NZ709872B2

NZ709872B2 - Spiro-lactam nmda receptor modulators and uses thereof

Info

Publication number: NZ709872B2
Application number: NZ709872A
Authority: NZ
Inventors: M Amin Khan; John A Lowe Iii
Original assignee: Aptinyx Inc
Priority date: 2013-01-29
Filing date: 2014-01-29
Publication date: 2021-01-06

Abstract

Disclosed are compounds having enhanced potency in the modulation of NMD A receptor activity. Such compounds are contemplated for use in the treatment of conditions such as depression and related disorders. Orally available formulations and other pharmaceutically acceptable delivery forms of the compounds, including intravenous formulations, are also disclosed. pounds, including intravenous formulations, are also disclosed.

Description

SPIRO-LACTAM NMDA RECEPTOR MODULATORS AND USES THEREOF CROSS REFERENCE TO RELATED APPLICATIONS This application claims the beneﬁt of United States Provisional Application No. ,920, ﬁled on January 29, 2013, which is incorporated by reference in its ty.

BACKGROUND An yl-d-aspartate (NMDA) receptor is a postsynaptic, ionotropic receptor that is responsive to, inter alia, the excitatory amino acids glutamate and glycine and the synthetic compound NMDA. The NMDA receptor controls the ﬂow of both divalent and monovalent ions into the postsynaptic neural cell h a receptor associated channel (Foster et 61]., Nature 1987, 329:395—396; Mayer et al, Trends in Pharmacol. Sci. 1990, 112254—260).

The NMDA receptor has been implicated during pment in ying neuronal architecture and synaptic tivity, and may be involved in experience—dependent synaptic modifications. In addition, NMDA receptors are also thought to be involved in long term potentiation and central nervous system disorders.

The NMDA receptor plays a major role in the synaptic plasticity that underlies many higher cognitive functions, such as memory acquisition, retention and learning, as well as in certain cognitive pathways and in the perception of pain (Collingridge et al., The NMDA Receptor, Oxford University Press, 1994). In addition, n properties ofNMDA receptors suggest that they may be involved in the information-processing in the brain that underlies consciousness itself.

The NMDA receptor has drawn particular interest since it appears to be involved in a broad spectrum of CNS ers. For instance, during brain ischemia caused by stroke or traumatic injury, excessive amounts of the excitatory amino acid glutamate are released from damaged or oxygen deprived neurons. This excess glutamate binds to the NMDA receptors which opens their ligand-gated ion ls; in turn the calcium inﬂux es a high level of intracellular calcium which activates a biochemical cascade resulting in protein degradation and cell death. This phenomenon, known as excitotoxicity, is also thought to be responsible for the neurological damage associated with other disorders ranging from hypoglycemia and cardiac arrest to epilepsy. In addition, there are preliminary reports indicating similar involvement in the chronic neurodegeneration of Huntington's, Parkinson's, and Alzheimer's diseases. Activation of the NMDA receptor has been shown to be responsible for post-stroke convulsions, and, in certain models of epilepsy, tion of the NMDA receptor has been shown to be necessary for the tion of seizures. Neuropsychiatric involvement of the NMDA receptor has also been recognized since blockage of the NMDA receptor Ca++ l by the animal anesthetic PCP (phencyclidine) produces a psychotic state in humans similar to schizophrenia (reviewed in Johnson, K. and Jones, S., 1990). Further, NMDA receptors have also been implicated in certain types of spatial learning.

The NMDA receptor is believed to consist of l protein chains embedded in the postsynaptic membrane. The first two types of subunits discovered so far form a large extracellular region, which ly contains most of the allosteric g sites, several embrane regions looped and folded so as to form a pore or channel, which is permeable to Ca++, and a carboxyl terminal region. The opening and closing of the channel is regulated by the binding of various ligands to domains (allosteric sites) of the protein ng on the ellular surface. The binding of the ligands is t to affect a conformational change in the overall structure of the protein which is ultimately reflected in the channel opening, partially opening, partially closing, or closing.

[0006] NMDA receptor compounds may exert dual st/antagonist) effect on the NMDA or through the allosteric sites. These compounds are typically termed “partial agonists”.

In the presence of the principal site ligand, a partial agonist will displace some of the ligand and thus decrease Ca++ flow through the receptor. In the absence of or lowered level of the principal site ligand, the l agonist acts to increase Ca++ flow through the or channel.

[0007] A need ues to exist in the art for novel and more specific/potent compounds that are capable of binding the glycine binding site of NMDA receptors, and provide pharmaceutical benefits. In addition, a need continues to exist in the medical arts for orally deliverable forms of such compounds.

SUMMARY [0007a] According to a first aspect, the present invention relates to a compound represented by formula I: or a pharmaceutically acceptable salt, a stereoisomer, or an e thereof, wherein Rb is selected from the group consisting of H, halogen, yl, cyano or C1-C6 alkyl; R1 is H or C1-C6 alkyl; R2 is H or C1-C6 alkyl; R3 is selected from the group consisting of H, C1-C6 alkyl and a nitrogen protecting group; wherein the nitrogen protecting group is selected from the group ting of 9- fluorenylmethyloxycarbonyl, tert-butoxycarbonyl, p-methoxybenzyloxycarbonyl, acetyl, trifluoroacetyl, benzoyl, benzyl, p-methoxybenzyl, p-methoxyphenyl, 3,4- dimethoxybenzyl, nylmethyl, p-toluenesulfonyl, –C(O)OR31 and 32; R31 is selected from the group consisting of: C1-C6 alkyl, C1-C6 haloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C10 cycloalkyl, -CH2-C3-C10 cycloalkyl, -CH2-phenyl, and –CH2- pyridyl, wherein any aforementioned cycloalkyl is optionally substituted with from 1-3 independently selected C1-C3 alkyl, and wherein the phenyl is optionally substituted with from 1-2 substituents independently selected from C1-C3 alkyl, C1-C3 kyl, C1-C3 alkoxy, C1-C3 haloalkoxy, nitro, halo, SO2Me, cyano, and -OC(O)CH3; and R32 is selected from the group consisting of: H, C1-C6 alkyl, C1-C6 haloalkyl, phenyl, and pyridyl, wherein the phenyl is ally tuted with from 1-2 substituents independently selected from C1-C3 alkyl, C1-C3 haloalkyl, C1-C3 alkoxy, C1-C3 koxy, nitro, halo, SO2Me, cyano, and -OC(O)CH3; R4 is H or C1-C6 alkyl; R5 is –C1-C6 alkylene-X, wherein X is selected from the group consisting of: (i) heteroaryl including from 5 to 6 ring atoms wherein 1, 2, or 3 of the ring atoms are independently selected from the group consisting of N, NH, N(C1-C3 alkyl), O, and S; (ii) heterocyclyl including from 3 to 6 ring atoms wherein 1, 2, or 3 of the ring atoms are independently selected from the group consisting of N, NH, N(C1-C3 alkyl), O, and S; R5 is X, wherein X is heterocyclyl including from 3 to 6 ring atoms wherein 1, 2, or 3 of the ring atoms are independently selected from the group consisting of N, NH, N(C1-C3 alkyl), O, and S; wherein R5 is unsubstituted or substituted with ; and R6 is selected from the group consisting of H, n, yl, cyano, -O-C(O)-C1-C6 alkyl, C1-C6 alkyl, and C1-C6 alkoxy. [0007b] According to a second aspect, the present invention relates to a pharmaceutical composition comprising a compound of the invention, and a pharmaceutically able excipient. [0007c] According to a third , the present invention relates to use of a compound of the ion in the manufacture of a medicament for treating depression, Alzheimer’s e, attention deficit disorder, schizophrenia, or anxiety.

Provided herein, at least in part, are compounds that are NMDA modulators, for example, partial agonists of NMDA. For example, disclosed herein are compounds represented by the formula: and pharmaceutically acceptable salts, stereoisomers, and N-oxides thereof, wherein Rb is ed from the group consisting of H, halogen, hydroxyl, cyano or C1-C6 alkyl; R1 is H or C1-C6 alkyl; R2 is H or C1-C6 alkyl; R3 is selected from the group consisting of H, C1-C6 alkyl and a nitrogen protecting group; R4 is H or C1-C6 alkyl; R5 is X or –C1-C6 alkylene-X, wherein X is selected from the group consisting of phenyl, a 4- to 6-membered heteroaryl ring having 1, 2, or 3 heteroatoms selected from O, S, or N, or a 3b followed by page 4 4- to 6-membered heterocyclyl ring having 1, 2, or 3 heteroatoms selected from O, S, or N, and wherein X is optionally substituted with ; and R6 is selected from the group consisting of H, halogen, yl, cyano, -O-C(O)-C1-C6 alkyl, C1-C6 alkyl, or C1-C6 alkoxy; or, in other ments, the variables set forth in formula (I) are defined as follows: Rb is selected from the group consisting of H, halogen, hydroxyl, cyano or C1-C6 alkyl; R1 is H or C1-C6 alkyl; R2 is H or C1-C6 alkyl; R3 is selected from the group consisting of H, C1-C6 alkyl and a nitrogen protecting group; R4 is H or C1-C6 alkyl; R5 is X or —C1—C6 alkylene—X, wherein X is selected from the group consisting of: (i) phenyl; (ii) heteroaryl including from 5 to 6 ring atoms wherein l, 2, or 3 of the ring atoms are independently selected from the group consisting ofN, NH, N(Cl-C3 alkyl), 0, and S; and (iii) cyclyl including from 3 to 6 ring atoms wherein l, 2, or 3 of the ring atoms are independently selected from the group ting ofN, NH, N(Cl-C3 alkyl), 0, and S; wherein R5 is optionally substituted with R6; and R6 is selected from the group consisting of H, halogen, hydroxyl, cyano, —O—C(O)—C1—C6 alkyl, C1—C6 alkyl, or C1-C6 alkoxy.

[0009] Also disclosed herein are compounds represented by the formula: N OH R3 0 (II) and pharmaceutically acceptable salts, isomers, and N—oxides thereof, wherein R3 is selected from the group consisting of H, C1—C6alkyl and a nitrogen ting group; and R7 is a 4- to 6-membered heteroaryl ring having 1, 2, or 3 heteroatoms selected from O, S, or N, ally substituted on a free carbon by a substituent selected from the group consisting of: halogen, C1-C5alkyl, hydroxyl, cyano, and .

Also provided herein are compounds represented by the formula: LN\ R4 N\XA R1 | 6 R3 0 (111) and pharmaceutically acceptable salts, stereoisomers, and N—oxides f, wherein Rb is selected from the group consisting of H, halogen, hydroxyl, cyano or C1-C6 alkyl; R1 is H or C1-C6 alkyl; R2 is H or C1-C6 alkyl; R3 is selected from the group consisting of H, C1—C6 alkyl and a nitrogen protecting group; R4 is H or C1-C6 alkyl, X is selected from the group consisting of phenyl, a 4- to 6-membered heteroaryl ring having 1, 2, or 3 heteroatoms ed from O, S, or N, or a 4- to 6—membered heterocyclyl ring having 1, 2, or 3 atoms selected from O, S, or N, wherein R6 is present on a free carbon of X; R6 is selected from the group ting of H, halogen, hydroxyl, cyano, )-C1-C6 alkyl, C1-C5 alkyl, 01' C1-C6 alkoxy.

Also provided herein are pharmaceutically acceptable compositions comprising a sed compound, and a pharmaceutically acceptable excipient. For example, such compositions may be suitable for oral or intravenous administration to a patient.

In another aspect, a method of treating a condition selected from the group consisting of , anxiety, depression, bipolar disorder, attention deﬁcit disorder, attention deficit hyperactivity disorder (ADHD), schizophrenia, a psychotic disorder, a psychotic symptom, social withdrawal, obsessive—compulsive disorder, phobia, raumatic stress syndrome, a behavior disorder, an impulse control disorder, a substance abuse disorder, a sleep er, a memory disorder, a learning disorder, urinary incontinence, le system atrophy, progressive supra-nuclear palsy, ich's ataxia, Down’s syndrome, fragile X syndrome, tuberous sclerosis, olivio—ponto-cerebellar atrophy, cerebral palsy, nduced optic neuritis, ischemic retinopathy, ic retinopathy, glaucoma, dementia, AIDS dementia, Alzheimer’s disease, Huntington’s chorea, city, myoclonus, muscle spasm, Tourette's syndrome, epilepsy, cerebral ischemia, , a brain tumor, traumatic brain injury, cardiac arrest, myelopathy, spinal cord injury, peripheral neuropathy, acute neuropathic pain, and chronic neuropathic, in a patient in need thereof is provided. Such methods may se administering to the patient a pharmaceutically effective amount of a disclosed compound or ceutically acceptable salts, stereoisomers, N—oxides, and es thereof.

In some embodiments, a contemplated method includes treating depression. For example, depression may include one or more of major depressive disorder, dysthymic disorder, psychotic depression, postpartum depression, seasonal affective disorder, bipolar disorder, mood disorder, or depression caused by a chronic medical condition. In other embodiments, a contemplated method may treat schizophrenia. Such schizophrenia may be, for example, paranoid type schizophrenia, disorganized type phrenia, catatonic type schizophrenia, undifferentiated type schizophrenia, residual type schizophrenia, post- schizophrenic depression, or simple schizophrenia.

BRIEF DESCRIPTION OF THE GS Figure 1 shows the potentiation of [3H]MK-801 binding in the presence of Compound Y.

ED DESCRIPTION This disclosure is generally directed to compounds that are capable of modulating NMDA, e.g., NMDA antagonists or partial agonists, and compositions and/or methods of using the disclosed compounds.

Deﬁnitions

[0016] “Treating” includes any effect, e. g., lessening, reducing, modulating, or eliminating, that s in the improvement of the condition, disease, disorder and the like.

The term “alkenyl” as used herein refers to an unsaturated ht or branched hydrocarbon having at least one carbon-carbon double bond, such as a straight or branched group of 2-6 or 3—4 carbon atoms, referred to herein for example as C2_C6 alkenyl, and C3_C4 alkenyl, respectively. Exemplary alkenyl groups include, but are not limited to, vinyl, allyl, butenyl, yl, etc.

The term “alkoxy” as used herein refers to a ht or branched alkyl group attached to an oxygen (alkyl-O—). Exemplary alkoxy groups include, but are not limited to, s of 1-6 or 2-6 carbon atoms, referred to herein as C1-C6 alkoxy, and C2-C6 alkoxy, respectively. Exemplary alkoxy groups e, but are not limited to methoxy, ethoxy, isopropoxy, etc.

The term “alkenyloxy” used herein refers to a straight or branched alkenyl group attached to an oxygen (alkenyl-O). Exemplary alkenoxy groupd include, but are not limited to, groups with an alkenyl group of 3-6 carbon atoms, (also e.g. referred to as C3—C6 alkenyloxy).

Exemplary oxy” groups include, but are not limited to allyloxy, butenyloxy, etc.

The term “alkynyloxy” used herein refers to a straight or branched alkynyl group attached to an oxygen (alkynyl-0)). ary alkynyloxy groups include, but are not d to, C3-C6 alkynyloxy, e. g., propynyloxy.

The term “alkyl” as used herein refers to a saturated straight or branched hydrocarbon, such as a straight or branched group of 1-6, 1-4, or 1—3 carbon atoms, referred to herein as C1-C5 alkyl, C1-C4 alkyl, and C1-C3 alkyl, respectively. Exemplary alkyl groups include, but are not d to, methyl, ethyl, propyl, isopropyl, 2-methyl-l-propyl, 2-methyl propyl, yl-l-butyl, 3-methylbutyl, ylbutyl, 2,2-dimethyl-l-propyl, 2- -l—pentyl, 3-methyl-l-pentyl, 4-methyl-l-pentyl, 2-methylpentyl, 3-methylpentyl, 4-methyl—2-pentyl, 2,2—dimethyl—l-butyl, 3,3-dimethyl—1—butyl, l—1—butyl, butyl, isobutyl, t—butyl, pentyl, isopentyl, neopentyl, hexyl, etc. The term “haloalkyl” as used herein refers to a saturated ht or branched alkyl groups, in which one or more hydrogen atoms of the alkyl group are replaced with one or more independently selected ns. The term “haloalkyl” encompasses alkyl groups in which all of hydrogen atoms of the alkyl group are replaced independently selected halogens imes referred to as “perhalo” alkyl groups. ary haloalkyl groups e, but are not limited to, CHZF, CHZCHZCI, CF3, CHFCHzCl.

The term “alkynyl” as used herein refers to an unsaturated straight or branched hydrocarbon having at least one carbon-carbon triple bond, such as a ht or branched group of 2-6, or 3-6 carbon atoms, referred to herein as C2-C6 alkynyl, and C3_C6 alkynyl, respectively.

Exemplary alkynyl groups include, but are not limited to, ethynyl, propynyl, butynyl, pentynyl, hexynyl, methylpropynyl, etc.

The term “bridged lkyl”, as used herein, is defined as a monocyclic 4— to 7— membered cycloalkyl group in which two non-adjacent atoms are linked by a CH2 or CH2CH2 group. A “bridged cycloalkyl” may be fused to one or more phenyl, partially unsaturated, or saturated rings. Examples of bridged carbocyclic groups include but are not limited to bicyclo[2.2. l]heptane etc. , bicyclo[2.2.2]octane, bicyclo[2.2.2]octene The term “carbonyl” as used herein refers to the radical -C(O)-. The term “cyano” as used herein refers to the radical -CN. The term “nitro” refers to the radical —N02. The term “H” refers to hydrogen.

The term “cycloalkoxy” as used herein refers to a cycloalkyl group attached to an oxygen (cycloalkyl—O).

The term “cycloalkyl” as used herein refers to a monocyclic saturated or partically unsatured hydrocarbon group of for example 3—6, or 4—6 s, referred to herein, e. g., as "C3. 6 cycloalkyl" or “C4_6 cycloalkyl," and derived from a cycloalkane. Exemplary cycloalkyl groups include, but are not limited to, cyclohexyl, cyclohexenyl, cyclopentyl, cyclobutyl, cyclopropyl or cyclopentyl.

The terms “halo” or “halogen” as used herein refer to F, Cl, Br, or I.

The terms “heteroaryl” as used herein refers to a clic aromatic 4—6 membered ring system containing one or more heteroatoms, for e one to three heteroatoms, such as nitrogen, oxygen, and sulfur. Where le, said heteroaryl ring may be linked to the adjacent radical though carbon or nitrogen. Examples of heteroaryl rings include but are not limited to furyl, thienyl, pyrrolyl, thiazolyl, oxazolyl, isothiazolyl, isoxazolyl, imidazolyl, pyrazolyl, triazolyl, oxadiazolyl (e.g., 1,2,4- oxadiazolyl or 1,3,4- oxadiazolyl), l, and pyrimidinyl.

The terms “heterocyclyl” or ocyclic group” are cognized and refer to saturated or partially unsaturated 4- to 7—membered ring structures, whose ring structures include one to three atoms, such as nitrogen, oxygen, and sulfur. A heterocycle may be fused to one or more phenyl, partially unsaturated, or saturated rings. Examples of heterocyclyl groups include but are not limited pyrrolidinyl, piperidinyl, morpholino, thiomorpholino, and zinyl.

The term ocyclylalkoxy” as used herein refers to a cyclyl- alkyl-O- group.

The term “heterocyclyloxyalkyl” refers to a heterocyclyl-O-alkyl- group.

[0032] The term “heterocycloxy” refers to a heterocyclyl-O- group. The term “cycloalkyloxy” refers to a lkyl—O— group.

The term “heteroaryloxy” referes to a heteroaryl-O— group.

The terms “hydroxy” and “hydroxyl” as used herein refers to the radical —OH.

The term “oxo” as used herein refers to the radical =0.

[0036] The term “nitrogen protecting group” or “amino protecting group” is art-recognized and as used herein refers to a chemical moiety that is covalently linked to a nitrogen atom of an amino (primary or secondary) group and that temporarily blocks the reactivity of the amino group during a synthetic step and is selectively removed once the synthetic step is complete.

Nitrogen protecting groups include, for example, 9-Fluorenylmethyloxycarbonyl (Fmoc), tert- butoxycarbonyl (Boc), carbobenzyloxycarbonyl (Cbz), p-methoxybenzyloxycarbonyl, acetyl, triﬂuoroacetyl, benzoyl, phthalimido, benzyl (Bn), p-methoxybenzyl, p-methoxyphenyl, 3,4— dimethoxybenzyl, triphenylmethyl, benzylidene, and p-toluenesulfonyl (Ts). In some embodiments, the nitrogen protecting group can have one of the following formulas: — C(O)OR31 or —C(O)R32 as defined herein. In certain embodiments, R31 is selected from the group consisting of: C1-C6 alkyl; C1-C6 haloalkyl; C2-C6 alkenyl; C2-C5 alkynyl; C3-C10 cycloalkyl, wherein the C3-C10 lkyl is ally substituted with from 1-3 independently selected C1-C3 alkyl; —CH2-C3—C10 cycloalkyl wherein the C3-C10 cycloalkyl is optionally substituted with from 1—3 independently selected C1-C3 alkyl, -CH2-phenyl, wherein the phenyl is optionally substituted with from 1-2 substituents independently selected from C1—C3 alkyl, C1-C3 haloalkyl, C1-C3 alkoxy, C1—C3 haloalkoxy, nitro, halo, SOzMe, cyano, and —OC(O)CH3; and —CH2-pyridy1. In certain embodiments, R32 is selected from the group consisting of: H; C1— C6 alkyl; C1-C6 haloalkyl; phenyl, wherein the phenyl is optionally substituted with from 1-2 substituents ndently selected from C1-C3 alkyl, C1—C3 haloalkyl, C1—C3 alkoxy, C1-C3 haloalkoxy, nitro, halo, SOzMe, cyano, and -OC(O)CH3; and pyridyl.

[0037] As used in the present disclosure, the term “partial NMDA receptor agonist” generally refers to a compound that is capable of binding to a glycine binding site of an NMDA receptor; at low trations a NMDA receptor agonist acts substantially as agonist and at high concentrations it acts substantially as an antagonist. These concentrations are experimentally ined for each and every “partial agonist.

[0038] “Pharmaceutically or pharmacologically acceptable” include molecular entities and itions that do not produce an adverse, allergic or other untoward reaction when administered to an , or a human, as appropriate. For human administration, ations should meet sterility, pyrogenicity, general safety and purity standards as required by FDA Ofﬁce of Biologics standards.

[0039] The term “pharmaceutically acceptable carrier” or “pharmaceutically able excipient” as used herein refers to any and all ts, dispersion media, coatings, ic and absorption delaying agents, and the like, that are ible with pharmaceutical administration. The use of such media and agents for pharmaceutically active substances is well known in the art. The compositions may also contain other active compounds providing mental, additional, or enhanced therapeutic functions.

The term “pharmaceutical composition” as used herein refers to a ition comprising at least one compound as sed herein formulated er with one or more pharmaceutically acceptable carriers.

“Individual,” “patient,” or “subject” are used interchangeably and include any animal, including mammals, preferably mice, rats, other rodents, s, dogs, cats, swine, cattle, sheep, horses, or primates, and most preferably humans. The compounds of the invention can be administered to a mammal, such as a human, but can also be administered to other mammals such as an animal in need of veterinary treatment, e.g., domestic animals (e.g., dogs, cats, and the like), farm animals (e. g., cows, sheep, pigs, horses, and the like) and laboratory animals (e. g., rats, mice, guinea pigs, and the like). The mammal treated in the methods of the invention is desirably a mammal in which ent e. g., of pain or depression is desired. ation” includes antagonism (e. g., inhibition), agonism, partial antagonism and/or partial agonism.

In the present speciﬁcation, the term “therapeutically effective amount” means the amount of the subject compound that will elicit the biological or medical response of a tissue, system, animal or human that is being sought by the researcher, veterinarian, medical doctor or other clinician. The compounds of the invention are administered in therapeutically effective amounts to treat a disease. Alternatively, a therapeutically effective amount of a compound is the ty required to achieve a desired therapeutic and/or prophylactic effect, such as an amount which s in lessening a symptom of sion.

The term aceutically acceptable salt(s)" as used herein refers to salts of acidic or basic groups that may be present in compounds used in the present compositions.

Compounds included in the present itions that are basic in nature are capable of forming a wide variety of salts with various inorganic and organic acids. The acids that may be used to prepare pharmaceutically able acid addition salts of such basic compounds are those that form non—toxic acid addition salts, i.e., salts containing pharmacologically acceptable anions, including but not d to malate, oxalate, chloride, e, iodide, nitrate, sulfate, bisulfate, phosphate, acid phosphate, isonicotinate, acetate, lactate, salicylate, e, tartrate, oleate, tannate, pantothenate, bitartrate, ascorbate, succinate, maleate, gentisinate, fumarate, gluconate, glucaronate, saccharate, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate and pamoate (i.e., l,l'-methylene-bis—(2—hydroxy naphthoate)) salts. Compounds included in the present compositions that are acidic in nature are capable of forming base salts with various pharmacologically acceptable cations. Examples of such salts include alkali metal or alkaline earth metal salts and, particularly, calcium, magnesium, sodium, lithium, zinc, potassium, and iron salts. Compounds included in the present compositions that include a basic or acidic moiety may also form pharmaceutically acceptable salts with various amino acids. The compounds of the disclosure may contain both acidic and basic groups; for example, one amino and one carboxylic acid group. In such a case, the compound can exist as an acid addition salt, a zwitterion, or a base salt.

The nds of the disclosure may contain one or more chiral s and/or double bonds and, therefore, exist as stereoisomers, such as geometric isomers, enantiomers or diastereomers. The term “stereoisomers” when used herein t of all geometric isomers, omers or diastereomers. These compounds may be designated by the symbols “R” or “S,” depending on the configuration of substituents around the stereogenic carbon atom. The present invention asses various stereoisomers of these compounds and mixtures thereof.

Stereoisomers e enantiomers and diastereomers. Mixtures of enantiomers or diastereomers may be designated “(::)” in lature, but the skilled artisan will recognize that a structure may denote a chiral center implicitly.

The nds of the disclosure may contain one or more chiral centers and/or double bonds and, therefore, exist as geometric isomers, enantiomers or diastereomers. The enantiomer and diastereomers may be ated by the symbols “(+),” “(—).” “R” or “S,” depending on the configuration of substituents around the stereogenic carbon atom, but the skilled artisan will recognize that a structure may denote a chiral center implicitly. ric isomers, resulting from the arrangement of substituents around a carbon—carbon double bond or arrangement of tuents around a lkyl or heterocyclic ring, can also exist in the compounds of the present invention. The symbol denotes a bond that may be a single, double or triple bond as described . Substituents around a carbon-carbon double bond are designated as being in the “Z” or “E” configuration wherein the terms “Z” and “E” are used in accordance with IUPAC standards. Unless otherwise speciﬁed, structures depicting double bonds encompass both the “E” and “Z” isomers. Substituents around a —carbon double bond alternatively can be referred to as “cis” or “trans,” where “cis” represents substituents on the same side of the double bond and “trans” represents substituents on opposite sides of the double bond. The arrangement of substituents around a carbocyclic ring can also be designated as “cis” or “trans.” The term “cis” represents substituents on the same side of the plane of the ring and the term “trans” ents substituents on opposite sides of the plane of the ring. es of compounds wherein the substituents are disposed on both the same and opposite sides of plane of the ring are designated “cis/trans.” The term “stereoisomers” when used herein consist of all geometric isomers, enantiomers or reomers. The present invention encompasses various stereoisomers of these nds and es thereof.

Individual enantiomers and diasteriomers of compounds of the present invention can be prepared synthetically from commercially available starting materials that contain asymmetric or stereogenic centers, or by preparation of racemic mixtures ed by resolution methods well known to those of ordinary skill in the art. These methods of resolution are exempliﬁed by (1) attachment of a mixture of enantiomers to a chiral auxiliary, separation of the resulting mixture of diastereomers by recrystallization or chromatography and liberation of the optically pure t from the auxiliary, (2) salt formation employing an optically active resolving agent, (3) direct separation of the mixture of optical enantiomers on chiral liquid tographic columns or (4) c resolution using steroselective chemical or enzymatic ts. Racemic mixtures can also be resolved into their component enantiomers by well—known methods, such as chiral-phase gas chromatography or crystallizing the compound in a chiral solvent. Stereoselective syntheses, a chemical or enzymatic reaction in which a single reactant forms an unequal mixture of stereoisomers during the creation of a new stereocenter or during the transformation of a pre-existing one, are well known in the art.

Stereoselective syntheses encompass both enantio— and reoselective ormations. For examples, see Carreira and Kvaemo, cs in Stereoseiective sis, Wiley—VCH: Weinheim, 2009.

The compounds disclosed herein can exist in solvated as well as unsolvated forms with pharmaceutically acceptable solvents such as water, ethanol, and the like, and it is ed that the invention e both solvated and unsolvated forms. In one embodiment, the compound is amorphous. In one embodiment, the compound is a single polymorph. In another embodiment, the compound is a mixture of polymorphs. In another embodiment, the compound is in a crystalline form.

The invention also embraces isotopically labeled compounds of the invention which are identical to those recited herein, except that one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Examples of isotopes that can be incorporated into compounds of the ion include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, ﬂuorine and chlorine, such as 2H, 3H, 13C, 14C, 15N, 18O, 17O, 31P, 32F, 35 S, 18F, and 36Cl, respectively. For example, a compound of the invention may have one or more H atom replaced with deuterium.

Certain ically—labeled disclosed compounds (e.g., those d with 3H and 14C) are useful in compound and/or substrate tissue bution assays. Tritiated (226., 3H) and carbon—l4 (i.e., 14C) isotopes are particularly preferred for their ease of preparation and detectability. Further, substitution with heavier isotopes such as deuterium (i.e., 2H) may afford certain therapeutic advantages resulting from greater metabolic stability (e.g., increased in viva half—life or reduced dosage ements) and hence may be preferred in some circumstances.

Isotopically labeled compounds of the invention can generally be prepared by following procedures ous to those disclosed in the e. g., Examples herein by substituting an isotopically labeled t for a non-isotopically labeled reagent.

The term “prodrug” refers to compounds that are transformed in vivo to yield a disclosed compound or a pharmaceutically able salt, hydrate or solvate of the compound.

The transformation may occur by various mechanisms (such as by esterase, amidase, phosphatase, oxidative and or reductive metabolism) in various locations (such as in the intestinal lumen or upon transit of the intestine, blood or liver). Prodrugs are well known in the art (for example, see Rautio, Kumpulainen, et al, Nature Reviews Drug Discovery 2008, 7, 255). For example, if a compound of the ion or a ceutically acceptable salt, hydrate or solvate of the compound contains a ylic acid functional group, a prodrug can comprise an ester formed by the replacement of the hydrogen atom of the acid group with a group such as (C1-C8)alkyl, (C2-C12)alkanoyloxymethyl, l—(alkanoyloxy)ethyl having from 4 to 9 carbon atoms, 1—methyl—l—(alkanoyloxy)—ethyl having from 5 to 10 carbon atoms, alkoxycarbonyloxymethyl having from 3 to 6 carbon atoms, l-(alkoxycarbonyloxy)ethyl having from 4 to 7 carbon atoms, 1—methyl—l—(alkoxycarbonyloxy)ethyl having from 5 to 8 carbon atoms, oxycarbonyl)aminomethyl having from 3 to 9 carbon atoms, l-(N-(alkoxycarbonyl)amino)ethyl having from 4 to 10 carbon atoms, 3-phthalidyl, 4-crotonolactonyl, gamma-butyrolacton-4—yl, di-N,N—(C1-C2)alkylamino(C2-C3)alkyl (such as B-dimethylaminoethyl), carbamoyl-(Cl—C2)alkyl, N,N—di(C1-C2)alkylcarbamoyl-(C1-C2)alkyl and dino-, pyrrolidino- or morpholino(C2-C3)alkyl.

Similarly, if a compound of the invention contains an alcohol functional group, a prodrug can be formed by the replacement of the hydrogen atom of the alcohol group with a 2014/013621 group such as (C1—C6)alkanoyloxymethyl, 1—((C1-C6)a1kanoyloxy)ethyl, 1 -methy1((C1-C6)alkanoyloxy)ethyl (C1-C6)alkoxycarbonyloxymethyl, N—(C1-C6)alkoxycarbonylaminomethyl, succinoyl, (C1-C6)alkanoyl, d-amino(C1-C4)alkanoyl, arylacyl and (x-aminoacyl, or (x-aminoacyl-(x-aminoacyl, where each or-aminoacyl group is independently selected from the naturally occurring L-amino acids, P(O)(OH)2, -P(O)(O(C1-C6)alkyl)2 or glycosyl (the radical resulting from the removal of a hydroxyl group of the hemiacetal form of a carbohydrate).

If a compound of the invention incorporates an amine functional group, a prodrug can be formed, for example, by creation of an amide or carbamate, an N—acyloxyakyl derivative, an (oxodioxolenyl)methyl tive, an N—Mannich base, imine or enamine. In on, a secondary amine can be metabolically cleaved to generate a bioactive primary amine, or a tertiary amine can metabolically cleaved to te a bioactive primary or ary amine. For examples, see Simplicio, et al., Molecules 2008, 13, 519 and references therein.

Compounds Disclosed compounds include those represented by formula I: R1 L%N_R5Rb\ N I R3 0 (I) and ceutically acceptable salts, stereoisomers, and es thereof, wherein Rb is selected from the group consisting of H, halogen, hydroxyl, cyano or C1-C6 alkyl; R1 is H or C1-C6 alkyl; R2 is H or C1-C6 alkyl; R3 is selected from the group consisting of H, C1-C6 alkyl and a nitrogen protecting group; R4 is H or C1-C5 alkyl; R5 is X or —C1-C6 alkylene—X, wherein X is selected from the group consisting of phenyl, a 4- to 6—membered heteroaryl ring having 1, 2, or 3 heteroatoms selected from O, S, or N, or a 4- to 6-membered heterocyclyl ring having 1, 2, or 3 heteroatoms selected from O, S, or N, and wherein R5 is optionally substituted with R6; and R6 is ed from the group ting of H, halogen, hydroxyl, cyano, -O—C(O)-C1-C6 alkyl, C1—C6 alkyl, or C1-C6 alkoxy; or, in other embodiments, the variables set forth in formula (I) are deﬁned as follows: Rb is selected from the group consisting of H, halogen, hydroxyl, cyano or C1-C6 alkyl (6%., H); R1 is H or C1-C6 alkyl; R2 is H or C1-C6 alkyl; R3 is selected from the group consisting of H, C1-C6 alkyl and a nitrogen protecting group; R4 is H or C1-C6 alkyl; R5 is X or —C1—C6 alkylene-X, wherein X is selected from the group consisting of: (i) phenyl; (ii) heteroaryl including from 5 to 6 ring atoms wherein 1, 2, or 3 of the ring atoms are ndently selected from the group consisting of N, NH, N(C1-C3 alkyl), 0, and S; (iii) cyclyl including from 3 to 6 ring atoms wherein 1, 2, or 3 of the ring atoms are independently selected from the group consisting ofN, NH, N(C1-C3 , 0, and S; wherein R5 is ally substituted with R6; and R6 is selected from the group consisting of H, halogen, hydroxyl, cyano, —O—C(O)—C1—C6 alkyl, C1—C6 alkyl, or C1-C6 alkoxy.

In certain embodiments, R1 is H.

In certain embodiments, R2 is H.

[0057] In certain embodiments, R3 is H.

In other embodiments, R3 is a nitrogen protecting group. In certain ments, R3 has formula —C(O)OR31, wherein R31 is selected from the group consisting of: C1-C6 alkyl; WO 20784 —16- C1—C6 haloalkyl; C2-C6 alkenyl; C2—C6 alkynyl; C3—C10 cycloalkyl, wherein the C3-C10 cycloalkyl is optionally substituted with from 1—3 independently selected C1—C3 alkyl; —CH2-C3— C10 lkyl wherein the C3-C10 cycloalkyl is optionally substituted with from 1-3 independently selected C1-C3 alkyl; -CH2-phenyl, wherein the phenyl is optionally substituted with from 1-2 substituents independently selected from C1-C3 alkyl; C1-C3 kyl; C1-C3 alkoxy; C1-C3 haloalkoxy; nitro; halo; SO2Me, cyano; and -OC(O)CH3; and -CH2-pyridyl. In n embodiments, R31 is C1—C6 alkyl (e.g., utyl). In other embodiments, R3 has formula —C(O)R32, wherein R32 is ed from the group consisting of: H; C1—C6 alkyl; C1—C6 haloalkyl; phenyl, wherein the phenyl is optionally substituted with from 1—2 substituents independently selected from C1—C3 alkyl; C1—C3 haloalkyl; C1-C3 alkoxy; C1—C3 haloalkoxy; nitro; halo; SO2Me, cyano; and -OC(O)CH3; and pyridyl. In certain embodiments, R32 is C1-C6 alkyl (e.g., —CH3 or iso—propyl).

In certain embodiments, R4 is C1—C6 alkyl. In some embodiments, R4 is methyl.

In certain embodiments, Rb is H.

[0061] In certain embodiments, R5 is X, and wherein X is a 5- to 6-membered heteroaryl ring ed from the group consisting of azetidine, pyrrolidine, pyrazolidine, pyridine, pyrimidine, isooxazolidine, imidazolidine, oxazolidine, thiazolidine, and isothiazolidine. In §—<\ 4 other embodiments, R5 is N or HD. In n other embodiments, R5 is X, and wherein X is ally substituted with R6 imes referred to herein as “— CH(R4)(R6)”)- In some embodiments, R5 is X. In certain embodiments, X is heteroaryl including from 5 to 6 ring atoms wherein l, 2, or 3 of the ring atoms are independently selected from the group consisting ofN, NH, N(C1-C3 alkyl), 0, and S. For example, X can be selected from the group consisting of 1,2,4— oxadiazolyl, 1,3,4- oxadiazolyl, pyridyl, and pyrimidinyl. In certain embodiments, R5 (here, X as deﬁned anywhere herein) is substituted with —CH(R4)(R6). In certain embodiments, R4 is C1-C6 alkyl (e.g., R4 is methyl). In certain embodiments, R6 is selected from the group consisting of hydroxyl, -O-C(O)—C1-C5 alkyl, and C1-C6 alkoxy (e.g., R6 is hydroxyl). Embodiments in which R5 is X can include one or more of the ing es: R1 is H or methyl; R2 is H or methyl; R3 is H, —C(O)OR31, or —C(O)R32; when present, R4 is C1—C6 alkyl (e. g., R4 is methyl), and R6 is selected from the group ting of hydroxyl, -O—C(O)-C1—C6 alkyl, and C1-C6 alkoxy (e. g., R6 is hydroxyl); Rb is H.

In some embodiments, R5 is —C1—C6 alkylene—X (e.g., —C1—C2 ne-X or —C1 alkylene—X). In n embodiments, X is heteroaryl including from 5 to 6 ring atoms wherein 1, 2, or 3 of the ring atoms are independently selected from the group consisting of N, NH, N(C1—C3 alkyl), 0, and S. For example, X can be selected from the group consisting of 1,2,4- oxadiazolyl, 1,3,4- oxadiazolyl, l, and pyrimidinyl. In certain embodiments, R5 is substituted on either the X portion or the alkylene chain portion with —CH(R4)(R6). In certain of these embodiments, R5 is substituted on the alkylene portion with )(R6), and R5 can have, for example, the formula —CH(CHR4R6)-X. In certain embodiments, R4 is C1-C6 alkyl (e. g., R4 is methyl). In certain embodiments, R6 is selected from the group consisting of hydroxyl, )—C1—C6 alkyl, and C1-C6 alkoxy (e.g., R6 is hydroxyl). ments in which R5 is —C1—C6 alkylene-X can include one or more of the following features: R1 is H or methyl; R2 is H or methyl; R3 is H, —C(O)OR31, or —C(O)R32; when present, R4 is C1—C6 alkyl (e. g., R4 is methyl), and R6 is selected from the group consisting of hydroxyl, —O—C(O)—C1—C6 alkyl, and C1-C6 alkoxy (e. g., R6 is hydroxyl); Rb is H.

[0064] In certain ments, R6 is hydroxyl.

In some embodiments, a sed compound includes those delineated in Table 1 and/or the Examples, e.g., one having the formula: O,N\> N \ N N—<—\N N.” N / 0 N N OH H ., H o OH O 'OH O 01‘ 7 7 ‘ Disclosed compounds also include compounds ented by formula II: Rs 0 (11) and pharmaceutically acceptable salts, stereoisomers, and N—oxides thereof, wherein R3 is selected from the group consisting of H, C1—C6alkyl and a nitrogen protecting group; and R7 is a 4- to 6-membered heteroaryl ring having 1, 2, or 3 heteroatoms selected from O, S, or N, optionally substituted on a free carbon by a substituent ed from the group consisting of: halogen, C1—C6alkyl, hydroxyl, cyano, and phenyl.

WO 20784 —18- Also disclosed are compounds represented by formula III: I‘ N X4R4 R 1 | R6 R3 0 (111) and pharmaceutically acceptable salts, stereoisomers, and N—oxides thereof, wherein Rb is selected from the group consisting of H, halogen, yl, cyano or C1-C6 alkyl; R1 is H or C1-C6 alkyl; R2 is H or C1-C6 alkyl; R3 is selected from the group consisting of H, C1-C6 alkyl and a nitrogen protecting group; R4 is H or C1-C6 alkyl; X is selected from the group consisting of phenyl, a 4- to 6—membered heteroaryl ring having 1, 2, or 3 heteroatoms selected from O, S, or N, or a 4— to 6—membered heterocyclyl ring having 1, 2, or 3 heteroatoms ed from O, S, or N, wherein R6 is present on a free carbon of X; and R6 is selected from the group consisting of H, halogen, hydroxyl, cyano, -O-C(O)-C1-C6 alkyl, C1-C6 alkyl, or C1-C6 alkoxy.

In certain embodiments, R1 is H.

In n ments, R2 is H.

In some ments, R3 is H.

In certain embodiments, R4 is C1-C6 alkyl. In other embodiments, R4 is methyl.

[0072] In some embodiments, Rb is H.

In certain embodiments, X is N—gﬂ In some embodiments, R6 is hydroxyl.

The compounds of the present disclosure and formulations thereof may have a plurality of chiral centers. Each chiral center may be independently R, S, or any mixture ofR and S. For example, in some embodiments, a chiral center may have an R:S ratio ofbetween WO 20784 about 100:0 and about 50:50, n about 100:0 and about 75:25, between about 100:0 and about 85:15, between about 100:0 and about 90: 10, between about 100:0 and about 95:5, between about 100:0 and about 98:2, between about 100:0 and about 99:1, between about 0:100 and 50:50, n about 0: 100 and about 25:75, between about 0:100 and about 15:85, between about 0: 100 and about 10:90, between about 0: 100 and about 5:95, between about 0: 100 and about 2:98, between about 0: 100 and about 1:99, between about 75:25 and 25:75, and about 50:50. Formulations of the disclosed compounds comprising a greater ratio of one or more s (i.e., R and/or 53 may possess enhanced therapeutic characteristic relative to racemic ations of a disclosed compounds or mixture of compounds. In some instances, chemical formulas contain the descriptor “-(R)-“ or “-(S)—“ that is further attached to solid wedge or dashed wedge. This descriptor is ed to show a methine carbon (CH) that is attached to three other substituents and has either the indicated R or S conﬁguration (see, e.g., Table 1).

Disclosed compounds may provide for efﬁcient cation channel opening at the NMDA receptor, e.g. may bind or associate with the glutamate site of the NMDA receptor to assist in opening the cation channel. The disclosed compounds may be used to regulate (turn on or turn off) the NMDA receptor through action as an agonist.

The compounds as described herein may be glycine site NMDA receptor l agonists. A l agonist as used in this context will be understood to mean that at a low concentration, the analog acts as an agonist and at a high concentration, the analog acts as an antagonist. Glycine g is not inhibited by glutamate or by itive inhibitors of glutamate, and also does not bind at the same site as glutamate on the NMDA receptor. A second and separate binding site for glycine exists at the NMDA receptor. The -gated ion channel of the NMDA receptor is, thus, under the control of at least these two distinct allosteric sites. Disclosed nds may be capable of binding or associating with the glycine binding site of the NMDA receptor. In some embodiments, disclosed compounds may possess a potency that is 10-fold or greater than the activity of existing NMDA receptor e site partial agonists.

The disclosed compounds may exhibit a high therapeutic index. The therapeutic index, as used herein, refers to the ratio of the dose that produces a toxicity in 50% of the population (i.e., TD50) to the minimum effective dose for 50% of the population (i.e., EDso).

Thus, the therapeutic index = (TDSO):(ED50). In some embodiments, a disclosed compound may have a therapeutic index of at least about 10:1, at least about 50:1, at least about 100:1, at least about 200: 1, at least about 500:1, or at least about 1000: l.

Compositions In other aspects, formulations and compositions comprising the disclosed compounds and optionally a pharmaceutically acceptable ent are provided. In some embodiments, a contemplated ation comprises a racemic mixture of one or more of the disclosed nds.

Contemplated formulations may be prepared in any of a variety of forms for use.

By way of example, and not limitation, the compounds may be prepared in a ation suitable for oral administration, subcutaneous injection, or other methods for administering an active agent to an animal known in the pharmaceutical arts.

Amounts of a disclosed compound as described herein in a formulation may vary according to factors such as the disease state, age, sex, and weight of the dual. Dosage regimens may be adjusted to provide the optimum therapeutic response. For example, a single bolus may be administered, several divided doses may be administered over time or the dose may be proportionally d or increased as indicated by the exigencies of the therapeutic ion. It is especially advantageous to formulate eral compositions in dosage unit form for ease of stration and uniformity of dosage. Dosage unit form as used herein refers to physically discrete units suited as unitary dosages for the mammalian subjects to be treated; each unit containing a predetermined quantity of active compound calculated to produce the desired therapeutic effect in association with the ed pharmaceutical carrier.

The speciﬁcation for the dosage unit forms of the invention are dictated by and directly ent on (a) the unique characteristics of the compound selected and the particular therapeutic effect to be achieved, and (b) the limitations inherent in the art of compounding such an active compound for the treatment of sensitivity in individuals.

Therapeutic compositions typically must be sterile and stable under the conditions of manufacture and e. The ition can be formulated as a solution, microemulsion, liposome, or other ordered structure suitable to high drug concentration. The carrier can be a solvent or sion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene , and the like), and suitable mixtures thereof. The proper ﬂuidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants. In many cases, it will be preferable to include isotonic agents, for example, WO 20784 sugars, polyalcohols such as mannitol, sorbitol, or sodium chloride in the composition.

Prolonged absorption of the inj ectable compositions can be brought about by including in the composition an agent which delays absorption, for example, monostearate salts and gelatin.

The compounds can be administered in a time release formulation, for e in a composition which includes a slow e polymer. The compounds can be prepared with carriers that will protect the compound against rapid release, such as a controlled release formulation, including implants and microencapsulated ry systems. Biodegradable, patible polymers can be used, such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, thoesters, polylactic acid and polylactic, polyglycolic copolymers (PLG). Many methods for the preparation of such formulations are generally known to those skilled in the art.

Sterile inj ectable solutions can be ed by orating the compound in the required amount in an appropriate solvent with one or a combination of ingredients enumerated above, as required, followed by ed sterilization. Generally, dispersions are prepared by incorporating the active compound into a sterile vehicle which contains a basic dispersion medium and the required other ingredients from those enumerated above. In the case of sterile powders for the preparation of sterile inj ectable ons, the preferred methods of preparation are vacuum drying and freeze-drying which yields a powder of the active ingredient plus any additional desired ient from a previously e-filtered solution thereof.

[0086] In accordance with an alternative aspect of the invention, a compound may be formulated with one or more additional compounds that enhance the solubility of the compound.

Methods Methods for treating a condition in a patient in need thereof by administering a therapeutically effective dose of a compound described herein are provided. In some embodiments, the ion may be a mental ion. For e, a mental illness may be treated. In another aspect, a nervous system condition may be d. For example, a condition that affects the central nervous system, the peripheral nervous system, and/or the eye may be treated. In some embodiments, neurodegenerative diseases may be treated.

[0088] In some embodiments, the methods include administering a nd to treat patients suffering from autism, anxiety, depression, bipolar disorder, attention deﬁcit disorder, attention deficit hyperactivity disorder (ADHD), schizophrenia, a psychotic disorder, a psychotic symptom, social withdrawal, obsessive—compulsive disorder (OCD), phobia, post— traumatic stress me, a behavior disorder, an impulse control disorder, a substance abuse disorder (e. g., a Withdrawal symptom, opiate addiction, nicotine addiction, and ethanol addition), a sleep disorder, a memory disorder (e. g., a deficit, loss, or reduced ability to make new memories), a learning disorder, urinary inence, multiple system atrophy, progressive supra-nuclear palsy, ich's ataxia, Down’s syndrome, fragile X me, tuberous sclerosis, olivio—ponto—cerebellar atrophy, cerebral palsy, drug-induced optic neuritis, ischemic retinopathy, ic retinopathy, ma, dementia, AIDS dementia, Alzheimer’s e, Huntington’s chorea, spasticity, myoclonus, muscle spasm, Tourette's syndrome, sy, cerebral ischemia, stroke, a brain tumor, traumatic brain injury, c arrest, athy, spinal cord , peripheral neuropathy, acute neuropathic pain, and chronic neuropathic pain.

In some embodiments, methods of treating a memory er associated with aging, schizophrenia, special learning disorders, seizures, post—stroke convulsions, brain ischemia, hypoglycemia, cardiac arrest, epilepsy, migraine, AIDS ia, Huntington’s chorea, Parkinson’s disease, early stage Alzheimer’s disease, and Alzheimer’s disease are contemplated.

In certain embodiments, methods for treating schizophrenia are provided. For example, paranoid type schizophrenia, disorganized type schizophrenia (i.e., hebephrenic schizophrenia), catatonic type schizophrenia, undifferentiated type schizophrenia, residual type schizophrenia, post-schizophrenic depression, and simple phrenia may be treated using the s and compositions contemplated herein. Psychotic disorders such as schizoaffective disorders, delusional disorders, brief psychotic disorders, shared psychotic disorders, and psychotic disorders with delusions or hallucinations may also be treated using the compositions contemplated herein.

Paranoid schizophrenia may be characterized where delusions or auditory hallucinations are present, but thought disorder, disorganized behavior, or affective ﬂattening are not. Delusions may be persecutory and/or grandiose, but in addition to these, other themes such as jealousy, religiosity, or somatization may also be present. Disorganized type schizophrenia may be characterized where thought disorder and ﬂat affect are t together.

Catatonic type schizophrenia may be characterized where the patient may be almost immobile or exhibit agitated, purposeless movement. Symptoms can include nic stupor and waxy ﬂexibility. erentiated type schizophrenia may be terized where psychotic symptoms are present but the criteria for id, disorganized, or catatonic types have not been met. Residual type schizophrenia may be characterized where positive symptoms are present at a low intensity only. Post-schizophrenic depression may be characterized where a depressive episode arises in the aftermath of a phrenic illness where some low—level schizophrenic symptoms may still be present. Simple schizophrenia may be characterized by insidious and progressive development of prominent negative symptoms with no history of psychotic episodes.

In some embodiments, methods are provided for treating psychotic symptoms that may be present in other mental ers, including, but not limited to, r disorder, line ality disorder, drug intoxication, and nduced psychosis. In another embodiment, methods for treating delusions (e. g., "non-bizarre") that may be present in, for e, delusional disorder are provided.

Also provided are methods for treating social withdrawal in conditions including, but not limited to, social anxiety disorder, avoidant personality disorder, and schizotypal personality disorder.

In some embodiments, methods are provided for ng neuropathic pain. The neuropathic pain may be acute or chronic. In some cases, the neuropathic pain may be associated with a condition such as herpes, HIV, traumatic nerve injury, stroke, schemia, ﬁbromyalgia, reﬂex sympathetic dystrophy, x regional pain syndrome, spinal cord injury, sciatica, phantom limb pain, diabetic neuropathy, and cancer chemotherapeutic—induced neuropathic pain. Methods for enhancing pain relief and for ing analgesia to a patient are also contemplated.

Further contemplated methods include a method of ng autism and/or an autism spectrum disorder in a patient need thereof, comprising administering an effective amount of a compound to the patient. In an embodiment, a method for reducing the symptoms of autism in a patient in need thereof is contemplated, comprising administering an ive amount of a disclosed compound to the t. For example, upon administration, the compound may se the incidence of one or more symptoms of autism such as eye contact avoidance, failure to socialize, attention deficit, poor mood, hyperactivity, abnormal sound sensitivity, inappropriate speech, disrupted sleep, and perseveration. Such decreased incidence may be measured relative to the incidence in the untreated individual or an untreated individual(s).

[0096] Also provided herein is a method of modulating an autism target gene expression in a cell comprising contacting a cell with an effective amount of a nd described herein.

The autism gene expression may be for example, selected from ABAT, APOE, CHRNA4, GABRA5,GFAP, GRINZA, PDYN, and PENK. In another embodiment, a method of modulating synaptic plasticity in a patient suffering from a ic plasticity related disorder is provided, comprising administering to the patient an effective amount of a compound.

In another ment, a method of ng Alzheimer’s disease, or e.g., treatment of memory loss that e.g., accompanies early stage Alzheimer’s disease, in a patient in need thereof is provided, comprising administering a compound. Also provided herein is a method of modulating an mer’s amyloid protein (e.g., beta amyloid peptide, e. g. the isoform A6142), in-vitro or in—vivo (e. g. in a cell) comprising contacting the protein with an ive amount of a compound is disclosed. For example, in some embodiments, a nd may block the ability of such amyloid protein to inhibit long-term potentiation in hippocampal slices as well as apoptotic neuronal cell death. In some embodiments, a disclosed compound may e neuroprotective properties to a Alzheimer’s patient in need thereof, for example, may provide a eutic effect on later stage Alzheimer’s —associated neuronal cell death.

In a further embodiment, a method of treating depression comprising administering a compound described herein is provided. In some embodiments, the treatment may e depression or a m of depression without affecting behavior or motor coordination and without inducing or promoting seizure activity. Exemplary sion conditions that are expected to be treated according to this aspect of the invention include, but are not limited to, major depressive disorder, dysthymic disorder, psychotic depression, postpartum depression, premenstrual syndrome, premenstrual dysphoric disorder, seasonal affective disorder (SAD), bipolar disorder (or manic sive er), mood disorder, and depressions caused by chronic medical ions such as cancer or c pain, herapy, chronic stress, and post traumatic stress disorders. In addition, patients suffering from any form of depression often experience anxiety. Various symptoms associated with anxiety include fear, panic, heart palpitations, shortness of breath, fatigue, nausea, and headaches among others. Anxiety or any of the symptoms thereof may be d by administering a compound as described herein.

Also provided herein are methods of treating a condition in treatment-resistant patients, e.g., patients suffering from a mental or central nervous system condition that does not, and/or has not, responded to adequate courses of at least one, or at least two, other compounds or therapeutics. For example, provided herein is a method of treating depression in a treatment resistant patient, comprising a) optionally identifying the patient as treatment resistant and b) administering an effective dose of a compound to said patient.

In some embodiments, a compound described herein may be used for acute care of a patient. For example, a compound may be administered to a patient to treat a particular episode (e.g., a severe e) of a condition contemplated herein.

Also contemplated herein are ation therapies sing a compound in combination with one or more other active agents. For example, a compound may be combined with one or more antidepressants, such as tricyclic antidepressants, MAO-1's, SSRI's, and double and triple uptake inhibitors and/or anxiolytic drugs. Exemplary drugs that may be used in combination with a compound include nil, , Aventyl, Elavil, Norpramin, Pamelor, Pertofrane, Sinequan, Surmontil, il, Vivactil, Parnate, , Marplan, , Lexapro, Luvox, Paxil, Prozac, Zoloft, Wellbutrin, Effexor, Remeron, Cymbalta, l (trazodone), and Ludiomill. In another e, a compound may be combined with an antipsychotic medication. Non-limiting examples of antipsychotics include butyrophenones, phenothiazines, thioxanthenes, clozapine, olanzapine, risperidone, quetiapine, ziprasidone, amisulpride, asenapine, paliperidone, iloperidone, zotepine, sertindole, lurasidone, and aripiprazole. It should be understood that combinations of a compound and one or more of the above eutics may be used for treatment of any suitable condition and are not limited to use as antidepressants or antipsychotics.

EXAMPLES The following examples are provided for illustrative purposes only, and are not intended to limit the scope of the disclosure.

[00103] Table 1 below shows some exemplary compounds of the disclosure and provides physiochemical characteristics of the compounds.

Table 1.

Compound Structure Molecular Weight (D3) Compound 252 nd 262 —0.5?’ 78.35 Y \> Compound —0. 15 78.35 C-16 252.2697 C—4 262.3076 -O.57075 «N N K ‘ a. .'>' M" ' C—15 352.3856 0.144989 109 WO 20784 -27_ C—20 276.3342 — 78.35 0.160256 C—36 276.3342 — 78.35 0.160256 C-33 290.3608 0.250239 78.35 C—10 252.2697 —1.83479 91.49 0.551597 362.4234 0.672806 95.86 366.4121 - 0.135022 1.0833 95.86 _ 29 _ C—26 366.4121 — 109 0.135022 C—29 1.0833 95.86 C-32 390.4766 1.4938 95.86 C—11 322.3596 — 99.77 ’08 0.981326 *6? -m:§$i3 “M“ Kw“) “Ti {if \gﬁ‘f .«~' '< E?! \M“ "J {3:3 ‘9‘“ gig “j? kt?” {a a} Wﬂx‘mﬂ' C-5 75 0.335914 86.63 C—37 346.424 0.746408 86.63 Exam 1e 1 — S nthesis of Com ound X Scheme 1. 0H Step 1 Step 2 OMe step 3 0 Step 4 OH N N N N N Boc o OMe Pd C/H- OMe H . 0 LiHMDS, O SOCIZ’ MeOH 2 O 8°C Bom-Cl 806/ O Boc/ O 8M1 1 2 3 4 ,N [N O O o—N \> \> Step 5 ‘0 Step 6 N “I, \ \> Step 7 ‘N Step 5 ‘N N N-., —. N Nu.

N BOG] I N —» OMe I—tF> ’ H DMP 8°C/ n- B0C o OMe ., 0 OH TFA O "'OH 0 ,OH 6 7 Compound X 0 o o H OMNHQ N_ NHACI. EDCI N.

Boc Boc Step E —> O AcOH Synthesis of 2-ggtert-butoxycarbonyl) amino )—3-hydroxybutanoic acid {A} To a stirring solution of 2-amino-3 -hydroxybutanoic acid (SM2) (10 g, 83.9 mmol) in oxane/water (100 mL, 1: 1)) was added NaHCO3 (21.1 g, 0.25 mol) followed by Boc— anhydride (21.9 mL, 0.101 mol) at 0°C. The reaction mixture was stirred at RT for 16 h. After consumption of the starting material (by TLC), the reaction mixture was diluted with water and washed with EtOAc. The aqueous layer was ed using citric acid solution (pH~3-4) and then extracted with CHZCIZ (2 x 150 mL). The separated organic extracts were dried over anhydrous NaZSO4, ﬁltered and concentrated under vacuum to afford A (15 g, crude). This material was directly used for the next step without further puriﬁcation.

S nthesis of 3- tert—butox carbon 1-2 2 5-trimeth lidinecarbox lic acid B To a stirring solution ofA (15 g, 59.28 mmol) in THF (150 mL) was added PPTS (1.47 g, 5.92 mmol) followed by 2,2—dimethoxy propane (21.79 mL, 0.17 mol) at 0°C under N2 here. The on mixture was stirred at RT for 16 h. The on mixture was again heated to reﬂux for 6 h. The reaction mixture was diluted with aqueous NaHCO3 solution and washed with EtOAc. The aqueous layer was acidiﬁed using citric acid solution (pH~2) and extracted with CHzClz (2x 100 mL). The organic layer was washed with brine, dried over anhydrous Na2S04 and concentrated under vacuum to afford B (18 g, crude). 1H-NMR: (400 MHz, DMSO—d6): 5 13.25 (br s, 1H), 4.11—4.05 (m, 1H), 3.79 (d, 1H), 1.50 (s, 3H), 1.67 (s, 3H), 1.45 (s, 9H), 1.29 (d, 3H).

Synthesis of tert-butyl 4-carbamoyl-2, 2, ethyloxazolidine—3-carb0xylate [C] To a stirring on of B (18 g, 69.4 mmol) in CHzClz (180 mL) was added HOBt (14.16 g, 0.104 mol), EDCI.HCl (19.88 g, 0.104 mol) followed by NH4C1 (5.56 g, 0.104 mol) and DIPEA (31.9 mL, 0.173 mol) at 0°C. The reaction mixture was stirred at RT for 16 h. After ption of the starting material (by TLC), the reaction mixture was washed with s citric acid, NaHCO3 followed by brine. The organic layer was dried over anhydrous NaZSO4 and concentrated under reduced pressure. The resulting crude material was purified by silica gel column chromatography eluting with 2% MeOH/CHzClz to afford C (13 g, 72.5%). 1H-NMR: (400 MHz, DMSO-ds): 5 7.51 (br s, 1H), 7.14 (br s, 1H), 3.97-3.95 (m, 1H), 3.71 (d, 1H), 1.51 (d, 6H), 1.34 (s, 9H), 1.24 (d, 3H).

LCMS (ESI): 159.1[(M++l)-Boc] S nthesis of Z -tert-but l4— dimeth lamino meth lene carbamo 1-2 2 5- trimethyloxazolidine—3-carboxylate ] D 1 A solution of C (13 g, 50.3 mmol) in DMF.DMA (130 mL) was stirred at reﬂux temperature for 3 h under N2 atmosphere. After consumption of the starting material (by TLC), the reaction mixture was concentrated under reduced pressure to afford D (15.7 g, . This crude material was directly taken for the next step without further purification.

S nthesis of tert-bu 12 2 5—trimeth l 1 2 iazol—5- l 0xazolidinecarbox late To a stirring on of D (15.7 g, 50.09 mmol) in ethanol (157 mL) was added hydroxylamine hydrochloride (6.96 g, 0.10 mol) under N2 atmosphere. The reaction e was heated to reﬂux and stirred for 2 h. After consumption of the starting al (by TLC), acetic acid (28.6 mL, 0.50 mol) was added to the on mixture and then reﬂuxed for 16 h.

Solvents from the reaction mixture were evaporated under vacuum. The resulting crude material was puriﬁed by silica gel column chromatography eluting with 10% EtOAc/Hexane to afford E (4.5 g, 32%). 1H-NMR: (400 MHz, DMSO—d6): 5 6.35 (s, 2H), 4.61 (d, 1H), 4.22—4.15 (m, 1H), 1.55 (s, 6H), 1.37 (s, 2H), 1.25 (d, 3H), 1.21 (s, 6H).

LCMS (E81): 284 [M++1] Mass (m/z): 283 [Ml] S nthesis of l-amino-l- 1 2 4-oxadiazol 1 r0 an-Z-ol Int-F To a stirring solution of E (5 g, 17.6 mmol) in water (25 mL) was added triﬂuoroacetic acid (25 mL). The reaction mixture was stirred at RT for 5 h. After consumption of the starting material (by TLC), the reaction mixture was concentrated under vacuum. The residue was dissolved in water and neutralized with aqueous NaHCO3. The solvents from the reaction mixture were evaporated under vacuum and extracted with 5% MeOH/CHzClz (4x 150 mL). The organic layer was concentrated under reduced pressure to afford Int-F (2.5 g, crude). 1H—NMR: (400 MHz, D20): 8 8.84 (s, 1H), 4.05 (d, 1H), 3.98—3.95 (m, 1H), 3.67 (s, 1H), 3.58 (d, 1H), 1.15 (d, 3H), 1.12 (d, 3H).

LCMS (ESI): 144.1 [M++1] S nthesis of meth l ine-Z-carbox late 1 To a stirring solution of idinecarboxy1ic acid (SMl) (100 g, 0.87 mol) in methanol (800 mL) was added thionyl de (76.9 mL, 104 mol) slowly drop wise at 0°C.

The reaction mixture was heated to reﬂux for 12 h. After consumption of the starting material (by TLC), the reaction was trated under vacuum. Obtained residue was washed with n— Hexane and distilled off the solvent to afford 1 (143.9 g, HCl salt). : (400 MHz, CDCl3) (Rotamers): 5 3.89 (s, 3H), 3.68—3.62 (m, 2H), 3.59—3.47 (m, 2H), 2.49—2.37 (m, 1H), 2.27—2.05 (m, 3H).

LCMS (E81): 166 [M++1] S nthesis of l-tert-but 12-meth l rrolidine—l 2-dicarb0x late 2 2014/013621 To a stirring solution of 1 (35 g, 0.22 mol) in CHzClz (175 mL) were added Et3N (90 mL, 0.65 mol) followed by hydride (56.9 mL, 0.26 mol) at 0°C. The reaction mixture was stirred at RT for 16 h. After consumption of the starting material (by TLC), the reaction was diluted with water (100 mL) and extracted with CHzClz (2x 100 mL). The organic layer was washed with water, brine, dried over NaQSO4 and concentrated. Obtained crude material was d by silica gel column chromatography eluting with 30% EtOAc/Hexane to afford 2 (41 g, 95%). 1H-NMR: (400 MHz, CDC13) (Rotamers): 5 4.25-4.21 (m, 1H), 3.75 (s, 3H), .26 (m, 2H), 2.29-2.10 (m, 1H), 1.99—1.75 (m, 3H), 1.45 (s, 9H).

LCMS (E81): 130 [(M++1)—Boc] S s of -but 12-meth 12- ben 10x meth l rrolidine—l rb0x late To a ng solution of 2 (100 g, 0.43 mol) in THF (800 mL) was added LiHMDS (873 mL, 0.87 mol) at —78°C and stirred for 1 h. To this BOM—chloride (93.2 mL, 0.65 mol) was added drop wise at -78°C and stirred for 2 h at —20 °C. After consumption of the starting material (by TLC), the reaction was quenched with NH4C1 at 0 °C. The separated organic layer was washed with water, dried over Na2S04 and concentrated to afford 3 (180 g, crude). This material was directly taken for the next step without further cation.

LCMS (E81): 250 [(M++1)—Boc] S nthesis of l-tert-bu 12-meth 12- h drox meth l rrolidine—l 2-dicarb0x late 4 ] To a stirring solution of 3 (74 g, 0.21 mol) in methanol (740 mL) was added 10% Pd/C (50% wet, 14.8 g) under N2 atmosphere and stirred for 6 h under H2 atmosphere (balloon pressure). The reaction mixture was filtered through celite pad and concentrated under reduced pressure to afford 4 (45 g, 82%) as crude.

S nthesis of l-tert-but l2-meth lZ-form l rrolidine—l 2-dicarb0x late 5 To a stirring solution of 4 (10 g, 38.5 mmol) in CHzClz (100 mL) was added Dess- martin periodinane (19.6 g, 46.27 mmol) at 0°C under N2 atmosphere and stirred for 3 h. After consumption of the starting material (determined by TLC), the reaction was quenched with aqueous NaHCO3 solution and extracted with CHzClz (2x 100 mL). The organic layer was dried over anhydrous NaZSO4 and concentrated under vacuum. The resulting crude material was puriﬁed by column chromatography g with 10% EtOAc/Hexane to afford 5 (7 g, S 'nthesis of l-tert-but ‘ ‘ ﬁrm 1 2 4-0xadiazol-5— propylg amino} methyl} pyrrolidine-l, 2—(licarboxylate (65 To a stirring solution of 5 (3 g, 11.6 mmol) in MeOH (30 mL) was added sodium acetate (1.91 g, 23.3 mmol) followed by Int-F (3.6 g, 13.9 mmol). The reaction mixture was heated to reﬂux for l h. The reaction mixture was slowly cooled to RT—OOC, to this sodium cyanoborohydride (1.465 g, 23.3 mmol) was added, and stirring was continued for another 6 h at RT. After consumption of the starting material (determined by TCL), methanol from the reaction was evaporated under reduced pressure and the residue was diluted with water and extracted with EtOAc (2x 50 mL). The separated organic layer was dried over ous NaZSO4 and concentrated under reduced pressure. The resulting crude material was puriﬁed by silica gel column chromatography eluting with 40% EtOAc/Hexane to afford 6 (2.5 g, 56%).

LCMS m/z: 385 [M++1] S nthesis of tert-but 12- IS 2R h drox 1 2 4-0xadiazol—5- 1 r0 l-l-oxo-Z 5- diazaspiro|3.4| octane-S-carboxylate 171 To a stirring solution of6 (1.5 g, 3.90 mmol) in THF (30 mL) was cooled to 0 °C and added t—BuMgCl (1M in THF, 15.6 mL, 15.6 mmol) and stirred for 15 min. After consumption of the starting material (by TLC), the reaction mixture was quenched with aqueous NH4C1 solution and d with water. Aqueous layer was extracted with EtOAc (2x mL). The separated organic layer was dried over anhydrous Na2S04 and concentrated under d re. The resulting crude material was puriﬁed by silica gel column chromatography eluting with 150% CHzClz to afford 7 (0.15 g, 11%). 1H-NMR: (400 MHz, DMSO-d6): 5 9.02 (s, 1H), 5.15 (s, 1H), .32 (m, 1H), 3.95 (d, 1H), 3.66-3.60 (m, 1H), 3.36—3.30 (m, 1H), 2.29-2.09 (m, 2H), 1.87-1.82 (m, 2H), 1.55 (s, 9H), 1.27 (d, 3H).

LCMS (ESI) m/z: 351 [Ml—l] UPLC Purity: 96% S nthesis of 2- IS 2R h drox 1 2 4-oxadiazol 1 r0 1-2 5-diazas iro 3.4 octan-l-one (Compound X2 To a ng solution of 7 (0.4 g, 1.13 mmol) in CH2C12 (4 mL) was added TFA (0.43 mL) at 0°C and stirred at RT for 30 min. The reaction e was concentrated under vacuum. The resulting crude material was puriﬁed by prep—HPLC to afford Compound X (65 mg) as TFA salt. 1H-NMR: (400 MHz, DMSO-d6): 5 9.89 (br s, 1H), 9.08 (s, 1H), 5.46 (d, 1H), 5.31 (s, 1H), 4.37—4.35 (m, 1H), 3.99 (d, 1H), 3.81 (d, 1H), 3.42—3.35 (m, 2H), 2.35—2.18(m, 2H), .03 (m, 2H), 1.24 (d, 3H).

LCMS (ESI) m/z: 253.4 [M++1] Prep-HPLC Purity: 95% Exam 1e 2 - S nthesis of Com ound Y Scheme 2.

NaBH3CN / \ Step-8 "N - Step-10 Mitsunobu Compound Y _ Step-C Step-D OHSOCIZ, EtOH N OCH (Boc)O OCH3 LIHMDS, | 2 .

H sec BOM-CI BooO 8M1 A B C Step-E Pd/C, H2 N S nthesis of -meth l rrolidine—Z-carbox late A —36- To a ng solution of L-proline (SMl) (100 g, 0.87 mol) in methanol (800 mL) was slowly added thionyl chloride (76.9 mL, 1.04 mol) at 0°C. The reaction mixture was slowly warmed to RT and heated to reﬂux for 12 h. After ption of the starting material (by TLC), the reaction was concentrated under reduced pressure to afford A (143.9 g, HCl salt). : (400 MHz, CDC13): 6 3.89 (s, 3H), 3.68-3.62 (m, 2H), 3.59-3.47 (m, 2H), 2.49-2.37 (m, 1H), 2.27—2.05 (m, 3H).

LCMS (m/z): 166 [M++l] S nthesis of tert-bu h l rrolidine—l 2-dicarbox late B ] To a stirring solution ofA (35 g, 0.22 mol) in CHzClz (175 mL) were added Et3N (90 mL, 0.65 mol) followed by Boc-anhydride (56.9 mL, 0.26 mol) at 0°C. The reaction mixture was stirred at RT for 16 h. After consumption of the starting material (by TLC), the reaction was diluted with water (100 mL) and extracted with CHZCIZ (2x 100 mL). The organic layer was washed with water, brine, dried over NaZSO4 and concentrated. The resulting crude material was puriﬁed by silica gel column chromatography eluting with 30% EtOAc/Hexane to afford B (41 g, 95%). 1H-NMR: (400 MHz, CDCl3): 5 4.25—4.21 (m, 1H), 3.75 (s, 3H), 3.57—3.26 (m, 2H), 2.29—2.10 (m, 1H), 1.99—1.75 (m, 3H), 1.45 (s, 9H).

LCMS (m/z): 130 )—Boc] S nthesis of l-tert-but 12-meth 12- ben 10x meth l rrolidine—l 2-dicarb0x late (£1 To a stirring solution of B (100 g, 0.43 mol) in THF (800 mL) was added LiHMDS (873 mL, 0.87 mol) at -78°C and stirred for 1 h. To this BOM-chloride (93.2 mL, 065 mol) was added drop wise at —78°C and stirred for 2 h at —20°C. After consumption of the starting material (by TLC), the reaction was quenched with sat.NH4C1 solution at 0°C. The separated organic layer was washed with water, dried over NaZSO4 and concentrated to afford C (180 g, . This material was directly taken for the next step without further puriﬁcation.

LCMS (m/z): 250 [(M++1)—Boc] S nthesis of 2- benz 10x meth l tert-butox carbon 1 rrolidine—2-carb0x lic acid ] To a stirring solution of C (100 g, 0.28 mol) in methanol (200 mL) was added 2N NaOH on (300 mL) at RT. The reaction mixture was heated to reﬂux for 4 h. After consumption of the starting material (by TLC), the solvent from the reaction was evaporated under reduced pressure and diluted with EtOAc (100 mL). The aqueous layer was acidiﬁed using citric acid solution and extracted with CH2C12 (2x 250 mL). The separated organic layer was washed with water, dried over Na2S04 and concentrated to afford D (60 g, 63%). 1H-NMR: (400 MHz, CDC13): 5 7.37-7.32 (m, 5H), 4.61 (s, 2H), .88 (m, 2H), 3.65-3.42 (m, 2H), 2.54—2.46 (m, 2H), 1.95 (br s, 2H), 1.57 (s, 9H).

LCMS (m/z): 334 [Mir] S nthesis of 1- tert—butox carbon 1 h drox meth l rrolidine—Z-carbox lic acid Int-

[00122] To a stirring solution of D (10 g, 29.81 mmol) in methanol (300 mL) was added 50% wet 10% Pd/C (5 g) at RT and stirred for 24 h under H2 atmosphere (balloon pressure).

After consumption of the starting material (by TLC), the reaction mixture was ﬁltered through a pad of celite and the pad was washed with methanol. The ﬁltrate was concentrated under reduced pressure to afford Int-E (6 g, 82%). 1H-NMR: (400 MHz, DMSO—ds): 5 12.55 (br m, 1H), 3.99 (d, 1H), 3.88 (d, 1H), 7.65—7.60 (m, 1H), 3.51-3.45 (m, 1H), 3.39—3.34 (m, 1H), 2.32—2.14 (m, 1H), 1.98—1.69 (m, 3H), 1.39 (s, 9H). 'nthesis 0f1~’ vrimidin-Z- 31%me '2 To a stirring solution of 1 (15 g, 0.14 mol) in THF (150 mL) was added agnesium bromide (1M in THF) (171.4 mL, 0.17 mol) at 0°C under nitrogen atmosphere slowly over a period of 15 min. After being stirred for 30 min, the reaction mixture was quenched with aqueous NH4Cl solution and the aqueous layer was extracted with EtOAc. The separated organic layer was dried over anhydrous Na2S04, ﬁltered and concentrated under reduced pressure to obtain crude product, which was d by silica gel column chromatography g with 50% EtOAc/hexane to afford 2 (8.5 g, 44%). 1H-NMR: (400 MHz, CDCl3): 5 8.95 (d, 2H), 7.47 (t, 1H), 3.26 (q, 2H), 1.25 (t, 3H).

S nthesis of 1- ut ldimeth lsil lox r0 1- l rimidine 3 To a ng on of 2 (8.5 g, 62.5 mmol) in THF (170 mL) was added LiHMDS (1M in THF) (93.7 mL, 93.7 mmol) and stirred for 1 h. To this was added TES—Cl (12.5 mL, 75 mmol) at 0°C under N2 atmosphere and further stirred for 1 h. After consumption of the starting al (by TLC), the reaction was quenched with aqueous NH4C1 solution and extracted with EtOAc. The separated c layer was dried over anhydrous NaZSO4, ﬁltered and 2014/013621 —38- concentrated under reduced re to afford 3 (10 g, 64%). The resulting crude al was directly used for the next step without further puriﬁcation.

S nthesis of o—1- rimidin-Z- 1 r0 an-l-one 4 To a stirring solution of 3 (10 g, 0.04 mol) in THF: H20 (100 mL, 4: 1) was added NBS (5.6 g, 0.05 mol) at RT and stirred for 1 h. After consumption of the starting material (by TLC), the reaction mixture was concentrated under reduced pressure and obtained residue was diluted with EtOAc and washed with water. The separated organic layer was dried over anhydrous Na2S04, d and concentrated under reduced pressure to obtain crude product, which was puriﬁed by silica gel column chromatography eluting with 30% EtOAc/hexane to afford 4 (6.5 g, 75.5%). 1H-NMR: (400 MHz, CDCl3): 5 8.97 (d, 2H), 7.52 (t, 1H), 5.92 (q, 1H), 2.79 (s, 1H), 1.99 (d, 3H).

S nthesis of 2-h drox rimidin-Z- 1 r0 an-l-one 5 To a stirring solution of 4 (6.5 g, 0.03 mol) in MeOH (65 mL) was added sodium formate (10.28 g, 0.15 mol) and heated to reﬂux for 8 h. After consumption of the starting material (by TLC), the reaction mixture was cooled to RT and solvent was removed under d pressure. The resulting crude material was puriﬁed by silica gel column chromatography eluting with 2% MeOH/CHzClz to afford 5 (3.5 g, 76%). 1H-NMR: (400 MHz, CDCl3): 6 9.01 (d, 2H), 7.52 (t, 1H), 5.92 (q, 1H), 3.73 (br s, 1H), 3.41 (s, 1H), 2.78 (s, 2H), 1.55 (d, 3H).

S nthesis of 2- tert-but ldimeth lsil 10x rimidin-Z- 1 r0 ne 6 To a stirring solution of 5 (3.5 g, 23 mmol) in CHzClz (70 mL) was added imidazole (3.91 g, 57 mmol) followed by DMAP (281 mg, 2.30 mmol) at RT. The reaction mixture was cooled to 0°C, tow which TBS—Cl (5.18 g, 0.03 mol) was added and stirred at RT for 4 h. After consumption of the ng al (by TLC), the reaction mixture was diluted with CHzClz and washed with water. The separated organic layer was dried over anhydrous NaZSO4, ﬁltered and concentrated under reduced pressure to obtain a crude product, which was puriﬁed by silica gel column chromatography eluting with 20% EtOAc/hexane to afford 6 (3 g, 49%). 1H-NMR: (400 MHz, CDC13: 5 8.93 (d, 2H), 7.45 (t, 1H), 5.59 (q, 1H), 1.52 (d, 3H), 0.89 (s, 9H), 0.09 (s, 6H).

S nthesis of 2- tert—but ldimeth lsil 10x rimidin-Z- 1 r0 an-l-amine 7 To a stirring solution of 6 (3.0 g, 11.27 mmol) in MeOH (60 mL) was added sodium acetate (1.84 g, 22.55 mmol) ed by ammonium carbonate (8.85 g, 56.3 mmol) and acetic acid (0.64 mL, 11.27 mmol) at RT. The reaction mixture was heated to reﬂux for l h, added NaCNBH3 (1.41 g, 22.5 mmol) and continued reﬂux for another 6 h. After consumption of the starting al (by TLC), the reaction mixture was cooled to RT and volatiles were evaporated. Obtained residue was diluted with water and extracted with EtOAc. The separated organic layer was dried over anhydrous NazSO4, ﬁltered and concentrated under reduced pressure to get crude product, which was puriﬁed by silica gel column chromatography eluting with 2% MeOH/CH2C12 to afford 7 (2 g, 66.4%). 1H-NMR: (400 MHz, DMSO-d6): 5 8.91 (d, 1H), 8.85 (d, 1H), 7.48 (t, 1H), 5.71 (br m, 2H), 4.24 (t, 1H), 4.05 (d, 1H), 1.29 (d, 1H), 1.12 (d, 2H), 0.74 (s, 9H), 0.03 (s, 3H), 0.02 (s, 2H), 0.01 (s, 1H).

LCMS (m/z): 268 [M++l] S nthesis of tert-but 12- 2- tert—bu ldimeth lsil 10x rimidin-Z- 1 r0 lcarbamo l h drox meth l ine—l-carbox late 8 To a ng solution of Int-E (2 g, 8.16 mmol) in CHzClz (60 mL) was added compound 7 (2.39 g, 8.97 mmol), EDCI. HCl (2.33 g, 12.2 mmol) followed by HOBt (1.66 g, 12.24 mmol) and DIPEA (4.5 mL, 24.4 mmol) at 0°C. The reaction mixture was warmed to RT and d for 16 h. After consumption of the starting material (by TLC), the reaction mixture was diluted with CHZClz. The separated c layer was washed with aqueous NaHCO3 on followed by aqueous NH4Cl. The organic layer was dried over anhydrous Na2SO4, ﬁltered and concentrated under reduced pressure to get crude product, which was puriﬁed by silica gel column chromatography eluting with 70% hexane to afford 8 (2.3 g, 57.5%). 1H-NMR: (400 MHz, DMSO—d6): 5 8.91 (d, 1H), 8.68 (d, 2H), 7.41 (br s, 1H), 5.74 (br t, 1H), .07—4.89 (m, 1H), 4.15—4.10 (m, 1H), 3.97—3.92 (m, 1H), 3.45-3.41 (m, 1H), 1.79-1.74 (m, 2H), 1.43-1.39 (m, 4H), 1.29-1.21 (m, 6H), 1.12 (d, 5H), 0.71 (s, 9H), 0.12 (t, 1H), 0.09 (s, 2H), 0.08 (s, 1H), 0.04 (s, 2H).

LCMS (m/z): 495.5 [M++1] Mass: 495.5 [M++l] S nthesis -but 12- 2- tert—bu ldimeth lsil 10x rimidin-Z- 2 5-diazas iro 3.4 octane-S-carbox late 9 To a stirring solution of 8 (2.3 g, 4.65 mmol) in THF (23 mL) was added TPP (1.34 g, 5.12 mmol) followed by DTAD (1.6 g, 6.98 mmol) at 0°C. The reaction mixture was warmed to RT and stirred for 16 h. After consumption of the starting material (by TLC), the reaction mixture was concentrated under reduced pressure to get crude product, which was puriﬁed by silica gel column tography eluting with 25% EtOAc/hexane to afford 9 (1.2 g, 54.2%).

LCMS (m/z): 477.4 [M++1] : (400 MHz, s): 5 8.82 (d, 2H), 7.49 (t, 1H), 4.72 (d, 1H), 4.31 (q, 1H), 3.62 (br s, 2H), 3.25—3.19 (m, 1H), 2.24—2.05 (m, 2H), 1.85—1.81 (m, 2H), 1.42 (br s, 1H), 1.25 (t, 3H), 0.92 (s, 8H), 0.75 (s, 9H), 0.02 (s, 3H).

S nthesis of tert-but 12- 2-h drox rimidin-Z- 1 r0 l-l-oxo-Z 5- diazaspiro|3.4[octane-S-carboxylate [10]

[00131] To a stirring solution of 9 (1.0 g, 2.10 mmol) in THF (20 mL) was added TBAF (1M in THF) (6.3 mL, 6.30 mmol) at 0°C under N2 atmosphere and stirred for l h. After consumption of the starting material (by TLC), the reaction mixture was quenched with ice water and extracted with EtOAc. The ted organic layer was dried over anhydrous Na2S04, ﬁltered and concentrated under reduced pressure to obtain crude product, which was d by silica gel column chromatography eluting with 5% MeOH/CHzClz to afford 10 (0.35 g, 46%). : (400 MHz, DMSO-d6): 5 8.81 (d, 2H), 7.49 (t, 1H), 4.81 (d, 1H), 4.65 (d, 1H), 4.25- 4.20 (m, 1H), 3.64-3.51 (m, 2H), 3.34 (s, 1H), 3.25-3.20 (m, 1H), 2.25—2.20 (m, 2H), 1.87—1.82 (m, 2H), 1.19 (d, 3H), 0.97 (s, 9H).

LCMS (m/z): 363.3 [M++1] S nthesis of 2- 2-h drox rimidin-Z- 1 r0 1-2 5-diazas iro 3.4 octan-l-one [Compound Y] To a stirring solution of 10 (0.3 g, 0.82 mmol) in CH2C12 (6 mL) was added molecular sieves (0.3 g) followed by BFg—etherate (0.31 mL, 2.48 mmol) at 0 OC and stirred at RT for l h. After ption of the starting material (by TLC), the reaction mixture was ﬁltered and obtained residue was dissolved in MeOH and washed with CHZClz. The volatiles were evaporated under reduced pressure to obtain crude product, which was puriﬁed by silica gel column chromatogrpahy eluting with 8% MeOH/CHZClz to afford Compound Y (0.12 g, 55%). : (400 MHz, DMSO—d6): 5 8.81 (d, 2H), 7.49 (t, 1H), 4.81 (d, 1H), 4.65 (d, 1H), 4.25— 4.20 (m, 1H), 3.64-3.51 (m, 2H), 3.12-3.01 (m, 2H), 2.15-2.10 (m, 2H), 1.87-1.82 (m, 2H), 1.19 (d, 3H).

LCMS (m/z): 263.1 [M++l] Pre aration of ke intermediates schemes L] to L7: Scheme I-1 OTES O A B \ N 0” OTBS OTBS Step D | Step E Step F —> f” C” N/JWKK N/ HCOONa TBDMS—CI NJﬁ/K o o NaBH3CN D E F S nthesis of 1- rimidin-Z- 1 r0 an-l-one A:

[00133] Referring to Scheme 1-1, to a stirring solution of pyrimidine—2-carbonitrile (25 g, 238 mmol) in THF (180 mL) was added ethyl magnesium bromide (1M in THF, 37.9 g, 285 mmol) at 0 OC and stirred for 2 h at RT. After completion of starting al (by TLC), the reaction mixture was diluted with saturated ammonium chloride solution and EtOAc (150 mL).

The separated c layer was washed with brine solution (2 x 100 mL). The extracted organic layer was dried over anhydrous NaZSO4, filtered and concentrated under reduced pressure to afford crude material which was puriﬁed by column chromatography eluting 30% EtOAc/hexane to afford A (18.3 g, 57%) as an ofi'lwhite solid. .lH-NMR: (400 MHz, 6): 5 9.00 ( d, J= 5.2 Hz, 2H), 7.70 (t, J= 4.8 Hz, 1H), 3.20- 3.15 (m, 2H), 1.09 (t, J: 7.2 Hz, 3H).

LCMS m/z: 137 [M++1].

S s of 1- trieth lsil 1 0X r0 en l rimidine B : To a stirring solution of A (14 g, 103 mmol) in dry THF (140 mL) was added LiHMDS (1M in THF, 206 mL, 206 mmol) slowly at 0 °C and stirred for 30 min. After added chloro triethylsilane (24.8 g, 165 mmol) in THF (50 mL) dropwise at 0 oC and d 1 h. After completion of starting material (by TLC), the reaction mixture was diluted with saturated ammonium chloride solution (100 mL) and EtOAc (150 mL). The separated organic layer was washed with brine solution (2 x 100 mL). The c layer was dried over anhydrous NazSO4, filtered and concentrated under reduced pressure to afford crude material which was puriﬁed by column chromatography eluting 20% EtOAc/hexane to afford B (23 g, 8949/61) as yellow thick syrup. 1H-NMR: (400 MHz, DMSO—dg): 5 8.75 (d, J= 4.8 Hz, 2H), 7.32 (t, J= 4.8 Hz, 1H), 6.36— 6.31 (m, 1H), 1.77 (d, J= 7.2 Hz, 3H), 0.95—0.87 (m, 9H), 0.71—0.65 (m, 6H).

S nthesis of 2-br0m0 rimidin-Z- 1 r0 an-l-one C: To a stirring solution of B (23 g, 92 mmol) in 0 (184 mL/46 mL) were added N—bromosuccinamide (18 g, 101 mmol) slowly at 0 OC and stirred for 3 h at RT. After tion of starting material (by TLC), the reaction mixture was diluted with H20 and EtOAc (100 ml/150 mL). The separated organic layer was washed with hypo solution (2 x 100 mL) followed by brine solution (2 x 100 mL). The organic layer was dried over anhydrous NaZSO4, d and trated under reduced pressure to afford crude material which was purified by column chromatography eluting 15% EtOAc/hexane to afford C (11.5 g, 58%) as yellow thick syrup. 1H—NMR: (400 MHz, DMSO—ds): 8 9.06 ( d, J: 4.8 Hz, 2H), 7.75 (t, J: 4.8 Hz, 1H), 5.97— .92 (m, 1H), 1.83 (d, J= 6.4 Hz, 3H).

S nthesis of 2-h drox rimidin-Z- r0 an-l-one D : ] To a stirring solution of C (11.5 g, 53.4 mmol) in MeOH (80 mL) was added sodium formate (14.5 g, 214 mmol) and stirred the reaction mass at 80 °C for 6 h. After completion of reaction (by TLC), the reaction mixture was ated under reduced pressure to give crude product, which was purified by column chromatography eluting 50% EtOAc/n- hexane to afford D (5.0 g, 61.6%) as colorless liquid. 1H—NMR: (400 MHz, DMSO—ds): 5 8.73 (d, J= 5.2 Hz, 2H), 7.55 (t, J: 4.8 Hz, 1H), 5.28— .26 (m, 1H), 1.24 (d, J: 6.4 Hz, 1H), 0.99 (d, J: 6.4 Hz, 3H) S 'nthesis of Z-tert—but ldimethvlsilvl oXVFl— wrimidin—Z—l Lone E-z To a stirring solution of D (5 g, 32.8 mmol) in DCM (50 mL) were added imidazole (5.5 g, 82.2 mmol), DMAP (800 mg, 0.65 mmol) at 0 0C and stirred for 10 min.

After added TBDMS—Cl (7.38 g, 49.2 mmol) at 0 0C and stirred at RT for 12 h. After completion of ng material (by TLC), diluted the reaction mass with H20 (50 ml.) The separated organic layer was washed with brine solution (2 X 50 niL). The organic layer was dried over ous Na2304, ﬁltered and concentrated under d pressure to afford crude material which was puriﬁed by column chromatography eluting 20% EtOAc/hexane to afford E (6 g, 68.8%) as an off—white solid. 1H—NMR: (400 MHz, CDC13): 5 9.00 ( d, J: 5.2 Hz, 2H), 7.71 (t, J: 4.8 Hz, 1H), 5.47—5.42 (m, 1H), 1.35 (d, J: 6.8 Hz, 3H), 0.79 (s, 9H), 0.05 (s, 6H).

S nthesis of 2- tart—but ldirneth lsilvl 0X —1- rimidin-Z— ' ro an-l-amine F :

[00138] To a stirring solution of E (6 g, 22.5 mmol) in MeOH (50 mL) were added sodium acetate (3.69 g, 45.1 mmol), ammonium carbonate (21 g, l35 mmol), AcOl-I (1.28 mL, 22.5 mmol) at RT and stirred the reaction mixture at 90 °C for 2 h. The temperature of the reaction was cooled to RT and added sodium cyanoborohydride (2.84 g, 45.1 11111101) slowly and stirred at 90 0C for 6 h. After completion of reaction (by TLC), evaporated MeOH under reduced re. The crude residue was diluted with water (20 mL) and ted with DCM (2 x 50 Hi). The organic layer was dried over anhydrous NagSO4, ﬁltered and concentrated under reduced pressure to afford crude material was puriﬁed by column chromatography eluting 5% MeOi—l/DCM to afford F (4.2 g, 69.6%) as semi solid. 1H-NMR: (400 MHz, CDC13): 5 8.83 (d, J= 4.8 Hz, 2H), 7.40 (t, J: 5.2 Hz, 1H), 4.13 (t, J: 6.4 Hz, 2H), 3.90 (d, J: 6.4 Hz, 2H), 1.12 (d, J= 6.4 Hz, 3H), 0.70 (s, 9H), 0.02 (s, 6H). _ 44 _ Scheme I-2 OBn OBn \ Step 3 | | Step 4 _> OH N O\ _> | LiHMDS N NaOH 'T' 3°C O 0 BOM_C| Boc 800 Step 5 0%(OHN Pd /C BOCI O S nthesis of 1-tert-bu lZ-meth 12- ben lox meth l rrolidine-l rb0x late (E); To a stirring solution of t-BOC proline methyl ester (25 g, 109 mmol) in THF (250 mL) was added LiHMDS (240 mL, 240 mmol) at -20 °C and stirred for 2 h. To this BOM- chloride (23 mL, 163 mmol) was added drop wise at -30 °C and stirred for 2 h. After consumption of the starting material (by TLC), the reaction was quenched with aqueous NH4Cl solution (100 mL) and extracted with EtOAc (2 x 200 mL). The combined organic layer was washed with water (2 x 150 mL) ed by brine solution (2 x 100 mL). The organic layer was dried over NaZSO4 and concentrated to obtain crude compound which was puriﬁed by column chromatography by eluting 10% n—hexane to afford compound G (30 g, 79%) as thick syrup. 1H-NMR: (500 MHz, DMSO-dg): 5 7.36—7.22 (m, 5H), 4.59—4.48 (m, 2H), 4.02—3.88 (m, 1H), 3.63 (s, 3H), 3.49-3.35 (m, 2H), 3.34—3.30 (m, 1H), 2.31—2.23 (m, 1H), 2.04-1.89 (m, 2H), 1.82— 1.78 (m, 1H).

LCMS (m/Z): 249.4 [(M++1)—Boc].

S nthesis of 2- benz lox meth l tert-butox carbon 1 rrolidine-Z-carbox lic acid (E); To a stirring solution of nd G (30 g, 86 mmol) in methanol (70 mL) was added NaOH on (6.88 g in 70 mL H20) at RT. The reaction mixture was heated to 70 °C for 16 h. After consumption of the starting material (by TLC), the solvent from the reaction was evaporated under reduced pressure and diluted with EtOAc (2 x 200 mL). The separated 2014/013621 aqueous layer was acidiﬁed using citric acid solution (pH~3) and extracted with EtOAc (2 x 250 mL). The combined organic layer was dried over NaZSO4 and concentrated to afford crude which was triturated with n—hexane to obtain compound H (25 g, 86.8%) as off—white solid. 1H-NMR: (400 MHz, DMSO-dg): 5 12.35 (br s, 1H), 7.37—7.29 (m, 5H), 4.56—4.48 (m, 2H), 4.06-4.00 (m, 1H), 3.92-3.89 (m, 1H), 3.66-3.45 (m, 1H), 3.37-3.28 (m, 1H), 2.31-2.20 (m, 1H), 2.05-1.97 (m, 1H), 1.87—1.75 (m, 2H), 1.38 (s, 9H).

LCMS (m/z): 335.3 [M++1].

S nthesis of 1- tert-butox carbon 1 h drox meth l rrolidine—Z-carbox lic acid I : ] To a ng solution of compound H (25 g, 74 mmol) in methanol (150 mL) was added 50% wet 10% Pd/C (7 g) at RT and stirred for 10 h under H2 atmosphere. After consumption of the starting material (by TLC), the reaction mixture was ﬁltered through a pad of celite and the pad was washed with methanol (100 mL). Obtained ﬁltrate was trated under reduced pressure to afford compound I (15 g, 82.8%) as white solid. 1H-NMR: (400 MHz, DMSO—dg): 5 4.66 (br s, 1H), 3.96—3.83 (m, 1H), 3.63—3.59 (m, 1H), 3.49—3.41 (m, 1H), .25 (m, 1H),2.30-2.17 (m, 1H), .72 (m, 3H), 1.38 (s, 9H).

Scheme I-3 OH 0 OTBSO OTBSO Step-C Step-D OMe —> 0M9 —> NHNH2 NHCbZ NHCbZ TBS-CI NHCbZ NH2NH2 J K L OTBS O/\ OTBS O/\ StepE ,N Step F (C2H50)3CH Pd——C/H2 NHCbZ )TghrN, S s of 2S ‘ ' ‘ . . tert—butvidimeth lsil l My; butanoate 5K}: Referring to Scheme 1—3, to a stirring solution of J (50 g, 187 mmol) in DMF (400 ml.) were added DIPEA (86 ng 468 mmol) TBDMS—Cl (33.66 mL, 224 mmol) at 0 OC and stirred at RT for 12 h. After completion of starting material (by TLC), diluted the reaction —46- mass with EtOAc (500 ml). The separated c layer was washed with (2x200 mL) of Water followed by brine solution (2x100 mL). The organic layer was dried over anhydrous , filtered and concentrated under reduced re to afford crude material was purified by column chromatography eluting 10% EtOAc/hexane to afford K (50 g, 70.1%) as colorless syrup. 1H-NMR: (400 MHz, CDCl3): 5 7.39—7.32 ( m, 5H), 5.43 (d, J= 9.6 Hz, 1H), 5.14 (s, 2H), 4.45—4.43 (m, 1H), 4.29—4.26 (m, 1H), 3.72 (s, 3H), 1.21 (d, J= 6.0 Hz, 3H), 0.83 (s, 9H), 0.09 (s, 6H).

LCMS m/z: 382.2[M++1].

S nthesis of hen 1 2S 3R tert-but ldimeth lsil 1 0X h drazin l-l-oxobutan-Z- yl) carbamate (L): A solution of K (50 g, 131 mmol) in EtOH (400 mL) was added hydrazine hydrate (32.8 g, 656 mmol), at RT and after stirred at 90 0C for 24 h. After tion of starting material (by TLC), evaporated ethanol under reduced pressure. The crude e was diluted with water (100 mL) and EtOAc (500 mL). After the separated organic layer was washed with (2x100 mL) of water followed by brine solution (1x100 mL). Dried over anhydrous NagSO4, ﬁltered and concentrated under reduced pressure. Obtained crude material was purified by column chromatography by g with 20% EtOAc/hexane to afford L (25 g, 50%) as colorless thick syrup. 1H-NMR: (400 MHz, DMSO—d6): 5 9.10 (s, 1H), 7.36—7.30 (m, 5H), 6.83 (d, J= 9.6 Hz, 1H), .02 ( s, 2H), 4.19 (s, 2H), 4.05—4.02 (m, 1H), 3.97—3.93 (m, 1H), 1.05 (d, J= 6.0 Hz, 3H), 0.81 (s, 9H), 0.01 (s, 6H).

S nthesis of ben 1 1S 2R -2— tert-but ldimeth lsil l ox 1 3 4-oxadiazol l propyl) ate {M}:

[00144] A solution of L (25 g, 65.6 mmol) in triethyl orthoformate (250 mL) was added p-TSA (catalytic, 250 mg) at RT and after stirred at 80 °C for 3 h. After completion of ng material (by TLC), evaporated triethyl orthoformate under reduced pressure. The crude residue was d by column chromatography eluting 10% EtOAc/hexane to afford M (15 g, 58%) as thick syrup. 1H-NMR: (500 MHz, DMSO—dg): 5 9.22 (s, 1H), 7.85 (d, J= 9.5 Hz, 1H), 7.36—7.31 (m, 5H), .05 (s, 2H), 4.96-4.93 (m, 1H), 4.25 (t, J= 6.0 Hz, 1H), 1.23 (d, J= 6.0 Hz, 3H), 0.80 (s, 9H), 0.10 (s, 6H).

LCMS m/z: M++1].

S nthesis of 1S 2R tert-bu ldimeth lsil 1 OX 1 3 4—0xadiazol—2- 1 r0 an-l- amine (N): To a stirring solution of M (15 g, 38.3 mmol) in methanol (200 mL) was added 50% wet 10% Pd/C (5 g) and stirred under H2 atmosphere (balloon pressure) for 4 h at RT. The reaction mixture was ﬁltered through a pad of celite and triturated with methanol (100 mL).

The ﬁltrate was trated under reduced pressure to afford N (10 g, crude) as thick syrup. 1H-NMR: (500 MHz, DMSO—d6): 5 9.15 (s, 1H), 4.11 (t, J= 5.0 Hz, 1H), 4.03 (d, J= 2.0 Hz, 1H), 2.05 (br s, 2H), 1.17 (d, J: 6.0 Hz, 3H), 0.76 (s, 9H), 0.02 (s, 6H).

LCMS m/z: 258.3 [M++1].

Scheme I-4 OH O O O Step8 St 0 St9 D OH —> OVOH i. OMNHZ _p.

,NH MeOy /)-—N NH N 300 4Cl EDCI BOG , X 300 DMF.DMA O P O \N/ O \ —N 4 Step E 73 0” O Step F 9 N —’ —’ o o \N )FN BOG NHZOHHCI )rN BoC TFA NH2 .TFA AcOH Q R S S nthesis of 3- tert-butox carbon 1-2 2 5-trimeth loxazolidinecarbox lic acid 0 To a stirring solution of N-BOC threonine ( 15 g, 59.28 mmol) in THF (150 mL) was added PPTS (1.47 g, 5.92 mmol) followed by 2,2—dimethoxy propane (21.79 mL, 0.17 mol) at 0 °C under N2 atmosphere. The reaction e was stirred at RT for 16 h. The reaction mixture was again heated to reﬂux for 6 h. The on mixture was diluted with aqueous NaHCO3 solution and washed with EtOAc (1x100 mL). Aqueous layer was acidiﬁed using citric acid solution (pH~2) and ted with CHzClz (2x100 mL). The organic layer was washed with brine, dried over anhydrous Na2SO4 and concentrated under vacuum to afford compound 0 (18 g, crude). 1H—NMR; (400 MHz, DMSO—d6): 513.25 (br s, 1H), .05 (m, 1H), 3.79 (d, 1H), 1.50 (s, 3H), 1.67 (s, 3H), 1.45 (s, 9H), 1.29 (d, 3H).

S nthesis of tert-bu l4-carbam0 l-2 2 5-trimeth loxazolidine—3-carbox late P : To a stirring solution of compound 0 (18 g, 69.4 mmol) in CHZCIZ (180 mL) was added HOBt (14.16 g, 0.104 mol), EDCI.HC1 (19.88 g, 0.104 mol) followed by NH4Cl (5.56 g, 0.104 mol) and DIPEA (31.9 mL, 0.173 mol) at 0 °C. The reaction e was d at RT for 16 h. After consumption of the starting material (by TLC), the reaction mixture was washed with aqueous citric acid, NaHC03 followed by brine. Organic layer was dried over anhydrous NaZSO4and concentrated under reduced pressure to give crude; which was puriﬁed by silica gel column chromatography eluting with 2% MeOH/CH2C12 to afford compound P (13 g, 72.5%). 1H-NMR: (400 MHz, DMSO—ds): 5 7.51 (br s, 1H), 7.14 (br s, 1H), 3.97—3.95 (m, 1H), 3.71 (d, 1H), 1.51 (d, 6H), 1.34 (s, 9H), 1.24 (d, 3H).

LCMS (ESI): 159.1[(M++1)—Boc] S nthesis of Z-tert-bu l 4- dimeth lamino meth lene carbamo l-2 2 5- trimethyloxazolidinecarboxylate ] S 2 2:

[00148] A on of compound P (13 g, 50.3 mmol) in DMF.DMA (130 mL) was d at reﬂux temperature for 3 h under N2 atmosphere. After consumption of the starting material (by TLC), the on mixture was concentrated under reduced pressure to afford compound Q (15.7 g, crude). This crude material was directly taken for the next step t further puriﬁcation.

S s of tert-bu 12 2 5—trimeth l 1 2 4-0xadiazol—5— l oxazolidinecarbox late 13).: ] To a stirring solution of compound Q (15.? g, 50.09 mmol) in ethanol (157 mL) was added hydroxylamine hydrochloride (6.96 g, 0.10 mol) under N2 atmosphere. The reaction mixture was heated to reﬂux and stirred for 2 h. After consumption of the starting material (by TLC), acetic acid (28.6 mL, 0.50 mol) was added to the reaction e and then reﬂuxed for 16 h. The solvents from the reaction e was evaporated under vacuum to give crude; which was puriﬁed by silica gel column tography eluting with 10% EtOAc/Hexane to afford compound R (4.5 g, 32%). 1H-NMR: (400 MHz, DMSO-ds): 5 6.35 (s, 2H), 4.61 (d, 1H), 4.22-4.15 (m, 1H), 1.55 (s, 6H), 1.37 (s, 2H), 1.25 (d, 3H), 1.21 (s, 6H).

LCMS (ESI): 284 [M++l] Mass (m/z): 283 [M+] S nthesis of l-amino-l- 1 2 4-oxadiazol 1 r0 an-Z-ol S :

[00150] To a stirring solution of compound R (5 g, 17.6 mmol) in water (25 mL) was added triﬂuoroacetic acid (25 mL). The on mixture was stirred at RT for 5 h. After consumption of the starting material (by TLC), the reaction mixture was concentrated under vacuum. The residue was dissolved in water and neutralized with aqueous NaHC03. The solvent from the reaction mixture was evaporated under vacuum and ted with 5% MeOH/CH2C12 (3x 100 mL). The organic layer was concentrated under reduced pressure to afford compound S (2.5 g, crude). 1H-NMR: (400 MHz, D20): 5 8.84 (s, 1H), 4.05 (d, 1H), 3.98-3.95 (m, 1H), 3.67 (s, 1H), 3.58 (d, 1H), 1.15 (d, 3H), 1.12 (d, 3H).

LCMS (ESI): 144.1 [M++l] Scheme I—5 3 Boo/HM“:Step 4AMZEt Step 5 o N. 0 MeMgBr Pd ”“2_ Boc COOEtT OEt OBn m Step 6 Step 7 N 0 M33 80020 LHMD Mon/19 H I 8,0 BOMCI V W x —’m(;Step 8 Step 9 MOI—I aqNaOH Pd/C H2 Y Z S nthesis of eth 12- tert—butox carbon 1 amino oxohexanoate T: To a stirring solution of l—tert-butyl 2-ethyl 5-oxopyrrolidine-1,2-dicarboxylate (12 g, 46.6 mmol) in THF (120 mL) under inert atmosphere was added MeMgBr (3M in ether) (20.2 mL, 60.6 mmol) at 0 °C and stirred for 2 h. After consumption of the starting material (by TLC), the reaction mixture was quenched with aqueous NH4C1 solution and the aqueous layer was extracted with EtOAc (2 x 200 mL). The combined organic extracts were dried over anhydrous Na2S04 and concentrated under reduced pressure. The crude residue obtained was puriﬁed by silica gel column chromatography eluting with 20% EtOAc/Hexane to afford compound T (10 g, 79%). 1H—NMR: (400 MHz, CDC13)I 8 5.14 (br s, 1H), 4.23 (q, 2H), 2.62-2.47 (m, 2H), 2.17 (s, 4H), 1.91-1.82 (m, 1H), 1.45 (s, 10H), 1.26 (t, 3H).

Synthesis of ethyl 5-methylpyrrolidine-Z-carboxylate (V): To a stirring solution of compound T (10 g, 36.7 mmol) in CHzClz (50 mL) was added TFA (14.89 mL, 194.6 mmol) at 0 °C. After being stirred for 2 h at RT, the reaction mixture was concentrated under reduced pressure to get compound U. Obtained material was dissolved in ethanol (100 mL) and 10% Pd/C (50% wet, 3 g) under N2 here. The on e was d under H2 atmosphere on pressure) for 16 h. The reaction mixture was d through a pad of celite and ﬁltrate was trated under reduced pressure to afford compound V (15 g, crude). This material was directly taken for the next step without further purification. : (500 MHz, DMSO-d6): 4.4 (m, 1H), 4.2 (m, 2H), 3.6 , 2.3 (m, 1H), 2.1 (m, 2H), 1.6 (m, 1H), 1.3 (d, 3H), 1.2 (t, 3H).

LCMS m/Z.‘ 158.1 [M++1].

S nthesis of 1-tert-but lZ-eth lS—meth l rrolidine-l 2-dicarb0x late To a stirring solution of compound V (30 g, 191 mmol) in CH2C12 (150 mL) was added DMAP (23.3 g, 191 mmol) followed by Et3N (79.8 mL, 573 mmol) and Boc—anhydride (104 mL, 477 mmol) at 0 °C. The reaction mixture was stirred at RT for 16 h. The reaction mixture was diluted with CH2C12 (50 mL) and washed with water (2x150 mL) followed by brine. The separated organic layer was dried over anhydrous NaZSO4 and concentrated under vacuum. Obtained crude material was purified by column chromatography eluting with 6% EtOAc/Hexane to afford compound W (12 g, 82%) as pale yellow . 1H-NMR: (500 MHz, DMSO—d6): 54.13-386 (m, 4H), 2.15 (d, J= 3.5 Hz, 1H), 1.99—1.82 (m, 2H), 1.52 (t, .1: 4.5 Hz, 1H), 1.38 (s, 9H), 1.24 (t, J= 5.5 Hz, 3H), 1.16 (d, J= 6.5 Hz, 3H) LCMS (m/Z): 258 [(M++1).

S nthesis of 1-tert-but l 2-eth l 2- benz 10x meth l meth l rrolidine—l 2- dicarboxylate (X): To a stirring on of compound W (8.0 g, 31.12 mmol) in THF (70 mL) was added LiHMDS (59 mL, 41.72 mmol) at -78 OC and stirred for 2 h. To this BOM—chloride (6.56 mL, 41.72 mmol) was added drop wise and stirred for 2 h at -30 0C. After consumption of the starting material (by TLC), the reaction was quenched with s NH4C1 solution (20 mL) and ted with DCM (30 mL). The separated organic layer was dried over Na2SO4 and concentrated to afford crude material was puriﬁed by column tography eluting with % EtOAc/Hexane to afford compound X (11 g, 94.2%) as pale yellow liquid. 1H-NMR: (500 MHz, DMSO-dg):5 7.33-7.25 (m, 5H),4.38 (d, J: 10.5 Hz, 2H), 4.08-3.98 (m, 1H), 3.88 (d, J: 9.5 Hz, 2H), 2.20—2.08 (m, 2H), 1.38 (s, 9H), 1.37—1.29 (m, 4H), 1.19 (t, J: 7.5 Hz, 3H), 1.14-1.10 (m, 3H).

LCMS (m/z): 378 (M++1).

WO 20784 S nthesis of 2- benz 10X meth l utox carbon lmeth l rrolidine—Z- carboxylic acid (Y): To a stirring solution of compound X (11 g, 29.17 mmol) in CH3OH/THF (22 mL/20 mL) were added 2N NaOH solution (33 mL) at RT. The reaction mixture was heated to 65 °C for 8 h. After consumption of the starting material (by TLC), the solvent from the reaction was evaporated under reduced pressure and diluted with EtOAc (50 mL). The aqueous layer was acidiﬁed using citric acid solution and extracted with CHZClz (2 x 100 mL). The separated organic layer was washed with water (1 x 50 mL), dried over Na2S04 and concentrated to afford compound Y (8 g, 80%). 1H-NMR: (400 MHz, DMSO—dg): 5 12.58 (s, 1H), 7.34—7.28 (m, 5H), 4.54—4.47 (m, 2H), 4.05— 3.87 (m, 2H), 3.70—3.62 (m, 1H), 2.28—2.08 (m, 3H), 1.46—1.37 (m, 1H), 1.28 (s, 9H).

LCMS (m/z): 350 [M++1].

Synthesis of 1-(tert-butoxycarbonyl)(hydr0xymethyl)methylpyrr0lidine—Z-carboxylic acid (Z): ] To a stirring solution of compound Y (8 g, 1.45 mmol) in methanol (40 mL) was added 50% wet Pd/C (4 g) under N2 atmosphere. The reaction mixture was stirred under H2 atmosphere (balloon re) at RT for 16 h. After consumption of the starting material (by TLC), the reaction mixture was ﬁltered through a pad of celite and the pad was washed with methanol. Obtained ﬁltrate was concentrated under reduced pressure to afford crude compound which was ated with ane to obtained nd Z (4.5 g, 75.2%) as white solid. 1H-NMR: (500 MHz, DMSO-dg): 512.37 (br s, 1H), 4.61 (br s, 1H), 3.95-3.85 (m, 3H), 2.18— 2.06 (m, 3H), 1.44—1.41 (m, 1H), 1.38 (s, 9H), 1.09 (d, J= 6.0 Hz, 3H); LCMS (ESI): 260 [M++1] Scheme I-6 m Step 3 B®O\ Step 4 $04 N 0 LiHMDS NaOH Boc Boc/ S nthesis of 1- tert—but l 2-meth l 2- 1-h drox eth l rrolidine—l 2-dicarbox late 1AA): To a ng solution of N—BOC proline methyl ester (30 g, 131 mmol) in THF (100 mL) was added LiHMDS (323 mL, 327 mmol) at -20 OC and d for 30 min. To this acetaldehyde (12.3 mL, 196 mmol) was added drop wise at —20 °C and stirred for 2 h. After ption of the starting material (by TLC), the reaction was quenched with s NH4Cl solution (100 mL) and extracted with EtOAc (2 x 200 mL). The combined organic layer was washed with brine solution (2 x 150 mL). The organic layer was dried over Na2S04 and concentrated to obtain crude compound which was puriﬁed by column chromatography by eluting 10% EtOAc/n—hexane to afford compound AA (30 g, 83.8%) as thick syrup. 1H-NMR: (400 MHz, : 8 5.82 (d, J= 10.0 Hz, 1H), 4.71—4.11 (m, 1H), 3.76 (s, 3H), 3.66—3.57 (m, 1H), 3.45-3.36 (m, 1H), 2.51—2.46 (m, 1H), 2.33-2.24 (m, 1H), 2.04—1.97 (m, 1H), 1.95—1.83 (m, 1H), 1.49 (s, 9H), 1.20-1.11 (m, 3H).

LCMS (m/Z): 274.3 [M++1].

S nthesis of 1- tert—butox carbon l 1-h drox eth l rrolidine—Z-carbox lic acid jBB): To a stirring solution of compound AA (30 g, 109 mmol) in methanol (50 mL) was added NaOH solution (8.72 g in 10 mL H20, 218 mmol) at RT. The reaction mixture was heated to 70 0C for 12 h. After consumption of the starting material (by TLC), the solvent from the on was evaporated under reduced pressure and diluted with DCM (200 mL). The separated aqueous layer was acidiﬁed using citric acid solution (pH~3) and extracted with DCM (2 x 250 mL). The combined organic layer was washed with brine solution (1 x 100 mL).

The organic layer was dried over NaZSO4 and concentrated to afford compound BB (9.5 g, 33.6%) as brown solid. 1H-NMR: (400 MHz, DMSO-d6): 5 4.53—4.45 (m, 1H), .48 (m, 1H), 3.25—3.19 (m, 1H), 2.33-2.16 (m, 1H), 1.90-1.78 (m, 3H), 1.38 (s, 9H), 0.95 (d, J= 6.4 Hz, 3H).

LCMS (m/z): 258.2 [M'—1]. _ 54 _ Scheme I—7 CHacHO N N a .NaOHq Bod! 0 BOCI O W CC DD S nthesis of -but l 2-eth l 2- 1-h drox eth th l rrolidine-l 2- dicarboxylate (CC 2: To a stirring solution of compound W (18.0 g, 70 mmol) in THF (200 mL) was added LiHMDS (84 mL, 84 mmol) drop wise at -20 °C and stirred for 30 min. To this acetaldehyde (4.2 mL, 77 mmol) was added drop wise and stirred for 45 min at -20 °C. After ption of the starting material (by TLC), the reaction was quenched with aqueous NH4Cl solution (100 mL) and extracted with EtOAc (2 x 150 mL). The separated organic layer was dried over NazSO4 and concentrated to afford crude material was purified by column chromatography eluting with 30% EtOAc/Hexane to afford compound CC (15 g, 71.4%) as colorless syrup. 1H-NMR: (500 MHz, DMSO—d6)28 .25 (m, 5H),4.38 (d, .1: 10.5 Hz, 2H), 4.08—3.98 (m, 1H), 3.88 (d, J= 9.5 Hz, 2H), 2.20-2.08 (m, 2H), 1.38 (s, 9H), 1.37—1.29 (m, 4H), 1.19 (t, J= 7.5 Hz, 3H), 1.14—1.10 (m, 3H); LCMS 378 (M++l) Synthesis of 1-(tert-butoxycarbonyl)(1-hydr0xyethyl)methylpyrrolidine-Z-carboxylic acid (DD): To a stirring solution of compound CC (15 g, 49 mmol) in CH3OH/THF (10 mL/40 mL) were added NaOH (3.98 g, 99 mmol) in water (10 mL) at RT. The reaction mixture was heated to 70 0C for 16 h. After consumption of the starting material (by TLC), the solvent from the reaction was evaporated under reduced pressure and acidiﬁed by using citric acid (pH~4). The aqueous layer was extracted with EtOAc (2 x 200 mL). The ed organic layer was dried over NaZSO4 and concentrated to obtained crude compound, which was puriﬁed by column chromatography g 40% EtOAc to afford compound DD (4 g, 29.4%) as brown syrup. : (500 MHz, DMSO—dg): 8 12.15 (br s, 2H), 4.54—4.50 (m, 1H), 4.03—4.02 (m, 1H), 2.17—1.77 (m, 3H), l.4l(s, 9H), 1.39—1.09 (m, 3H), 0.99—0.94 (m, 3H).

LCMS (m/z): 272.4 [M'-1].

Scheme 1 HO \ OH 69 H \ 69 —’ N N —’ N —> N N N EDCI 7 o / DTAD , TBAF 3°C 3°C 0 0TBS Bee 0 0TBs Int-F C-l C 2 Step 9 N 0 N N '. N OH OH BF3(OEt)2 N OH Isobutyryl chlorlde O BocO H o o -3 C—4 C-5 S nthesis of tert-but 12- 2- tert—but ldimeth lsil 10X carbamo l h drox meth l rrolidine—l-carbox late C-l : ] Referring to Scheme 1, to a stirring solution of I (2 g, 8.16 mmol) in CH2C12 (60 mL) was added intermediate F (2.39 g, 8.97 mmol), EDCI. HCl (2.33 g, 12.2 mmol) followed by HOBt (1.66 g, 12.24 mmol) and DIPEA (4.5 mL, 24.4 mmol) at 0°C. The reaction e was warmed to RT and stirred for 16 h. After consumption of the starting material (by TLC), the reaction mixture was diluted with CHZClz. The separated organic layer was washed with aqueous NaHC03 solution ed by aqueous NH4Cl. The organic layer was dried over anhydrous NaZSO4, ﬁltered and concentrated under reduced pressure to get crude product, which was purified by silica gel column chromatography eluting with 70% EtOAc/hexane to afford compound C-l (2.3 g, 57.5%). 1H-NMR: (400 MHz, DMSO—dg): 88.91 (d, 1H), 8.68 (d, 2H), 7.41 (br s, 1H), 5.74 (br t, 1H), .07—4.89 (m, 1H), 4.15—4.10 (m, 1H), 3.97—3.92 (m, 1H), 3.45—3.41 (m, 1H), 1.79—1.74 (m, 2H), .39 (m, 4H), 1.29—1.21 (m, 6H), 1.12 (d, 5H), 0.71 (s, 9H), 0.12 (t, 1H), 0.09 (s, 2H), 0.08 (s, 1H), 0.04 (s, 2H).

LCMS (m/z): 495.5 [M++1]. —56- S nthesis of tert—but 12- 2- tert—bu ldimeth lsil 10x rimidin 1 r0 l-l-oxo- 2 5-diazas iro 3.4 octane-S-carbox late C-2 : To a ng solution of compound C-l (2.3 g, 4.65mmol) in THF (23 mL) was added TPP (1.34 g, 5.12 mmol) followed by DTAD (1.6 g, 6.98 mmol) at 0 °C. The reaction mixture was warmed to RT and stirred for 16 h. After consumption of the starting material (by TLC), the reaction mixture was concentrated under reduced re to get crude product, which was puriﬁed by silica gel column chromatography eluting with 25% EtOAc/hexane to afford nd C-2 (1.2 g, 54.2%) 1H-NMR: (400 MHz, DMSO—dg): 88.82 ((1, 2H), 7.49 (t, 1H), 4.72 (d, 1H), 4.31 (q, 1H), 3.62 (br s, 2H), .19 (m, 1H), 2.24—2.05 (m, 2H), 1.85—1.81 (m, 2H), 1.42 (br s, 1H), 1.25 (t, 3H), 0.92 (s, 8H), 0.75 (s, 9H), 0.02 (s, 3H).

LCMS (m/z): 477.4 [M++1].

S nthesis of tert-but 12- 2-h drox -l- rimidin-Z- 1 r0 l-l-oxo-Z 5 diazas iro 3.4 octane-S-carboxylate gC-3 [: To a stirring solution of compound C-2 (1.0 g, 2.10 mmol) in THF (20 mL) was added TBAF (1M in THF) (6.3 mL, 6.30 mmol) at 0 0C under N2 atmosphere and stirred for 1 h. After consumption of the starting material (by TLC), the reaction mixture was quenched with ice water and extracted with EtOAc. The separated organic layer was dried over anhydrous NaZSO4, filtered and concentrated under reduced pressure to obtain crude t, which was purified by silica gel column chromatography eluting with 5% MeOH/CHZCIZ to afford compound C-3 (0.35 g, 46%). 1H-NMR: (400 MHz, DMSO-ds): 58.81 (d, 2H), 7.49 (t, 1H), 4.81 (d, 1H), 4.65 (d, 1H), 4.25— 4.20 (m, 1H), 3.64—3.51 (m, 2H), 3.34 (s, 1H), 3.25—3.20 (m, 1H), .20 (m, 2H), 1.87—1.82 (m, 2H), 1.19 (d, 3H), 0.97 (s, 9H).

LCMS (m/z): 363.3 [M++1].

S s of tert—bu 12- 2-h drox rimidin 1 r0 loxo—2 5-diazas iro 3.4 octane-S-carboxylate [04]

[00164] To a stirring on of compound C-3 (200 mg, 0.42 mmol) in THF (5 mL) was added TBAF (1M in THF) (0.84 mL, 0.84 mmol) at 0 °C under N2 here and stirred for 2 h. After consumption of the starting material (by TLC), the reaction mixture was concentrated under reduced pressure to obtain crude product, which was puriﬁed by silica gel column chromatography eluting with 2% MeOH/CHzClz to afford C-4 (125 mg, . 1H-NMR: (400 MHz, CD3OD): 8.78 (t, J: 3.2 Hz, 2H), .38 (m, 1H), 4.92—4.87 (m, 1H), 4.79-4.56 (m, 1H), .38 (m, 1H), 4.33-3.97 (m, 1H), 3.88-3.80 (m, 1H), 3.48—3.44 (m, 1H), .30 (m, 1H), 2.31-2.12 (m, 2H), 1.96-1.84 (m, 2H), 1.42 (s, 9H), 1.32-1.28 (m, 3H); LCMS (m/z): 363.3 [M++1].

HPLC: 94.5%.

S nthesis of 2- 2-h drox n-Z- 1 r0 1-2 5-diazas iro 3.4 octan-l-one C-5 :

[00165] To a stirring solution of compound C—4 (0.3 g, 0.82 mmol) in CHzClz (6 mL) was added molecular sieves (0.3 g) ed by BF3—etherate (0.31 mL, 2.48 mmol) at 0 °C and stirred at RT for 1 h. After consumption of the starting material (by TLC), the reaction mixture was ﬁltered and obtained residue was dissolved in MeOH and washed with CHZClz. The volatiles were evaporated under reduced re to obtain crude product, which was puriﬁed by silica gel column tography eluting with 8% MeOH/CHZClzto afford C—5 (0.12 g, 55%). 1H—NMR: (400 MHz, DMSO—d6): 88.81 (d, 2H), 7.49 (t, 1H), 4.81 (d, 1H), 4.65 (d, 1H), 4.25— 4.20 (m, 1H), 3.64—3.51 (m, 2H), 3.12—3.01 (m, 2H), .10 (m, 2H), 1.87-1.82 (m, 2H), 1.19 (d, 3H).

LCMS (m/z): 263.1 [M++1].

S nthesis of 2- 2-h drox rimidin-Z- 1 r0 l-5—is0but r 1-2 as iro 3.4 octan-l-one gC-6): To a stirring solution of compound C-5 (500 mg, 1.9 mmol) in DCM (10 mL) was added TEA (0.79 mL, 5.72 mmol) followed by isobutyryl chloride (241 mg, 2.28 mmol) at 0 °C under N2 atmosphere and stirred for 2 h at RT. After consumption of the starting material (by TLC), the reaction mixture was diluted with water (10 mL) and extracted with CHzClz (2 x mL). The combined organic layer was washed with citric acid solution (1 x 20 mL). The separated organic layer was dried over anhydrous Na2S04, ﬁltered and concentrated under reduced pressure. Obtained crude material was puriﬁed by silica gel column chromatography eluting 2% MeOH/DCM to afford C-6 (85 mg, 13.4%) as white solid. —58- 1H—NMR: (400 MHz, CD30D): 5 4.91—4.75 (m, 1H), 4.46-4.40 (m, 1H), 3.99-3.85 (m, 1H), 3.70-3.64 (m, 1H), 3.60—3.45 (m, 3H), 2.80-2.69 (m, 1H), 2.29—2.22 (m, 2H), 2.07—1.96 (m, 2H),1.45—1.33(m,3H),1.16—1.10(m,3H), 1.06-1.01 (m, 3H).

Scheme 2 OH 0A[‘1 Step-6 Step-7 ’N \ Int-N ,HATU DIAD PPh3 N --IOTBS O ”I TBAF Boc OTBS I 3000 I C-7 C-8 CA[‘1 O oA!“ ’l\1 ’N Step--9 ’N WM” Step-10 SPERM“), N...

N “CH N "'OH '0' | SObU yryt I BF3<OE02 éocO chloride \(KOO C-9 C- C-11 S s of tert—butvl 2— 1R 2 —Z— tert—butvldimeth 15111 '1 ox ' —l— 1 3 4-oxadiazol-Z— To a stirring solution of compound I (2.1 g, 8.57 mmol) in CHzClz (20 mL) were added DIPEA (3.81 mL, 21.4 mmol), intermediate F (2.64 g, 10.28 mmol), HATU (3.90 g, 10.28 mmol) at 0 °C and stirred to RT for 16 h. After consumption of the ng material (by TLC), the reaction mixture was diluted with water (30 mL). The organic layer was washed with citric acid (1 x 50 mL) followed by brine on (1 X 50 mL). The organic layer was dried over anhydrous Na2S04, ﬁltered and concentrated under reduced pressure. Obtained crude material was d by silica gel column chromatography eluting with 50% n-hexane to afford compound C-7 (3.5 g, 85.15%) as yellow thick syrup. 1H-NMR: (400 MHz, DMSO-dg): 5 9.24 (s, 1H), 8.17 (d, J: 8.4 Hz, 1H), .54 (m, 1H), 4.36—4.28 (m, 1H), 4.12-4.00 (m, 1H), 3.93—3.66 (m, 1H), 3.43-3.39 (m, 1H), 2.69—2.66 (m, 2H), 2.02—1.94 (m, 2H), 1.78—1.68 (m, 2H), 1.40 (s, 9H), 1.23—1.15 (m, 3H), 0.75 (s, 9H), —0.08 (s, 6H); Mass (ESI): m/z 484.7 [M+ +1] S nthesis of tert—bu 12- IR 2 tert—but ldimeth lsil 1 0X 1 3 4-oxadiazol 1 r0 1 carbamo 1 h drox meth l ine-l-carbox late C-8 : To a ng solution of triphenylphosphine (4.05 g, 15.4 mmol) in THF (30 mL) was added DIAD (3.03 g, 15.4 mmol) at RT and stirred for 15 min. After added compound C—7 (3 g, 6.19 mmol) in (20 mL) THF slowly and reaction mixture was stirred at RT for 3 h.

After consumption of the starting material (by TLC), the reaction was concentrated under reduced pressure. The crude material was triturated with 30% Ether/n—hexane (2 X 50 mL). The ﬁltered organic solvent was concentrated and crude residue was d by column chromatography by eluting 30% n—hexane to afford compound C-8 (2 g, 69.4%) as thick syrup. 1H-NMR: (400 MHz, DMSO-dg): 5 9.28 (s, 1H), 5.30-5.21 (m, 1H), 4.84—4.81 (m, 1H), 4.49— 4.29 (m, 1H), 4.05—3.91 (m, 1H), 3.65—3.61 (m, 1H), 3.39—3.24 (m, 1H), .12 (m, 2H), 1.81-1.76 (m, 2H), 1.41 (s, 9H), 1.20-1.14 (m, 3H), 0.75 (s, 9H), -0.22 (s, 6H); Mass (ESI): m/Z 467.6 [M++1] S s of tert—bu 12- IR 2 h drox 1 3 4-oxadiazol 1 r0 l-l-oxo—Z 5- piro |3.4| octane-S-carboxylate [C-9): To a stirring solution of compound C-8 (2 g, 4.29 mmol) in THF (20 mL) was added TBAF (4.3 mL, 6.43 mmol) slowly at 0° C and stirred at RT for 30 min. After completion of reaction (by TLC), the reaction mixture was evaporated under reduced pressure.

The crude residue was puriﬁed by column chromatography by eluting 70% EtOAc/n—hexane to afford C-9 (400 mg, 26.5%) as white solid. 1H-NMR: (400 MHz, DMSO-dg): 9.29 (s, 1H), 5.30-5.21 (m, 1H), 5.08-4.95 (m, 1H), 4.08- 4.03 (m, 1H), 3.91—3.71 (m, 1H), 3.75—3.71 (m, 1H), 3.65—3.52 (m, 1H), 3.36—3.22 (m, 1H), 2.22—2.09 (m, 2H), 1.83-1.77 (m, 2H), 1.41 (s, 9H), 1.10—1.04 (m, 3H); LCMS: 353.4; HPLC: 99.82%.

S nthesis of 2- 1R 2S h drox 1 3 iazol—2- 1 r0 1-2 5-diazas iro 3.4 octan-l-one C-10 :

[00170] To a stirring on of compound C-9 (500 mg, 1.07 mmol) in DCM (5 mL) was added BF3(OEt)2 (0.26 mL, 2.14 mmol) slowly at 0 °C and stirred at RT for 30 min. After completion of reaction (by TLC), the reaction mixture was evaporated under reduced pressure. —60- The crude e was triturated with di ther/n-pentane (5 mL/5 mL) to obtain solid, C- , which was taken into the next step without further puriﬁcation.

S nthesis of 2- 1R 2 h drox 1 3 4-0xadiazol 1 r0 l-5—isobut r 1-2 5- diazas iro 3.4 octan-l-one C-ll :

[00171] To a stirring solution of compound C-10 (700 mg, 2.77 mmol) in DCM (10 mL) was added TEA (0.94 mL, 6.92 mmol), SM-3 (440 mg, 4.15 mmol) at 0 °C and stirred to RT for 1 h. After tion of reaction (by TLC), diluted with water (20 mL). The organic layer was washed with brine solution (1 x 50 mL). The organic layer was dried over anhydrous NaZSO4, ﬁltered and concentrated under reduced pressure. The crude residue was puriﬁed by column chromatography ed by preparative HPLC puriﬁcation to afford C-11 (150 mg. 16.8%) as white solid. 1H-NMR: (400 MHz, DMSO-dg): 9.26 (s, 1H), .04 (m, 1H), 5.03—4.93 (m, 1H), 4.27— 3.91 (m, 1H), 3.90—3.64 (m, 1H), 3.49—3.45 (m, 3H), 2.73—2.61 (m, 1H), 2.17-2.14 (m, 2H), 2.10-2.06 (m, 2H), 1.25—1.07 (m, 3H), 0.95—0.93 (m, 6H).

LCMS m/z: 323.3.

HPLC: 97.89%.

Scheme 3 \0 E, OH Step 6 N Step5 '}‘ OMe pd_C,H2 , OMe 800 800 Int S O 0 G C-12 C-l3 o’ ‘7 Step 8 Step 7 HN\I N BOCON ”OH TFAHO "OH | OMe C-l4 C-15 S nthesis of l-tert-bu lZ-meth 12- h drox meth l rrolidine-l 2-dicarbox late C-12 : To a stirring solution of compound G (74 g, 0.21 mol) in methanol (740 mL) was added 10% Pd/C (50% wet, 14.8 g) under N2 atmosphere and stirred for 6 h under H2 2014/013621 —61- atmosphere (balloon pressure). The reaction mixture was ﬁltered through celite pad and trated under reduced re to afford compound C-12 (45 g, 82%) as crude.

S nthesis of l-tert-but lZ-meth lZ-form l rrolidine—l 2-dicarb0x late C-13 : ] To a stirring solution of compound C-12 (10 g, 38.5 mmol) in CHzClz (100 mL) was added Dess—Martin periodinane (19.6 g, 46.27 mmol) at 0 °C under N2 atmosphere and stirred fro 3 h. After consumption of the starting material (by TLC), the reaction was quenched with aqueous NaHCO3 solution and extracted with CH2C12 (2x 100 mL). The organic layer was dried over anhydrous NaZSO4 and concentrated under vacuum. The crude was puriﬁed by column chromatography g with 10% EtOAc/Hexane to afford compound C-13 (7 g, 70.5%).

S s of l—tert-butvi Z—meth 1 2— z. —hvdmx -1—-12 4—0xadiazoi—5— '1; Ho amino meth l 'rrolidine-l Z-dicarbox 'late C-14 :

[00174] To a stirring solution of compound C-13 (3 g, 11.6 mmol) in MeOH (30 mL) was added sodium acetate (1.91 g, 23.3 mmol) followed by intermediate S (3.6 g, 13.9 mmol).

The reaction mixture was heated to reﬂux for 1 h. The on mixture was slowly cooled to RT—O °C, to this sodium cyanoborohydride (1.465 g, 23.3 mmol) and stirring was continued for another 6 h at RT. After consumption of the starting material (by TCL), methanol from the reaction was evaporated under reduced pressure and the residue was diluted with water and extracted with EtOAc (2x 50 mL). The separated organic layer was dried over anhydrous Na2S04 and concentrated under reduced pressure to give crude; which was puriﬁed by silica gel column chromatography eluting with 40% EtOAc/Hexane to afford nd C-14 (2.5 g, 56%).

LCMS m/z: 385 [M++1].

S nthesis of ut l 2- 1S 2R h drox 1 2 4—0xadiazol—5- 1 r0 l-l-oxo-Z 5- piro |3.4| octane-S-carboxylate gC-lS]: To a stirring solution of compound C-14 (1.5 g, 3.90 mmol) in THF (30 mL) was cooled to 0 °C and added t—BuMgCl (1M in THF, 15.6 mL, 15.6 mmol) and stirred for 15 min. After consumption of the starting material (by TLC), the reaction mixture was quenched with aqueous NH4Cl solution and diluted with water. Aqueous layer was extracted with EtOAc (2x 25 mL). The separated organic layer was dried over anhydrous NaZSO4 and concentrated 2014/013621 under reduced pressure. Obtained crude material was puriﬁed by silica gel column tography eluting with 150% EtOAc/CHZCIZto afford C-15 (0.15 g, 11%). 1H—NMR: (400 MHz, DMSO—d6): 5 9.02 (s, 1H), 5.15 (s, 1H), 4.37-4.32 (m, 1H), 3.95 (d, 1H), 3.66—3.60 (m, 1H), 3.36—3.30 (m, 1H), 2.29—2.09 (m, 2H), 1.87—1.82 (m, 2H), 1.55 (s, 9H), 1.27 (d, 3H).

LCMS (ESI) m/z: 351 [Mt—1].

HPLC Purity: 96%.

S s of 2- 1S 2R h drox 1 2 4-oxadiazol 1 r0 1-2 5—diazas iro |3.4|0ctan0ne gC-162:

[00176] To a stirring solution of C—15 (0.4 g, 1.13 mmol) in CHzClz (4 mL) was added TFA (0.43 mL) at 0 OC and stirred at RT for 30 min. The reaction e was concentrated under vacuum. Obtained crude material was puriﬁed by prep-HPLC to afford C-16 (65mg) as TFA salt. 1H—NMR: (400 MHz, DMSO-d6): 5 9.89 (br s, 1H), 9.08 (s, 1H), 5.46 (d, 1H), 5.31 (s, 1H), 4.37-4.35 (m, 1H), 3.99 (d, 1H), 3.81 (d, 1H), 3.42-3.35 (m, 2H), 2.35-2.18 (m, 2H), 2.10-2.03 (m, 2H), 1.24 (d, 3H).

LCMS (ESI) m/z: 253.4 [M++1].

HPLC Purity: 95%.

Scheme 4 Ste 10 N:\ Ste 11 NP) \BOCO Int F,EDC| | _ DTAD: PPh3 BOCO OTBS BOCO OTBS Step 12 Step 13 ‘N —> > \N —> THF/TBAF I BF OEt M3 Boco H OH C—19 C-20 S nthesis of tert—but l 2- 2- tert-but ldimeth lsil l ox rimidin-Z-l r0 1 carbamo 1 h drox meth lmeth l rrolidine-l-carbox late C-17 : —63- To a stirring solution of compound Z (500 mg, 1.93 mmol) in DCM (10 mL) were added isopropylethylamine (1.0 mL, 5.79 mmol), intermediate F (566mg, 2.12mmol), followed by EDCI (737mg, 3.86 mmol), HOBT (521mg, 3.86mmol) at 0 °C and stirred at RT for 16 h. After consumption of the starting material (by TLC), the reaction e was diluted with water (20 mL). The separated organic layer was washed with saturated brine solution (1x30 mL). The separated organic layer was dried over anhydrous NaZSO4 and concentrated under reduced pressure to afford crude compound which was purified by column chromatography to obtained compound C-17 (300 mg, 30.6%) as pale yellow liquid. 1H-NMR: (500 MHz, CDC13): 8 .68 (m, 3H), 5.30 (s, 2H), 4.40—4.15 (m, 1H), .61 (m, 1H), 3.18-3.13 (m, 1H), 2.18-2.13 (m, 4H), 1.46 (s, 9H), 1.32 (d, J= 5.5 Hz, 3H), 1.24 (d, J = 6.5 Hz, 3H), 1.08 (d, J= 6.5 Hz, 1H), 0.79 (s, 9H), 0.74 (t, J= 10.5 Hz, 1H), 0.12 (s, 6H) S nthesis of tert—but l 2- 2- tert-but ldimeth lsil l ox n-Z- 1 r0 1 methyl-l-oxo-Z, aspiro |3.4| octane-S-carboxylate gC-182:

[00178] To a stirring solution of triphenylphosphine (3 80 mg, 1.47 mmol) in dry THF (10 mL) was added DTAD (339 mg, 1.47 mmol) at RT and stirred for 10 min. After added compound C—17 (3 00 mg, 0.59 mmol) and the reaction mixture was stirred at RT for 16 h. After consumption of the starting al (by TLC), the reaction e was concentrated under reduced pressure. The crude material was purified by silica gel column chromatography eluting 30% EtOAc/hexane to afford compound C-18 (80 mg, 27.5%) as yellow liquid. 1H-NMR: (400 MHz, CDC13):57.73-7.64 (m, 3H), 5.26 (s, 2H), 4.30 (dd, J: 4.4 Hz, 4.4 Hz, 1H), 4.17-4.12 (m,1H), 2.76 (t,J= 6.8 Hz, 1H), 2.32—2.28 (m,2H), 2.05—2.00 (m, 2H), 1.49 (s, 9H), 1.45 (d, J: 5.2 Hz, 3H), 1.42 (d, J: 4.8 Hz, 3H), 0.89 (s, 9H), 0.15 (s, 6H).

S nthesis of tert—but l 2- 2-h drox rimidin 1 r0 lmeth loxo-2 5- diazaspiro |3.4| octane-S-carboxylate gC-l9]: To a stirring on of nd C-18 (700 mg, 1.42 mmol) in dry THF (10 mL) was added TBAF (744 mg, 2.85mmol) at 0 OC. The reaction mixture was stirred at RT for 3 h. After consumption of the starting material (by TLC), the reaction mixture was concentrated under reduced pressure. The crude al was purified by silica gel column chromatography eluting 80% EtOAc/hexane to afford C-19 (0.16 g, 30%) as yellow thick syrup. —64- 1H-NMR: (500 MHz, DMSO-da): 5 8.77 (d, J= 5.0 Hz, 2H), 7.42(t, J= 4.5 Hz, 1H), 4.76-4.63 (m, 2H), 4.14—4.11 (m, 1H), 3.85-3.83 (m,lH), 3.54—3.34 (m, 1H), 2.25-2.21 (m,2H), 2.08—1.93 (m, 1H), 1.53—1.47 (m, 1H), 1.39 (s, 9H), 1.15(d, J= 6.0 Hz, 3H), 1.03-0.80 (m, 4H).

LCMS (ESI): 377.4 .

HPLC: 94.6% (both isomers).

S nthesis of 2- 2-h drox n-Z- 1 r0 lmeth 1-2 5-diazas iro 3.4 octan- l-one 10201: To a stirring solution of compound C-19 (260 mg, 0.69 mmol) in DCM (10 mL) was added 4A molecular sieves (70 mg), BF3'OEt2 (0.08 mL) at 0° C under N2 here. The reaction mixture was stirred at RT for 30 min. After ption of the starting material (by TLC), the reaction mixture was concentrated under d pressure to afford crude, which was puriﬁed by preparative HPLC method to afford C-20 (65 mg, 34%) as white solid. 1H-NMR: (400 MHz, D20):8 8.88—8.86 (m, 2H), 7.63—7.57 (m, 1H), 4.92—4.89 (m, 1H), 4.61— 4.56 (m, 1H), 4.03—3.91 (m, 3H), 2.61—2.41 (m, 3H), 1.92—1.84 (m,lH), 1.46 (d, J: 6.8 Hz, 3H), 1.37 (d, J: 4.8 Hz, 3H); LCMS (ESI): 295.3 [M++H20].

HPLC: 91.2% (both enantiomers).

Scheme 5 OH 0AI," Step-10 Step11 ’N \ 0 Int-N EDCI DTAD PPh3 N 0TBs 3°C ores Elmo Z C-21 C-22 Step—120_NN THF / TBAF BocN C-23 S nthesis of ut 12- 2- tert—bu ldimeth lsil 1 0x 1 3 4-oxadiazol 1 r0 1 carbamo l h drox meth lmeth l rrolidine—l-carbox late C-21 : ] To a ng solution of compound Z (4 g, 15.4 mmol) in DCM (30 mL) were added AZN—diisopropylethylamine (8 mL, 46.33mmol), intermediate N (4.74 g, 18.48 mmol) followed by HATU ( 7 g, 18.48 mmol) at 0 OC and stirred at RT for 12 h. After consumption of the ng material (by TLC), the reaction mixture was diluted with water (20 mL). The separated organic layer was washed with 10% citric acid solution (1 x 50 mL) followed by saturated brine solution (1x50 mL). The separated c layer was dried over anhydrous Na2S04 and concentrated under reduced pressure to afford crude compound which was puriﬁed by column chromatography g 30% n—hexane to obtained compound C-21 (5.5g, 71.8%) as pale yellow liquid. 1H-NMR: (500 MHZ, DMSO-dg): 59.25 (s, 1H), 5.50-5.41 (m, 1H),5.19—5.06 (m, 2H), 4.38— 4.36 (m,1H), 4.05—3.90 (m, 3H), 2.69—1.98 (m, 2H), 1.39(s, 9H), 1.37—1.26 (m, 2H), 1.23—1.16 (m, 6H), 0.76 (s, 9H), -0.01 (s, 6H).

To a ng solution of triphenylphosphine (5.5 g, 11.04 mmol) in THF (55 mL) was added DTAD (5.07 g, 22.05 mmol) at RT and stirred for 10 min. After added compound C—21 (5.5g, 11.04mmol) and the reaction mixture was stirred at RT for 16 h. After consumption of the starting material (by TLC), the reaction mixture was concentrated under reduced pressure. The crude material was puriﬁed by silica gel column chromatography eluting % EtOAc/hexane to afford compound C-22 (4 g, 75.4%) as yellow liquid. 1H-NMR: (400 MHz, DMSO-dg): 5 9.36 (s, 1H), 5.30—4.48 (m, 1H), 4.47—4.31 (m, 1H), 4.03— 3.88 (m,2H), 3.62—3.58(m,1H), 2.38—1.98 (m,3H), 1.57—1.53 (m, 1H), 1.41 (s, 9H), 1.38—1.15 (m, 6H), 0.69 (s,9H), -0.01 (s, 6H).

S nthesis tert—but l 2- 2-h drox 1 34-oxadiazol 1 r0 1 meth loxo-2 5- diazaspiro |3.4| octane-S-carboxylate [C-23]: ] To a ng solution of compound C-22 (3.5 g, 7.29 mmol) in THF (35 mL) was added TBAF (1M THF) (14.5 mL, 14.58 mmol) at 0 oC. The reaction mixture was stirred at RT for 1 h. After consumption of the starting material (by TLC), the reaction mixture was concentrated under reduced pressure. The crude material was puriﬁed by silica gel column chromatography eluting 60% EtOAc/hexane to afford C-23 (800 mg, 30.7%) as an off—White solid. 1H-NMR: (400 MHz, DMSO-dg):89.30 (s, 1H), .2l(m, 1H), 5.13-5.02 (m, 1H), 4.13- 4.03 (m, 1H), 3.90-3.83 (m,1H), 3.74-3.68 (m, 1H), .49 (m,lH), 2.29 (m, 3H), 1.56-1.52 (m, 1H), 1.40 (s, 9H), l.25—l.l4(m, 6H).

LCMS (ESI) (m/z): 377.4 [M++1].

HPLC: .

Scheme 6 Step 5 Step 6 OH —> BOON EDCI DIAD, PPh "IOTBS ’IOTBS 3 Int-N BOCO BB C-24 -C25 Step 7 L N II TBAF N BocO ] To a stirring solution of compound BB (3 g, 11.5 mmol) in CHzClz (30 mL) were added DIPEA (6 mL, 34.5 mmol), intermediate N (2.95 g, 11.5 mmol), HATU (5.24 g, 13.8 mmol) at 0 °C and stirred to RT for 12 h. After consumption of the starting material (by TLC), the reaction mixture was diluted with water (30 mL) and extracted with CHZClz (50 mL).

The combined organic layer was washed with citric acid (1 x 50 mL) followed by brine solution (1 x 50 mL). The organic layer was dried over anhydrous NaZSO4, ﬁltered and concentrated under reduced pressure. Obtained crude material was puriﬁed by silica gel column chromatography g with 35% EtOAc/n—hexane to afford compound C-24 (3.6 g, 62.5%) as yellow thick syrup. 2014/013621 1H-NMR: (500 MHz, DMSO—dg): 5 9.26 (s, 1H), 8.74 (d, J: 8.5 Hz, 2H), 5.30-5.12 (m, 1H), .58 (m, 1H), 4.38—4.35 (m, 2H), 3.57—3.54 (m, 1H), 2.68—1.98 (m, 2H), 1.97—1.69 (m, 2H), 1.40 (s, 9H), 1.19-1.00 (m, 6H), 0.85 (s, 6H), —0.25 (s, 9H).

Mass (ESI): m/z 497.6[M‘—l].

S nthesis of tert-but 12- IR 2S tert-bu ldimeth lsil 1 0x 1 3 4-oxadiazol 1 r0 l-l-meth loxo-2 as iro 3.4 octane-S-carbox late C-25: To a stirring solution of triphenylphosphine (3.6 g, 14.05 mmol) in THF (20 mL) was added DIAD (2.8 g, 14.05 mmol) at RT and stirred for 30 min. After added nd C-24 (3.5 g, 7.02 mmol) in (15 mL) THF slowly and reaction mixture was stirred at RT for 6 h.

After consumption of the starting material (by TLC), the reaction was trated under reduced pressure. The crude material was puriﬁed by column chromatography by eluting 30% EtOAc/n—hexane to afford compound C-25 (3 g, 89.2%) as thick syrup. 1H-NMR: (400 MHz, DMSO—d6): 8 9.33 (s, 1H), .29 (m, 2H), 4.79—4.75 (m, 1H), 4.50— 4.45 (m, 1H), 4.06-3.96 (m, 1H), 2.16-2.08 (m, 1H), 2.01-1.89 (m, 1H), 1.81-1.76 (m, 2H), 1.40 (s, 9H), 1.24—1.15 (s, 6H), 0.80 (s, 9H), —0.08 (s, 6H).

Mass (ESI): m/Z 319.3 [M++1].

S nthesis of tert—but 12- IR 2S h drox 1 3 4-oxadiazol 1 r0 l-l-meth l oxo—Z, 5-diazaspiro |3.4| octane-S-carboxylate 1C-26): To a stirring solution of compound C—25 (3 g, 6.25 mmol) in THF (25 mL) was added TBAF (12.5 mL, 12.5 mmol) at 0 °C and stirred for 30 min. After consumption of the starting material (by TLC), the reaction mixture was concentrated under reduced pressure.

Obtained crude material was puriﬁed by silica gel column tography eluting with 60% EtOAc/n-hexane to afford C-26 (0.8 g, 35%) as white solid. 1H-NMR: (400 MHz, g): 8 9.29 (s, 1H), 5.28—5.15 (m,lH), 5.08—4.89 (m, 1H), 4.43- 4.35 (m, 1H), 3.97-3.79 (m, 1H), 3.65—3.40 (m, 1H), .25 (m, 1H), 2.15—2.00 (m, 2H), 1.93—1.86 (m, 1H), 1.80—1.68 (m, 1H), 1.40 (s, 9H), 1.18—1.04 (m, 6H); Mass (ESI): m/z 367.4 [M++1].

HPLC:95.85% (both isomers). _ 68 _ Scheme 7 HO HO \ Step 5 \ / Step 6 N¢\\> H \ OH —. N _. N N N N EDCI ,N DIAD /N 300/ 0 Int-F B00 0 OTBS B00 0 OTBS BB C-27 C-28 Step 7 \ / TBAF 5" B00 0 OH C-29 To a stirring solution of nd BB (200 mg, 0.77 mmol) in CHzClz (20 mL) were added DIPEA (298 mg, 2.31 mmol), EDCI (221 mg, 1.15 mmol), HOBt (177 mg, 1.15 mmol) followed by intermediate F (206 mg, 0.77 mmol) at 0 OC and stirred for 16 h at RT.

After consumption of the starting material (by TLC), the reaction mixture was concentrated under reduced pressure. ed crude material was puriﬁed by silica gel column chromatography eluting with 4% MeOH/DCM to afford compound C-27 (210 mg, 53.6%) as pale yellow thick syrup. 1H-NMR: (500 MHz, DMSO-dg): 5 8.77 (d, J= 4.5 Hz, 2H), 7.40 (t, J= 5.0 Hz, 1H), 4.95— 4.92 (m, 2H), 4.21—4.05 (m, 2H), 3.34-3.25 (m, 2H), 3.20—3.12 (m, 1H), 1.81-1.63 (m, 4H), 1.48 (s, 9H), 1.12—1.06 (m, 3H), 0.99—0.85 (m, 3H), 0.70 (s, 9H), —0.06 (s, 6H); LCMS (m/z): 509.4 [M++l] S nthesis of tert—but l 2- 2- tert-but ldimeth lsil 1 0X n-Z- 1 r0 l To a stirring solution of triphenylphosphine (206 mg, 0.78 mmol) in THF (6 mL) was added DIAD (179 mg, 0.78 mmol) at RT and stirred for 30 min. To this added compound C—27 (200 mg, 0.39 mmol) in (5 mL) THF slowly and reaction e was stirred at RT for 2 h. After consumption of the starting material (by TLC), the reaction was concentrated under d pressure. The crude material was triturated with diethylether/n— pentane (10 mL/10 mL). The ﬁltered solvent was concentrated and puriﬁed by silica gel column chromatography eluting 30% EtOAc/n—hexane to afford compound C-28 (80 mg, 41.8%) as pale yellow liquid. : (400 MHz, g): 5 8.81—8.77 (m, 2H), 7.72 (d, J= 5.5 Hz, 1H), 4.92—4.80 (m, 1H), 4.70-4.65 (m, 2H), 4.46-4.20 (m, 1H), 3.30-3.16 (m, 1H), 1.86-1.65 (m, 2H), 1.64-1.43 (m, 2H), 1.40 (s, 9H), 1.20—1.01 (m, 3H), 0.92—0.85 (m, 3H), 0.74 (s, 9H),—0.06 (s, 6H); LCMS (m/z): 491.4 [M++l].

S nthesis of tert-but l 2- 2-h drox rimidin-Z- 1 r0 l-l-meth l0X0-2 5- diazaspiro |3.4| octane-S-carboxylate 10292:

[00190] To a stirring solution of compound C-28 (450 mg, 0.91 mmol) in THF (10 mL) was added TBAF (1.8 mL, 1.83 mmol) in 5 mL THF at 0 °C and stirred to RT for 4 h. After consumption of the starting material (by TLC), the reaction was concentrated under reduced pressure. The crude material was puriﬁed by silica gel column tography eluting 2% CM followed by preparative HPLC puriﬁcation to afford C-29 (100 mg, 29%) as a semi solid. 1H-NMR: (400 MHz, CD3OD): 5 8.82—8.79 (m, 2H), 8.78-7.43 (m, 1H), .89 (m, 1H), 4.63—4.44 (m, 1H), 3.99-3.73 (m, 1H), 3.56—3.36 (m, 2H), 2.29—2.22 (m, 2H), 1.99-1.73 (m, 2H), 1.40 (s, 9H), .20 (m, 3H), 1.13—1.05 (m, 3H).

LCMS(m/s): 377.5 [M++1].

HPLC: 94.4% (both isomers).

Scheme 9 OH N/\> 0H Step 9 \/\ Step 10 N4\\>\ OH —> N —> N N EDCI N 800’ 0 DTAD Int-F Bocl /N OTBS 800 o OTBS DD C— C—31 \ \ N N Step 11 \ N/ Step 12 \ N/ —> —> N BF ()OEt2 TBAF BocN 3( H 0 OH 032 C-33 S nthesis of tert—but l 2- 2- ut ldimeth lsil l ox rimidin-Z-l r0 1 carbamo l 1-h drox eth lmeth l rrolidine-l-carbox late C-30 : To a stirring on of compound DD (500 mg, 1.83 mmol) in DCM (10 mL) were added iisopropylethylamine (0.84 mL, 4.57 mmol), intermediate F (586 mg, 2.19 mmol), followed by EDCI (296 mg, 2.19 mmol), HOBT (338 mg, 2.19 mmol) at 0 OC and d at RT for 16 h. After consumption of the starting material (by TLC), the reaction e was diluted with water (20 mL). The separated organic layer was washed with brine solution (1 x 20 mL). The separated organic layer was dried over anhydrous NaZSO4 and concentrated under reduced pressure to afford crude compound which was puriﬁed by column chromatography by eluting 2% MeOH/DCM to obtained compound C-30 (800 mg, 83.6%) as pale yellow liquid. 1H—NMR: (500 MHZ, DMSO-d6)258.78-8.75 (m, 2H), 7.42—7.39 (m, 1H), 5.01-4.92 (m,lH), 4.21—4.09 (m, 1H), 4.05-4.02 (m, 2H), 1.98 (s, 2H), 1.44—1.31 (m, 7H), 1.29—1.11 (m, 10H), 1.02-0.96 (m, 4H), 0.70-0.63 (m, 9H), -0.25 (s, 3H), -0.32 (s, 3H).

LCMS (ESI) : 523.6 [M++1].

S nthesis of tert—bu l 2- dimeth loxo-2 5-diazas iro 3.4 octane-S-carbox late C-3l : To a stirring solution of triphenylphosphine (1.23 g, 4.87 mmol) in dry THF (10 mL) was added DTAD (1.09 g, 4.77 mmol) as n-wise and stirred for 15 min at RT. To this precipitated solution, added compound C-30 (1.0 g, 1.91 mmol) in dry THF (10 mL) slowly at RT and stirred for 16 h. After consumption of the starting material (by TLC), the reaction mixture was concentrated under reduced pressure. The crude material was triturated with n—pentane (20 mL) and ﬁltered solid (TPPO). The ﬁltrate was concentrated under reduced pressure to obtained crude compound was puriﬁed by silica gel column chromatography eluting % EtOAc/hexane to afford compound C-31 (400 mg, 41.4%) as yellow liquid. 1H-NMR: (400 MHz, g): 5 8.82-8.79 (m, 2H), .42 (m, 1H), 3.88-3.70 (m, 2H), 1.82—1.78 (m, 2H), 1.49—1.41 , 1.40 (s, 9H), 1.21—1.08 (m,9H), 0.57 (s, 6H), — 0.05 (s, 9H); LCMS (E51): 505.5 [M++1].

S nthesis of tert-but l 2— 2-h drox rimidin-Z- 1 r0 l-1 6-dimeth loxo-2 5- diazaspiro |3.4| octane-S-carboxylate 10321: 2014/013621 To a stirring solution of compound C-31 (220 mg, 0.43 mmol) in dry THF (3 mL) was added TBAF (227 mg) slowly at 0 OC and stirred at RT for 16 h. After consumption of the starting material (by TLC), the reaction mixture was concentrated under reduced pressure to afford crude, which was puriﬁed by silica gel column chromatography eluting 80% EtOAc/hexane to afford compound C-32 (150 mg, 88.2%) as yellow liquid. 1H-NMR: (500 MHz, DMSO-dg): 8881—879 (m, 2H), 7.43—7.41 (m, 1H), 4.36—4.33 (m, 1H), 4.27-4.24 (m, 1H), 3.86—3.80 (m, 2H), 2.15-1.86 (m, 4H), 1.53—1.50 (m, 1H), 1.43 (s, 9H), 1.19— 1.03 (m, 9H). octan-l-one C-33 : To a stirring on of compound C-32 (120 mg, 0.25 mmol) in DCM (10 mL) was added molecular sieves (100 mg), BF3(OEt)2 (72 mg, 0.51 mmol) at 0 °C under N2 atmosphere. The reaction mixture was stirred at RT for 10 min. After consumption of the starting material (by TLC), the obtained precipitate was ated with n—pentane/diethylether (5 mL/5 mL) and the ﬁltered solid was dried on vacuum to afford C—33 (80 mg, 89.8%) as white solid. : (400 MHz, D20): 8 8.90 (t, J: 4.8 Hz, 2H), 7.64—7.60 (m, 1H), 4.72-4.62 (m, 2H), 4.24—4.15 (m, 1H), .00 (m, 1H), 2.60—2.32 (m,3H), 2.02—1.91 (m, 1H), .51 (m, 3H), 1.48-1.45 (m, 3H), 1.39-1.25 (m, 3H).

LCMS (ESI): 581.3 [2M++1].

Scheme 10 pew iSt 5 “1ij St6 6 NW\ N a N N HATU DTAD IN Bog 0 Int-F Boc OTBS B00 0 OTBS BB C-34 C-35 Step7 \ / Step8 N N —> N BF30<02H5>2 0“ OHYkCI O C-36 To a stirring solution of compound BB (2.5 g, 9.65 mmol) in CHZCIZ (25 mL) were added DIPEA (5 mL, 28.95 mmol), intermediate F (2.57 g, 9.65 mmol) followed by HATU (4.4 g, 11.58 mmol) at 0 °C and stirred for 12 h at RT. After consumption of the starting material (by TLC), the reaction mixture was the reaction was diluted with water (20 mL). The ted organic layer was washed with citric acid solution (1 x 50 mL) followed by brine solution (1 x 50 mL). The organic layer was dried over anhydrous , ﬁltered and concentrated under reduced re. ed crude material was ed by silica gel column chromatography g 50% EtOAc/n—hexane to afford compound C-34 (3.1 g, 63.2%) as an off—white solid. 1H—NMR: (500 MHz, DMSO—dg): 5 8.76 (d, J= 13.5 Hz, 2H), 8.42 (d, J: 9.0 Hz, 1H), 7.41 (d, J: 9.0 Hz, 1H), .41 (m, 1H), 4.23-4.11 (m, 1H), 3.58—3.40 (m, 3H), 2.37—1.62 (m, 4H), 1.39 (s, 9H), 1.28—1.19(m, 3H), 1.18-1.15 (m, 3H), 1.09 (s, 9H), —0.04 (s, 6H); LCMS (ESI): m/z 509.7 [M++1] S nthesis of tert—but l 2- 2- tert-but ldimeth lsil 1 OX rimidin-Z- 1 r0 l methyl-3—oxo—2a 5-diazaspiro |3.4| octane-S-carboxylate gC-352:

[00196] To a stirring on of triphenylphosphine (1.6 g, 6.29 mmol) in THF (15 mL) was added DIAD (1.44 g, 6.29 mmol) at RT and stirred for 30 min. To this added compound C— 34 (1.6 g, 3.14 mmol) in (5 mL) THF slowly and reaction mixture was stirred at RT for 3 h.

After consumption of the starting material (by TLC), the reaction was concentrated under reduced pressure. The crude material was triturated with diethyl ether/n-pentane (10 mL/ 10 mL). The filtered t was concentrated and purified by silica gel column chromatography eluting 20% EtOAc/n—hexane to afford compound C-35 (1 g, 65.3%) as pale yellow liquid. 1H-NMR: (500 MHz, DMSO—dg): 8 8.80 (d, J= 14.5 Hz, 2H), 7.44 (d, J= 9.0 Hz, 1H), 4.72- 4.46 (m, 2H), 3.36—3.23 (m, 3H), 2.06—1.61 (m, 4H), 1.40 (s, 9H), 1.35—1.23 (m, 3H), 1.16—1.12 (m, 3H), 1.00 (s, 9H), —0.01 (s, 6H); LCMS (ESI): m/z 491.4 [M++1] S nthesis of 2- 2-h drox rimidin-Z- 1 r0 lmeth 1-2 5—diazas iro 3.4 octan- l-one gC-361: 2014/013621 To a stirring solution of compound C—35 (1 g, 2.04 mmol) in DCM (5 mL) was added 2H5)2 (0.05 mL, 4.08 mmol) followed by molecular sieves (50 mg) at 0 °C and stirred to RT for 3 h. After consumption of the starting material (by TLC), the reaction was diluted with n-pentane and ed the obtained solid. The ﬁltered solid was puriﬁed by silica gel column chromatography eluting 5% MeOH/DCM to afford C-36 (180 mg, 32%) as an off- white solid. 1H-NMR: (400 MHz, D20): 8 8.89 (d, J= 4.8 Hz, 2H), 7.61 (t, J= 4.8 Hz, 1H), 4.85-4.73 (m, 2H), 4.18—4.13 (m, 1H), 3.54—3.41 (m, 2H), 2.43—2.36 (m, 1H), 2.31—2.12 (m, 3H), 1.43-1.37 (m, 6H) LCMS (ESI): m/z 277.3 [M++l]; HPLC: 98.85%. diazas iro 3.4 octan-l-one C-37 :

[00198] To a stirring solution of nd C-36 (600 mg, 2.17 mmol) in CH2C12 (10 mL) was added TEA (1.05 mL, 759 mmol) at 0 0C. After added isobutyryl chloride (253 mg, 2.39 mmol) slowly and stirred for l h at RT. After consumption of the starting material (by TLC), the reaction e was diluted with water (10 mL). The separated organic layer was washed with brine on (1 x 50 mL). The organic layer was dried over anhydrous Na2S04, ﬁltered and concentrated under reduced pressure. Obtained crude material was purified by silica gel column chromatography eluting 3% MeOH/DCM followed by preparative HPLC puriﬁcation to afford C-37 (120 mg, 16%) as an off-white solid. 1H-NMR: (500 MHz, DMSO—dg): 8 8.81 (d, J= 7.5 Hz, 2H), 7.43 (d, J: 11.5 Hz, 1H), 5.05— 4.34 (m, 2H), 3.64-3.44 (m, 3H), 2.01—1.81 (m, 5H), 1.35—1.13 (m, 6H), 1.10-1.04 (m, 6H); LCMS (ESI): m/z 347.4 [M++l] HPLC: 91.1% Preparation of key intermediates, schemes I-8 to I-10: Scheme I—8 WO 20784 _ 74 _ OBn Step 1 Step 2 CE OE Pd--C/H2 t BoxcO H 0 EE FF Step 3 Step 4 chogensr 1}] NaOH 66 HH S nthesis of eth 12- ben 10x meth th l rrolidine-Z-carbox late EE : To a stirring solution of X (8.5 g, 22.51 mmol) in DCM (50 mL) was added TFA (8.6 mL, 112.58 mmol) at 0 °C. The reaction mixture was stirred at RT for 4 h. After consumption of the starting material (by TLC), the reaction mixture was concentrated under reduced pressure to afford EE (14 g, crude) as pink liquid (TFA salt). 1H-NMR: (400 MHz, DMSO—dg): 5 9.71 (br s, 1H), 7.45-7.30 (m, 5H), 4.62 (s, 2H), 4.27—4.22 (m, 2H), 3.90—3.73 (m, 3H), 2.26-2.08 (m, 2H), 2.01—1.95 (m, 1H), 1.59—1.51 (m, 1H), 1.36— 1.31 (m, 3H), 1.22—1.18 (m, 3H); LCMS (ESI): m/z 278.36 [M++l] S nthesis of eth 12- h drox meth th l rrolidine—Z-carbox late FF : To a stirring solution of EE (14 g (crude), 35.80 mmol) in methanol (100 mL) was added (50% wet) 10% Pd/C (5 g) under N2 atmosphere. The reaction mixture was d under H2 atmosphere (balloon pressure) at RT for 16 h. After consumption of the starting material (by TLC), the reaction mixture was ﬁltered through a pad of celite and the pad was washed with ol (100 mL). Obtained te was concentrated under reduced pressure to afford FF (8 g, crude) as yellow syrup. 1H-NMR: (400 MHz, DMSO—dg): 8 9.52 (s, 1H), 8.74 (s, 1H), 4.28—4.20 (m, 2H), 3.94—3.69 (m, 3H), 2.22-2.16 (m, 2H), .92 (m, 1H), 1.60—1.53 (m, 1H), 1.32-1.18 (m, 6H); LCMS (ESI): m/z 188.24 [M++1] S nthesis of eth ll-ben 1 h drox meth lmeth l rrolidine-Z-carbox late GG : To a stirring solution of FF (8 g, 26.57 mmol) in CH3CN (50 mL) was added K2CO3 (11.02 g, 79.73 mmol) ed by benzyl bromide (3.78 mL, 31.89 mmol) at RT and stirred for 16 h. After consumption of the starting material, filtered the reaction mass through celite bed and ﬁlterate was washed with EtOAc (250 mL). The ﬁlterate was concentrated under reduced pressure to afford crude. ed crude material was puriﬁed by column chromatography eluting with 10% EtOAc/Hexane to afford CG (5 g, 68%) as brown syrup. 1H-NMR: (500 MHz, DMSO—dg): 5 .15 (m, 5H), 5.17 (t, J= 5.5 Hz, 1H), 4.75 (d, J= 5.5 Hz, 1H), 4.50 (d, J: 6.0 Hz, 1H), 4.14—3.93 (m, 2H), 3.93 (s, 2H), 3.81—3.60 (m, 1H), 3.13—3.09 (m, 1H), 2.20—2.17 (m, 1H), 1.99-1.88 (m, 2H), 1.40—1.36 (m, 3H), 1.14 (t, J= 7.0 Hz, 3H) LCMS (ESI): m/z 278.36 [M++1] S nthesis of 1-ben 1 h drox meth th l rrolidine-Z-carbox lic acid HH: To a stirring solution of GG (5 g, 18.05 mmol) in EtOH/HZO (25 mL/25 mL) were added NaOH (1.44 g, 36.1 mg) at RT. The reaction e was heated to 100 °C for 2 h.

After consumption of the starting material (by TLC), the solvent from the reaction was evaporated under reduced pressure and extracted with di ethylether (2 X 75 mL). The aqueous layer was acidiﬁed by using 1N HCl and extracted with 20% MeOH/CHzClz (2 x 100 mL). The organic layer was dried over anhydrous NaZSO4, ﬁltered and concentrated under reduced pressure to afford HH (3.5 g, 78%) as brown syrup. 1H-NMR: (400 MHz, 6): 6 11.17 (s, 1H), 7.49—7.27 (m, 5H), 4.55 (d, J: 14.0 Hz, 1H), 4.06 (d, J: 14.4 Hz, 1H), 3.83 (s, 2H), 3.51—3.41 (m, 2H), 2.18—1.90 (m, 3H), 1.45-1.39 (m, 1H), .45 (m, 3H) LCMS (ESI): m/z 250.13 [M++1] Scheme I—9 OWOH Step 1 (EWO\ Step 2 ” SOCIZ HHClo BnBr O 4;}0\ LiHMDS NaOH CH3CHO Bn 0 Bn 0 S nthesis of meth l rrolidinecarbox late 11 : To a stirring solution of L-Proline (50 g, 434 mmol) in methanol was added thionyl chloride (37.5 ml, 521 mmol) at 0 OC and heated to 70 °C for 16 h. The reaction —76- mixture was brought to RT and concentrated under vacuum to afford II as (70 g, 99 %) as thick syrup chloride salt). 1H—NMR: (500 MHz, DMSO—d6): 5 4.15—4.13 (m, 1H), 3.65 (s, 3H), 3.35—3.30 (m, 2H), 2.23— 2.15 (m, 1H), 1.86—1.78 (m, 3H), 1.41 (s, 9H) LCMS (ESI): m/Z 129 [M++1] sis of methylbenzylprolinate [JJ 2: To a stirring solution of compound II (18 g, 108 mmol) in DCM (200 mL) was added TEA (45.35 mL, 326 mmol) followed by benzyl bromide (15.5 mL, 130 mmol) at 0 °C and stirred at RT for 24 h. After completion of the reaction (by TLC) was diluted with water (75 mL) and EtOAc (500 mL). The organic layer was washed with water (2 X 100 mL), brine solution (2 x 50 mL). The organic layer was dried over anhydrous Na2SO4 and concentrated under vacuum. Obtained crude material was d by column chromatography eluting with % EtOAc/Hexane to afford JJ (5 g, 21%) as oily liquid. lH-NMR: (400 MHz, DMSO—d6): 5 7.32—7.22 (m, 5H), 3.86-3.80 (m, 1H), 3.58 (s, 3H), 3.48- 3.24 (m, 2H), 2.85—2.80 (m, 1H), 2.38—2.32 (m, 1H), 2.10-1.98 (m, 1H), 1.85-1.69 (m, 3H) LCMS (ESI): m/z 220.28 [M++1] S s of meth ll-benz 1 1-h drox eth l rrolidine—2-carb0x late KK: To a stirring solution of JJ (5 g, 22.80 mmol) in THF (50 mL) was added LiHMDS (46 mL, 45.60 mmol) dropwise at -20 oC and stirred for 45 min. To this acetaldehyde (2.33 mL, 45.60 mmol) was added drop wise and d for 3 h at —20 °C. After consumption of the starting material (by TLC), the reaction was quenched with aqueous NH4Cl on (100 mL) and extracted with EtOAc (2 x 150 mL). The separated organic layer was washed with water (200 mL) and brine solution (200 mL).The separated c layer was dried over Na2S04 and concentrated to afford crude which was puriﬁed by column chromatography eluting with 10% EtOAc/Hexane to afford K (3.5 g, 58%) as yellow syrup.

LCMS (ESI): m/z 264.3 [M++1] S nthesis of l-ben 1 1-h drox eth l rrolidine—2-carb0x lic acid LL : To a stirring solution of KK (4.5 g, 17.11 mmol) in MeOH/THF/HZO (10 mL/ 10 mL/10 mL) were added NaOH (1.02 g, 25.66 mmol) at RT. The reaction mixture was heated to 95 0C for l h. After consumption of the starting al (by TLC), the solvent was evaporated under reduced re. The aqueous layer was washed with EtOAc (100 mL). The separated aqueous layer was acidiﬁed by using 1N HCl (pH~3). The aqueous layer was extracted with % MeOH/DCM (2 x 100 mL). The combined organic layer was dried over Na2S04 and concentrated under reduced pressure to afford LL (2.7 g, 63%) as an off—white solid. 1H-NMR: (400 MHz, DMSO-d6): 5 7.31-7.13 (m, 5H), .78 (m, 2H), 3.61-3.57 (m, 1H), 2.75—2.64 (m, 2H), 1.90—1.78 (m, 2H), 1.69—1.65 (m, 1H), 1.53—1.43 (m, 1H), 1.06 (d, J= 6.4 Hz, 3H); LCMS (ESI): m/z 250.4 [MT-1] Scheme I-10 OH o OH o OH 0 OTBSO Step 1 Step 2 Step 3 OH —> OMe —> OMe —> OMe SOC'Z CbZ'CI TBS'C' NH2 NH2 NHCbz NHCbz MM NN 00 OTBS OTBS ores O’\ O’\\ Step 4 Si Step 5 \ \,N Step 6 \ ,N —’ —’ NHNH2 N _’ N NH NH (02H50)3CH Pd'C/Hz 2 2 ”HOW NHCbz NH2 PP QQ RR S nthesis of meth lL-serinate MM : To a stirring solution of L—serine (12 g, 114 mmol) in CH3OH (100 mL) was added thionyl chloride (10 mL, 137 mmol) at 0 °C and stirred at 80 °C for 16 h. After completion of ng material (by TLC), the reaction mixture was concentrated under reduced pressure to afford MM (16 g, crude, HCl salt) as white solid. This material was directly used for the next step without further puriﬁcation. 1H-NMR: (400 MHz, DMSO—d6): 8 8.61 (s, 2H), 4.50 (s, 1H), 4.20—4.16 (m, 1H), 3.82 (d, J= 3.6 Hz, 2H), 3.73 (s, 3H) 0f meth'l benz'ldx carbon'l—L—serinate NN: To a stirring solution of MM (16 g, 103 mmol) in 1, 4 dioxane (130 mL/65 mL) were added added N32CO3 (27.3 g, 257 mmol) at RT. After added Cbz—Cl (17.6 mL, 123 mmol) was added at 0 °C drop wise and d for 16 h at RT. After completion of starting material (by TLC), diluted the reaction mass with EtOAc (300 ml). The separated c layer was dried over anhydrous , ﬁltered and concentrated under reduced W0 20784 —78- pressure to afford crude material which was d by column chromatography to afford NN (23 g, 88 %) as thick syrup. 1H—NMR: (500 MHZ, DMSO—ds): 5 7.50 (d, J: 8.0 Hz, 1H), 7.38—7.31 (m, 5H), 5.04 (s, 2H), 4.95—4.92 (m, 1H), 4.17—4.13 (m, 1H), 3.66 (s, 3H), 3.65—3.56 (m, 2H).

LCMS (ESI): m/z 254.2[M++1] —L—serinate 205'.

To a. stirring solution of NN (23 g, 91 mmol) in DCM (700 mL) were added imidazole (12.37 g, 182 mmol), DMAP (2.22 g, 18.2 mmol) followed by TBDMS—Cl (20.4 g, 136 mmol) at 0 °C and stirred at RT for 16 h. After completion of ng material (by TLC), diluted the reaction mass with water (200 ml). The separated organic layer was washed with brine solution (2 x 200 mL). The organic layer was dried over anhydrous NagSO4, filtered and concentrated under reduced pressure to afford crude material was purified by column chromatography eluting 20% EtOAc/hexane to afford 00 (32 g, 97%) as thick syrup. : (400 MHz, DMSO-d6): 5 7.56 ( d, J= 8.0 Hz, 1H), 7.38-7.30 (m, 5H), 5.07 (s, 2H), 4.24—4.19 (m, 1H), 3.81 (d, J= 5.2 Hz, 2H), 3.63 (s, 3H), 0.84 (s, 9H), -0.04 (s, 6H) LCMS (ESI): m/z 368.5 [M++1] S nthesis of hen l S - 3- tert-but ldimeth lsil 1 0X h drazin l-l-oxo r0 an-Z- yllcarbamate {PP}: To a d on of 00 (20 g, 54.5 mmol) in methanol (120 mL) was added hydrazine hydrate (27 g, 545 mmol) at 0 0C and after stirred at 80 °C for 2 h. After completion of starting material (by TLC), ethanol was evaporated under reduced pressure. The crude residue was triturated with n-pentane (100 mL) to afford PP (17 g, 85%) as an off—white solid. 1H-NMR: (500 MHz, g): 5 9.17 (s, 1H), 9.10 (s, 1H), 7.35-7.14 (m, 5H), 5.02 ( s, 2H), 4.48 (s, 2H), 4.16—4.07 (m, 2H), 3.73—3.58 (m, 1H), 0.81 (s, 9H), —0.04 (s, 6H).

S nthesis of hen 1 IS 2R tert-but ldimeth lsil l ox 1 3 4-0xadiazol—2— l nropyl) carbamate 10$ 22: A solution of PP (17 g, 46.3 mmol) in triethylorthoformate (68.5 g, 463 mmol) was added p—TSA (8.8 g, 4.63 mmol) at RT and after stirred at 80 °C for 2 h. After completion of starting material (by TLC), ylorthoformate was evaporated under reduced pressure. The crude e was purified by column chromatography eluting 10% EtOAc/hexane to afford QQ (5 g, 29%) as thick syrup. 1H-NMR: (500 MHz, DMSO—dg): 5 9.22 (s, 1H), 8.05 (d, J= 7.5 Hz, 1H), 7.36-?.30 (m, 5H), .08 (s, 2H), 5.05-4.97 (m, 1H), 4.03—3.91 (m, 2H), 0.84 (s, 9H), 0.03 (s, 6H); LCMS (ESI): m/z 378.5 [M++1] S nthesis of S tert-bu ldimeth lsil 1 0X 1 3 4-oxadiazol l ethan-l-amine 1RR1: To a stirring solution of QQ (5 g, 13.2 mmol) in methanol (30 mL) was added 50% wet 10% Pd/C (1.2 g) and stirred under H2 atmosphere (balloon re) for 2 h at RT.

After completion of reaction, the reaction mixture was filtered through a pad of celite and triturated with methanol (20 mL). The ﬁltrate was concentrated under reduced pressure to afford RR (2 g, 63%) as thick. syrup. 1H-NMR: (400 MHz, DMSO—dg): 8 9.16 (s, 1H), 4.19—4.16 (m, 1H), 3.85—3.75 (m, 2H), 2.11 (s, 2H), 0.77 (s, 9H), —0.04 (s, 6H) LCMs (ESI): m/z 244.3 [M++1] Scheme 11 N/:N\> / / N \ %ﬁH Step1 W /:N> _, N . N |sobutry|chlorlde Yg OH C-20 C-38 S nthesis of 2- 2-h drox -l- rimidin 1 r0 lisobu r lmeth l-2 5- diazas iro 3.4 octan-l-one C-38: To a stirring solution of C-20 (500 mg, 1.81 mmol) in DCM (10 mL) was added DIPEA (0.94 mL, 5.43 mmol) at 0 °C. After added iso lchloride (578 mg, 5.43 mmol) at 0 OC and stirred at RT for 4 h. After consumption of the starting material (by TLC), the reaction mixture was diluted with water (10 mL). The separated organic layer was dried over anhydrous NaZSO4 and concentrated under reduced pressure to afford crude compound which was purified by column chromatography by eluting 2% MeOH/DCM to ed C-38 (150 mg, 24%) as pale yellow solid. 1H-NMR: (400 MHz, CD30D): 8 8.82-8.77 (m, 2H), 7.39 (t, J= 5.2 Hz, 1H), 4.76 (d, J= 8.0 Hz, 1H), .44 (m, 1H), 4.20—4.14 (m, 1H), 3.82-3.74 (m, 1H), 3.51—3.40 (m, 1H), 2.78— 2.72 (m, 1H), 2.46-2.37 (m, 1H), 2.18—2.12 (m, 2H), .72 (m, 1H), .29 (m, 3H), .22 (m, 3H), 1.06-1.02 (m, 6H) LCMS (ESI): m/z 347.2 [M++1]; HPLC: 99.7% Scheme 12 N/ﬁy N/\\N\>/ N MNf Step 1 MN \ {g 0 OH 3,3- dimethylbutanoyl chloride, TEA C-20 S nthesis of 5- 33-dimeth lbutano 1 2-h drox rimidin-Z- 1 r0 lmeth l- 2 5-diazas iro 3.4 l-one C-39 : To a stirring solution of C-20 (150 mg, 0.54 mmol) in DCM (10 mL) was added TEA (137 mg, 1.35 mmol) at 0 °C. After added 3, 3—dimethylbutanoyl chloride (87 mg, 0.65 mmol) at 0 °C and stirred at RT for l h. After consumption of the starting material (by TLC), the reaction mixture was diluted with water (10 mL). The ted organic layer was dried over anhydrous Na2S04 and trated under reduced pressure to afford crude compound which was puriﬁed by column chromatography by eluting 2% MeOH/DCM to ed C-39 (150 mg, 24%) as sticky solid. 1H-NMR: (400 MHZ, DMSO-dg): 5 8.80—8.75 (m, 2H), 7.43—7.37 (m, 1H), 4.73-4.65 (m, 2H), 4.24—4.17 (m, 2H), 3.78—3.59 (m, 1H), 3.23—3.16 (m, 1H), 2.36 (s, 2H), 2.32—2.08 (m, 3H), 1.58— 1.50 (m, 1H), 1.20-1.08 (m, 6H), 0.86 (s, 9H) LCMS (ESI): m/z 375.49 [M++l]; HPLC: 96.9% Scheme 13 Nq ”5) Mi” LNN W”? OH H entyl chloro oi O formate, TEA g 0 C-20 C-40 S nthesis of c clo en 1 2- 2-h drox rimidin-2— 1 r0 lmeth l0X0-2 5- diazas iro 3.4 octane-S-carbox late C-40: To a stirring solution of C-20 (150 mg, 0.54 mmol) in DCM (5 mL) was added TEA (0.23 mL, 1.63 mmol) at 0 °C. After added cyclopentyl chloroformate (88 mg, 0.59 —81- mmol) at 0 OC and stirred at RT for 1 h. After consumption of the starting material (by TLC), the reaction mixture was diluted with water (10 mL). The separated organic layer was washed with saturated brine solution (20 mL). The organic layer was dried over anhydrous Na2S04 and concentrated under reduced pressure to afford crude compound which was puriﬁed by column chromatography by eluting 3% MeOH/DCM to obtained C-40 (110mg, 52%) as sticky solid. 1H-NMR: (400 MHz, g): 8 8.81—8.76 (m, 2H), .42 (m, 1H), 5.20—4.91 (m, 1H), 4.77—4.64 (m, 2H), 4.30—3.63 (m, 1H), 3.57—3.31 (m, 3H), 2.16—1.86 (m, 4H), 1.77—1.32 (m, 8H), 1.18—1.08 (m, 6H) LCMS (ESI): m/z 389.4 [M++1]; UPLC: 97.6% Scheme 14 MN{if Step1 N N?OH Cyclohexyl chloro 0&0 O formate, TEA 0 (3-20 C-41 S nthesis of c clohex 12- 2-h drox rimidin-Z- 1 r0 th l-l-oxo-Z 5- diazas iro 3.4 octane-S-carbox late C-41 : ] To a stirring solution of C-20 (150 mg, 0.54 mmol) in DCM (5 mL) was added TEA (0.2 mL, 1.35 mmol) at 0 °C. After added cyclohexyl chloroformate (105 mg, 0.65 mmol) at 0 OC and stirred at RT for 1 h. After ption of the starting material (by TLC), the on mixture was diluted with water (10 mL). The ted organic layer was washed with citric acid solution (20 mL), saturated brine solution (20 mL). The organic layer was dried over anhydrous Na2S04 and concentrated under reduced pressure to afford crude compound which was puriﬁed by column chromatography by eluting 2% MeOH/DCM to obtained C-41 (90 mg, 41%) as sticky solid. : (400 MHz, DMSO-dg): 8 8.82-8.78 (m, 2H), 7.48—7.41 (m, 1H), 4.78—4.61 (m, 2H), 4.24—4.13 (m, 2H), 3.94—3.87 (m, 1H), 3.76—3.53 (m, 2H), 2.27—1.99 (m, 3H), 1.97—1.92 (m, 1H), 1.56—1.32 (m, 10H), 1.29—1.10 (m, 2H) LCMS (ESI): m/z 403.5 [M++1]; UPLC: 98.06% Scheme 15 _ 82 _ OH HO 0A?! 0A“ ‘N N OH Step Step 2 —> HNII- —> N..- | "IOTBS o Int-N. HATU I}! DIAD. TPP I}! "'OTBS Bn 0 Bn 0 HH C-42 C-43 CAN OAN Step 3 —l\l Step 4 —,\j —> Nu. N‘“ THF / TBAF N N Bn, ..,OH H2/ Pd-C H ..,OH 0 0 C 44 C-45 S nthesis of l-ben l-N- 1S 2R tert-bu ldimeth lsil 1 0x 1 3 4-oxadiazol 1 r0 1 h drox meth l-S-meth l rrolidine-Z-carboxamide C—42 : To a stirring solution of HH (2.5 g, 10.04 mmol) in DMF (15 mL) were added N, N-diisopropylethylamine (5.4 mL, 30.12 mmol), Int N (2.58 g, 10.04 mmol) followed by HATU (4.57 g, 12.04 mmol) at 0 OC and stirred at RT for 16 h. After consumption of the starting material (by TLC), the reaction e was diluted with water (100 mL) and EtOAc (100 mL). The organic layer was washed with water (100 mL), saturated sodium bicarbonate solution (50 mL) ed by brine solution (50 mL). The organic layer was dried over anhydrous NaZSO4, filtered and concentrated under reduced pressure to afford crude compound which was puriﬁed by column tography by eluting 20% EtOAc/n-hexane to obtained C- 42 ( 3.2 g. 65%) as brown syrup. 1H-NMR: (500 MHz, DMSO—d6): 5 9.26 (s, 1H), 8.78 (d, J= 10.0 Hz, 1H), 7.30—7.21 (m, 5H), .26 (d, J: 10.0 Hz, 1H), 4.43 (s, 2H), .02 (m, 2H), 3.87—3.72 (m, 1H), 3.15—3.10 (m, 1H), 2.81—2.65 (m, 1H), 1.39—1.26 (m, 4H), 1.25—1.16 (m, 6H), 0.05 (s, 6H) LCMS (ESI): m/z 489.70 [M++1] S nthesis of 5-ben 1 1S 2R tert-but ldimeth lsil l ox 1 3 4-0xadiazol—2— 1 r0 lmeth l-2 S-diazas iro 3.4 octan-l-one C-43 : To a ng on of triphenylphosphine (4.02 g, 15.36 mmol) in dry THF (30 mL) was added DIAD (2.48 g, 12.29 mmol) at RT and stirred for 15 min. After added C-42 (3 g, 6.14 mmol) in THF (30 mL) was added dropwise and the reaction mixture was stirred at RT for 4 h. After consumption of the starting material (by TLC), the reaction mixture was concentrated under d pressure. The crude material was purified by column chromatography by eluting 20% EtOAc/rz—hexane to afford C-43 (1.3 g, 45%) as pale green syrup. —83- 1H—NMR: (400 MHz, DMSO—ds): 8 9.28 (s, 1H), .19 (m, 5H), 5.24—5.19 (m, 1H), 4.73 (s, 2H), 4.47—4.41 (m, 1H), 4.02 (t, .1: 7.2 Hz, 1H), 3.68 (s, 2H), 2.49—2.28 (m, 1H), 2.27—2.10 (m, 1H), 1.98—1.94 (m, 1H), 1.47—1.40 (m, 1H), 1.23—1.15(m, 6H), 0.87 (s, 9H), 0.02 (s, 6H) LCMs (ESI): m/z 491.6 [M++1] Synthesis of 5-benzyl((1S, hydr0xy(1, 3, 4-oxadiazolyl) propyl)methyl—2, —diazaspiro [3.4] octan-l-one (C-44): ] To a stirring solution of C-43 (1.3 g, 2.76 mmol) in dry THF (30 mL) was added TBAF (1M in THF) (4.13 mL, 4.14 mmol) at 0 °C and stirred for 2 h at RT. After consumption of the starting material (by TLC), the reaction mixture was trated under d pressure. The crude material was diluted with water (100 mL) and EtOAc (100 mL). The c layer was washed with water (100 mL), brine on (50 mL). The organic layer was dried over anhydrous Na2S04, ﬁltered and concentrated under reduced pressure to afford crude compound which was puriﬁed by column chromatography by eluting 2% MeOH/CHZClz to obtained C-44 (350 mg, 35%) as thick syrup. 1H-NMR: (400 MHz, DMSO—ds): 5 9.18 (s, 1H), 7.34-7.16 (m, 5H), 5.21 (t, J: 5.6 Hz, 1H), .01—4.83 (m, 1H), .18 (m, 1H), 3.88 (s, 2H), 3.84—3.67 (m, 1H), 3.54-3.35 (m, 1H), 2.98- 2.89 (m, 1H), 2.31-1.89 (m, 3H), 1.45-1.41(rn, 1H), 1.39-1.14 (m, 3H), 0.90-0.81(s, 3H) LCMS (ESI): m/z 357.43 [M++1] HPLC: 92.46% S nthesis of 5-ben l 1S 2R h drox 1 3 4-0xadiazol—2- 1 r0 lmeth l-2 —diazaspiro |3.4| octan-l-one gC-451: To a stirring solution of C-44 (1.3 g, 2.76 mmol) in dry THF (30 mL) was added TBAF (1M in THF) (4.13 mL, 4.14 mmol) at 0 °C and stirred for 2 h at RT. After consumption of the starting material (by TLC), the reaction mixture was concentrated under reduced pressure. The crude material was diluted with water (100 mL) and EtOAc (100 mL). The organic layer was washed with water (100 mL), brine solution (50 mL). The organic layer was dried over anhydrous Na2S04, ﬁltered and concentrated under reduced pressure to afford crude compound which was puriﬁed by column chromatography by eluting 2% MeOH/CHzClz to obtained C-45 (350 mg, 35%) as thick syrup. —84- 1H—NMR: (400 MHz, s): 8 9.18 (s, 1H), 7.34—7.16 (m, 5H), 5.21 (t, J: 5.6 Hz, 1H), .01—4.83 (m, 1H), .18 (m, 1H), 3.88 (s, 2H), 3.84—3.67 (m, 1H), 3.54—3.35 (m, 1H), 2.98— 2.89 (m, 1H), 2.31—1.89 (m, 3H), 1.45—1.41(m, 1H), .14 (m, 3H), 0.90—0.81 (s, 3H) LCMs (ESI): m/z 357.43 [M++1] HPLC: 92.46% Scheme 16 CAN CAI,“ o "N Step 1 "N Step 2 N...

NII- —> N“! N "'OH I}! --IOH 3:3032 N -"OH isobutyryl O BocO o chloride 0 C-26 C-46 C-47 S nthesis of 2- IS 2R h drox 1 3 4-oxadiazol 1 r0 1 meth 1-2 5- diazas iro 3.4 octan-l-one C-46 : To a stirring solution of C-26 (800 mg, 2.18 mmol) in DCM (10 mL) were added BF30Et2 (620 mg, 4.37 mmol), molecular sieves (150 mg) at 0 oC and stirred for 3 h at RT. After consumption of the starting material (by TLC), the reaction mixture was diluted with n-pentane (5 mL). The precipitated solid was ﬁltered and ﬁltrate was concentrated under d pressure. Obtained crude material was purified by silica gel column tography eluting with 3% MeOH/DCM to afford C-46 (700 mg, crude) as yellow liquid.

Mass (ESI): m/Z 267.3 [M++1] S nthesis of 2- 1S 2R h drox l 3 4-0xadiazol 1 r0 lisobut r lmeth l- 2,5—diazaspiro|3.4|octan0ne (C-47):

[00222] To a stirring solution of C-46 (700 mg (crude), 2.63 mmol) in DCM (10 mL) was added TEA (0.53 mL, 3.94 mmol) at 0 °C. After 10 min added isobutyryl chloride (0.41 mL, 3.94 mmol) at 0 °C and stirred for 2 h at RT. After consumption of the starting material (by TLC), the reaction mixture was diluted with water (5 mL). The separated organic layer was dried over ous NaZSO4, ﬁltered and concentrated under reduced pressure. Obtained crude material was puriﬁed by silica gel column tography eluting with 3% MeOH/DCM followed by ative HPLC purification to afford C-47 (150 mg, 17%) as thick syrup. 1H—NMR: (400 MHz, DMSO—dg): 5 9.27 (s, 1H), .17 (m, 1H), 4.87—4.85 (m, 1H), 4.30— 4.25 (m, 1H), .79 (m, 1H), 3.64—3.50 (m, 2H), 2.72—2.66 (m, 1H), 2.13—2.09 (m, 1H), 2.01—1.79 (m, 3H), 1.37—1.19 (m, 3H), 1.10—0.94 (m, 9H) Mass (ESI): m/z 337.4 [M++l] 9.5% Scheme 17 0%;0H A Step 1 Step 2 O- IN OH —.

N NH.

| Int-N, HATU 02;“: I N Bn 0 TPP, DIAD \ fN.., ’OTBS Bno OTBS LL C-48 C-49 Step 3 < O?N N, Step 4 —> N Nln.

TBAF Elmo Pd/C C-50 C-51 S nthesis of l-ben l-N- 1S 2R tert—bu ldimeth lsil 1 0X 1 3 4-0xadiazol—2- 1 r0 l 1-h drox eth l rrolidine—2-carboxamide C-48 :

[00223] To a stirring solution of LL (3.9 g, 15.66 mmol) in DMF (10 mL) were added N, N—diisopropylethylamine (8.16 mL, 46.98 mmol), Int-N (4.03 g, 15.66 mmol) followed by HATU (7.14 g, 18.79 mmol) at 0 °C and stirred at RT for 16 h. After consumption of the starting al (by TLC), the reaction mixture was diluted with water (100 mL) and EtOAc (150 mL). The ted organic layer was washed with water (100 mL) followed by brine solution (100 mL). The organic layer was dried over anhydrous NazSO4 and concentrated under reduced pressure to afford crude compound which was puriﬁed by column chromatography by eluting 30% EtOAc/n—hexane to obtained C-48 (1.8 g, 24%) as yellow syrup. 1H-NMR: (400 MHz, DMSO—dg): 5 9.25 (s, 1H), 8.74 (d, J= 8.8 Hz, 1H), 7.34-7.21 (m, 5H), .47-5.27 (m, 1H), 5.26-5.23 (m, 1H), 4.42-4.40 (m, 1H), 4.13-4.01 (m, 2H), 3.82-3.77 (m, 1H), 2.70-2.60 (m, 2H), 1.99—1.82 (m, 2H), 1.69-1.64 (m, 2H), 1.29—1.21 (m, 6H), 0.72 (s, 9H), -0.01 (s, 3H), -0.02 (s, 3H); LCMS (ESI): m/z 489.6 [M++l] S nthesis 0f 5—ben l 1S 2R tert-but ldimeth lsil 1 0X 1 3 4-oxadiazol 1 r0 lmeth l-2 5—diazas iro 3.4 l-one C-49:

[00224] To a stirring on of triphenylphosphine (2.41 g, 9.22 mmol) in dry THF (20 mL) was added DIAD (1.49 g, 7.37 mmol) as portionwise and stirred for 15 min at RT. To this —86- itated solution added C-48 (1.8 g, 3.68 mmol) in dry THF (15 mL) dropwise at RT and d for 16 h. After consumption of the starting material (by TLC), the reaction mixture was concentrated under reduced pressure. The obtained solid was triturated with 20% di ehylether/n—hexane (2 x 100 mL). The ﬁlterate was concentrated under reduced pressure to obtained crude compound which was puriﬁed by silica gel column chromatography eluting % EtOAc/hexane to afford C-49 (400 mg, crude) as pale green syrup. 1H-NMR: (400 MHz, DMSO—d6): 5 9.30 (s, 1H), .24 (m, 5H), 5.26-5.22 (m, 1H), 4.80- 4.73 (m, 2H), 4.62-4.48 (m, 1H), 4.15-4.00 (m, 1H), 3.81—3.75 (m, 1H), 2.85 (t, J= 8.0 Hz, 1H), .39 (m, 3H), 2.26-2.11 (m, 1H), 1.71-1.64 (m, 3H), 1.25-1.15 (m, 3H), 0.78 (s, 9H), 0.02 (s, 6H); LCMS (ESI): m/z 471.6 [M++1] S nthesis of 5-ben l 1S 2R h drox 1 3 iazol—2- 1 r0 lmeth l-2 -diazaspiro |3.4| octan-l-one gC-SO): To a stirring solution of C-49 (1.5 g, 3.19 mmol) in dry THF (30 mL) was added TBAF (1M in THF) (4.78 mL, 1.5 mmol) at 0 OC and d at RT for l h. After consumption of the starting al (by TLC), the reaction mixture was concentrated under reduced pressure. The crude material was diluted with water (75 mL) and EtOAc (100 mL). The organic layer was washed with water (2 x 50 mL), brine solution (2 X 50 mL). The organic layer was dried over anhydrous NaZSO4, filtered and concentrated under reduced pressure to afford crude compound which was puriﬁed by column chromatography by eluting 30% EtOAc/n—hexane to obtained C-50 (500 mg, 44%) as white solid. 1H-NMR: (400 MHz, DMSO—dg): 5 9.26 (s, 1H), 7.37—7.22 (m, 5H), 5.29-5.21 (m, 1H), 4.96— 4.75 (m, 1H), 4.33—4.02 (m, 1H), 4.00—3.94 (m, 2H), 3.77—3.70 (m, 2H), 2.91—2.80 (m, 1H), 2.42-2.38 (m, 1H), .08 (m, 2H), 1.80-1.63 (m, 1H), 1.30 (d, J= 6.4 Hz, 3H), 1.12 (d, J= 6.0 Hz, 3H); LCMS (ESI): m/z 357.43 [M++1] HPLC: 99.61% S nthesis of 2- 1S 2R h drox 1 3 4-oxadiazol 1 r0 lmeth l-2 5- diazaspiro|3.4|octanone gC-Sl 1:

[00226] To a stirring solution of compound C-50 (420 mg, 1.17 mmol) in methanol (10 mL) was added 10%Pd/C (400 mg) under N2 atmosphere. The reaction mixture was stirred under H2 atmosphere at RT for 3 h. After consumption of the ng material (by TLC), the —87- on mixture was ﬁltered through a pad of celite and the pad was washed with methanol (10 mL). Obtained ﬁltrate was concentrated under reduced pressure to afford C-51 (220 mg, 70%) as an off—white solid. 1H-NMR: (500 MHz, DMSO—d6): 8 9.26 (s, 1H), 5.25—5.17 (m, 1H), 4.89—4.69 (m, 1H), 4.28— 4.21 (m, 1H), 3.84-3.56 (m, 2H), 2.95-2.79 (m, 2H), 1.99-1.84 (m, 2H), 1.77-1.59 (m, 2H), 1.24—1.10 (m, 6H); LCMS (ESI): m/z 260 [M++1] HPLC: 97.94% Scheme 18 N N‘ N/: \ / Step 1 N N N N H Iso butyryl O OH O chloride C-52 S nthesis of 2- 2—h drox rimidin-Z- 1 r0 1-3 6-dimeth 1-2 5-diazas iro 3.4 octan-l-one C-52 : To a stirring on of C-33 (500 mg, 1.72 mmol) in dry DCM (10 mL) was added DIPEA (556 mg, 4.31 mmol) ed by isobutyryl chloride (273 mg, 2.58 mmol) slowly at 0 OC and stirred at RT for 3 h. After ption of the starting material (by TLC), the reaction mixture was concentrated under reduced pressure. The crude residue was d with water (10 mL). The separated organic layer was washed with brine solution (20 mL). The separated organic layer was dried over anhydrous NaZSO4 and trated under reduced pressure to afford crude compound which was purified by column chromatography by eluting 1% MeOH/DCM to obtained C-52 (90 mg, 14.5%) as pale brown syrup. 1H-NMR: (400 MHz, : 8 8.81—8.77 (m, 2H), 7.41—7.37 (m, 1H), 4.63—4.53 (m, 2H), 4.24—4.18 (m, 2H), 2.87—2.72 (m, 1H), 2.33—2.27 (m, 1H), 2.16—2.08 (m, 2H), 1.73—1.69 (m, 1H), 1.40—1.35 (m, 3H), 1.28—1.20 (m, 3H), 1.15—1.02 (m, 9H); LCMS (ESI): m/z 361.3 [M++1]; HPLC:94.85% Scheme 19 WO 20784 _ 88 _ mo:Et sTtep1m; OH Step2 Step3 OHﬂ) ”CO #Iﬁﬂ N30“ K2C03,BnBr '3 Int-N, HATU C-503 0 BC-54 Bn C-55 HNi-IIOAN of ’N Step5 Step6 0:1?! "OTBS DIAD, TPP HIOTBS TBAF Bn 0 Bn 0 C-56 C-57 C-58 S nthesis of eth 12- 1-h drox eth l-S-meth l rrolidine—Z-carbox late C-53 : To a stirring on of CC (5 g, 16.61 mmol) in CH2C12 (50 mL) was added TFA (6.34 mL, 83.05 mmol) at 0 °C. After being d at RT for 4 h, the reaction e was concentrated under reduced pressure to afford crude compound which was triturated with n— pentane (50 mL) to obtained C-53 (5 g, crude) as black syrup was directly taken for the next step without further purification. 1H-NMR: (500 MHz, DMSO-d6): 5 8.71 (br s, 1H), 4.29-4.25 (m, 1H), 4.25-4.21 (m, 3H), 4.14—4.03 (m, 1H), 2.31—2.10(m, 4H), 1.36—1.18 (m, 6H), 1.18—1.10(m, 3H) LCMS (ESI): m/z 202.38 [M++1] S s of eth l l-ben 1 l-h drox eth l-S-meth l rrolidine—Z-carbox late C-54: CA & C-54—B) : To a stirring solution of C-53 (5 g (crude), 15.87 mmol) in acetonitrile (50 mL) was added K2CO3 (6.57 g, 47.61 mmol) followed by benzyl bromide (2.82 mL, 23.80 mmol) at RT and stirred for 16 h. After completion of the reaction, diluted the reaction mixture with EtOAc (150 mL) and water (75 mL). The organic layer was washed with brine solution (2 x 150 mL). The organic layer was dried over anhydrous NaZSO4 and concentrated under vacuum.

Obtained crude material was puriﬁed by column tography eluting with 10% EtOAc/Hexane to afford 1 g of CA and 1.2 g of CB (separated isomers of C—54) (48%) as yellow syrups. 1H-NMR (C—54—A isomer): (500 MHz, s): 8 7.35—7.14 (m, 5H), 4.55 (s, 2H), 4.24— 4.19 (m, 1H), 4.13—4.02 (m, 2H), 3.96—3.91 (m, 2H), 2.14—1.99 (m, 1H), 1.95—1.82 (m, 3H), 1.25—1.18(m,3H), 1.10-1.06 (m, 3H), 0.75 (d, J: 6.0 Hz, 3H) LCMS (ESI): m/z 292.3 [M++1] 1H-NMR B isomer): (500 MHz, DMSO—d6): 8 7.34—7.18 (m, 5H), 5.15 (t, J= 5.5 Hz, 2H), .49 (m, 1H), 4.24—4.22 (m, 1H), 4.10-4.06 (m, 2H), .10 (m, 1H), 2.39—2.33 (m, 1H), 1.89—1.83 (m, 3H), 1.42-1.39 (m, 3H), 1.20-1.13 (m, 3H), 0.76 (d, J: 6.0 Hz, 3H) LCMS (ESI): m/z 292.3 [M++l] S nthesis of l-ben 1 1-h drox eth th l rrolidine—Z-carbox lic acid CA : To a stirring solution of CA (l g, 3.43 mmol) in MeOH/THF/H20 (5 mL/5 mL/5 mL) were added NaOH (206 mg, 5.14 mmol) at 0 °C. The reaction mixture was heated to 90 °C for 6 h. After consumption of the starting material (by TLC), the solvent was evaporated under reduced pressure. The aqueous layer was washed with di ethylether (50 mL). The separated aqueous layer was acidiﬁed by using 2N HCl (pH~3). The aqueous layer was extracted with 10% MeOH/DCM (2 x 100 mL). The combined c layer was dried over Na2S04 and concentrated under reduced pressure to afford compound CA (1 g, crude) as yellow solid. 1H-NMR: (400 MHz, DMSO—dg): 5 9.84 (s, 1H), 7.60 (d, J= 6.8 Hz, 2H), 7.50—7.30 (m, 3H), .12 (br s, 1H), 4.48-4.35 (m, 2H), 4.09-3.81 (m, 1H), 2.39-2.22 (m, 2H), 2.16-2.08 (m, 2H), 1.66—1.57 (m, 1H), 1.16—0.86 (m, 6H); LCMS (ESI): m/z 264.3 [M++1] S nthesis of l-ben 1 1-h drox eth l-S-meth l rrolidine—2-carb0x lic acid CB :

[00231] To a stirring solution of CB (1.2 g, 4.12 mmol) in HF/HZO (5 mL/5 mL/5 mL) were added NaOH (206 mg, 5.14 mmol) at 0 °C. The reaction mixture was heated to 90 °C for 6 h. After consumption of the starting material (by TLC), the solvent was evaporated under reduced pressure. The aqueous layer was washed with di ethylether (50 mL). The ted aqueous layer was acidiﬁed by using 2N HCl . The aqueous layer was extracted with 10% MeOH/DCM (2 x 100 mL). The ed organic layer was dried over NaZSO4 and concentrated under reduced pressure to afford compound C-55—B (780 mg) as yellow solid. 1H-NMR (C-55—B isomer): (400 MHz, DMSO—dg): 8 9.14 (s, 1H), 7.64 (d, J = 6.0 Hz, 2H), 7.49-7.44 (m, 3H), 4.99 (d, J= 12.8 Hz, 1H), 4.48 (s, 2H), 4.45-4.41 (m, 1H), 3.98-3.93 (m, 1H), 2.32—2.13 (m, 4H), 1.40 (d, J= 6.4 Hz, 3H), 0.76 (d, J= 6.4 Hz, 3H); LCMS (ESI): m/z 272.4 [M11] To a stirring solution of C-55 (1.78 g, 6.75 mmol )(mixture of two isomers C- 55—A & C-SS-B) in DMF (10 mL) were added N, N—diisopropylethylamine (3.51 mL, 20.27 mmol), Int N (1.74 g, 6.75 mmol) followed by HATU (3.08 g, 8.11 mmol) at 0 °C and d at RT for 16 h. After consumption of the starting material (by TLC), the reaction mixture was diluted with water (100 mL) and EtOAc (150 mL). The separated organic layer was washed with water (100 mL) followed by brine on (100 mL). The organic layer was dried over anhydrous Na2S04 and concentrated under reduced pressure to afford crude compound which was puriﬁed by column chromatography by g 30% EtOAc/n—hexane to obtained C-56 (1.2 g, 35%) as brown syrup. 1H-NMR: (500 MHz, DMSO—ds): 5 9.31 (s, 1H), 8.42 (s, 1H), 7.44-7.19 (m, 5H), 5.26-4.95 (m, 1H), 4.36 (s, 2H), 4.33—4.18 (m, 2H), 4.09—3.94 (m, 1H), 2.32-2.22 (m, 1H), 2.15—1.86 (m, 4H), 1.37-1.05 (m, 9H), 0.85 (s, 9H), 0.04 (s, 6H); LCMS (ESI): m/z 503.7 [M++l] S nthesis 0f l-ben l-N- 1S 2R tert-bu ldimeth lsil 1 0X 1 3 iazol 1 r0 1 1-h drox eth l-5—meth l rrolidine—Z-carboxamide C—57 : To a stirring solution of triphenylphosphine (1.56 g, 5.97 mmol) in dry THF (15 mL) was added DIAD (967 mg, 4.78 mmol) as portionwise and stirred for 15 min at RT. To this precipitated solution added C-56 (1.2 g, 2.39 mmol) in dry THF (15 mL) slowly at RT and stirred for 4 h. After consumption of the starting material (by TLC), the reaction mixture was concentrated under d pressure. The obtained solid was triturated with 20% di ehylether/n—hexane (2 x 100 mL). The ﬁlterate was concentrated under d pressure to obtained crude compound which was d by silica gel column chromatography g 20% EtOAc/hexane to afford C-57 (400 mg, crude) as brown thick syrup. 1H-NMR: (400 MHz, DMSO—ds): 6 9.27 (s, 1H), 7.43—7.18 (m, 5H), 4.91—4.73 (m, 2H), 4.42— 4.02 (m, 2H), 3.96-3.75 (m, 2H), 2.11—1.84 (m, 3H), 1.42—1.37 (m, 1H), 1.35 (d, J= 6.0 Hz, 3H), 1.23—1.11 (m, 6H), 0.70 (s, 9H), 0.02 (s, 6H); LCMS (ESI): m/z 485.7 [M++l] S nthesis of 5-benz l 1S 2R h drox 1 3 4-oxadiazoll r0 1-3 6- dimethyl—Z, aspiro |3.4| octan-l-one 1C-58): To a stirring solution of C-57 (400 mg, 0.82 mmol) in dry THF (5 mL) was added TBAF (1M in THF) (1.23 mL, 1.5 mmol) at 0 °C and d for l h at RT. After consumption of the starting material (by TLC), the reaction mixture was concentrated under reduced re. The crude material was diluted with water (30 mL) and EtOAc (50 mL). The organic layer was washed with water (2 x 20 mL), brine solution (2 x 20 mL). The organic layer was dried over anhydrous NaZSO4, ﬁltered and concentrated under d pressure to afford crude compound which was purified by column tography by eluting 30% EtOAc/n—hexane to obtained C-58 (90 mg, 29%) as thick syrup. 1H-NMR: (400 MHz, DMSO-dg): 5 9.25 (s, 1H), 7.41-7.18 (m, 5H), 5.28-5.17 (m, 1H), 4.97- 4.62 (m, 1H), 4.43—4.20 (m, 1H), 4.02—3.89 (m, 1H), 3.77-3.64 (m, 2H), 3.20-3.01 (m, 1H), 2.09-1.80 (m, 3H), 1.44—1.40 (m, 1H), 1.38—1.30 (m, 1H), 1.28—1.21 (m, 4H), 1.19—1.06 (m, 1H), 0.90—0.85 (m, 3H) LCMS (ESI):m/z 371.45 [M++l] HPLC: 94.53% Scheme 20 /\\5“ /\ OH HO O O \E“ DAN Step 1 TN Step 2 ”N OH Step 3 —[\i —> HN|.. —> N... —> Nu. "i N o Int—RR, HATU DTAD THF/TBAF , 3°C I}: I}! B°° BocOTBSO BocOTBSO O Z C-S9 C-60 C-61 S nthesis of ut 12- tert-but ldimeth lsil 1 0x 1 3 4-oxadiazol leth lcarbamo 1 h drox meth lmeth l ine—l-carbox late C-59 : To a stirring solution of Z (2 g, 7.72 mmol) in DCM (25 mL) were added N. 1 7— diisopropylethylamine (4 mL, 23.16 mmol), Int RR (2.06 g, 8.49 mmol), followed by HATU (3.52 g, 9.26 mmol) at 0 °C and stirred at RT for 16 h. After consumption of the starting material (by TLC), the reaction mixture was diluted with water (50 mL). The separated organic layer was washed with citric acid on (1 x 50 mL), saturated brine solution (1 x 50 mL).

The separated organic layer was dried over ous Na2SO4 and concentrated under reduced pressure to afford crude compound which was puriﬁed by column chromatography by eluting 50% EtOAc/n—hexane to obtained C-59 ( 2.5 g, 67%) as thick syrup. 1H-NMR: (500 MHz, DMSO—d6): 8 9.25 (s, 1H), 7.99 (d, J= 3.5 Hz, 1H), 5.29—5.26 (m, 2H), 4.05-3.99 (m, 3H), 3.57-3.54 (m, 2H), 2.28-1.91 (m, 3H), 1.42 (s, 9H), 1.38-1.31 (m, 1H), 1.18- 1.11 (m, 3H), 0.84 (s, 9H), —0.01 (s, 6H) ] To a stirring solution of triphenylphosphine (2.7 g, 10.33 mmol) in dry THF (20 mL) was added DTAD (2.37 g, 10.33 mmol) at RT and stirred for 15 min. After added C-59 (25 g, 5.16 mmol) and the reaction mixture was stirred at RT for 3 h. After consumption of the starting material (by TLC), the reaction mixture was concentrated under reduced pressure. The crude material was triturated with di ethylether/n—pentane and obtained solid was ﬁltered. The ﬁltrate was evaporated, ed by silica gel column chromatography eluting 30% EtOAc/hexane to afford C-60 (2.1 g, 87.5%) as yellow liquid. 1H-NMR: (500 MHz, DMSO—ds): 5 9.32 (s, 1H), 5.19—5.08 (m, 1H), 4.12—3.87 (m, 3H), 3.58— 3.45 (m, 2H), 1.56—1.42 (m, 3H), 1.40 (s, 9H), 1.38—1.20 (m, 1H), 1.18—1.12 (m, 3H), 0.80 (s, 9H), 0.02 (s, 6H) S nthesis of tert—but l 2- 2-h drox rimidin 1 r0 lmeth l0xo-2 5- diazaspiro |3.4| -S-carboxylate gC-612: ] To a stirring solution of C—60 (2.1 g, 4.5 mmol) in dry THF (10 mL) was added TBAF (2.34 g, 9 mmol) at 0 °C. The reaction mixture was stirred at RT for 3 h. After ption of the starting material (by TLC), the on mixture was concentrated under reduced pressure. The crude material was purified by silica gel column chromatography g 2% MeOH/DCM to afford C-61 (800 mg, 51%) as thick syrup. 1H-NMR: (400 MHz, DMSO—d6): 8 9.25 (s, 1H), 5.33—5.08 (m, 2H), 3.94—3.75 (m, 3H), 3.67— 3.51 (m, 1H), 3.39—3.31 , 2.13—1.93 (m, 3H), 1.55-1.50 (m, 1H), 1.40 (s, 9H), 1.15-1.08 (m, 3H), LCMS (ESI): m/z 353.3 [M++1]; HPLC: 96.6% Scheme 21 /\ /\ HO HO o/SN o \u o ‘91 Step1 I "N "N ‘ Step 2 Step 3 OH —’ —’ N, NI" —> N' N HATU N DIAD TBAF N OTBS N OH oc’ O Int—RR Boo 0 was Bog 0 ml 0 BB C-62 C-63 C-64 S nthesis of tert—bu l 2- tert-but ldimeth lsil l ox 1 3 4-0xadiazol—2- l eth l carbamo l l-h drox eth l rrolidine-l-carbox late C-62 : To a stirring solution of BB (3.5 g, 13.51 mmol) in CHzClz (40 mL) were added DIPEA (7.0 mL, 40.53 mmol), RR (3.2 g, 13.51 mmol), HATU (5.6 g, 14.85 mmol) at 0 OC and d for 12 h. After consumption of the starting material (by TLC), the reaction mixture was diluted with water (30 mL) and extracted with CHzClz (2 x 100 mL). The separated organic layer was washed with brine, dried over anhydrous Na2S04, ﬁltered and concentrated under reduced pressure to afford C-62 (3.5 g, crude) as colorless liquid.

Mass (ESI): m/z 485.67 [M++1] S nthesis of tert—bu l 2- tert-but ldimeth lsil l ox 1 3 4-oxadiazol l eth l-l-meth loxo-2 5-diazas iro 3.4 octane-S-carbox late C-63 :

[00239] To a stirring solution of triphenylphosphine (2.03 g, 7.75 mmol) in THF (25 mL) was added DIAD (1.59 g, 7.75 mmol) at 0 °C and stirred for 30 min. C-62 (1.5 g, 3.10 mmol) in THF (10 mL) was added dropwise and the reaction e was stirred at RT for 16 h. After consumption of the starting material (by TLC), the on was trated under reduced pressure. The crude material was puriﬁed by silica gel column chromatography eluting 30% EtOAc/hexane to afford C-63 (600 mg, 43%) as pale yellow liquid. : (500 MHz, DMSO—d6): 5 9.24 (s, 1H), 5.93—4.75 (m, 2H), 4.16-3.91 (m, 2H), 3.78— 3.70 (m, 2H), 3.91 (d, J= 7.0 Hz, 1H), 3.77 (d, J= 7.0 Hz, 2H), 3.44—3.34 (m, 4H), 2.01-1.91 (m, 2H), 1.85—1.68 (m, 6H), 2.11—1.68 (m, 4H), 1.40 (s, 9H), 1.38—1.18 (m, 3H), 0.80 (s, 9H), — 0.02 (s, 6H).

Mass (ESI): m/z 467.6 [M++1] S nthesis of ut l 2- h drox 1 3 4-0xadiazol—2- l eth l meth lox0-2 -diazaspiro |3.4| octane-S-carboxylate gC-64]: To a stirring solution of C-63 (0.5 g, 1.07 mmol) in dry THF (10 mL) was added TBAF (1M in THF) (1.07 mL, 1.91 mmol) at 0 OC and d at RT for 3 h. After consumption of the starting material (by TLC), the on mixture was concentrated under reduced pressure. The crude material was diluted with water (15 mL) and EtOAc (30 mL). The organic layer was washed with water (2 x 15 mL), brine on (2 x 10 mL). The organic layer was dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to afford crude compound which was d by column chromatography by eluting 70% EtOAc/n-hexane to obtained C-64 (95 mg, 25%) as white solid. 1H-NMR: (400 MHz, DMSO-d6): 5 9.26 (s, 1H), 5.32-5.04 (m, 2H), 3.98-3.72 (m, 4H), 2.14- 1.66 (m, 4H), 1.39 (s, 1H), 1.37—1.09 (m, 3H) LCMS (ESI): m/z 353.3 [M++1] _ 94 _ HPLC: 88.9% Scheme 22 OH o/ZN 0&1“ CAN Step 1 Step 2 'N Step 3 "N OH —> N, N... N...

Boo/N EDCI OTBS 300/ 0 B000 C 65 C-66 C 67 S nthesis of ut 12- tert—but ldimeth lsil 1 0X 1 3 4-oxadiazol leth lcarbamo l 1-h drox eth lmeth l rrolidine-l-carbox late C—65 : To a stirring solution of DD (1 g, 3.66 mmol) in DCM (20 mL) were added N, N—diisopropylethylamine (1.9 mL, 10.98 mmol), Int RR (978 mg, 4.02 mmol), EDCI (1.04 g, 5.49 mmol) followed by HOBT (741 mg, 5.49 mmol) at 0 °C and stirred at RT for 16 h. After consumption of the starting material (by TLC), the reaction mixture was diluted with water (20 mL). The separated organic layer was washed with brine solution (30 mL). The separated organic layer was dried over ous Na2S04 and concentrated under d pressure to afford crude nd which was purified by column chromatography by eluting 40% EtOAc/n—hexane to obtained C-65 (700 mg, 38.4%) as thick syrup. 1H-NMR: (500 MHz, DMSO-dg): 8 9.26 (s, 1H), 5.22-5.18 (m, 1H), 4.58—4.50 (m, 2H), 4.04— 3.92 (m, 3H), 1.47—1.41 (m, 1H), 1.38 (s, 9H), 1.29—1.16 (m, 6H), 1.14—1.11 (m, 3H), 0.83 (s, 9H), —0.02 (s, 6H); LCMS (ESI): m/z 523.6 [M++1] S nthesis of tert-but l 2- S tert-but ldimeth lsil 10x 1 3 4-0xadiazol l eth l-1 6-dimeth loxo-2 5-diazas iro 3.4 octane-S-carbox late C-66 : To a stirring on of triphenylphosphine (736 mg, 2.81 mmol) in dry THF (10 mL) was added DTAD (646 mg, 2.81 mmol) as portionwise and d for 15 min at RT.

To this precipitated solution added C-65 (700 mg, 1.4 mmol) in dry THF (15 mL) slowly at RT and d for 6 h. After consumption of the starting material (by TLC), the reaction mixture was trated under reduced pressure. The crude material was triturated with n-pentane (20 mL) /di ehylether (20 mL). The ﬁlterate was concentrated under reduced pressure to obtained crude compound which was purified by silica gel column chromatography eluting 25% EtOAc/hexane to afford C-66 (430 mg, 64%) as thick syrup. 1H—NMR: (400 MHz, DMSO—ds): 5 9.25 (s, 1H), 5.20—5.16 (m, 1H), 4.16—4.00 (m, 1H), 3.88— 3.70 (m, 3H), 2.17—2.08 (m, 1H), 1.96—1.85 (m, 1H), 1.40 (s, 9H), 1.26—1.07 (m, 8H), 0.82 (s, 9H), —0.02 (s, 6H); LCMS (ESI): m/z 505.5 [M++1] S nthesis of tert—bu 12- h drox 1 3 4-oxadiazol l eth l-1 th 1 oxo—Z, aspiro |3.4| octane-S-carboxylate (C-67): To a stirring solution of C-66 (430 mg, 0.89 mmol) in dry THF (10 mL) was added TBAF (1.8 mL, 1.79 mmol) slowly at 0 °C and d at RT for 6 h. After ption of the starting material (by TLC), the reaction mixture was concentrated under reduced pressure. The crude residue was puriﬁed with column chromatography by eluting 90% EtOAc/n—hexane to afford C-67 (80 mg, 24.5%) as thick syrup. 1H-NMR: (400 MHz, DMSO-ds): 5 9.25 (s, 1H), 5.20-5.03 (m, 2H), 3.98—3.72 (m, 3H), 2.50- 1.85 (m, 3H), 1.56—1.50 (m, 1H), 1.40 (s, 9H), 1.27—1.11 (m, 6H) LCMS (ESI): m/z 291.3 [M++1]; HPLC: 91.4% Example 3 - 3H MK-801 binding assay Methods Assays were conducted as bed in Moskal et al. (Moskal, J.R., Kuo, A.G., Weiss, C., Wood, P.L., O'Connor Hanson, A., Kelso, S., Harris, R.B., Disterhoft, J.F., 2005.

GLYX-13: a monoclonal dy—derived peptide that acts as an N—methyl—D-aspartate receptor modulator. Neuropharmacology. 49, 1077—87) The potentiation of [3H]MK—801 binding (5 nM; 22.5 Ci / mmol) to well washed rat cortical nes (200 pg) was measured under uilibrium conditions (15 min @ 25 0C) in the presence of increasing concentrations of test compounds and 50uM glutamate. Zero levels were determined in the absence of any glycine ligand and in the presence of 30uM 5,7 DCKA. Maximal stimulation was measured in the presence of 1 mM glycine, and 5011M glutamate was present in all s. The tation of [3H]MK—801 binding by tests compounds was calculated by using a 3 parameter log agonist vs. response equation (Graph pad Prism, USA) and potency (EC50, expressed in pM) and maximal activity (% maximal stimulation) were calculated for the test compound. —96- The potency and maximal activity for Compound Y is 288 pM, 14%.

Table 2. Additional Biological Data Compound Uniﬁed Uniﬁed Uniﬁed Uniﬁed Uniﬁed Uniﬁed Uniﬁed MK—801 Activity Data: Activity Data: Activity Activity Activity Activity Activity e LTP LTP Data: LTP, Data: Data: Data: Data: Site Augmentation Concentration Signiﬁcant Porsolt Porsolt Porsolt Porsolt Binding (Percent) (uM) (S) or Non- Floating Dose Dose, Time Post Assay: Rat signiﬁcant Time (mg/kg) route Dose Cortex (NS) Inhibition (Hours) EC50 (M) ——————nm— Table 3. Additional Biological Data Compound MK-801 Glycine Site LTP: LTP LTP: LTP LTP: LTP Binding Assay: Rat Augmentation (%) Concentration (uM) Significance, S or NS Cortex EC50 712913-12 ——— 1.0961240 ——— mm ——— 135312-10 ——— EQUIVALENTS Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the speciﬁc ments of the invention described herein. Such equivalents are intended to be assed by the following claims.

INCORPORATION BY REFERENCE The entire contents of all patents, published patent ations, websites, and other references cited herein are hereby expressly orated herein in their entireties by reference.

Claims

What is Claimed is:

1. A compound represented by formula I: or a pharmaceutically acceptable salt, a stereoisomer, or an N-oxide thereof, n Rb is selected from the group consisting of H, halogen, hydroxyl, cyano or C1-C6 alkyl; R1 is H or C1-C6 alkyl; R2 is H or C1-C6 alkyl; R3 is selected from the group consisting of H, C1-C6 alkyl and a nitrogen protecting group; wherein the nitrogen protecting group is selected from the group consisting of 9- fluorenylmethyloxycarbonyl, tert-butoxycarbonyl, oxybenzyloxycarbonyl, acetyl, trifluoroacetyl, benzoyl, benzyl, p-methoxybenzyl, p-methoxyphenyl, 3,4- dimethoxybenzyl, triphenylmethyl, p-toluenesulfonyl, –C(O)OR31 and –C(O)R32; R31 is selected from the group consisting of: C1-C6 alkyl, C1-C6 haloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C10 cycloalkyl, -CH2-C3-C10 cycloalkyl, -CH2-phenyl, and –CH2- pyridyl, n any aforementioned cycloalkyl is optionally substituted with from 1-3 independently selected C1-C3 alkyl, and wherein the phenyl is optionally substituted with from 1-2 substituents independently selected from C1-C3 alkyl, C1-C3 haloalkyl, C1-C3 alkoxy, C1-C3 koxy, nitro, halo, SO2Me, cyano, and -OC(O)CH3; and R32 is selected from the group consisting of: H, C1-C6 alkyl, C1-C6 haloalkyl, phenyl, and pyridyl, wherein the phenyl is optionally substituted with from 1-2 substituents ndently selected from C1-C3 alkyl, C1-C3 haloalkyl, C1-C3 alkoxy, C1-C3 haloalkoxy, nitro, halo, SO2Me, cyano, and -OC(O)CH3; R4 is H or C1-C6 alkyl; R5 is –C1-C6 ne-X, wherein X is selected from the group ting of: (i) aryl including from 5 to 6 ring atoms wherein 1, 2, or 3 of the ring atoms are independently selected from the group consisting of N, NH, N(C1-C3 alkyl), O, and S; (ii) heterocyclyl ing from 3 to 6 ring atoms n 1, 2, or 3 of the ring atoms are independently selected from the group consisting of N, NH, N(C1-C3 alkyl), O, and S; R5 is X, wherein X is heterocyclyl including from 3 to 6 ring atoms wherein 1, 2, or 3 of the ring atoms are independently selected from the group consisting of N, NH, N(C1-C3 alkyl), O, and S; wherein R5 is unsubstituted or substituted with ; and R6 is selected from the group ting of H, halogen, hydroxyl, cyano, -O-C(O)-C1-C6 alkyl, C1-C6 alkyl, and C1-C6 alkoxy.

2. The nd of claim 1, wherein R1 is H.

3. The compound of claim 1 or 2, wherein R2 is H.

4. The compound of any one of claims 1 to 3, wherein R3 is H.

5. The compound of any one of claims 1 to 3, wherein R3 is a nitrogen protecting group.

6. The compound of claim 5, n R3 has formula –C(O)OR31.

7. The compound of claim 6, wherein R31 is C1-C6 alkyl.

8. The compound of claim 7, wherein R31 is utyl.

9. The compound of claim 5, wherein R3 has formula –C(O)R32.

10. The compound of claim 9, wherein R32 is C1-C6 alkyl.

11. The compound of claim 10, wherein R32 is –CH3 or iso-propyl.

12. The compound of any one of claims 1 to 11, wherein R5 is X.

13. The compound of claim 10, wherein X is heteroaryl including from 5 to 6 ring atoms wherein 1, 2, or 3 of the ring atoms are independently selected from the group consisting of N, NH, N(C1-C3 alkyl), O, and S.

14. The compound of claim 13, wherein X is selected from the group consisting of 1,2,4- zolyl, oxadiazolyl, pyridyl, and pyrimidinyl.

15. The compound of any one of claims 12 to 14, wherein R5 is substituted with – CH(R4)(R6).

16. The compound of claim 15, wherein R4 is C1-C6 alkyl.

17. The compound of claim 15 or 16, wherein R4 is methyl.

18. The compound of any one of claims 15 to 17, wherein R6 is selected from the group consisting of hydroxyl, )-C1-C6 alkyl, and C1-C6 alkoxy.

19. The compound of claim 18, wherein R6 is hydroxyl.

20. The compound of any one of claims 1 to 11, wherein R5 is –C1-C6 ne-X.

21. The compound of any one of claims 1 to 11, wherein R5 is –C1-C2 alkylene-X.

22. The compound of claim 21 or 22, wherein R5 is substituted with –CH(R4)(R6).

23. The compound of claim 22, wherein R5 is substituted on the alkylene portion with – CH(R4)(R6).

24. The compound of claim 23, n R5 has the a –CH(CHR4R6)-X.

25. The compound of any one of claims 22 to 24, wherein R4 is C1-C6 alkyl.

26. The compound of any one of claims 22 to 25, wherein R4 is methyl.

27. The compound of any one of claims 22 to 26, wherein R6 is selected from the group consisting of hydroxyl, -O-C(O)-C1-C6 alkyl, and C1-C6 alkoxy.

28. The compound of claim 27, wherein R6 is hydroxyl.

29. The compound of claim 20 or 21, wherein X is heteroaryl including from 5 to 6 ring atoms wherein 1, 2, or 3 of the ring atoms are independently selected from the group ting of N, NH, N(C1-C3 alkyl), O, and S.

30. The compound of any one of claims 20 to 22, wherein X is selected from the group ting of 1,2,4-oxadiazolyl, 1,3,4-oxadiazolyl, pyridyl, and pyrimidinyl.

31. The compound of any one of claims 1 to 30, wherein Rb is H.

32. The compound of claim 1, wherein the compound is selected from the group consisting , , , , , , , , , , , , , , , , , , and

33. The compound of claim 1, n the compound is or a pharmaceutically acceptable salt, a stereoisomer, or an N-oxide thereof.

34. The compound of claim 1, wherein Rb is H; R1 is H; R2 is H; R3 is –C(O)R32, wherein R32 is –CH3; R4 is methyl; R5 has the formula –CH(CHR4R6)-X, wherein X is pyrimidinyl; and R6 is hydroxyl.

35. A ceutical composition comprising a nd of any one of claims 1 to 34, and a pharmaceutically acceptable excipient.

36. The pharmaceutical composition of claim 35, suitable for oral administration.

37. The pharmaceutical composition of claim 35, suitable for enous administration.

38. Use of a compound of any one of claims 1 to 34 in the manufacture of a medicament for treating depression, Alzheimer’s disease, ion deficit disorder, schizophrenia, or anxiety.

39. The compound of claim 1, ented by formula I: or a pharmaceutically acceptable salt, a stereoisomer, or an N-oxide thereof, wherein Rb is selected from the group consisting of H, halogen, hydroxyl, cyano or C1-C6 alkyl; R1 is H or C1-C6 alkyl; R2 is H or C1-C6 alkyl; R3 is selected from the group consisting of H, C1-C6 alkyl and a nitrogen protecting group; R4 is H or C1-C6 alkyl; R5 C6 alkylene-X, wherein X is selected from the group consisting of a 5- to 6- membered heteroaryl ring having 1, 2, or 3 heteroatoms selected from O, S, or N, or a 4- to 6-membered cyclyl ring having 1, 2, or 3 heteroatoms ed from O, S, or N, and wherein R5 is unsubstituted or substituted with ; and R6 is selected from the group consisting of H, halogen, hydroxyl, cyano, -O-C(O)-C1-C6 alkyl, C1-C6 alkyl, or C1-C6 alkoxy.

40. The compound of claim 39, wherein R1 is H.

41. The compound of claim 39 or claim 40, wherein R2 is H.

42. The compound of any one of claims 39 to 41, wherein R3 is H.

43. The compound of any one of claims 39 to 41, wherein R4 is C1-C6 alkyl.

44. The compound of any one of claims 39 to 42, wherein R4 is methyl.

45. The compound of any one of claims 39 to 44, wherein Rb is H.

46. The compound of any one of claims 39 to 45, wherein R5 is X, and n X is a 5- to 6-membered heteroaryl ring selected from the group consisting of azetidinyl, pyrrolidinyl, pyrazolidinyl, pyridinyl, pyrimidinyl, isooxazolidinyl, imidazolidinyl, oxazolidinyl, thiazolidinyl, and isothiazolidinyl.

47. The compound of any one of claims 39 to 45, wherein R5 is or .

48. The compound of any one of claims 39 to 45, n R5 is X, and wherein X is tituted or substituted with .

49. The compound of any one of claims 39 to 48, wherein R6 is hydroxyl.

50. The compound of claim 39, wherein the compound is or , or a pharmaceutically acceptable salt, a stereoisomer, or an N-oxide thereof.

51. The compound of claim 39, wherein Rb is H; R1 is H; R2 is H; R3 is –C(O)R32, wherein R32 is –CH3; R4 is methyl; R5 has the formula –CH(CHR4R6)-X, wherein X is 1,3,4-oxadiazolyl; and R6 is hydroxyl.

52. The compound of claim 1, represented by formula II: (II) or a pharmaceutically acceptable salt, a isomer, or an N-oxide thereof, wherein R3 is selected from the group consisting of H, C1-C6alkyl and a nitrogen protecting group; R7 is a 5- to 6-membered aryl ring having 1, 2, or 3 heteroatoms selected from O, S, or N, tituted or substituted on a free carbon by a substituent selected from the group consisting of: halogen, C1-C6alkyl, hydroxyl, cyano, and phenyl.

53. The compound of claim 1, represented by a III: (III) or a pharmaceutically acceptable salt, a isomer, or an N-oxide thereof, wherein Rb is selected from the group consisting of H, halogen, hydroxyl, cyano or C1-C6 alkyl; R1 is H or C1-C6 alkyl; R2 is H or C1-C6 alkyl; R3 is selected from the group consisting of H, C1-C6 alkyl and a nitrogen protecting group; R4 is H or C1-C6 alkyl; X is a 4- to 6-membered heterocyclyl ring having 1, 2, or 3 heteroatoms selected from O, S, or N, wherein is present on a free carbon of X; and R6 is selected from the group consisting of H, halogen, hydroxyl, cyano, -O-C(O)-C1-C6 alkyl, C1-C6 alkyl, or C1-C6 .

54. The compound of claim 53, wherein R1 is H.

55. The compound of claim 53 or claim 54, n R2 is H.

56. The compound of any one of claims 53 to 55, n R3 is H.

57. The compound of any one of claims 53 to 56, wherein R4 is C1-C6 alkyl.

58. The nd of any one of claims 53 to 56, wherein R4 is methyl.

59. The compound of any one of claims 53 to 58, wherein Rb is H.

60. The compound of any one of claims 53 to 59, wherein X is .

61. The compound of any one of claims 53 to 60, wherein R6 is hydroxyl.

62. The compound of claim 53, wherein the compound is .

63. A pharmaceutical composition comprising a compound of any one of claims 39 to 62, and a pharmaceutically acceptable excipient.

64. The pharmaceutical composition of claim 63, suitable for oral administration.

65. The pharmaceutical composition of claim 63, suitable for intravenous administration.

66. Use of a compound of any one of claims 39 to 62 in the manufacture of a medicament for treating sion, Alzheimer’s disease, attention deficit disorder, schizophrenia, or anxiety.

67. A compound according to any one of claims 1 to 34 or claims 39 to 62, substantially as herein described with reference to any one or ore of the examples but ing ative examples.

68. A pharmaceutical composition according to any one of claims 35 to 37 or claims 63 to 65, substantially as herein described with reference to any one or more of the examples but excluding comparative examples.

69. Use of claim 38 or claim 66, substantially as herein described with reference to any one or more of the examples but excluding comparative examples.