosterol (24(S)-hydroxycholesterol) levels. SLOS is an autosomal recessive genetic condition resulting from deficiency in the final enzyme of the cholesterol synthesis pathway, and causes low or low-normal plasma cholesterol levels and increased 7- and 8-dehydrocholesterol (DHC; 7DHC and 8DHC) levels. Common therapies currently used include dietary cholesterol supplementation, treatment with 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors (HMG CoA reductase inhibitors, also known as statins), and treatment with agents that enhance cholesterol production and/or accretion; and to decrease the accumulation of 7DHC and 8DHC, the potentially toxic precursors of cholesterol.

Desmosterolosis is a deficiency in desmosterol reductase and has a similar phenotype to SLOS. In one aspect, described herein are methods for treating desmosterolosis with compounds described herein.

Sitosterolemia is a rare autosomal recessive disorder caused by mutations in two ATP-binding cassette (ABC) transporter genes (ABCG5 and ABCG8). Sitosterolemia enhances the absorption of plant sterols and cholesterol from the intestines. Patients typically present with tendon and tuberous xanthomas and premature coronary artery disease. In one aspect, described herein are methods for treating sitosterolemia with compounds described herein.

In one aspect, described herein are methods for treating cerebrotendinous xanthomatosis (also referred to as cerebral cholesterosis, or Van Bogaert-Scherer-Epstein syndrome) with compounds described herein. CTX can be caused by a mutation in the CYP27A1 gene, which produces the sterol 27-hydroxylase enzyme. Sterol 27-hydroxylase metabolizes cholesterol into bile acids (e.g., chenodeoxycholic acid) that are important in the absorption of fats in the intestine. Enzyme dysfunction can lead to cholesterol accumulation in tissues. CTX is characterized by childhood diarrhea, cataracts, tendon xanthomas, reduced mental capability and abnormal movements in adults.

Mevalonate Kinase Deficiency (also referred to as mevalonic aciduria (a more severe form of MKD), or Hyper IgD Syndrome (HIDS, or hyperimmunoglobulinemia D) with period fever syndrome (a more benign form of MKD)) causes an accumulation of mevalonic acid in the urine as a result of insufficient activity of mevalonate kinase. MKD can result in developmental delay, hypotonia, anemia, hepatosplenomegaly, dysmorphic features, mental retardation, and overall failure to thrive. Mevalonic aciduria is characterized by delayed physical and mental development, failure to thrive, recurrent episodes of fever with vomiting and diarrhea, enlarged liver, spleen and lymph nodes, microcephaly (small head size), cataract, low muscle tone, short statute, distinct facial features, ataxia, and anemia. HIDS is characterized by recurrent episodes of fever associated with swollen lymph nodes, joint pain, gastrointestinal issues and skin rash. In one aspect, described herein are methods for treating MKD with the compounds described herein.

SC4MOL gene deficiency is a genetic disorder in the cholesterol biosynthesis pathway (e.g., mutations in the SC4MOL gene encoding a novel sterol oxidase). SC$MOL deficiency is characterized by the accumulation of dimethyl and monomethyl sterols that can be detected in blood, skin flakes or primary skin fibroblasts. In one aspect, described herein are methods for treating SMO deficiency with compounds described herein.

Niemann-Pick disease is a lysosomal storage disease resulting from a genetic mutation that affects metabolism. Niemann-Pick disease leads to abnormal accumulation of cholesterol and other fatty substances (lipids) due to an inability of the body to transport the substances. The accumulation damages the affected areas.

Autism

In one aspect, described herein are methods for treating autism spectrum disorder or autism. Autism spectrum disorder (ASD) and autism refer to a group of complex disorders of brain development. Autism is typically characterized by difficulties in social interaction, for example in verbal and nonverbal communication. Repetitive behaviors are also often seen in individuals having autism. Autism can be associated with intellectual disability, difficulties in motor coordination and attention and physical health issues, e.g., sleep and gastrointestinal disturbances. Individuals having autism can also excel in visual skills, music, math and art. Autism can refer to autistic disorder, childhood disintegrative disorder, pervasive developmental disorder-not otherwise specified (PDD-NOS), and Asperger syndrome. Autism also refers to monogenetic causes of autism such as synaptophathy's, e.g., Rett syndrome, Fragile X syndrome, Angelman syndrome.

Disorders Associated with Phenylketonuria

In one aspect, described herein are methods for treating disorders associated with phenylketonuria (e.g., cognitive disorders) with compounds described herein. Phenylketonuria can lead to hypochesterolemia and lowered vitamin D status. Total and low-density cholesterols and 25-hydroxy vitamin D have been found to be decreased in subjects suffering from phenylketonuria as compared with subjects not suffering from phenylketonuria (Clin. Chim. Acta2013, 416: 54-59). 24S-hydroxycholesterol and 27S-hydroxycholesterol and 7α-hydroxycholesterol (e.g., representing peripheral and hepatic cholesterol elimination, respectively) have been shown to be significantly decreased in subjects suffering from phenylketonuria, while 7β-hydroxycholesterol (e.g., reflecting oxidative stress) was increased significantly in subjects suffering from phenylketonuria. Changes in the levels of 24S—OHC and 71-hydroxycholesterol correlate with phenylalanine level, and 27S-hydroxycholesterol levels may correlate with the 25-hydroxy vitamin D level in subjects suffering from phenylketonuria.

EXAMPLES

In order that the invention described herein may be more fully understood, the following examples are set forth. Synthetic methods or intermediates may be found, for example in WO2014/160480*. The synthetic and biological examples described in this application are offered to illustrate the compounds, pharmaceutical compositions, and methods provided herein and are not to be construed in any way as limiting their scope.

The stereochemistry assigned herein (e.g., the assignment of “R” or “S” to the C24 position of the steroid) may be tentatively (e.g., randomly) assigned. For example, a C24 position may be drawn in the “R” configuration when the absolute configuration is “S.” A C24 position may also be drawn in the “S” configuration when the absolute configuration is “R.”.

Abbreviation List

Synthetic Methods

Example 1. Synthesis of Compound 1

Step 1. Synthesis of Intermediate A-1

To a suspension of PPh3MeBr (2.13 kg, 5.97 mol) in THF (3000 mL) was added t-BuOK (688 g, 6.14 mol) at 20° C. The color of the suspension was turned to yellow. After stirring at 50° C. for 1 h, Pregnenolone (630 g, 2.05 mol) was added at 50° C. The reaction mixture was stirred at 50° C. for 2 h. After cooling to 20° C., the mixture was treated with NH4Cl (10% aq., 5 L) and heptane (3.5 L), stirred for 15 minutes. The organic layer was separated, concentrated in vacuum to give a crude material as a thick oil, which was poured into MTBE (10 L) with violent stirring and the mixture was stirred at room temperature for 16 hours. An off-white solid was formed and collected by filtration and washed with MTBE (3 L). The combined filtrate was mixed with MeOH (10 L) and concentrated to 6 L in vacuum. An off-white solid was formed, which was collected by filtration, washed with MeOH (3 L), dried in air to give 700 g of wet off-white solid. The combined MeOH filtrate was concentrated in vacuum to give a thick oil. The oil was poured into MTBE (3 L) with violent stirring and the mixture was stirred for 3 hours. An off-white solid was formed and collected by filtration, washed with MTBE (1 L). The combined filtrate was mixed with MeOH (3 L) and concentrated to 1.5 L in vacuum. An off-white solid was formed which was collected by filtration, washed with MeOH (500 mL), dried in air to give 150 g of a wet off-white solid. The previous 700 g and 150 g batch were combined, dried in vacuum to give A-1 (552 g, 88%) as an off-white solid.1H NMR (400 MHz, CDCl3) δ 5.40-5.30 (m, 1H), 4.85 (s, 1H), 4.71 (s, 1H), 3.60-3.50 (m, 1H), 2.36-2.18 (m, 2H), 2.08-1.96 (m, 2H), 1.92-1.78 (m, 3H), 1.76 (s, 3H), 1.73-1.48 (m, 9H), 1.38-1.03 (m, 4H), 1.01 (s, 3H), 1.00-0.91 (m, 1H), 0.58 (s, 3H).

Step 2. Synthesis of Intermediate A-2

To a solution of A-1 (184 g, 585 mmol) in DCM (2000 mL) was added DMP (496 g, 1.17 mol) at 25° C. in portions, followed by adding water (42 mL). The mixture was stirred at 25° C. for 30 min the water (1500 mL) and NaHCO3(750 g) were added in portions with gas evolvolution. The mixture was then filtered through a pad of celite and the solid was washed with DCM (500 mL). The organic layer of the filtrate was separated, washed with Na2S2O3(1000 mL, sat.), dried over Na2SO4, filtered and concentrated in vacuum below 30° C. to give A-2 (250 g, crude) of a light yellow gum. The crude was used in the next step directly.

Step 3. Synthesis of Intermediate A-3

Step 4. Synthesis of Intermediate A-4

Step 5. Synthesis of Intermediate A-5

Step 6. Synthesis of Compound A-6

Step 7. Synthesis of Compound 1-2

To THF (2 mL) under N2at −70° C. was added n-BuLi (1.69 mL, 4.24 mmol) and a suspension of A-6 (500 mg, 1.06 mmol) in THF (5 mL) was added drop-wise to give a light yellow suspension. After stirring at −70° C. for 30 mins, a solution of Compound 1-1 (212 mg, 2.12 mmol) in THF (2 mL) was added. The reaction was stirred at −70° C. for 10 mins and stirred at 25° C. for 16 hrs. The reaction mixture was quenched with water (10 mL) and extracted with EtOAc (3×50 mL). The combined organic phase was washed with brine (100 mL), dried over Na2SO4, filtered and concentrated under vacuum to give the crude product, which was purified by a silica gel column (PE/EtOAc=10/1) to give Compound 1-2 (500 mg, crude) as a yellow solid, which was used directly.

Step 8. Synthesis of Compound 1

Example 2. Synthesis of Compound 2

Synthesis of Compound 2-2

To THF (6 mL) was added n-BuLi (2.5 M, 2.65 mmol, 1.05 mL, 2.5 eq) under N2at −70° C. and a suspension of A-6 (1.06 mmol, 500 mg, 1.0 eq.) in THF (3 mL) was added dropwise to give a light yellow suspension. After stirring at −70° C. for 30 mins, a solution of Compound 2-1 (1.27 mmol, 127 mg, 1.2 eq.) in THF (1 mL) was added dropwise. The reaction was stirred at 15° C. for 12 hrs. The reaction was quenched with sat.NH4Cl (30 mL) and extracted with EtOAc (3×10 mL). The combined organic layers were dried over Na2SO4, filtered and concentrated to give Compound 2-2 (560 mg, crude) as a light yellow foam, which was used directly in the next step.

Synthesis of Compound 2

Example 3. Synthesis of Compounds 3 and 4

Synthesis of Compound 3-2

Synthesis of Compound 3-3

Synthesis of Compounds 3 and 4

Example 4. Synthesis of Compounds 6 and 7

Example 5. Synthesis of Compounds 8 and 9

Example 6. Synthesis of Compounds 10 and 11

Synthesis of Compound 6-2

Synthesis of Compound 6-3

Synthesis of Compounds 10 and 11

Example 7. Synthesis of Compounds 12 and 13

Example 8. Synthesis of Compounds 14 and 15

Example 9. Synthesis of Compounds 16 and 17

Synthesis of Compound 9-2

To a solution of 2,6-di-tert-butyl-4-methylphenol (220 g, 1.0 mol) in toluene (250 mL) was added AlMe3(250 mL, 501 mmol, 2 M in toluene) dropwise below 25° C. The solution was stirred at 25° C. for 1 h. Then a solution of Compound 9-1 (50 g, 167 mmol) in DCM (400 mL) was added dropwise at −78° C. After stirring at the −78° C. for 1 h, EtMgBr (167 mL, 501 mmol, 3M in ethyl ether) was added dropwise at −78° C. The resulting solution was stirred at −78° C. to −50° C. for 3 h. The reaction was quenched by saturated citric acid (100 mL) at −78° C. After stirring at 25° C. for 0.5 h, the resulting mixture was filtered and the filtrate was extracted with DCM (3×100 mL). The combined organic layer was dried over Na2SO4, filtered and concentrated in vacuum. The reaction was conducted in parallel for 2 times. The crude product was combined and purified by a silica gel column (PE/EtOAc=5/1) to give 38 g of the crude product as white solid, which was recrystallized from PE to give a Compound 9-2 as an off white solid (13.5 g, 13%).1H NMR (CDCl3) 400 MHz δ 5.33-5.26 (m, 1H), 5.23-5.10 (m, 1H), 2.45-1.90 (m, 6H), 1.78-0.70 (m, 28H).

Synthesis of Compound 9-3

Synthesis of Compound 9-4

Synthesis of Compound 9-5

Synthesis of Compound 9-6

Synthesis of Compound 9-7

Synthesis of Compounds 16 and 17

Example 10. Synthesis of Compounds 18 and 19

Synthesis of Compound 10-2

To a solution of Compound 10-1* (2 g, 5.01 mmol) and Pd/C (200 mg, 10%) in THF (30 mL) was hydrogenated under 15 psi of hydrogen at 25° C. for 3 h. The mixture was filtered through a pad of celite and the filtrate was concentrated in vacuum to afford Compound 10-2 (1.8 g, crude) as an off-white solid.

Synthesis of Compound 10-3

To a solution of Compound 10-2 (1.8 g, 4.47 mmol) in THF (25 mL) was added a solution LiAlH4(339 mg, 8.94 mmol) in THF (5 mL) drop wise below 15° C. The solution was stirred at 15° C. for 2 h. The reaction was quenched by the addition of saturated aqueous NH4Cl (20 mL) at 0° C. The resulting mixture was extracted with EtOAc (2×50 mL). The combined organic layer was washed with brine (2×30 mL) and concentrated in vacuum to afford Compound 10-3 (1.6 g, crude) as a light yellow solid.

Synthesis of Compound 10-4

A mixture of Compound 10-3 (1.6 g, 4.27 mmol) in DCM (10 mL) and THF (10 mL) was added PCC (2.27 g, 10.6 mmol) at 25° C. The reaction was stirred at 25° C. for 3 hrs. The solution was filtered and the filter cake was washed with DCM (25 mL). The combined filtrate was concentrated in vacuum. The residue was purified by silica gel column, eluting with PE/EtOAc=8/1 to give Compound 10-4 (0.9 g, 54%) as an off-white solid. Synthesis of Compound 10-5. To a solution of Compound 10-4 (0.9 g, 2.41 mmol) in THF (30 mL) was added drop wise isopropyl magnesium chloride (3.61 mL, 7.23 mmol, 2M in THF) at −78° C. The mixture was stirred at −78° C. for 2 hrs. Then, the mixture was allowed to warm up to 25° C. and stirred for 3 hrs. The reaction was poured into water (100 mL) and extracted with EtOAc (2×30 mL). The combined organic layer was washed with brine (50 mL), dried over Na2SO4and concentrated in vacuum. The residue was purified by silica gel column, eluting with PE/EtOAc=5/1 to afford Compound 10-5 (0.6 g, 57%) as an off-white solid.

Synthesis of Compounds 18 and 19

Example 11. Synthesis of Compound 20

Example 12. Synthesis of Compound 21

Example 13. Synthesis of Compounds 22 and 23

Synthesis of Compounds 22 and 23

Further Purification of Compound 23

Example 14. Synthesis of Compounds 24, 25, and 26

Synthesis of Compound 24

Synthesis of Compounds 25 and 26

Example 15. Synthesis of Compounds 27, 28, and 29

Synthesis of Compounds 28 and 29

Example 16. Synthesis of Compound 30

Synthesis of Compound 16-2

Synthesis of Compound 16-3

Synthesis of Compound 16-4

Synthesis of Compound 16-5

Synthesis of Compound 16-6

Synthesis of Compound 30

Example 17. Synthesis of Compounds 31 and 32

Synthesis of Compound 17-2

To a solution of Compound 30 (440 mg, 1.05 mmol) in pyridine (10 mL) was added benzoyl chloride (295 mg, 2.10 mmol) at 25° C. Then the reaction was stirred at 50° C. for 16 h. The reaction was quenched by adding water (10 mL) and extracted with EtOAc (2×10 mL). The combined organic layers were dried over Na2SO4, filtered and concentrated in vacuum. The crude product was purified by a silica gel column (PE/EtOAC=40/1) to give Compound 17-2 (420 mg, 64%) as yellow oil.

Synthesis of Compounds 17-3 and 17-4

Synthesis of Compound 31

Synthesis of Compound 32

Example 18. Synthesis of Compound 33

Synthesis of Compound 18-2

To a solution of 2,6-di-tert-butyl-4-methylphenol (17 g, 77.1 mmol) in toluene (50 mL) was added trimethylaluminum (19.2 mL, 2M in toluene) at 10° C. The mixture was stirred at 20° C. for 1 h. To the solution was added a solution of Compound 16-3 (5 g, 12.8 mmol) in toluene (20 mL) dropwise at −70° C. dropwise under N2. The mixture was stirred at −70° C. for 1 hour. A solution of EtMgBr (12.7 mL, 3M) was added dropwise at −70° C. The mixture was stirred at −70° C. for another 3 hours and then the reaction mixture was quenched with citric acid (150 mL, sat. aq.). The reaction was warmed to 25° C. The organic layer was separated and concentrated in vacuum. The crude product was purified by column chromatography on silica gel (PE/EtOAc=200/1 to 10/1) to give Compound 18-2 (3.8 g) as an off-white solid.

Synthesis of Compound 18-3

Synthesis of Compound 18-4

To a solution of Compound 18-3 (100 mg, crude) in DCM (5 mL) was added Dess Martin reagent (215 mg) at 0° C. under N2. The mixture was stirred at 20° C. for 2 h. A solution of NaHCO3(215 mg) and Na2S2O3(348 mg) in water (10 mL) was added. The mixture was extracted with petroleum ether (3×10 mL). The organic layer was separated, washed with water (20 mL), dried over Na2SO4, filtered and concentrated in vacuum to afford the crude product, which was purified by column chromatography on silica gel (PE/EtOAc=100/1 to 8/1) to give Compound 18-4 (52 mg) as an off-white solid.

Synthesis of Compound 33

Example 19. Synthesis of Compound 34

Synthesis of Compound 34

Example 20. Synthesis of Compounds 35 and 36

Synthesis of Compound 20-2

To a solution of Compound 34 (64 mg, 0.153 mmol) in pyridine (3 mL) was added benzoyl chloride (32.2 mg, 0.229 mmol) and triethylamine (23.1 mg, 0.229 mmol). The mixture was stirred at 25° C. for 5 h and then the reaction mixture was quenched with saturated NH4Cl (6 mL). The mixture was extracted with EtOAc (3×6 mL), washed with Sat. NaCl (2×15 mL), dried over Na2SO4, filtered, concentrated in vacuum to give a crude product, which was purified by column chromatography on silica gel (PE/EtOAc=50/1 to 10/1) to give Compound 20-2 (70 mg, crude). LCMS Rt=1.257 min in 1.5 min chromatography, 5-95AB, MS ESI calcd. for C28H47[M−BzOH−H2O+H]+383, found 383.

Synthesis of Compounds and 20-3 and 20-4

Synthesis of Compound 35

Synthesis of Compound 36

Example 21. Synthesis of Compound 37

Example 22. Synthesis of Compound 38

Example 23. Synthesis of Compounds 39 and 40

Synthesis of Compound 23-2

To a solution of Compound 38 (80 mg, 0.186 mmol) in pyridine (3 mL) was added benzoyl chloride (39.1 mg, 0.279 mmol) and triethylamine (28.1 mg, 0.279 mmol). The mixture was stirred at 25° C. for 5 h. The reaction mixture was quenched with saturated NH4Cl (6 mL). The mixture was extracted with EtOAc (3×6 mL), washed with saturated NaCl (2×15 mL), dried over Na2SO4, filtered, concentrated in vacuum to give the crude product, which was purified by column chromatography on silica gel (PE/EtOAc=50/1 to 10/1) to give Compound 23-2 (80 mg, crude). LCMS tR=1.247 min in 1.5 min chromatography, 5-95 AB_E, MS ESI calcd. for C29H47[M−BzOH−H2O+H]+395, found 395.

Synthesis of Compounds 23-3 and 23-4

Synthesis of Compound 39

Synthesis of Compound 40

Example 24. Synthesis of Compound 41

To a solution of Gla-1 (1 g, 7.4 mmol) in THF (7 mL) under N2was addedt-BuLi (9.25 mL, 1.6 M in pentanes, 14.8 mmol) dropwise at −60° C. After addition, the mixture was warmed to −40° C. slowly for 0.5 h to give a solution of Gla in THF, which was used for the next step directly.

To THF (2 mL) under N2was added Gla (0.87 mL, 0.46 M in THF and pentanes, 0.402 mmol) at −70° C. After stirring at −70° C. for 5 min, a solution of Compound 10-4 (50 mg, 0.314 mmol) in THF (3 mL) was added. The reaction mixture was warmed gradually to 15° C. for 10 hrs. The mixture was quenched with 10 mL of sat.NH4Cl and extracted with 50 mL of EtOAc. The separated organic phase was washed with 10 mL of brine, dried over Na2SO4, filtered and concentrated. The residue was purified by silica gel chromatography eluting with PE/EtOAc=10/1˜2/1 to give Compound 41 (7.3 mg, 13%) as an off-white solid.

Example 25. Synthesis of Compound 42

To THF (1 mL) under N2was added cyclopentylmagnesium chloride (0.402 mL, 1.0 M in THF, 0.402 mmol) at −70° C. After stirring at −70° C. for 5 min, a solution of Compound 10-4 (50 mg, 0.134 mmol) in THF (4 mL) was added. The reaction mixture was gradually warmed to 15° C. for 18 hrs. The mixture was quenched with 10 mL of saturated NH4Cl and extracted with 50 mL of EtOAc. The separated organic phase was washed with 10 mL of brine, dried over Na2SO4, filtered and concentrated. The residue was purified by flash column eluting with PE/EtOAc=10/1-2/1 to give Compound 42 (4.1 mg, 7%) as an off-white solid.

Example 26. Synthesis of Compound 43

To THF (1 mL) under N2was added cyclohexylmagnesium chloride (0.402 mL, 2.0 M in THF, 0.804 mmol) at −70° C. After stirring at −70° C. for 5 min, a solution of Compound 10-4 (100 mg, 0.268 mmol) in THF (1 mL) was added. The reaction mixture was gradually warmed to 15° C. for 18 hrs. The mixture was quenched with 10 mL of sat.NH4Cl and extracted with 50 mL of EtOAc. The separated organic phase was washed with 10 mL of brine, dried over Na2SO4, filtered and concentrated. The residue was purified by flash column eluting with PE/EtOAc, 10/1 to 2/1 to give Compound 43 (20 mg, 16%) as an off-white solid.

Example 27. Synthesis of Compound 44

To a vigorously stirred suspension of Mg turnings (602 mg, 24.8 mmol) and iodine (31.3 mg, 0.124 mmol) in THF (5 mL) under N2was added G5-1 (0.15 g, 1.24 mmol). The mixture was heated to 60° C. After the reaction was initiated, G5-1 (1.35 g, 11.16 mmol) in THF (6 mL) was added slowly. After addition, the mixture was stirred at 60° C. for 2 hrs to give a gray suspension of G5 in THF, which was used for the next step directly.

To THF (2 mL) under N2was added G5 (0.402 mL, 1.0 M in THF, 0.402 mmol) at −70° C. After stirring at −70° C. for 5 mins, a solution of Compound 10-4 (50 mg, 0.314 mmol) in THF (3 mL) was added. The reaction mixture was gradually warmed to 15° C. and stirred for 18 hrs. The mixture was quenched with 10 mL of saturated NH4Cl and extracted with 50 mL of EtOAc. The separated organic phase was washed with 10 mL of brine, dried over Na2SO4and concentrated. The residue was purified by flash column eluting with PE/EtOAc=10/1 to 2/1 to give Compound 44 (10 mg, 16%) as an off-white solid.

Example 28. Synthesis of Compounds 45 and 46

Synthesis of Compound 28-1

To THF (5 mL) under N2at −70° C. was added n-BuLi (2.96 mL, 2.5 M in hexane, 7.42 mmol). After that, a suspension of A-6 (1 g, 2.12 mmol) in THF (8 mL) was added dropwise to give a light yellow suspension. After stirring at −70° C. for 30 mins, a solution of 2-(2,2,2-trifluoroethyl)oxirane (320 mg, 2.52 mmol) in THF (5 mL) was added. The reaction mixture was stirred at −70° C. for 10 mins, warmed to 15° C. and stirred for 16 hrs. The reaction was quenched with saturated NH4Cl (40 mL), extracted with EtOAc (100 mL×3). The combined organic layers were dried over Na2SO4, filtered and concentrated to give Compound 28-1 (1.15 g, crude) as a light yellow solid, which was used in the next step directly.

Synthesis of Compounds 45 and 46

To a solution of Compound 28-1 (1.15 g, 1.92 mmol) in 25 mL of dry MeOH was added under N2magnesium turnings (0.2 g, 8.22 mmol) (activated with 0.5% aqueous HCl, water, dry EtOH, and MTBE) and NiCl2(49.7 mg, 0.384 mmol) with stirring at 50° C. to initiate continuous hydrogen generation. After ten batches of 0.2 g of magnesium turnings were added, the reaction mixture was quenched by 2M HCl (250 mL) at 10° C. until the solid was dissolved. The mixture was extracted with EtOAc (400 mL). The organic layer was washed with saturated NaHCO3(50 mL), brine (50 mL), dried over Na2SO4, filtered and concentrated. The residue was purified by flash column, eluting with PE:EtOAc=20:1-5:1 to give 300 mg of impure product as an off-white solid. The impure product was further purified by SFC (Column: Chiralpak AD-3 150×4.6 mm I.D., 3 um Mobile phase: A: CO2B: methanol (0.05% DEA) Gradient: from 5% to 40% of B in 5 min and hold 40% for 2.5 min, then 5% of B for 2.5 min Flow rate: 2.5 mL/min Column temp.: 35° C.) to give Compound 45 (99.9 mg, 11%) and Compound 46 (84 mg, 10%).

Example 29. Synthesis of Compound 47

To a solution of Compound 45 (140 mg, 0.307 mmol) in MeOH/THF (10 mL/1 mL) was added Pd/C (dry, 10%, 350 mg) under Ar. After degassing for three times with N2, the reaction mixture was degassed for three times with H2. The reaction mixture was stirred for 16 hrs at 55° C. in H2atmosphere (50 Psi). The catalyst was removed by suction, and the filtrate was concentrated to give crude product, which was purified by a silica gel column (EtOAc in PE, 10%-15%) to give Compound 47 (30 mg, 21%) as a white solid.

Example 30. Synthesis of Compound 48

To a solution of Compound 46 (102 mg, 0.223 mmol) in MeOH/THF (10 mL/1 mL) was added Pd/C (dry, 10%, 350 mg) under Ar. After degassing for three times with N2, the reaction mixture was degassed for three times with H2. The reaction mixture was stirred for 16 hrs at 55° C. under H2atmosphere (50 Psi). The catalyst was removed by suction, and the filtrate was concentrated to give crude product, which was purified by a silica gel column (EtOAc in PE, 10%-15%) to give Compound 48 (25 mg, 24%) as a white solid.

Example 31. Synthesis of Compounds 49, 50 and 51

Synthesis of K2

To 150 mL of 30% H2O2in water was added MoO3(3 g, 208 mmol). The mixture was stirred at 40° C. for 5 hrs to form a yellow solution containing a suspended white solid. After cooling to 20° C., the suspension was filtered through a 1-cm mat of Celite. The yellow filtrate is cooled to 10° C. (with an ice bath and magnetic stirring) and HMPA (37.2 g, 208 mmol) was added dropwise. A yellow crystalline precipitate was produced. After filtration, the yellow product was recrystallized from 100 mL of EtOH at −20° C. to give 52 g of crude K2 as a yellow solid.

Synthesis of K3

K2 (52 g, 138 mmol) was dried over P2O5in vacuum for 6 hrs to give 50 g of a yellow solid. The yellow solid was dissolved in 150 mL of THF at 20° C. Pyridine (11.1 g, 140 mmol) was added. After stirring at 20° C. for 10 mins, a yellow crystalline solid was obtained. After filtration, the filtered cake was washed with THF (50 mL), MTBE (200 mL) and dried in vacuum to give 48 g of crude K3 as a yellow solid, which was used directly.

Synthesis of Compound 29-2

To a solution of diisopropylamine (1.81 mL, 12.9 mmol) in THF (80 mL) under N2at −70° C. was added a solution of n-BuLi (5.15 mL, 12.9 mmol, 2.5 M in hexane) dropwise. After stirring at −70° C. for 10 mins, the reaction mixture was warmed to 10° C. gradually for 0.5 h. After cooling to −70° C., a solution of Compound 29-1 (2 g, 10.8 mmol) in THF (20 mL) was added. The reaction mixture was stirred for 0.5 h. K3 (7.06 g, 16.2 mmol) was added. After stirring at −20° C. for 3 hrs, the mixture was quenched with 200 mL of saturated Na2SO3and extracted with MTBE (2×200 mL). The combined organic phase was washed with 100 mL of brine, dried over NaSO4, filtered and concentrated to give 2.1 g of crude product as brown oil.

Synthesis of Compound 29-3

To a solution of Compound 29-2 (2.1 g, 10.4 mmol) in THF (100 mL) was added LiAlH4(789 mg, 20.8 mmol) in portions at −10° C. under N2. The reaction mixture was stirred at 15° C. for 2 hrs. The reaction was quenched with water (1 mL), 15% NaOH aqueous solution (1 mL) and water (3 mL) dropwise at 0° C. After stirring at 15° C. for 15 mins, 2 g of MgSO4was added at 15° C. The mixture was stirred at this temperature for 1 h. After filtering through celite under vacuum and washing with DCM (2×100 mL), the organic layer was concentrated under vacuum to give 2 g of crude product as a yellow oil.

Synthesis of Compound 29-4

To a solution of Compound 29-3 (2 g, 12.6 mmol) in pyridine (15 mL) was added TsCl (2.87 g, 15.1 mmol) in portions during 5 minutes at 0° C. The reaction solution was stirred at 15° C. for 16 hrs. The reaction mixture was quenched with 2N HCl (95 mL) to pH=1-2 at 0° C. The inner temperature was maintained below 30° C. and the mixture was extracted with MTBE (2×250 mL). The combined organic layer was dried over Na2SO4, filtered, concentrated and purified by column (0-20% of EtOAc in PE) to give Compound 29-4 (2.1 g, 53%) as light yellow oil.

Synthesis of Compound 29-5

To THF (3.5 mL) under N2at −70° C. was added diisopropylamine (2.35 mmol, 237 mg), followed by an addition of n-BuLi (2.22 mmol, 0.89 mL, 2.5M in hexane). The reaction was allowed to warm to 15° C. and re-cooled to −70° C. A suspension of A-6 (0.637 mmol, 300 mg) in THF (1.5 mL) was added dropwise to give a light yellow suspension. After stirring at −70° C. for 30 mins, a solution of Compound 29-4 (700 μmol, 218 mg) in THF (1.5 mL) was added over 5 min (slightly exothermic, keeping internal T<−70° C.). The reaction was stirred at 15° C. for 12 hrs. The reaction was quenched with saturated NH4Cl (30 mL) and extracted with EtOAc (3×10 mL). The combined organic layers were dried over Na2SO4, filtered and concentrated to give Compound 29-5 (400 mg, crude) as light yellow foam, which was used in the next step directly.

Synthesis of Compound 49

To a solution of Compound 29-5 (400 mg, 0.654 mmol) in MeOH (5 mL) was added Mg powder (940 mg, 39.2 mmol) at 55° C. The mixture was stirred at 55° C. for 16 hrs. The mixture was quenched with HCl (30 mL, 1N) until the mixture became clear and was extracted with DCM (3×10 mL). The combined organic phase was washed with saturated NaHCO3(20 mL), dried over Na2SO4, filtered, concentrated and purified by combi-flash (0-15% of EtOAc in PE) to give Compound 49 (80 mg, 26%) as an off-white solid.

Synthesis of Compounds 50 and 51

Example 32. Synthesis of Compounds 52, 53, and 54

To a solution of diisopropylamine (781 mg, 7.72 mmol) in THF (8 mL) was added a solution of n-BuLi (2.8 mL, 2.5 M in hexane, 7.10 mmol) dropwise under N2at −78° C. The mixture was warmed to 0° C. To a suspension of 32-1 (1.5 g, 3.09 mmol) in THF (15 mL) was added the fresh prepared LDA solution dropwise under N2at −78° C. The mixture was stirred at −78° C. for 30 mins. A solution of 2-(2,2,2-trifluoroethyl)oxirane (583 mg, 4.63 mmol) in THF (6 mL) was added. The mixture was stirred at −78° C. for 30 mins and allowed to warm to 25° C. and stirred for 48 hrs. The reaction mixture was quenched by water (100 mL) and HCl (1 M, aq.) until pH=5 at 15° C. The mixture was extracted with EtOAc (500 mL). The separated organic phase was washed with brine, dried over Na2SO4, filtered and concentrated. The crude product was purified by flash column (10-50% of EtOAc in PE) to give 32-2 (1.4 g, 74%) as an off white solid, which was used directly.

To a solution of 32-2 (1.4 g, 2.29 mmol) in 20 mL of dry MeOH was added Mg powder (1.64 g, 68.7 mmol) under N2at 60° C. The reaction mixture was quenched by 2 M HCl (250 mL) at 10° C. until the solid was dissolved. After extracted with EtOAc (400 mL), the organic layer was washed with Sat. NaHCO3(50 mL), brine (50 mL), dried over Na2SO4, filtered and concentrated. The residue was purified by flash column eluted with PE/EtOAc=20:1-5:1 to give Compound 52 (510 mg, 47%) as an off white solid.

Example 33. Synthesis of Compound 55

Example 34. Synthesis of Compound 56

To a solution of Compound 53 (183 mg, 0.389 mmol) in MeOH (20 mL) was added Pd/C (dry, 10%, 350 mg) under Ar. After degassing for three times with N2, the reaction mixture was degassed for three times with H2. The reaction mixture was stirred for 16 hrs at 55° C. under H2atmosphere (50 Psi). The catalyst was removed by suction, and the filtrate was concentrated to give the crude product which was purified by a silica gel column (EtOAc in PE, 10%-15%) to give Compound 56 (20 mg, 10%) as an off-white solid.

Example 35. Synthesis of Compounds 57 and 58

To a vigorously stirred suspension of Mg (1.76 g, 72.8 mmol) turnings and iodine (46.1 mg, 0.182 mmol) in THF (2 mL) under N2was added 1,2-dibromoethane (68.3 mg, 0.364 mmol) and 10% of a solution of 4-chlorotetrahydro-2H-pyran (4.4 g, 36.4 mmol) in THF (18 mL). The mixture was heated to 60° C. and as the reaction mixture turned clear and Grignard initiated took place, the remainder of the solution of 4-chlorotetrahydro-2H-pyran in THF was added slowly over 30 min. The reaction mixture was stirred at 65° C. for 2 h to give a solution of (tetrahydro-2H-pyran-4-yl)magnesium chloride in THF (˜2M). The Grignard solution was used without any further purification. The solution of 36-4 (800 mg, 2.06 mmol) in THF (150 mL) under N2was added to Grignard reagent at 15° C. in one portion. After stirring at 15° C. for 2 min, the mixture was quenched by 200 mL of sat.NH4Cl and extracted with 200 mL of EtOAc. The separated organic phase was washed with 200 mL of brine, dried over Na2SO4, filtered and concentrated. The residue was purified by Combi-flash (0%-30% of EtOAc in PE/DCM (v/v=1/1)) to afford 36-5 (550 mg, 56%) as off-white solid, and 50 mg of 36-5 was delivered.1H NMR (400 MHz, CDCl3) δ 5.32-5.25 (m, 1H), 4.06-3.96 (m, 2H), 3.42-3.29 (m, 3H), 2.39-2.33 (m, 1H), 2.07-1.79 (m, 6H), 1.77-1.60 (m, 7H), 1.51-1.38 (m, 10H), 1.35-1.21 (m, 4H), 1.16-1.01 (m, 8H), 0.97-0.90 (m, 4H), 0.85 (t, J=7.4 Hz, 3H), 0.71-0.66 (m, 3H). LCMS Rt=1.212 min in 2 min chromatography, 30-90AB_2MIN_E.M, MS ESI calcd. for C31H51O2[M+H−H2O]+455, found 455.

To a mixture of DMP (539 g, 1271 mmol) in DCM (800 mL) was added 37-1 (200 g, 636 mmol) in DCM (2.2 L) at 30° C. The reaction mixture was stirred at 40° C. for 1 h. The reaction mixture was quenched with saturated NaHCO3aqueous (1.2 L) at 10° C. The mixture was filtered. The DCM phase in filtrate was separated and washed with saturated NaHCO3/Na2S2O3aqueous (1:1, 2×1 L), brine (1 L), dried over Na2SO4, filtered and concentrated under vacuum to give a yellow solid, which was triturated in MeCN (700 mL) to give 37-2 (115 g, 58%) as a off-white solid.

To a solution of diisopropylamine (7.28 g, 72.1 mmol) in THF (20 mL) under N2at −70° C., was added n-BuLi (27.1 mL, 2.5 M, 67.9 mmol). The resulting mixture was stirred at 0° C. for 30 min. The mixture was re-cooled to −70° C. To the mixture was added 32-1 (10 g, 20.6 mmol) in THF (50 mL) at −70° C. The reaction mixture was stirred at −70° C. for 1 h. 2-isopropyloxirane (2.12 g, 24.7 mmol) in THF (10 mL) was added at −70° C. The reaction mixture was warmed to 15° C. slowly, and stirred at 15° C. for 16 hrs. The reaction mixture was quenched with saturated NH4Cl aqueous (100 mL) at 0° C. The mixture was extracted with EtOAc (2×200 mL). The combined organic phase was washed with brine (150 mL), dried over Na2SO4, filtered and concentrated under vacuum to give 37-6 (12 g, crude) as a yellow solid. LCMS Rt=3.784 & 3.859 min in 7 min chromatography, 30-90AB_7MIN_E.M, MS ESI calcd. for C35H53O3S [M+H−H2O]+553, found 553.

To a solution of 37-6 (12 g, 21.0 mmol) in 200 mL of anhydrous MeOH was added Mg powder (30.6 g, 1260 mmol) and NiCl2(27.0 mg, 0.21 mmol) with stirring under N2at 50° C. to initiate continuous hydrogen generation. The reaction mixture was quenched by 2 M HCl (100 mL) until solid was dissolved. The mixture was extracted with EtOAc (3×200 mL). The combined organic layer was washed with Sat. NaHCO3(50 mL), brine (50 mL), dried over Na2SO4, filtered and concentrated. The residue was purified by silica gel chromatography eluted with PE/EtOAc=20/1-8/1 to give 5.6 g of off-white solid, which was purified by SFC (Column: Chiralpak AD 250×30 mm I.D., 5 um Mobile phase: A: CO2 B: methanol (0.1% NH3H2O) Gradient: from 35% to 35% of B, Flow rate: 60 mL/min) to give Compound 10 (2.2 g, 24%), Compound 11 (2.2 g, 24%) as off-white solid.

A solution of 38-1 (0.6 g, 15.4 mmol) in MeOH (20 mL) was purified by prep-SFC to give the Compound 66 (180 mg, 30%) and Compound 67 (240 mg, 40%). The absolute configuration of Compound 66 and Compound 67 was confirmed by Mosher method.

To a solution of compound Compound 66 (140 mg, 0.36 mmol) in EtOAc (5 mL) was added 5% Pd/C (56 mg) under N2. The suspension was degassed under vacuum and purged with H2several times. Then the mixture was stirred under H2(50 psi) at 50° C. for 12 hours. The mixture was filtered through a pad of celite and the pad was washed with EtOAc (2×5 mL). The combined filtrates were concentrated to dryness to give a crude product, which was purified by column chromatography on silica gel (PE/EtOAc/EActOAc=12/1 to 10/1) to afford the Compound 68 (80 mg, 57%) and Compound 69 (18 mg, 13%) as off-white powder.

To a solution of compound Compound 67 (120 mg, 0.30 mmol) in EtOAc (5 mL) was added 5% Pd/C (48 mg) under N2. The suspension was degassed under vacuum and purged with H2several times. Then the mixture was stirred under H2(50 psi) at 50° C. for 12 hours. The mixture was filtered through a pad of celite and the pad was washed with EtOAc (2×5 mL). The combined filtrates were concentrated to dryness to give a crude product, which was purified by column chromatography on silica gel (PE/EtOAc/EtOAc=12/1 to 10/1) to afford the Compound 70 (78 mg, 65%) and Compound 71 (26 mg, 21%) as off-white powder.

nBuLi (2.06 mL, 2.5 M in hexane, 5.15 mmol) was added to THF (3 mL) dropwise under N2at −70° C., followed by adding a suspension of 32-1 (1 g, 2.06 mmol) in THF (5 mL). After stirring at −70° C. for 30 min, a solution of 2-(tert-butyl)oxirane (309 mg, 3.09 mmol) in THF (2 mL) was added dropwise. The mixture was stirred at −70° C. for 30 min and allowed to warm to 20° C. and stirred at 20° C. for 16 hrs. The reaction mixture was quenched by adding 30 mL of sat.NH4Cl at 20° C. The organic layer was separated. The aqueous phase was extracted with EtOAc (2×30 mL). The combined organic layer dried over Na2SO4, filtered, concentrated and purified by combi-flash (0-15% of EtOAc in PE) to give impure 39-1 (700 mg, impure) as an off-white solid.

Step 4. Synthesis of Compounds 74-A and 74-B.

To a solution of Compound 72 (110 mg, 0.247 mmol) in MeOH (30 mL) was added Pd/C (dry, 200 mg). The mixture was stirred at 50° C. for 72 hrs under H2(50 psi). The mixture was filtered, concentrated and purified by combi-flash (0-15% of EtOAc in PE) to give Compound 74-A (19 mg, 17%) as a off-white solid and Compound 74-B (18 mg, 16%) as an off-white solid.

Step 5. Synthesis of Compounds 75-A and 75-B.

To a solution of Compound 73 (70 mg, 0.157 mmol) in MeOH (30 mL) was added Pd/C (dry, 150 mg). The mixture was stirred at 50° C. for 72 hrs under H2(50 psi). The mixture was filtered, concentrated and purified by combi-flash (0-15% of EtOAc in PE) to give Compound 75-A (10 mg, 14%) as an off-white solid and Compound 75-B (12 mg, 17%) as a off-white solid.

Materials and Methods

Additionally, as will be apparent to those skilled in the art, conventional protecting groups may be necessary to prevent certain functional groups from undergoing undesired reactions. The choice of a suitable protecting group for a particular functional group as well as suitable conditions for protection and deprotection are well known in the art. For example, numerous protecting groups, and their introduction and removal, are described in T. W. Greene and P. G. M. Wuts,Protecting Groups in Organic Synthesis, Second Edition, Wiley, New York, 1991, and references cited therein.

The compounds provided herein may be isolated and purified by known standard procedures. Such procedures include (but are not limited to) recrystallization, column chromatography, HPLC, or supercritical fluid chromatography (SFC). The compounds provided herein may be prepared from known or commercially available starting materials and reagents by one skilled in the art of organic synthesis. Exemplary chiral columns available for use in the separation/purification of the enantiomers/diastereomers provided herein include, but are not limited to, CHIRALPAK® AD-10, CHIRALCEL® OB, CHIRALCEL® OB-H, CHIRALCEL® OD, CHIRALCEL® OD-H, CHIRALCEL® OF, CHIRALCEL® OG, CHIRALCEL® OJ and CHIRALCEL® OK.

1H-NMR reported herein (e.g., for the region between δ (ppm) of about 1 to about 4 ppm) will be understood to be an exemplary interpretation of the NMR spectrum (e.g., exemplary peak integratations) of a compound. Exemplary general method for preparative HPLC: Column: Waters RBridge prep 10 μm C18, 19*250 mm. Mobile phase: acetonitrile, water (NH4HCO3) (30 L water, 24 g NH4HCO3, 30 mL NH3.H2O). Flow rate: 25 mL/min

NMDA potentiation was assessed using either whole cell patch clamp of mammalian cells which expressed NMDA receptors.

Whole-cell Patch Clamp of Mammalian Cells (Ionworks Barracuda (IWB))

The whole-cell patch-clamp technique was used to investigate the effects of compounds on GlunN1/GluN2A glutamate receptors expressed in mammalian cells. The results are shown on Table 1.

HEK293 cells were transformed with adenovirus 5 DNA and transfected with cDNA encoding the human GRIN1/GRIN2A genes. Stable transfectants were selected using G418 and Zeocin-resistance genes incorporated into the expression plasmid and selection pressure maintained with G418 and Zeocin in the medium. Cells were cultured in Dulbecco's Modified Eagle Medium/Nutrient Mixture (D-MEM/F-12) supplemented with 10% fetal bovine serum, 100 μg/ml penicillin G sodium, 100 μg/ml streptomycin sulphate, 100 μg/ml Zeocin, 5 μg/ml blasticidin and 500 μg/ml G418.

Test article effects were evaluated in 8-point concentration-response format (4 replicate wells/concentration). All test and control solutions contained 0.3% DMSO and 0.01% KOLLIPHOR (polyethoxylated castor oil) EL (C5135, Sigma). The test article formulations were loaded in a 384-well compound plate using an automated liquid handling system (SciClone ALH3000, Caliper LifeScienses). The measurements were performed using Ion Works Barracuda platform following this procedure:

a) Intracellular solution (mM): 50 mM CsCl, 90 mM CsF, 2 mM MgCl2, 5 mM EGTA, 10 mM HEPES. Adjust to pH 7.2 with CsOH.b) Extracellular solution, HB-PS (composition in mM): NaCl, 137; KCl, 1.0; CaCl2, 5; HEPES, 10; Glucose, 10; pH adjusted to 7.4 with NaOH (refrigerated until use).c) Holding potential: −70 mV, potential during agonist/PAM application: −40 mV.
Recording Procedure:a) Extracellular buffer will be loaded into the PPC plate wells (11 μL per well). Cell suspension will be pipetted into the wells (9 μL per well) of the PPC planar electrode.b) Whole-cell recording configuration will be established via patch perforation with membrane currents recorded by on-board patch clamp amplifiers.c) Two recordings (scans) will be performed. First, during pre-application of test article alone (duration of pre-application—5 min) and second, during test articles and agonist (EC20L-glutamate and 30 μM glycine) co-application to detect positive modulatory effects of the test article.

Test Article Administration: The first pre-application will consist of the addition of 20 μL of 2× concentrated test article solution and, second, of 20 μL of 1× concentrated test article and agonist at 10 μL/s (2 second total application time).

TABLE 1GluN2A PCA IWB EphysStructure% potentiation at 3 μM1B1-AB1-BC2A3C4C6C7C8A9B10C11C12A13C14A15C16C17B18C19C20C21C22C23C23C25C26C28C29B30C31C32C33A34C35B36C37C38C39A40C41C41-AC41-BC42B43B44C44-AC44-BB45C46C47C48C50C51C52C53C54C55C56C66B67A68C70CFor Table 1, “A” indicates 10 to 75%, “B” indicates potentiation of >75% to 150%; “C” indicates potentiation of >150%; and “ND” indicates not determinable or not determined.

OTHER EMBODIMENTS