AU2016231532B2 - Crystalline peptide epoxy ketone protease inhibitors and the synthesis of amino acid keto-epoxides - Google Patents

Abstract

The invention relates to crystalline peptide keto epoxide compounds, methods of their preparation, and related pharmaceutical compositions. This invention also relates to methods for the preparation of amino acid keto-epoxides. Specifically, allylic ketones 5 are stereoselectively converted to the desired keto epoxides.

Description

CRYSTALLINE PEPTIDE EPOXY KETONE PROTEASE INHIBITORS AND THE SYNTHESIS OF AMINO ACID KETO-EPOXIDES

The present application is a divisional application of Australian Application No. 2013203566, which is incorporated in its entirety herein by reference.

Related Applications

This application claims the benefit of priority of U.S. Provisional Patent Application Serial No. 60/997,613 filed on October 4, 2007 and U.S. Provisional Patent Application Serial No. 61/008,987 filed on December 20, 2007, the teachings of which are incorporated by reference in their entirety.

Background of the Invention

In eukaryotes, protein degradation is predominately mediated through the ubiquitin pathway in which proteins targeted for destruction are ligated to the 76 amino acid polypeptide ubiquitin. Once targeted, ubiquitinated proteins then serve as substrates for the 26S proteasome, a multicatalytic protease, which cleaves proteins into short peptides through the action of its three major proteolytic activities. While having a general function in intracellular protein turnover, proteasome-mediated degradation also plays a key role in many processes such as major histocompatibility complex (MHC) class I antigen presentation, apoptosis, cell growth regulation, NF-κΒ activation, antigen processing, and transduction of pro-inflammatory signals.

The 20S proteasome is a 700 kDa cylindrical-shaped multicatalytic protease complex comprised of 28 subunits organized into four rings. In yeast and other eukaryotes, 7 different a subunits form the outer rings and 7 different β subunits comprise the inner rings. The a subunits serve as binding sites for the 19S (PA700) and 1 IS (PA28) regulatory complexes, as well as a physical barrier for the inner proteolytic chamber formed by the two β subunit rings. Thus, in vivo, the proteasome is believed to exist as a 26S particle (“the 26S proteasome”). In vivo experiments have shown that inhibition of the 20S form of the proteasome can be readily correlated to inhibition of 26S proteasome. Cleavage of amino-terminal prosequences of β subunits during particle formation expose amino-terminal threonine residues, which serve as the catalytic nucleophiles. The subunits responsible for catalytic activity in proteasomes thus possess an amino terminal nucleophilic residue, and these subunits belong to the family of N- terminal nucleophile (Ntn) hydrolases (where the nucleophilic N-terminal residue is, for example, Cys, Ser, Thr, and other nucleophilic moieties). This family includes, for_ example, penicillin G acylase (PGA), penicillin V acylase (PVA), glutamine PRPP amidotransferase (GAT), and bacterial glycosylasparaginase. In addition to the ubiquitously expressed β subunits, higher vertebrates also possess three interferon-y-inducible β subunits (LMP7, LMP2 and MECLI), which replace their normal counterparts, X, Y and Z respectively, thus altering the catalytic activities of the proteasome. Through the use of different peptide substrates, three major proteolytic activities have been defined for the eukaryote 20S proteasome: chymotrypsin-like activity (CT-L), which cleaves after large hydrophobic residues; trypsin-like activity (Τι), which cleaves after basic residues; and peptidylglutamyl peptide hydrolyzing activity (PGPH), which cleaves after acidic residues. Two additional less characterized activities have also been ascribed to the proteasome: BrAAP activity, which cleaves after branched-chain amino acids; and SNAAP activity, which cleaves after small neutral amino acids. The major proteasome proteolytic activities appear to be contributed by different catalytic sites, since inhibitors, point mutations in β subunits and the exchange of y interferon-inducing β subunits alter these activities to various degrees.

What is needed are improved compositions and methods for preparing and formulating proteasome inhibitor(s).

Any discussion of the prior art throughout the specification should in no way be considered as an admission that such prior art is widely known or forms part of common general knowledge in the field.

Summary of the Invention

According to a first aspect, the present invention provides a pharmaceutical formulation comprising a pharmaceutically acceptable carrier and a crystalline compound of Formula (II)

wherein the crystalline compound has 2Θ x-ray powder diffraction values 6.10; 8.10; 9.32; 10.10; 11.00; 12.14; 12.50; 13.64; 13.94; 17.14; 17.52; 18.44; 20.38; 21.00; 22.26; 23.30; 24.66; 25.98; 26.02; 27.84; 28.00; 28.16; 29.98; 30.46; 32.98; 33.22; 34.52; and 39.46

According to a second aspect, the present invention provides use of the formulation of the first aspect for the manufacture of a medicament for treating a disease or condition selected from multiple myeloma, Waldenstrom’s macroglobulinemia, mantle cell lymphoma, diffuse B cell lymphoma, mediastinal (thymic) large B cell lymphoma, intravascular large B cell lymphoma, cancer of the lung, and cancer of the pancreas.

According to a third aspect, the present invention provides a method for treating a disease or condition selected from multiple myeloma, Waldenstrom’s macroglobulinemia, mantle cell lymphoma, diffuse B cell lymphoma, mediastinal (thymic) large B cell lymphoma, intravascular large B cell lymphoma, cancer of the lung, and cancer of the pancreas, comprising administering to a subject in need thereof the formulation of the first aspect.

Unless the context clearly requires otherwise, throughout the description and the claims, the words “comprise”, “comprising”, and the like are to be construed in an inclusive sense as opposed to an exclusive or exhaustive sense; that is to say, in the sense of “including, but not limited to”.

The invention generally relates to the synthesis of proteasome inhibitors and the preparation and purification of intermediates useful therefor.

Another aspect of the invention relates to crystalline compounds having a structure of Formula (I) or a pharmaceutically acceptable salt thereof,

wherein X is Ο, NH, or N-alkyl, preferably O; Y is NH, N-alkyl, O, or C(R9)2, preferably N-alkyl, O, or C(R9)2; Z is O or C(R9)2, preferably C(R9)2; R\ Rf, R:\and' R4 are hydrogen; •each of R\ R*. R", R% and R9 is independently selected from hydrogen, Ci.salkyi, Gi^hydroxyalkyl, 0M8lkoxyalky!:! aryh and Gi ^aralkyk each of which is optionally substituted with a gmup isetecfed from alkyl,: amide, annne, carboxylic add or a pharmaceutically acceptable salt thereof» carboxyl ester, thiol, and thioether, preferably Rs, R*, R7, and Rs are independently sdeeted front Gcsatkyl, Co^hydroxyaikyl, and gatallyl andeaeh is hydrogen, more preferably, R^: and R:8 are iTxlependeniiy Cy^alkyi, R5 and R7 are:independently Cj^amlkyl and eachR* is M;: sn is an integer from 0 to 2; and rs is an integer from:0 to 2, preferably 0 or 1.

Another: aspect of the invention relates to a crystalline compound of Poromla (111)

wherein X Is any suitable counterion.

Ano ther aspect of this i nvention relates to methods for the synthesis of amino acid kelmepoxides accordmg; to scheme (1}

wherein R1 is selected from a protecting group or a further chain of anuoo acids, which Itself «ray be optionally substituted, preferably a protecting grodp. most preferahly an electron withdrawing protecting group: R2 Is selected from hydrogen and C;.{>elkyJ; R5 is selected heterocyclyL aryl, heteroaryl C^feeteroaralMyb and C ^aralkyl; and wherein the method comprises a stereoselective epoxidation under epoxMisring conditions, preferably an aqueous sodium hypochlorite (hieach) or calcium hypochlorite solution in the presence of a eosoivent selected; #om pyridine, acetonitrile, dimeihyifdrmatnlde fDMF), dimethyisulfoxide (DMSO), N~ methylpyirolidme IhlMP)s dmteihyiaceiamide (DMA), tetrahydrofuran (TMP)S and nitroaaethane.

Brief Description of the Figures figure 1 shows a DSC: {di-ifefa&tial scanning calorimetry} thermogram of crystalline compound '1.

Figure 2 shows; anDiftPD (X-ray powder diffraction) pattern of erpialllne compound I...

Figure 3 shows a TG themtogram of crystalline compound 1. f igure 4 shows;; a DSC thermogram of amorphous compound 1 Compared to a BSC thermogram of erystalitne compound 1,

Figure 5 shows an XRFD pattenvof amorphous compound I compared to the XRPD pattern of crystalline compound 1.

Figure 6shows a TG thermogram of amorphous compound 1 compared to the TO pattern of crystalline compound F figure ? shows a BSC curve of an amorphous sample of eotnpound 1.

Figure S shows the XRPD pattern of amorphous compound I .

Figure 9 shows a BSC curve of a crystalline compound F,

Fiprro 10 shows an XRFD pattern of a crystalline compound f:

Figure 1 1 shows a DSC curve of a crystalline citrate salt of compound 1.

Figure 12 shows an XRPB partem of a crystalline citrate sait of compound 1.

Detailed Description of the Invention

In certain embodiments, the invention relates to crystalline compounds having a stmcture of Formul a (I) or a pharmaceutically acceptable salt thereof,

wherein X is Ο, ΝΉ, or N-alkyl, preferably G; Y IsfNH, id-alkyl, O, or €(1¾¾ preferably N-alkyl( Q, or ¢(11¾ Z is O or €(11¾¾ preferably 0¾¾ R*:> Ry RJ.j and R4 are hydrogen; each of Ry, R6, R:\ R8 and R^ is independently selected horn hydrogeu, G^alkyl, Gj^hydroxyafkyh Ci.?,aikoxyaikyl, aryl, and G^aralkyi, each of which is optionally :S.«i]bsitltMted--alkyl, amide, amine, carboxylic acid or a pharmaceutically acceptable salt thereof: carhoxyl ester, thiol, and thioether, preferably R3, R6, R7, and R* are ind^!mdenUy.^eetc4:.ib®tlR:C|.6alkyl9 C^hydroxyalky|,,and Cn $aralky! and each R$ is hydrogen, more preferably, Rfe and Rs are independently Ci^alkyl, Rs and R7 are Independently C^araikyl add each R9 is H; m is an integer frpfn 0 to 2; and n Is an integer Ront 0 to 2, preferably 0 or I ,

In certain embodiments,. Xis 0 and R^ R^ Rfe and R4 afe all the Sathe, preferably R!,: Ry R3, end R4 are all hydrogen. In certain such embodiments, E-s R^, Ry and R* arc independcntl y selected ifom C j ..6aikyh. € j^hydroxyalkyh and €fyam!kyl, more preferably, R6 and R^ are independently Chalky I and R3 and R7 are Independently Gs. garalkyl.

In certain preferred embodiments, X is 0, Rf, Ry R^ and R4 are ail hydrogen, R* and R8 are both isobutyl, Rs is phenylethyl, and R7 is phenyhnethyi. Ιό certain embodiments, R'\ R* R', and R* are independently selected from hydrogen, €nsldkyl5 C^ghydroxyaikyl, Ci-ealkox^lfe^li^yi^i^d^-Ci^aralky!, each of which is optionally substituted with a group selected fern alkyl, amide, amine, carboxylic acid or a iritarmaseuticaily ueeeptahlesait thereof, carboxyl ester, thiol, and tbioether. In certain embodiments, at least one of It5 and K! is Cj^aralkyl substituted with alkyl, more preferably substituted with perhaioalkyl. In certain such embodiments, R ' is C^aralkyi substituted With trilluororaethyl.

In certain embodiments, Y is selected from N-alkyl. O, and €,¾ In certain such embodiments, Z is €%, and m and n are both 0. In certain £ is 0¾ m Is % and n is 2 or 3. In yet another alternative Such embodiments. £ is 0, m is i, and n is 2.

In ccrtain embodiments, the Invention relates to a crystalline compound of Formula (II)

In certain embodiments, the invention relates to a method Idr the preparation, of a crystalline compound of formula (I) or f I E), comprising one Or mbm ofr (i) preparing the amorphous compound, e.g,, according tp U.S. Patent bln.. 7,232,BIS; (li) dissolving the amorphous .compound in an organic so!vein; ins) bringing the solution to supersaturation to cause ibrirtation of crystals: and (iy) isolating the crystals, elg,, by filtering the crystals, by decanting fluid from the crystals, or by any other suitable separation technique. In certain.: embodiments. preparation further comprises:adducing crystalfixation. In certain embodiments, 'preparation further comprises washing the filtered crystals, e.g,, with a solvent or non-solvent fluid In certain: emhodimems,:prepamtion fttrther cnmprises drying, preferably under reduced pressure, such as under vacuum pressure, ϊή certain embodinteuts, the invention mlafos fo a method for the preparation of a crystalline compound of Formula (1} or (11), eoTOprismg one or more of (?) preparing a solution of the amorphous compound, which compound may be prepared according to, for example, U.S. PatcniNo. 7,23.2,818, ip an organic solvent; (ii) bringing the solution to supersaturation to cause formation of crystals; and (in) isolating the crystals, e,;g., by filtering the crystals, by decanting fluid from the crystals, or by any other suitable separation technique, In .certain embodiments, preparation further comprises inducing crystallization, in certain embodiments, preparation fortber comprises washing the filtered crystals, e.g., with a solvent or non-solvent fluid. In certain embodimests, preparation further comprises drying, preferably under reduced pressure. Such as under vacuum pressure. in certain emfeodimeuts, the amorphous compopidmayhe dissolved in an organic solvent selected from acetonitrile, methanol ethanol, ethyl acetate, isopropanol isopropyl acetate, isohutyi acetate, butyl acetate, propyl acetate, metbylethyl ketone, tnethyhsobniyi ketone, and acetone, or any combination thereof In certain embodiments, the amorphous compound may be dissolved in an organic sol verb selected from acetonitrile, methanol, ethanol,: ethyl acetate, isopropyl acetate, mcthylethyi ketone, arid acetone, or hrsy combination thereof In certain embodiments, the amorphous compound may be dissolved inari Ofganie solvent selected from acetonitrile, methanol, ethanol, ethyl acetate, methyleihyiketone, or any combination thereof In certain embodimentss the organic solvent or solvents may he Combined with water.

In certain embodiments, bringing the solution to supersaturation comprises tlie addition of an anti-solvent, such as water or another polar liquid miscible Withthe Otiahic solvent, allowing the solution to cool, reducing the volume of the solution, or any combination, thereof in certain embodiments, bringing the solution to stipersatufation comprises adding an anti-sol vent, cooling the solution to ambsem temperature or lower, and reducing the; volume of foe solution, e.g., by evaporating solvent from the solution, lit certain embodiments. allowing the solution to cool may be passive |e,g., alibwing the solution to stand at ambient temperature) or active (e g., cooling the solution in an ice bath or freezer).

In certain embodiments, the method further comprises inducing precipitation or crystallization. In certain embodiments inducing precipitation or crystallization comprises secondary sucleation, wherein nucleation occurs in the presence of seed crystals or interactions with the environment (crystallizer walls, stirring impellers, sonioaiim etc,}.

In certain embodiments, washing the crystals comprises washing:with a liquid selected from Mil-solvent, acetonitrile; methanol, ethanol ethyl acetate,: methylethyl ketone, acetone, or a comhiriation thereof! Preferably the crystals are washed: with a 'combination of anti-sol vent and the organic solvent;, It? certain embodiments, the anti-solvent is water.

In certain embodiments, washing the crystals comprises washing the crystalline Compound of Formula (Π) with methanol and: Water.

In certain embodiments,. a crystalline Compound of Formula (II) is substantially pnre. In certain embodiments, the melting point of the crystalline compound of Formula |11) is in the range of about 200 to about 220 °€, about 205 to about 215 °C, about 211 to about 213 *€1 or even about 212-G,

In: Certain embodiments, the DSC of a crystalline eornpound of Formula {11} has a sharp endothermic maximum at about 212 °G, e.g.> resulting from melting and deCqmposriioriofthe ays%jfinefomt,#.shoiiMn,:in Figure 1,

In certain embodiments, the X-ray powder pattern of a crysiailine compound of Formula (ίί) is {6-2θ°}:: 6riS; 8 JO; 9.32; I0,1Qy 11.00; 12; 14: 122.50; 13.64; 13.94; 17.14; I7J2; 313,44; 20.38; 21.00; 22.26; 23.30; 24.66; 25.98, 26.02: 27.84; 28.00; 2S.16; 29.98; 30.46; 32.9S; 33.22; 34.52; 39.46 as shown in Figure 2. ip certain embodiments, the TG thermogram of a crystal ling; compound; of Formula pi) exhibits from 0.0 to OP weight loss in the temperature range of 25 to 200 °C as shown in Figure 3.

In certain embodiments,: a crystalline compound of Formula (II) is not solvated. :(e.:g,, the: Crystal lattice dobs pot comprise molecules of a solvent). In certain alternative embodiments, a crystalline compound of formula (II) Is solvated.

In certain embodiments, the invention relates to a method for the preparation of an amorphous compound of Formulafl!) comprising one or more of (i) dissolving the crystalline compotlud:In an organic solvent; (h) bringing the solution to supersaturahon to (ιΐί) isolating the crystals, e,g., by filtering the crystals, or byany other suitable separation teclmirtne. ίο certain embodiments, pregurarion further comprises ioditeihg precipitation. In certain embodiments, preparation farther comprises washing the amorphous compound. In certain embodiments, the method forther comprises drying, preferably under reduced pressure, such as under vacuum pressure. In certain embodiments, the invention relates to a crystalline salt of a compound of Formula (I) or (II), wherein the salt counterion is ^elected ^Ιοπ^'ΙηΙ^^.ρ^ό^ρΗδίο, nitrate; acetate, trifluoroaeetate, •citrate, methanesulfonate, valerate, oieate, pah«itate, steamte, jaurate, benzoate, lactate, succinate, tosylate, maldnate, maleate, fhmarate; succinate, tartrate, mesylate, 2~ hydroxyethansuifbnate, and the like. In certain such embodiments, the salt counterion is selected fmm citrate,, tartrate, trifluoroaeetate, methaoesulfonste, tolnencsulthnate, chloride, and bromide, preferably citrate.

In certain embodiments, the invention relates to a method tor the preparation of a crystalline salt of a compound of Formula (II), comprising one or more of: (i) preparing lite amorphous compound e.g., according to U.S. Eatent No. 7,232.81 S; (ii) dissolving the amorphous compound in an organic solvent; (in) bringing the solution to supersaturation to cause formation of crystals; and (i v) isolating the crystals, e.g., by filtering the crystals, by decanting fluid from the crystals, or by any other suitable separation technique. In certain embodiments, preparation forther comprises inducing crystal 1 ination. In certain embodiments. preparation further:comprises wasfeipg-the crystals, e.g., with a solvent or non-solvent fluid. In certain embodiments, preparation further comprises drying, preferably under reduced pressure, snob as under vacuum pressure.

In certain embodiments, the invention relates to a method for the preparation Of a crystalline salt of a compound of Fonnok (II), comprising one or more of (i) preparing a solution of a compound of Formula (I S) in an organic solvent; (it) adding a suitable acid; (iii) bringingthe solution to supersaturation to cause formation of crystals; and (tv) isolating the crystals, e.g., by Altering the crystals^ by decanting fluid from the crystals, or by any other suitable separation teehnigue. hr certain embodiments, preparation further comprises inducing crystallization,. In certain embodiments, preparation further comprises washing the crystals, e. g., with a solvent or non-solvent fluid. In certain embodiments, preparation further comprises drying, preferably under reduced pressure, such as uuder vacuum pressure.. In certain embodithents where the salt is less soluble in a solveM than the tree base, adding the acid to a solution may itsdf he sufficient to bring the solution to supersaturatiom

In certain embodiments; thesalt eoimterion is selected from selected from bromide, chloride, sulfate, phosphate, nitrate, acetate, Mfluoroaeetate, citrate, methanesol formic; valerate, oleate, palmitate, stearate, fduraie, benzoate, lactate, succinate, tosylaie, tnalonate, maieatUj flmtarate, succinate,:tartrate, mesylate,;2~ hydro&yedtansuifoMie, and the lihe, in certain such embodiments, the salt counterion is selected fkim citrate, tartraiepiriOuoroacstate, methanesuffonate, toluenesuffehaie, Chloride, and:bromide, preferably citrate. iu eertain embodiments, the:organic soivent is selected from THF, acetonitrile, ether, and IVITBE, or any eombinatiorr thereof, preferably TMF or acetonitrile,: or a eonibination thereof:

In certainiembodimcuts, a cryslaliineciu'atesali ofaeompound of Formula (li)is substantially pure.: .In certain embodiments, the melting: point of the crystalline citrate salt of a compound Of Formul a (II) is in the range of about 180 to about 190 °C or even about 184 to about 188 °G,

In certain embodiments, the DSC of a crystalline citrate salt of a compound of Formula pi) has a sharp eudothermie maximum at about 1S7 ftC, e.g., resulting #om melting and decomposition of the crystalline form as shown in figure 11, la certain embodiments, the X-ray powder pattern of a crystalline citrate salt of a compound of Fonuula (11) is {6-20*); 4.40; 7.22- 9,12: 12.3b· 13.35; 14.34; 13,54; l b. 14; 16,54; 17.00, 18.24; 18.58; 19.70, 19.90; 20.30; 20.42; 21.84; 22.02; 23,34; 23 J4; 24:04; 24.08; 2448; 2:4.76;: 254S; 26.,18; 28,14; 28¾ 28.04; 29,64; 31.04; 31.84; 33:;00; 3:3.20; 34.06; 34:.301 34.50; 35.18; 3748; 37.90; 39.48 as shown in Figure: 12, 3ln certain embodiments, theinvemion relates to a crystalsiue compound of Formula fill)

wherein X is: any sui table counterion.

In certain embodiments, X is a counterfoil·selected from bromide, eHIoride, sulfate, plmsphate, nitrate, acetate, frifluoroacetatej citrate, nmthanesulfonatej valerate, oleate, pahnitate, Stearate, daurate, benzoate, lactate, succinate, iosyltde, malonate, maleate, fomarate, succinate, tartrate, mesylate, '2-hydroxyethansulfoaate, and the like. (See, forexample, Berge etal. (199:7) 'Thanoaeeuucal Salts'5, ,1. PMarm. $ci.#6: 1-19.) in certain embodiments X isselected fmm trffluomacetate, methanesuiibnate, to! uenesulfonate, acetate, chloride, and bromide, preferably trifluoroaceiate.

In certain embodiments, tlminvention relates to a method for the preparation of a crystalline compound of Formula (III) composing one or more of: (I) preparing a compound of Formula (IV). e.g,, according to Bioorg, Med. Chem. Letter 1999, 9, 22Sa-88 or U ,S. Fatent Application 20()54)256324, wherein PO is a suitable protecting group (e,g.}: Bocor Gbz)

(ii) dissolving the compound of Formula (IV) in an organic soiventj (iii) adding a suitable acid) (tv) bringing the solution to supersatnratiOn to cause fermatipriof crystals: and (9) isolating the crystals, eig., by filtering the crystals, by decanting fluid from the crystals,oi' by any other suitable separation technique. In cejtain embodiment% preparation further comprises inducing crystallization, in certain: embodiments:, preparation fufther comprises washing the crystals, e.g., with a solvent or nun-solvent fluid. In certain embodiments, preparation further comprises drying, preferably under reduced pressure: such as Under vacuum pressure.

In eertain emfcodiinenis, the invention relates to a method for the preparation of a crystalline compound of formula {illX comprising one or more of ft) preparing a solution ofan anfo^hous compound of Formula (IV), «.g,, according to Bioofg, Med, Otero, loiter 1999» 9,2283-S8 or U.S, Patent Application 2005-0256324» in an organic solvent, wherein PG is a suitable protecting group (e.g., Boo or Cbz),

(ii) bringing the solution to supersaiuraiiOu to cause formation of crystals; and (lii) isolating the crystals» e.g., by filtering the crystals, by decanting fluid Icom the crystals, or by any other suitable separation technique. In certain embodiments, preparation further comprises inducing crystallization, in certain embodiments, preparation further comprises washing;foe crystals, e.g , with a solvent or non-solvent fluid. In certain embodiments, preparation further comprises drying, preferably under reduced pressure, such as under vacuum pressure.

In cert ain embed intents the acid is selected from hydrobromic, hydrochloric, sulfuric, phosphoric, nitric, acetic, Irifiuorpaeetjo, citric, methanesul feme, valeric, oieaic, palmitic, stearic, lanric, b^zoic, lactic, succinic, p-tolnenesuitonic, citric, malonic, maleic, fomaric, succinic, tartaric, methanesnifonic, 2-hydroxyetbanesulfonic, and the like. Preferably the acid is trifluoroacetic acid.

In certain embodiments, X t$ a counterion selected from hydrobromi.de, hydrochlor i de. m I fate,: phosp hate, nitrate, aceta te, iri fi uoroaeet ate, citrate, meihanesutibnate, valerate, olcate, paltnitate, stearate, laurate, benzoate, lactate, succinate, tosylate, malonatc, maleafe, fornarate, succinate, iartrafe»:mesylate»:,2~ hydrexyethansulfonatc, and the like. {See, for example»: Bergeet al. (1977) “Pharmaceutical Salts", 3, Phams. Set 66: 1-19.) In certain embediments, X is selected from trifluoroacetate, methanesn donate. to! uenesul lunate, acetate, cbloride, and bromide, preferably trifluoroacetate.

In certain embodiments, the compound of fortmila (1¾ he dissolved in an organic solvent selected from dichloromelhans, ethyl acetate, isopropyl acetate, isobutyl acetate, butyl acetate, propyl acetate, diethyl ether, methyl teft-butyi ether (PTBE), or any combination thereof In certain embodiments, the organic solvent is selected from dichloromethane, ethyl acetate, MT8E, or any combination thereof, preferably either dichloromethane and MTBE or ethyl acetate and ΜΤϋΒ,

In certain embodiments, bringing the solution to sopersaturation comprises the addition of an anti-solvent, such as hexanes or heptanes oratmther l iquid miscible with the organic solvent, allowing the solution to cool, reducing the vohime of the solution, or any combination thereof. In certain embodiments, bringing the solution to supereaturatiohcomprises adding an anti-solvent, cooling the solution to ambient temperature or lower, and reducing the volume of the solution, e.g., by evaporating solvettt from the solution. In certain embodiments, the anti-solvent is hexanes or heptanes, preferably heptanes.

In certain embodiments, washing the crystals comprises washing with a liquid selected front anti-solvent, ethyl acetate, dichloromethane, or a combination thereof Preferably the crystals are washed with antfrsotveni, 'preferably heptanes.

In certain embodiments, the: BSC of a crystalline compound of Formula (Ml) has a sharp endbfrienhiem^ °C, e.g., resulting from melting and decomposition of the crystalline lerm as; shown in Figure 9,

In certain embodiments, the X-ray powder pattern of a crystal line compound of Fonmdafll) is 10202: 8,84; lllS; 15.32; 10.20; 16.82; I 7.66; IS 26; 19.30; 21.20; 22.58; 23.0b; 23.32; 2S,32; 26158f 28-60; 30.08; .30.48; 30.84; 32 20; 3624; 37.12 as showniin Figure 30,

In certain embodiments, a crystalline compound of Formula (111) is not solvated (e.g,, the crystal lattice does not comprise molecules of a solvent). In certain alternative embodiments, a crystalline compound of Formula (11 1) is sol vated.

In certain embodiments, the invention relates to a method for the preparation of a crystalline compound of Formula (II). composing one or more of fi) preparing a solution of compound of Formula fW) wherein FG is a suitable protecting group {e.g,? Boe or Cbx), in a first organic solvent

(n) adding a suitable acid; (in) bringing the solution to supersaturation to cause fonnation of crystals; (iv) i solating the crystals to provide a crystalline compound of Formula (ill): {v) reacting the crystalling compound of Formula (ill)

wherein X is any suitable couittermn, with a compound of Formula (V)

to provide a compound of Formula (II); (vi) preparing a solut ion of the compound ot Formula (If) in a second organic soivrstt; |vii) bringing the solution to supematwation to cause formation of crystals; and (viii) isolating the crystals to provide a crystalline compound of Formula (II), e,g<, by filtering the crystals, by decanting, w by any Othes' suitable separation technique. In certain embodiments, preparation furiher comprises inducing crystallization, hi certain embodiments, preparation further comprises washing the Crystals, e.g., with a solvent or oomsolveut fluid. In certain-embodiments, preparation riatber comprises drying, preferably under reduced pressure, such as under vacuum pressure.

In certain embodiments, the acid, is Selected Irom bydrobmmic, hydrochloric, sulfuric,, phosphoric, nitric, acetic, trifluoroacetie, citric, methanesu! tonic, valeric, ofearc, palmitic, stearic, lauric, benzoic, Ibptic, succinic, pdofuenssulfbmo, citric, malocic, mafeie, fbmaric,, succinic, tartanc^ methancsniibnjc, 2-hydroxyethansulfomei and the like. Preferably the add is trifruaroaCetiebcid,

In certain embodiments, Xfs aieounterion selected item hydrobmmiie, hydrochloride, sulfate, phosphate, nitrate, acetate, triffeoroaeetafe, citrate, mctiranesul&amp;naie, valerate, ideate, palmitate, stearate, laurate, benzoate, lactate,

Succinate, tosyiaie, malonate, maleate, fiimaratc, succinate, tartrate, mesylate, % bydmxyethaucsulfbnate, and the like, (See,for example, Berge et af (19:77) "PltStmaceaticaf Saits’5, J< Pharra, Set. 66; 1-19,) In certain embodiments, X is selected from inffeoroacetate, nmthanesuifenaie, tofnenesuHbnafe, acetate, chloride, and bromide, preferably mfluoroacoiate.

Ih certain embodiments,: the first organic solvent is selected from dichlorofeeihape, ethyl acetate, isppropyi acetate, isofetttyl acetate, butyl acetate, propyl acetate, diethyl ether, methyl tert-butyf ether (MTEEf or any combination thereof! 1« certain embodiments, the organic solvent is selected from dieblorpmetbane, ethyl acetate, MTBB, or any combination thereof’ p'efeeabiy either diehloromethane and MTBE or ethyl acetate and Mf BE,

In certain embodiments, the second organic solvent is sefected lions acetonitrile, rnethahoi, ethanol, ethyl acefete, isopropahol, isopropyl acetate, isohutyl acetate, btityl acetate, propyl acetate, methvlethyl ketone, methylisohntyl ketone, and acetone, or any contbination thereof! In certain embodiments, the amorphous componnd may he dissolved in an organic solvent selected from acetonitrile, rneihanoi. ethanol, ethyl acetate, acetone, or any combination thereof! In certain embodiments, the organic solvent or sol vents may be combined with wafer,

In certain embodiments, preparation furth^ Comprises washing the crystals of either or both of formula (If) or (Iff). In certain embodiments, washing the crystals of a compound of Formula fl!) cort^rtses^washing with a iipuM selected from: anti-solvent, acetonitrile, methanol, ethanol, ethyl acetate, acetone, or a combination thereof!

Preferably the crystals of a compound of Formula (I!) are washed with a combination of anti-solvent and the- organic solvent, in certain embodiments, washing the crystals comprises washing the crystalline compound of Formula (II) with methanol and water. In certain embodiments, 'Washing the crystals of a compound of Formula (III) comprises washing with a liquid selected from anti-so! vent, ethyl acetate, dichloromethane, ora combination thereof. Prefemhly the crystals of a compound ofFonnuis (III) are washed with anti-solvent, preferably heptanes.

In certain embodiments, preparation further comprises drying the crystals of either or both of Formula ft !) or (111.), preferably under reduced pressure, such as under vacuum pressure. tn certain embodiments, the Invention relates to a phaouaceuticai contpoMhort comprising a crystalline compound of Formula 0} or (II) and a pharmaceutically acceptable carrier, IP certain enrbodimenis, the pharmaceutical composition is selected from tablets,, capsules, and injections.

This invention also relates to methods for the:synthesis of epoxyketones, such as fennufeeffll) and (W) above. Thus, in another aspect, the invention provides a method fer preparing amino acid keto-epoxjdesuecardiug"to Scheme (I),

wherein R! is selected from a protecting group or a further chain of amino acids, which itself may be optionally substituted, preferably a protecting group, most preferably an electron withdrawing protecting group; R2 Is selected irom hydrogen and Cj.ealkyl; and R3 is selected from hydrogen, Chalky!, Cj,§alkoxyalkyI, heterocyclyh aryl, hetemaryl, Cr^heteroaralkyl, and G^aralkyi; and wherein the method comprises a stereoselective epoxidation under epOxidixing conditions, preferably art aqueous sodium hypochlorite(hleaeh) or calcium hypochlorite solution in the presence of a cosolvent selected from pyridine, acetonitrile, DMF, DMSO, NMP, DMA, THF, and riitr^dihpnex

In certain embodiments, the eosolvent is selected from jNMP and pyridine, preferably pyridine. in certain embodiments, the epoxidaiton is performed using acfoeous sodium hypochlorite in the presence of a cpshlveot selected from pyridine, acetonitrile, DMF, DMSO, NMP, ΟΜΑ, ΤΗΡ, and nitromethane, preferably NMP or pyridine, more preferably pyridine, in pertain embodiments, the epoxidation is performed using a 10% aqueous sodium hypochlorite solution. In certain embodiments, the epoxidation is performed using a 10% aqueous sodium hypochlorite soluti on in the presence of pyridine. In certain embodiments, the epoxidation is performed using a calcium, hypochlorite solution in the presence of NMF.

In certain embodiments, R· i&amp; selected, from a; protecting group or a further chain: df amino acids, fehich itseif'may be optionally substituted, In certain such embodiments, R* is a protecting group, preferably an electron withdrawing protecting group.

In certain bnlbridiments,,ϋ* Is selected from nbutoxy earbonyl (Boo), benzoyl {8x):> riU0ren~9~yhnethoxyearbonyi (:Frnoc), triehloroetbaxyearbonyl (Tree), and benzylpxy carbonyl (Cte).. In certain such embodiments, R1 is beleeted from n-buioxy carbonyl (Boe), benzoyl (Bg), triehloroethoxycarbonyl (Troc), and benxyloxv carbonyl (Cfe), preferably Chz or Boo; In certain preferred embodiments, B* is Boc,

In certain embodiments, R3 is selected from hydrogen, Cj^atkyi, 0;.$alkoxyalfcyi, heterocyciyf, aryi, hetcroarvl, Gusheteroaralkyl, and Cfe«aralkyl. In preferred embodiments, EJ is C^alkyl., preferably isobutyl. In certain preferred embodiments, R3 is ©^aralkyl. preferably phenylmethyl, 4-hydrdxyphenylmethyi, or 2-phepylethyi

In certain embodiments, the Stereoselective epoxidation is performed under conditions that do not result In signs II cam epimerizailon of the carbon bearing Re such that there is less than 10%, less than S%, less than 2%, or even less than 1% epimerixation of the carbon bearing EG in certain embodiments, the stereoselective epoxidation is performed such that the product is greater than about 90%. greater than 9S%, greater than 98%, or even greater than 99% diastereomerieally pure.

In certai n embodiments, the cpoxidation is perlormed at a temperature In the range of about -1S °C to about 10 °C, about -10 :iC to about 5 °C, or even about -5 °C to about 0 **C.

Iltefcrtain the compounds in scheme I have the following stereochemistry

1« certain embodimeMsythe stereoselective epoxidation is performed such that the product is greater than about 90%, greater than 95%, greater than 98%, or even greater than 99% diastereomencal 1 y pure.

The useof various N-proteeting groups, e.g., the benxyioxy carbonyl group or the t-hutyfoxycarhouyi group (Boe), various coupling reagents, e,g,, dicydohexylearbodilmide 0CC), 1 ^-diisopropyloarhodiimide (D1 C),-..i :-(3-dimethylam:in:Opropyi)-3-ethylcarhodiimiile (EDG)vN-hydroxyazabenzotrtazoie (HMlj), carbonyldiraddassole, or l-hydroxybeuzetfiazole mdnohydrate (HOST), and various: cleavage conditions; tor exauipje,:trtfiuoracetfc aeid(TFA}s HC1 in dioxane* hydrogenation on Pd/C in organic: solvents (such as methanol or ethyl acetate!, boron trisCtriluoroacCtate), and cyanogen bromide, and reaction in solution with isolation and purification ofintennediaies are well-known in the art of peptide synthesis, and are equally applicable to the preparation of the subject compounds (fureene, T.W.; Wuts, 9 .G$d.:Froteciim Cmups im Organic Synthesis, ed>; Wiley: New York, 1999),

In certain embodiments, the amino acid koto-epoxide rnay be further modified by deprotection of the amine, if applicable, and coupling with a chain of amino acids. Methods for the coupling of such fragments are well known in the art (EloMon, M., et ah (1999) Chemistry &amp; Bmkigy, 6:811-82% BloNson, M,, et at (1999) Chemistry H'ffloipgy, 6:811-822), In a preferred embodiment, the chain of anuno acids com prises one to three amino acids, in certain embodiments, the chain of amino acids has a structure of formula (Vi) or a pharmaceutically acceptable salt thereof

wherein each A is independently selected Item C~D, €-$, and SQb preferably C::sCkof A is optionally a covalent bohd when adjacent to an eccurrenee of Z·; L i$ absent or is selected :frpm:C”:0, C-S* and SO?, preferably L is absent or 0*01- M is absent or is Cj .stalky!, preferably C^alkyl; Q is absent or is selected from O, MM, and H^Cj^alkyl, preferably <5 Is absents D, or.N-H, most preferably Q is absent or O; X is CQOH or an activated form thereof preferably X is GGOM, COOL or €QM(Me)(OMe), most preferably X is GOOH or COC1; Y is absent oris selected from O, MM , M Xifrsaikyl, S, SO, S02, GHORr\ and GHGOaR17; each Zis independently selected from O, S, NH, and N-<fr.<>aikyL preferably O; dr Z is optionally a covalent bond when adjacent to an occurrence of A; R5, ,Ry and R( are each independently selected from Cj.«alkyl* Ct.«hydmxyalkyls Cfe ^alkoxyaflcyl,.,;]^!, and G^*afalkyl, any of which is optioinaliy substituted with one or more of amide, amine, carboxylic acid (or a salt thereof^, ester (including Co «alky! and Gj.$aikyl ester and aryl ester}, thiol, or thioetbcr substituents’ ft* is Ν(Ε1{ί}Ζ®Η; I's Ri2, and IIs 3 are independently selected from hydrogen, OH, and Gi^aikyi, preferably, Rm is selected from hydrogen, OH, and C^alkyl, and Ri2 and E*5 are independently seieeted from hydrogen and Chalky!, preferably hydrogen; RM is selected from hydrogen, Gj^aikyl, €|-«alke»yl, C^alkynyl, aryl, heteroaryl, Crdretemaralkyfr Ri5ZAZ~Ci,salkyl*·, RfSZXZ^aikyH {Rs sO)( R!sO)Pfr"O)0~Cs .galkyi 'ZAZ-Cs-salkyl-, R i5ZAZ--C j ^al kylZtAZXlj. 8alkyl·, heteroeyclylMZAZ-Cj^alkyl', (R!>O}(Riy0)Pfe:Q)O-C:l^alky!-, (R!7)aH-Ci-ualkyJ-, (R5 ^jN^GM^alkyl*, ItetefeCyclyJM-, eafeoeyelylM-, E^SGbC|,xalkyl~ * wtf R^SOsMH; preferablyGr^afkyfGi.«alkenyl, Cv^aikynyi, aryl, C{.«aralkyl, heteroaryl, G|,sheteroara!kyl, RiSZA~C#.salkyb, R^Z-C^aikyfr, (R!sO)(R.’60)Ρ(~Ο)Ο“·ϋ( -salkyl-ZAZ-Cj.galkyi-, (RL\1)(RS60}P(^0)0-Ci,salkyl·' 2-Cnsatkyt-, RbZA”C|..ta.ikyI.~ZiAZ'-C*.ialkylr, betemeyelylMZAZrCpaaikyi-, (Rί5O)(Rl·&amp;0}F(™0)0”Ci.8alky|>,:(Rs¾N-Cs,salkyl-·, (Rs7)sN*-Cs.safky!:v heteroeyclylM-, earbocydyllVK R^SOiCi-sallcyK and R!%0->NH, wherein each occurrence of Z and A is independently other than a covalent bond; or

Rm and Rn together are Cs^kyl-Y-C^alkyl, Q.^ikyiriZAZ«C,.«alkyl, ZAZ-C^alkyl· ZAZ~Cj.«alkyI, ZAZ-Cj^aikyl-ZAZ, or C^alkyl-A, thereby forming a ring; preferably Ci-aalkyl-Y-C $ dalkyl, Ci .salkyl-Z A~Cj -2alkyl, A-Ci.aalkyt-ZA-Cj. jalkyl, A-Q .jalkyl-A, or CMalkyi~A, wherein each occurrence of Z and A is independently other than a covalent bond; R*5 and Rlf> are independently selected from hydrogen, metal cation. Chalky!, €j. «alkenyl, C^alkynyl, aryl, heteroaryl, C^arslkyh and Cnsheteroaralkyl*· preferably from hydrogen, metal cation, and Cj .«alkyl, or R° and Rift together are C^alkyl, thereby forming a .ring; each R 1 'f is independently selected from hydrogen and C^alkyl, preferably C?.«alkyl;

Rts is independently selected from hydrogen, OH, Cusalkyl, € ^alkenyl, Cj.«alkynyl, carbocyclyl, hetorocyclyl, aryl, heteroairyf, Cj.«aralkyl,-and Cj^heteroaralkyl; pro vided that in any occurrence of the seq uence ZAZ, at least one member of the. sequence must be other than a covalent bond.

In some embodiments, R\ R°s and R7 arc selected from-Chalky! or €h .«aralkyl.

In preferred embodiments, Rfi is Chalky! and R5 and FZ are C^aralkyl. In the most preferred embodiment, Rb is isobutyl, Rs is 2-phenyl ethyL and R; is phenylmethyl in certain embodiments, L and Q are absent and Rn is selected from Chalky! C*. «alkenyl, €j.«alkynyl, C^aralkyl, and C^heteroaralkyl. In certain such embodiments, Ri0 is Cs.«alkyl and Rh is selected from butyl, ally!, propargyl, phenylmethyl, 2-pyridyl, 3-pyridyl, and 4-pyridyk in other embodiments.. L is SO?, Q is absent, and RM is selected from Cf..«alkyl and aryl. In certain such embodiments, R * is selected from methyl .and phenyl.

In certain embodiments, L is C~G and R5* is selected from Chalky!, C; .«alkenyl, C^alkynyl, aryl, C^aralfeyl, heteroaryl, C^heteroaralkyl, R^ZA-Cpttalky!-, RiSZ-Cn galkyfr^R^OKR^DjPf-OJG-Cs.salkyb.iR^OKR^OIPC-OlO-Ci.salkyl-ZAZ-C^gajkyl-, (R! 50)(R1 ή0)Ρ(™0)0-€ s .galky! -Z- CI .&amp;a 1 ky I-, Κ^ΖΑ^ί^ΙΙίνΙ-ΖΑΖ-ϋι^Ι^Ι., heterocydylMZAZ-C^alkyl-, (R4 7) ? N - C 5a Ik y 1 ~,s (R^XsbR-Ci.fcalkyl-j heterocyciylM-, carbocyciyiM^ RlsSO^C|.$alkyi~s andwherein each occurrence of Z and A is independently other than a covalent bond, in certain embodiments, L is 0:::0, Q is absent, and Rn is H.

In certain embodiments, RKt is C^alkyl, R!i is Cj^alkyh Q is absent, and Lis €=0. In certain such emfeodimeols, R11 is ethyl, isopropyl, S^^^iftlinoroethyi, or 2·· (meth y 1 sal ion y !)cthyi.

In other embodiments, I, is 0=0, Q is absent, and Rn is C| .^aralkyl. In certain such embodiments, R1* is selected#om 2-pbenylethyl, phenyimsthyl, (4-methoxyphenyi|med^yl,f4~eMdrophenyi)inetby!, and {44\ norophcnyl }methvl. in other embodiments, L is CM), Q is absent Ri0 is Chalky I, and Rf s is aryl. In certain such embodiments, Efl is substituted or unsubstituted phenyl

In certain embodiments, L is 0=0, Q is absent or 0, n is 0 or l, and Ru is -{CHa)„carboeyclyL in certain such embodiments, Κί! is cyelopeopyl or cyclohexyl

In certain embodiments, L and A are OO, 0 is absent, Z is Ο, n is an integer from I to 8 (preferably 1), and Ru is selected from lli5ZA-Cs.#IkyI~, R'%~0j.8aikyl-S R'^A-Ci^alkyLZAZ-CcsalkyL, {Ε^ίΕ^Ρί^ΟΑΖ^ 4!l,50)(Ri60}P(=0)iC^Ci.pIkyLZ~di^aIltyt^:8nd:betersdyeiylMZAZiCfii9lkyli wherein each occurrence of A is independently other than a covalent bond, In certain such embodiments, R7 is heterocycIylMZAZ-Ci-saikyi" where beteroeyclyl is substituted or unsuhstituied oxodioxolenyl or N(R52){Rs7), wherem '11u and Ru together are Chalky!-Y-Ci.saikyl, preferably Co^a3kybY-Cl^.3alkyli themby forming a ring, in certain preferred embodiments, L is O:::0, Q is absent, n is an integeriroui 1 to 8, and Rn is selected fumUR^OKR^PC^ and heterocyclyf-M·-. In certain such embodiments, Ef ‘ is -C5. «alkyl Nf R1 y)j or -0 * ,sglky1bO( R1 %, where R'7 i s € *. ,$a! kyl. In cetf ai n other such embodiments, Ri: is hetemcpIylM-, where heterocyclyi is selected ;irom morpholine, piperidino, piperaxmo, and pyrrblidino.

In certain envhodiments, L is i> O, RK! is Chalky!, Q is selected from 0 and Hid and Eu is selected from tSi^alkyl, cycloalkyl-M, Ci,^aralkyl, and Ct ishefenaaraikyL in other embodiments, L is 0=0, R*Ns Gg^alkyi Q is selected fenm 0 and Nil, and R -* is

Ct «alkyl where Gtfekyl is selected It® methyL ethyl,and isopropyl. In ferther embodiments. Lis€>0* R!0 is Chalky!. Q is selected from 0 and NH and Rn is Cj. «.aralkyl, where aralkyl is phenylmethyl. In oiberembodiments, L is C-O, R10 is CY «alkyl, Q is selected from 0 and Hf-f, and R"51 is (¾«Mtemaialfeyl* where heteroaralkyl is (4~pyridyi)m ethyl ln.:j^aidi#|ii>bdimed^y)lv|si absent or is C"0, and Ri0 and liu together are .Op plkyl-Y-Ci.salkyl, Gi.«a!ky3-ZAYfeaIkyl, or Cj^alkvl-A, wherein each occnrrenee of Z and A is independently other than a covalent bond, thereby forming a ring. In certain prefeed: emhodlments, L is 0=-0, 0 andΎ are absent, and Ri0 and Ru together are C ( ..: wlkyl-Y-'Cs.ialkyi. In another preferred embodiment, L and Q are absent, and R10 and Rn tdgOthendre CKjalkyl-YAfealkyl. In another preferred embodiment, L is.0==0, Q is: absent, Y is selected from NH and N-Cs «alkyl, and R>y and Ru together are Ct.^alkyl-Y-Cmalkyl. In another preferred embodiment, L is 0-0, Y is absent, and R;!,'} and R!i together are Cj«alkyjy Y «Ο i «alkyl In another preferred embodiment, L and A are C~0, and Rf0 and R ! 1 together are Cj^alkyREACi^alkyl.. In another preferred:embodiment, L and A are OO and R^ and Ru together: ard Cy-jalkyLA.

In certain embodiments, the chain of amino acids has a stmclnre of fomt«k..(YR}:

wherein each A is independently selected from 0==0, 0-3, and SO?, preferably C-O; or A is optionally a Covalent bond when adjacent to an occurrence of Z; each B is independently selected from 0=0, C-S, and $Cb, preferably 0-0; D is absent or is Cs. «alkyl; G is selected from Ο. NH, and N-Cjwalkyl; K is absent or is seieeted iffom 0-0,0*$, and SO?, preferably K is absent or is C-O; L is absent: or is selected fern C~Oi C:ssS5 and SOj, preferably L is absent or 0-0; M is absent, or is €t.salkvl; Q is absent or is selected from O, MB, ant^Cj.^alityt preferably Q is absent, O, or NH, most preferably Q is absent; X is COQH or an activated form thereof, preferably X is C<X>B, COG!, or CON(Me)(OMe), most preferably X is COOH or COCI' each V is independently absent or is selected from O, $, MB, and N-Cngalkyl, preferably V is absent or O; W is absent or is indepepdb&amp;tly selected feemO, S, BE, and N~Cj.<;alkyk preferably 0* ¥ is absent or is selected from Ο, MB, M~Ci.salkyl, $, SO, SCb, OBOR’and CBOOiRi?j each X is independently selected from O, S, MB, and B-Cs.«alkyl, preferably O; or Z is optionally a covalent bond when adjacent to an occurrence of A ; R*, R6, and R' are each independently selected from CMa!|y!, Cs «bydroxyafky!, €5.. salkoxyalkyl,aryl, Ci.saralkyf, and R*H) VKOC*„.?alkyl-, wherein at least one of R* and R? is R'^eVROCj^amyl- ^isNCR^LQRS}; R!S iS selected from hydrogen, Olii and C^alkyl, preferably hydrogenor € s ,&amp;alkyl; R!i is a further chain of amino acids, hydrogen, a protecting group, aryl, Or beteroaryl, any Ofwhieh is optionally substituted with halogen, carbonyl, nitro, hydroxy, aryl, C^alkyl; or Rli is selected from Gj^alkyl, Ccftalkenyl, Ci^alkyoyl, C^aralkyl, G^heteroaralkyl, R*2ZAZ~€f .salkyk R^ZAZ-C^aikyi··, {Ri;?0)(Fd50)F(-t)}C)^ C^alkyl-ZAZ-G^alkyh R!3ZAZ-Oi.ealkybZAZ-C^alkyi-, heterocyciylMZAZ-C^alkyk {Rs%XRl3G)k{-0)0~€psalkyl~, «alkyl-, heteroeyclvlM-, CarbocyclylM-,^S-QaClij.^lkyl», and R^SOaMM; or RS0: and R *] together are C s .«alkyl - Y.-G i -salkyl,: Cs ^aikyl-Z AZ-G u«al kyl, ZAZ-C r^alkyl ZAZ-Cr-salkyl, ZAZ-Ci.6alkybZAZ, or Ci.satkyf-ZAZ;

Ru and R1'4 are independently selected from hydrogen, metal cation, CcsalkyL Ck gaikenyi, Cvi^alkynyf, aryl, heteroaryk C ^aralkyl, and Gi^heteroaralkyl,·. pfef0*$$X cation, and Cj.&amp;alkyi* or R52 and R*3 together are

Cf ^alkyi, thereby fenmng a ring; each RH ss pd#eniesit!y selected from hydrogen apd Chalky!, preferably Cj..6alkyl; each RS;s ss independently selected from hydrogen, ORM, Chalky!, Cswalkenyh C}.. ga!kynyls:carhoeyelyk heterocycly), aryl heieroary I, Cs^aralkyl, and Cfe «heteroaraikyl; ΐ:^:ΐ$:(R^{RssO)Pf~G)W», R!?GB-; heteroeydyj·, (R%M~, {Rw)iN7*>, R1 %020ΒΟ-, and Ri7GBCs .salkyl· where the Chalky! moiety Is optionally substituted with OB; C=^alky 1W {option a\\y su bstit uted with halogen, preferably fluotiTie), aryl, heteroaryl,:.earbocyeiyl, heterocyclyl,.and Cs ^aralkyl, preferably at least one occurrence of as other than hydrogen' R- and R‘* are mdependeoily selected from hydrogen, metal cation, Cpaalkyl, Cs. 6alkenyii Cs>#lky.nyi aryl, heieroary!, G;.«aralkyl,, and Godictefeaf alkyl, preferably from hydrogen, metal cation, and C^sikyl, or:Rf' and Rig together are Cstalky!, therebyi fenning a ring; and each R,$ Is Independently selected from hydrogen, OR^YCj.gaiky!, Ci^aikenyl, Cj. $alkynyl, carbocyelyl, heterocyclyl, aryl, hetcroatylj Cgsaralkyk and C*. ^hcteroaralkyl; and D, G, V, K, and W am selected such that there are no O-G, IN-O, S-N, or S-0 bonds.

In certain embodiments, R*\ Rfe, and R7 are each independently selected from Cfe ,$alkyl, Ci.fthydroxyalkyi, Otwalkoxyalkyi, aryl, Cj^araikyl, and R!6DVKOCs^alkyl· wherein at least one OFlG and R' is RitJDVKOCs^alkyh. In preferred embodiments, one of R’’ and R7 is Cj ^aralky! and the other is R^DVROCs-jalky!-, and R6 is independently C^alfcyk In the most preferred embodiment, one of R” and R; is 2-phenyiethyi or phenylmethyiand the other is RS6DVKOCHr or R':>I)Vi< ()(Clfs)Cl'l·, and R6 is isobutvl.

Inpertain embodiments, each, Rbl;S:independently selected from hydrogen, Co <ρ1kylr:1¾,i$alkenyl, .· C.; .§alkynyf: carbocyclyl, betemcyelyl, aryl, heferoaryi,f^aralkyh and Ci,aheteroaralkyf in certain embodiments, each Ri9 Is independently selected from hydrogen, Cj. salkyl, C\ ^alkenyl, Cs,$atkynyf, carbocyclyl, heterocyclyl, aryl, heteroaryl- Q^aralky!, and C* ..fcheteroaralky!. 1« L and Q are absent and R1S is selected font hydrogen, a twite chain of amino acids, C^acyl, a protecting group* aryl, beteroaryi, C*. 6alIceoyl, C^IkynyL Ceearalkyl, and Gpgheteroaralkyk In certain such embodiments,, R1® Is Chalky! and RM is selected from butyl* ally!, propargyl, phenytaetbyl, 2-pyridyi, 3-pyridyl, and 4-pyridyi

In other embodiments, L· is SCkyQdsah^t,' aod'.:R,'l: is seleeted fom Cj.sa!kyl and aryl in certain such embodiments, Ru is selected fom methyl and phenyl. in certain embodiments, L is C™0 and R1-1 is selected from Ci^al'lyl* Cj galkenyl, Gj^alkynyi, aryl* G^aralkyl, bcteroaryb e^ibeteroaralkyl, R^ZA-Gj-galkyl-, R1:!Z~€j. sally!-, fR i20}{R%lPf-0)0~ei,saIMyl-, (R ^OXR^OiPp-Op^salkyhZAZ^saikyl·, (R* ~G)C3l'5O)P|“O|O-0i ,saikyl-B-G s .pikyl -, RiaZA~£ j.ptkyl-Z A2LG s ..*alkyl -, hetero^yciylMZA^^ (R^JiN^Ci-saikyir, fR'4);d^~Ab.aaikyK heteroeyeijdM-,

carbocvdylM--, R5 5 SO>C5 ..salky!~, and RSS$G>NH~. In certain embodiments, L is G“€k Q is absent, and Ri:* is BL

In certain embodiments, Ri0 is G^alkyl, R!i Is Ci-salkyL Q is absent, and Lis C=0. Sn eertairr such embodiments, R* * is ethyl, isopropyl, 2,2,2~tri fluoroethyi, or 2-(methylsul f«nyi}ethyl.

In other embodiments, L is O-O, Q is absent, and RH is Cr-^aralkyl, In certain such embodiments, R!! is selected from 2-pbmylethyk phenylmetbyk (4~ meiboxyphenyOmethyl, (4-chforophenyi )methyi, and {4-fl uoroph enyl )rnet by!. in other embodiments, L is €~0, Q Is absent, Rf0 is Chalky!, and Rn is aryl, in certain such embodiments, Ril is substituted or unsubsli tuted phenyl.

In certain embodiments, L is O-O, Q is absent ot O, and Rn is -(GB2},,carbocycIyI. In certain such embodiments, R5* is eyclopropyl or-cyclohexyl

In certain embodiments. L and A are G^O, Q Is absent, E is O, and Ru is selected from R! “ZA- C, .«alkyl, R!5Z-GS .«alkyl-, R! 2Z A~Ct .«alky! · ZAZ-GS.sa!ky I-, (R!20}(R! '0)P{ -0)0-Ct .«alkyl-Z.AZ.-Cs saikyl··, {R'^OXR'HdlFC-OIO-Gj.salkyl-Z-Ci. «alkyl-, and heteroeyciylΜΖΛΖ -C* .«al kyl -. In certain suciv embodiments. Ri5 is heterocyclylMZAE-Gi-salkyb where hetcrocyclyl Is substituted or unsubstituted oxodioxolenyl or N(lP°){R2i), wherein R20 and R25 together are Cj^alkyl-Y-Chsalkyl, preferably €j oalkyh Y~€5 .mlky), thereby femiing a nog.

In certain preferred embodiments* L Is G“0, Q is absent, and Ru is selected from {R*2ΟKR'3Q)P{-0)0-C;.;<a 1 kyi-, (R ^NCfemlkyL (R’V^iCHj)»-, and heterocydyl-M-. In certain snobembodiments* 'R!- is -C; or -Ch^aikylN ’ {R * *)R4 is C {^alkyt. M certain other such embodiments» R. *1 is heferecyelyllv! -, where h eterocyefy! is selected from morpholino, piperidine, piperazine, and pyrrolidine,

In certain embodiments,:this :C~DS R10 is Chalky!, Q is selected Rent 0 and NH andRu Is selected from Cs^alkyl, cycloalkyi-Mj Cosaraalkyl, and Gj^heteroaraalkyl, id-other embodiments, L is C~0, RH} is Gp^allcyl, 0 i$ selected froth 0 add NH, and Ru is Cosalkyly where€}„6a!kyl Is selected from methyl, ethyl, and isopropyl:, ll«: further embodimOntspL is 0=0, R,ft is C^alkyl, Q is selectedafrdm O and NH and RH is Gp. 5aralkyt, where aralkyl is phenyl methyl. In other embodiments, L is OCX Rl° is Cn &amp;aikyj, Q is selected from Oand NH, and Rsi is C^hetefoaralkyl, where heteroaralkyl is :(4~pyridyii)ittethyl.

In Certain emhodrinentSili is absent dr is C::::D, and RiS arid R1 * togethH' are C>. 6alkyl-Y~C| Aalkyi, Cnftalkyl-EA-Cg&amp;alkyl, orO-^lkyLA* thereby forming a ring. In certain preferred embodiniejits, L is C:::0, Q and Y are absent, and R*° and RIJ together are CMSikyl-Y-Gsoalkyl. In another preferred embodiment, L and Q are absedt, and Rfe and R * together are C^pikyl-Y-Cjoaikyh In another preferred embodiment, L is C~€), Q is absent, Y is selected from NH and N-Cj^aikyt, and Rr> and ili! together are Cp jaiky|-Y~Gt sdikyL In another preferred emhodiment, 1, is C™0, Y Is absent, and R^ and •R *'* -«r«::€ik?hlk^RY»-Cj nalkyik In another prefened embodiment, L and A are €=0, and R!i>and R together are Giaalkyl-ZA-Cj^alkyl. In another preferred embodiment, Land A areC~0 and Riy and R!^ togetlier are Gs^alkyl-A, in certain embodiments, Rki is (R* ?0}(R sfr>}Pfr=0)W- In certain such emhodimfents, 0, V, R, and W are absent. In other such embodiments, V and K. are absent, P is C^gStkyL and W is O, In yet other such embodiments, D is Cj^alkyL R is 0-0, and V and W are O.

In certain embodiments, R’* is R!?GB··. in preferred embodiments, B is 0=0, 0 is 0, D is Chalky!, V is O, and K is C O. in certain mbodimenfs, R'6 ss heieroeyelyl·. In preferred such embodiments, β is Qi^iikyL .In certain such embodiments, V is 0, Kis (>-0, and heteroeyelyi is oxodioxolenyl. In other stmh embodiments, V is absent. K is absent or is C-0, and heteroeyelyi is N(R20}{ll23}9 where R20 and I3' together are)0rd,;l~WB-fs or B~)~Tel, T is absent or is selected from O, NR,#, S, §0,: SQs, OHORt<3, CHCOiR^, 0-0, CP2, add CHF, and l is absent or is CYyalkyi, in certain embodiments, Rifi is (Rs<ihN- or (RSA.-;N% and preferably V is absent, Ibipreierred such embodiments, D is C^alkyl and KBs absent Of C“€i. In certain embodiments where V is absent and R!i> A(R3^N',0 is absent K; is absent obis GAO, preferably K is D-O,

In cortairrembodiments, RIs R^SdjGBG-, In preferred such embodiments, B Is C~0, D, V,and K are absent, and G is NB Or MCupiltyl,

In certain embodiments, R**% Ri7Ci8C· .^ikyk In preferred embodiments, Bits 0-0, G is O. and the CYsaikyi moiety is optionally substituted with OB, C stalky I (optionally substituted with halogen, preferably iinorme}, Gi.8olkylW:, aryl, het eroaryl, carbocyclyl, heterocyelyl, and Cj ^aralkyl. In certain such embodimeets, the Ch#lkyi •moiety is an unsubstitnted, mono-, or disubstituted Cu alkyl

In certain embodiments, the chain of amino acids has a structure offemula (VIII) or (IX) or a pharmaeeutieally acceptable salt thereof

wherein

each Af is independently an aromatic or hetempromatie group optionally substituted with I to 4 substituents; L is absent or is selected from C“0. <!>$, and S0a> preferably S0a orC~0; X is GQ0FI or an actiyuied form thereof, preferably Xis GO0M, C0CI, or CON{Me)(0Me), most preferably X is COOB or COGI; Y is absent or Is selected from 0“Q and S0a; Z is absent or is € stalky!;

Es endK6' are each independently selected from Ch-salky!, C.YfrtydiOxyaikyt,

Ch^alkoxyaikybaryl, and e^araikyi, .any-of which is optionally substituted with one or mol* of amide, amine, carboxylic acid for a Salt thereof), ester {including Chalky! esteiy Ci -salkyi ester, and aryl ester), thiol,.or fhioether substituents; R^ is selected from hydrogen, OH, Ci^aralkyhY», and Cpsalkyl-Y-, preferably hydrogen;

Ru is selected from hydrogen, CM52, Oj^alkenyl, Ar-Υ-, cnrbocydyk seim! heteroeyelyk and

Ri2 is seleeted from hydrogen, O^aikyl, and 0( aralkyl, preferably hydrogen.

In certain embodiment^"L.&amp; selected from 0===0, G:::S, and SOj, preferably 3(¾ dr CM),

In certain embodiments, RUi is selected from hydrogen, OH, Ci^aastlkyl, sod C*. galkyk preferably hydrogen.

In certain embodiments, R* 5 is seleeted: front hydrogen, Cj^alkeuyl, Ar~Y-} carbocyolyl, and heterocyclyl

In certain embodiments, Rs and R* are each independently seleeted from Ci^alkyi, Cs^hydroxyalkyl, and Cj^aralkyl In preferred; such embodiments, Rs is Chalky! and R* is €j.<;araikyL 1« more preferred such embodiments, R’ is isohulyl and E/1 is phenyhnefhyl.

In certain embodiments, RliJ is hydrogen, L taC^O or: S% R! 5 is Ar-Y~, and each Ar Is Independently::seleeted from phenyl, iodolyl, beneofhranyi, :naphthyl, quinolinyl, pulnolonyl, thienyl, pyridyl, pyrazyl, and the like. In certain such embodiments, Ar may be substituted wsth AfrQo where Q Is selected from a direct bond,-Ο-, and Chalky!. In certain other such embodi ments where Z mih^alkyl, Z may he substituted,. preferably with Ar, e,g„ phenyk in:certain embodiments, R!0 is hydrogen, Z is absent, L is· C::::0 or S0j. and R** is seleeted front Ar-Y and heteroeyciyl. In certain preferred suefremhodiments. heferoeydyl is selected from chromonyi, cbrntnanyl, morpholine, and piperidinyl. In certain other preferred such embodiments, Ar is selected from phenyl, indolyl, bemrofimahyi, naphthyl, ^uinolinyh quinolonyl, thienyl, pyridyl, pyraxyl, and the like.

In certain embodiments. Ri0 is hydrogen, L is 0=0 or SOj, 1 is absent, and R! ^ is t$:a substituted vinyl group where the substituent is pret^bly'MxaryJiM'feetmPntyl group* more preferably a phenyl group optionally substituted with one to four substituents, in aatain ^bodirnentSj Ru is selected from hydrogea and Cf .«alkyl. la certain .-11¾ preferred sudi eiabodliaentSv is selected from hydrogen and methyl. In more preferred such embodiments, R‘” is hydrogen.

In certain pretetTed embodiments, the chain of amino acids has a structure Of formula < X)

X is COOli or an activated form thereof preferably X is €{X)H, CDC1, or CONlMeXOMe), most preferably: X is COOK or COCh

Rf R6, and R7 are independently selected frotfeCf^tfey^ eplkoxyalkyh aryl, and if.«aralkyl, each of which is optionally; substituted with a group .selected from amide, amine, carboxylic acid or a pharmaceutically acceptable salt thereof; carboxyl ester, thiol, and: ibioether, preferably R6 is eii ^alkfl and Es and R7 are Gf. «aralkyl, most preferably, R6 is isobutyl, Rvis 2“phenyl.^liyi,iPddB:?:'j&amp; phenyljpethyl; R9 is a further chain of am ino acids, hydrogen, Gi^acyl , a protecting group, aryl, or heteroaryL where substituents include halogen, carbonyl, nifro, hydroxy, aryl, and €*, jslkyl, preferably R9 is G«.«acy|; most preferably R*' is acetyl

In certain preferred embodiments* the chain of amino acids has a structure of formula (XI) or a pharmaceuticall v aceeptabie salt thereof, wherein

L is absent ar is selected from or ~CC S}0; X is COOH or an activated form thereof preferably X is COOH, COCi, or CONCMelfOMe), most preferably X is COOH or COQ; Y is NB, N-alkyl, O, or 0(10¾ preferably M-alkyl, Θ, or €{10>2; Z is O or 0(1¾¾. preferably 0(8¾; R\ R\ and are imiepewlently selected from hydrogen and a group of formula (XII), preferably, 10, R2, and R' are all the same, more preferably R\ R", and R3 are all hydrogen;

each Rs, R&amp;, Rfr and R^is independently selected from hydrogesg C^alkyl, Op diydroxyalkyl, C, .,ydkoxyalfcyL aryl., and C*^aralkyl, each of wbieb; is optionally: sebstitMted with a group,selected from alkyl,,amide, amine, carboxylic acid or a: phannaeeytically: acceptable salt thereof! earboxyl ester, thiol, and fhioetber, pfeferably R5, R6, and R7 are independently selected from C^salkyl, C^ftbydfosyalkyi, and Co «aralkyl and each.R9 is hydrogen, more preferably,: R* is Cf^ikyl, R5 and R? are independently C|^aralkyl and each R9 is H;

Rs0 and Rn are independently selected from, hydrogen: and Chalky!, or R- and Rh together :fbrm a 3·- to frmembered carbocydic or heterocyclic ring; R-2 and Ru are tbdepepdently selected from hydrogen, a metal cation, CMslhyl, and Ci^aralkyl, or R1” and Ri3 together represent Q.«a|kyt:! thereby fermiog a ring; in is an integer from t) to 2: and n is m integer from 0 to 2, preferably 0: or I. in certain::embodimentS,IX:dS::® and R\,;Ra,: and R* are all: the: same, preferably R*, Rl and Rl are all hydrogen, in certain such embodiments, R\ R&amp;, and R’ are independently selected Rom C^alkyl, Ch.fr ydroxyaikyl and Q ^aralkyl, more preferably, Rft is Chalky! and Rs and R' are independently iSs^aralkyi,

in certain preferred embodiments, Rs, Rt, and RJ are all hydrogen, R° and R8 are both isobntyl,: R5 is phenyiethyl, and R; is phiJiylmethyL

In certain embodiments, .:1¾ .R**, and R.f ere independently selected Rom. hydrogen, C\ ^alkyl, <fr ^hydroxyalkyi, Cr,fial koxyal ky I,: aryl, and C t .^aralkyl, each of which is:: optionally substituted with a groop selected from:.alkyl, amide, amine, carboxylic add: dr a pimnnaceniicaily aceepiable isaR thereof, carboxyl ester, thiol,land thioether, in certain embodiments, alleast one of R^ and R' ;s (Ife^aralkyl substitnfed with alkyl, more preferably substituted with perhaloaikyl. In certain: such embodiments, R J is Cj^aralkyl substituted with tri0uoromcihyl.

In certain ernbodiniehts,: Yiis'Selected: Romihi^dlkyl, O, and (¾. In certain such embodiments, 2 IpCHj, anti m and n are both 0- in certain alternative such embodiments, 2 is CHj, m is 0, and n is I or f. In yet another alternative such embodiments, 2 is O, m IS : i,: and π is 2,

In certain preferred embodiments, the chain of amino acids has a structure of formula (XIII)

each Ar is independently an aromatic or heteroaromatic group optionally substituted with 1 to 4 substituents, each Λ is independently selected Rom OO, £=¾ and SQh, preferably GXX or A is optionally a covalent bond when adjacent to an oceunmme ofZ; B is absent or is N{R9)It5i>s. .preferably absent;: B: is absent or is selected' horn (X), C::::S:y and Sd*, preferably S€>> or OO; M is absent or is €,. ^askyh preferably C$.g alkyl; Q is absent or is selected from Ο, NH, andiM-C^alkyl; X is CDOH or an activated form thereof, preferably X is CQ0H, C0C1. or GQMCMejfOMe), most preferably X is COOH or COCi; Y is absent or is selected from OO and SCb; each Z is independently selected from Q,% NH, and N^^kyl, preferably O; or Z is optionally a covalent bond when adjacent to an occurrence of A;

Rr is selected fromlf, *C j .ralkybR,:C; ydroxyalkyl, G| .ralkoxyalkyl, aryl, and €|^afaikyl;:: R2 i s selected from aryl, C [ ^aralkyL hetetoaryl, and Ge«beteroaralkyi; R<! is N(R5)L-Q-R6;:; R:<> is selected from hydrogen, OH, C^aaralkyl, arid GMalkyk preferably hydrogen;

Ris Is selected fiOffi hydrogen, Chalky!, Gj^alkenyi, C^aikynyi, Xr-Y~, earhoeyelyl, heterocyellyk an NXermihal protecting gronp, aryl, Oossralkyl, hetcroaryl, Oi^eteroaraikyi, RnZAZ-C,.«alkyl·, ft|4Z-C,.8alkyh i R M0)(RS''0}P(-:0){>C,..saikyl-ZXZ-Cj.salkyi~,Ri^XZ-0i.«aikyt~ZAZ-G5-salkyi~, heterocyclylMZAZ-Ci^alkyl-, {Rrs0)CRsa0)FC^0^8#kyi-? (R0)2N-€M2alkyS-, (RB)3NGCMaslkyfe hoteroeye!yiM--vc8rbooyeIylM-¥ E^SOjCraaikyl--, and R!%0>NM; preferably an bi-capping group, nroro pfefersijly t-butoxycarbonyl or benxyloxycafeonyl’ or R^ and R6 fegefber are Gi,6alkyl~Y-Cv.salkyl5 Ci^alkykZAZ-Ci^aikyi, ZAZ~Ci-(salkyi-ZAZ-Cj.galkyi, ZAZ-Cj^alkyl-ZAZ, or Cj^alkyl-A, thereby fenning a ring; R7 is selected from hydrogen,. Chalky!, and Cj^aralkyi, preferably hydrogen; R:^ is selected from hydrogen, OB, and <31^alkyl, preferably Ct,*aiky 1; and

Ri0 is an N-tenninai protecting group; R** aiid R!i are independently selected from hydrogen, metal cation, Cj .^alkyl, Co «alkenyl, Ci ^alkynyl, aryl, heieroaryl, Ci &amp;araikyl, and Ci^beteroaralkyl, preferably from hydrogen, Metal cation, and Cppiky!, or R H and Ru together ate Cnplkyl* thereby fomting a ring; each Ri3 is independently selected horn hydrogen and Chalky!, preferably Cn pfkyl; and

Ri4 is independently seleetedtfom hydrogen. Ci^alkyl, Cj-aeikeoyb Cj-salkynyl, carfeoeyolyb heterocyelyl, aryl, hetsraaryl, C^aralkyl, and Cosheteroaralfeyl; pro vided that in any occurrence of the sequence ZAZ, at least one member of the sequence must be other than a covalent bond.

In certain embbdimentsv R5 is selected frbm~-0t.pfkybB and Cj^aralkyk in certain such embodiments, II! is substituted with one or more substituents selected from hydroxy* halogen, amide, amine,: carboxylic acid (or a: salt thmeofR ester (including C|„ §aikyl ester, C> ..jalkyi ester, and aryl ester),d!eol:, or thioether. In certain preferred such emhodiments, R is; substituted with: one or more substituents selected Ifonit carboxylic acid and ester. In certain embodiments, Rs is selected if dm methyl* ethyl,; isopropyl, baffooxymethyl* and benzyl, In certain phtbbdim^tklR;V:ia-Cnftal:ky|:*'B and C ^aralkyl. In certain prefened sneh embodiments, B is absent,

In certain embodiments, R2 is seiecied from Ci^amlkyl and Cj^heteroaralkyl. in certain such embodiments* R3 is selected Item .Gr^Ik|j-pteftyl>: Clidal kyWndoiyl* Cm $alkyl"thienyl, CusalkylrtluazolyS, and €i plkyl-isolhiaxolyl, wherein the alkyl moiety may contain six, fi ve, four, three* two, or one carbon atoms, preferably one or two, In certain such embodiments, Rz is substituted with one or more substituents selected from hydroxy, halogen, amide, amine, carboxylic add (or a salt thereof), ester ^including C\. plky! ester, Ci-salkyl esteh and aryl ester), thiol, or thioether. In certain such embodiments, R";is substituted with a substituent selected from alkyl, frihaioalkyl, alkoxy, hydroxy* or eyano. In certain such embodiments* E" is selected Rom Cfeplkyb; phenyl and Cpplkyl-mdolyl, In certain preferred sueh emhodimeutSi R2 is selected from

R ~ H or any suitable pr otect teg group wherein DIs. selected from Ji OM^.^B'UViDHV^'KCFj and CFb, In certain embodiments 0 is selected fern H, O^e, 0¾ ChkCFj and 0¾..

In certain preferred such embodiments where 0 is attached to a sixonembefed ring, B is attached at the 4~positioo relative to the point of attachment» preferably excluding embodiments where the 4-posifion· of the ring is occupied by the nitrogen of a pyridine ring.

In certain embodiments, R3 is hydrogen, L is O::0 or SQ-j, Rb is Ar~¥~, md each :A.r is independently selectedfrom phenyl, indelyl, benzofnranyl, naphthyl, qninoMnyl, quinolonyl, thienyl, pyridyl, pyraay!, and the like. In certain such embodiments, Ar may be substituted with ArtE*, where E is selected from a direct bond, -0-, and Chalky!. In certain other such embodiments where 0 is C;»salkyi, Q may be substituted, preferably with Ar, e.g., phenyl.

In certain eanbodimentSj R5 is hydrogen, Qis absent, L is €-0 or SOb, and is selected from Aw¥ and heterpcyciyl, In certain pretorred such embodiments, hetcrocyclyl is; selected from ehromonyl, cbromanyl, morpholine, and piperidinyl, In certain other preferred such embodiments, Ar is selected from phenyl, indolyk benzofuranyj, naphthyl, qomolistyL qumolonyl, thienyl, pyridyl, pyraayl, and the like.

In certain embodiments, R5 is hydrogen, L is C::::0 or SO2, Q is absent, and R6 is Ci^aikenvl, where 0r.£aikenyl is a substituted vinyl group where the substituent is preferably an ary! or betemaryl group, more preferably a phenyl group optionally substituted with one to four substituents.

In certain ernlmdimentSjX arai § are absent and E* is selected from Chalky!, Ci^aikeny!, Cnsaikynyi, Ci^aralkyl, and Cnshefemaralkyl. in certain such embodiments, Rs is O^Iky! and is selected from butyl, aliyi, propargyi, phenylmethyl, 2-pyridyl. 3-pyridyl, and 4~pyridyl. i«•ofli^r'etn'bodinidntSi.'iL.is.§0&amp;;·'$is absent, and E^ is selected from Chalky! and aryl, in certain sncb embodiments, R6 is selected from methyl and phenyl

In certain embodiments, L is 0=0 and R* is selected from Oi^alky1, 05.^alkenyl, Cs,saikynyh aryl,·Co4«ikyi teteroaryl, C|.i;beteroaraIkyl.s En:2A-Cs^aikyl-5 Ri4Z-(R,suikyK (RαΟ)(Η:\)}Ρ(-0)Ο·(:'ί.κ3^ν!.., (R5O)iRnO>Plrt>;<>C, ndkyi-ZAZ-€j..§aikyl~, (RM0<R,2O)P(-O)a<^#^^ heiemcyclylMZAZ-Ci^alkyb, Cl^lp^C^salkybaft'^sH- carbocyc3yiy -?: Ri4S02€}..8aikyb, and Il^SQjldB-, wherein each occurrence of Z add A is independently ctlver than a covalent: frond. In certain:embodiments. L is CX), Q is absent, and Rf' is H. in certain embodiments^ R® isOj^alkyl, E4 is GrAalkyb Q is absent, and L is 0=0. In certain such embodiments, R6 is ethyl. isopropyl, 2,2;2driftuoroethyl, or 2-:(met:hylsu!tbny!}etbyi.

In otter embodinients, L is 0=0, Q is absent, and Rb is Cj^aralkyl In certain such embodiments, R° is selected lorn 2-phenyIethyi, phenylmetfryi, (A -methoxypheny!}methyi, {4-chk>rophenyl)rnethyl, and (4-tluorophenyl)methyl.

In other embodiments, I. is OQ, Q is absent, R5 is Ct ^alkyL and R’s is aryl. Id certain such embodiments, R‘' is substituted or nnsnbstitnted phenyl.

In certain emhodiments, R is; C>0, Q is absent, and Rtl is selected from heteroaryl and Cs sheferoaraikyL In certain such embodiments, R6 is heteroaryl selected trotn pyrrole, fwan, thiophene, imidazole, isox azole, ox azole, Oxadi azole, thiazole, thiadiazole, triazole, pyrazole, pyridine, pyrazipe, pyridazine and pyrimidine. In certain alternative such embodiments, R'1 is Ct .«.heteroas alkyl selected from pyrrolylmethyl, ffrranylmetfryI. thicuyltneibyh imidazolylmethy!, isoxazolylmethyl, oxazolylmeibyi, ox;adiazol ylmethyl, thiazolylmetbyl, ihiadiazolylsnethyl, tnazolylmethyh/pyrazolylnteihyl, pyridyinm:thyl,; pyrazinyhnethyhpyrsdazihy and pyri m id inyl met h y I, in certain embodiments, L is €=0, Q is absent or O, and R6 is earbocyciyIM', wherein M is C<m alkyl. In certain such embodiments, R6 is cyclopropyl or cyelohexyl.

In certain embodiments, L and A are OO, Q is absent, X is Ο, M is €s .«alkyl, preferably methylene, and K6 is selected from R‘ ’ZA-Ci^k^.'E^^CngalkyK.;^ {^A-Ci^alfeyl-^ZrCi^aiksd'S.CR^^K^^PC-OJO-Ci^alk^E^^Malfcyi^ (Ru0)(R520}l>{:::0}0~(l's .saikybZ*Ci..55alkyl··, and heterocydylMZAZ-C i.^alkyl-, wherein each occurrence of A is independently other than a covalent bond, in certain such embodiments, R6 ishemmcyclylMZAZ-C^alkyi- wbere hetemcyclyl is substitnted or unsubstituted oxodioxolenyl or N{Ru>)(Ri?}, wherein ll!6 and Ri? together are C u,a Iky L ¥-€;.^alkyk preferably^Gi^aikyl-Y-C^jaikyi, therdsy fermiog a ring.

In certain absent, M is Gi^alkyh and Rfc is selected from {RH0iCRl20iP(-0)0-€i,ga!kybi iR'^Ci.saikyi, iR^sN^alkyK and beterocycIyl-M-. hi certain such embodiments, R6 is fRi3)gN€r,8aikyl or (R^'N^Cj. «alkyl-, where R*3 is Gr^alkyi in certain other such embodiments, R^ is heterocyclylM-, where betefocyciy! is selected from morpho 1 ino, piperidine, piperazine, and pyrrolidino.

In certain embodiments, L is OO, R5 is C|.ealkyls Q is selected from 0 and NH and R6 is selected from :C i ..«alkyl cycloatkybM, Cppraikyl, and €ΰ ^beteroaralkyl. in other embodiments, Lis 0=0, R* is Cis^alkyL Q is selected from O and Nil, and R6 is 0¾. «alkyl, where Oydkyl is selected from methyl, ethyl, arid isopropyl In further embodiments, L is C-D;, 1G is Gnplkyl, Q is selected from O and Mil and Rf> isCv. ^aralkyl, where aralkyl is: phenylmeihyL in other embodiments, L is €-0, R5 isGpgalkyl Q is selected from 0 and NIL and R6 is Ci.&amp;heteroaralkyl, where hetcroaralkyl is (4-pyridyfrmethyl.

In certain embodiments, L is absent or is €=0, and R* and R6 together are €]., .ftalkylO^-Cusaikyh Gr.saikyl'ZA-Cj.salkyl^ Or C^alfcyl-Ά, wherein each occorrenee of Z and A is independently other than a covalent bond, t hereby forming a ri ng. In certain prelened embed knentsi L is €=0, Q and Y are absent, and R5 and R^ together are Cfr jalkyi-Y-Cpjaifeyi, In another prefen-ed: embodiment, L and Q are absehl and R^ and R6 together are CiaalkyiLY-Ci .«alkyl, In another preferred embodiment, l is OD, Q is absent, Y Is selected front NH and N-Gi^alkyl, and R3 and R^ together are G^yalfeyl-Y-C*.«alkyl lb another preferred embodiment.,ill.: is €=G, Y is absent, and R3 and R6 together are C|,3Mkyl-¥“Qi 33lkyL In another preferred embodiment, L and A are € O, and R* and Εδ together are G^lk^fiSA-Ci.jalkyi. In another preferred embodiment, L· and A are C^O and R5 ami R6 together are C-j^alkyl-A.

In certain embodiments, E7 is selected front hydrogen and Oi^alkyl In certain preferred such embodiments, E7 is selected from hydregenand methyl, in mote preferred shell embodiments, R? is hydrogen.

In certain embodiments, R2 and R3 are cadi independently Gi,saralkyl, and R1 is selected from Clpgalkyl, C^fihydroxyaikyl, Ci ^alknxyalkyl, aryl, and Cr-praikyl, any of which is optionally substituted with one or mote of amide, amine, carboxylic acid (or a; salt thereof), esterfincluding C^alkyf ester, C^alkyl ester, and aryl ester), thiol, or ihioeiher substituents.

In certain: preferred enrbodhnenlsdtts chain of amino acids has a structure of formula (XI¥)

each Ar is independently an aromatic or heteroaromatic group optionally substituted with ! to 4 substituents;. each A ts independently selected from O O, D::S, and SO>, preferably C::::0; or A is optionally a covalent bond when adpeent to urr occurrence of X; L is absent or is selected fix>m C:::0, C^S, and SLA, preferably SOy or O~0; ;M Is absent or is Ci.ijaikyl, preferably G-^aikyl; Q is absent or is selected from Ο, MB, and N-Ci^alkyl; X is COOH or an activated form thereof) preferably X is COOH, COC1, or GOM(Me)(GMe|, moM preferably X is COOM or COGt: Y is absent or Is selected from CfeQ and SCL: each Z is independently selected from O, S. RH, and N-Cf.f,alkyl, preferably O; or % is optionally a covalent bond when adjbpmi to an occurrence of A; R2 is'selected, from aryl, C^araikyL heteK>aryI,^,0{^heioroa^|kyU; R4 is N(RS)L"Q-R6;

Rs1sMec^:ft^R,^progen, OH, Q^aralkyl, and Chalky!, preferably hydrogen; R6 is selected fern hydrogen, C}..6a!kyh 6j.ea3kenyl, Cs.6alkyny1, Ar-Y-S catfeocyelyl, heterocyelyl, an ^-terminal proteetiog group, aryl, Cr^alkyl, heteroaryl, ZAZ-Cfealkyl·, .R^Mi^alkyl-ZAZ-Cualkyi-, hrgeroeyclyMZMXh .*alkyK (R^XR^OlP^OX^alky)-, (Ru hN-Cf.t2alkyk (R’^bRC, .i;?alkyS··, R^SOjCi^alkyl», and R^SOaNIfc prefer#ly an.N-eapping ifenp, more preferably Pbutoxycarbonyl or btmyloxyearhnny!; or R1 and R'1 fogether are C^.6aikyl-Y~Ci.salkyl, C^6alkyl-ZA2~C>o6alkyl, 2A2-€p galkyREAE-Qi^alkyi, 2AS»C|^aikyl-ZAZ, or €| xsalkyl - A, thereby forming a ring; .R*ifc.^ected-feto.&amp;yifegeni, OB, and Ci,6a!kyk preferably Ci^alkyl; and R*is is an NAetrnina! protecting group;

Rl i and Ru are independently selected fern hydrogen, metal cation, Chalky!, On alkenyl, C.i^analfcyl, and 0|.glteterdara|kyl, preferably from liydmgen,: metal cation, and Cj^alky I, or Rf and R*2 together are th-salkyl, thereby: fermiPg a ring; each R° is independently selected from hydrogen and Cualkyl, preferably Co calky!; and

Ri4 is independently selected from hydrogen, C^alkyl. Cs^alkenyl, Oi^alkynyl, earbocyciyl, heterocycSyl, aryi,: heteroaryl, OtAaralkyl, andCi^eteroaralkyl; selected from hydrogen, Cfealkyl, CYghydroxyalkyl, Cs Aalkoxy, -C{0)OC(.<,alkyl, YhX))NH€s.(,aikyi; and Cj.^aralkyl, preferably Cj^alkyl and C$. fj h yd ro x y a 1 k y 1, m o re preferably methyl, ethyl, hydroxymethyl, and 2 - hvdroxyethy 1; provided that in any occurrence of the sequence ZAZ, at least one member of the sequence most be other than: a covalent bond.

In Gpssiralkyl and GswheteroaralkyL In certain such enibodiments, R2 is selected ^ιη.Οι^Ι^ί-ρΗ^Ι,.όΐΛ^ιΙ^-'ΐβίίοΙγΙ, C|.· j^'k^^pNiy^'C{^a!kyl4hlax#yi, and Civgalkyl^sotlnazoiyi, wherein the alky! moiety may contain: six, five, four, three»:*wo, nr one carbon atoms, preferably one or two. In certain such embodiments,. R* is substituted with one of more substituents selected from hydroxy, halogen, amide, amine, carboxylic add (or a saltlhereaf}, ester (including (¾. <j«alfeyl ester, Chalky! ester, and aryi ester}, thiol, orithioether. In certain such embodiments, R2 is substituted with a substituent selected from alkyl, trihaloalkyl, alkoxy, hydroxy, or eyano. Ipeeriain such embodiments, R3is selected from C<.,,alkyl-phenyl anti Ci.^alkyl~indolyl. In certain preferred such embodiments, R2 is selected irom

R Ώ H or any statable protecting group wbereid D is selected from H, OMe, OBu‘, did, GM, CFj and GHy In certain embodiments P is selected from H, OMe, OH, ON, Cld and Cfk in certain preferred such embodiments where D is attached to a six-membered ring, D t$ attached at the 4-position relative to fhe point of aitaehment, preferably excluding emhodimenis where the d-posihon of the ring is occupied by the nitrogen of a pyridine ring. in certain embodiments. is hydrogen, L is Θ-Θ or SO>,s R6 is Ατ-Υ-, and each Ar is independently selected from phenyl nidolyl, henxofuranyl, naphthyl, qumolmyl, quinolonyl, thienyl, pyridyl, pyrazyl, and the like. In certain such embodiments, Ar may be substitutea with:AmE~s where E is selected frqra a direct bond, -0% and Chalky!, In certain other such embodiments where Q is Ci^alkyl, Q may be substituted, preferably With Ar, e.g., phenyl.

In certain embodiments, R5 is hydrogen, Q iGabsent, L i s 0*0 or Mb* and R* is selected from Ar-Y and heierocyelyl. In certain preferred such embodiments, heierpeyefyi is selected jfefn chromonyh chromanyl, morpholine, and piperidinyi. In certain other preferred snch embodiments, At is selected from phenyl, indolyl, hit^ofnranyi, naphthyl, gtnnolinyi, qumolonyl, tMenyl, pyridyi, pyraxyi, and the like.

In certain embodiment», R" is hydrogen, L is 0=0 or SOa, is absent, and R6 is C^alkeuyl, where Cueallenyl isa;sdlN5iSd^-'yi;pyl group where the substituent is preferably an aryl or heteroaryl group, more preferably a phenyl group optionally substituted with one to four substituents,

In certain embodiments, R and Qiamubsen? and R6 is:seleetedifrci:m: Oj^aikyR Cj^aikenyl, Oi^alkynyl, Cb^aralkyl, and Cs^heteroaralkyL In certain such;embodiments, Rs is Ci^alkyi and R^ is selected from butyl, allyl, propargyl, phenyimethyi, :2*-pyridyI, 3-pyridyl, and dqjprldyL !n other enihpdimeniSi, L is SCn, Q is absent, and Rfj is selected from G{,#lkyland aryl, in certain such embodiments, R6 is selected from methyl: and phenyl.

In certain embodiments, L is C™0 and R-is selected from Cr^IRylj IGj^alkenyl, Gi^alkynyl, aryl, Gt^aralkyl, heteroaryl. Ci^hetemamlky!, R^EA-Cugalkyl··, R'H:2“C:i^alkyl-, (R5 !0)i RrO)P{:;:0)Q-C,,8aIkyK (Rs s0)(R ,3Ο)Ρ(Ο)0-€<.3alky!~ZAZ-GusalkyR, {R!i0)(Ri^)Fi-0)0CusalkyR^G14alkyh> RHM^aalkyI*ZAZ*Gp#%R, heterocycl y !MZA 2-G 5 ^alky 1 -, (R13 h N-Cj galfcyl ~s {R i3h.N*-Gj ,galk yl-, heterocydiytM", carboeydylM-, R M$0aCh -salkyR, and Rf %0?NH-, wherein each occurrence of 2 and A is independently other than a covalent bond, in certain embodiments, L is CA0, Q is absent, and R* is H.

In certain emhpdimeattSi Rs is C|.^alkyls Rs is Gnsafkyl, 0 is absent, and L is C”0. In certain such embodiments, R^ is ethyl, Isopopyl, 2,f,2-trifiuoroethyk or 2-(methylsul ionyi)ethy!.

In other embodiments, L isC-O,0 is absent, and R^5 is Ci^araikyh hr certain such embodiments, R6 is selected from 2-pbeny!ethyi, phenylmethyL (4-methoxypheny!)methyl, (4 chIorophenyl)tnethyl, and {4d)uofophenyl)methyL ίπ other embodiments, L is 0=0, Q is absent, R5 is Cs^alkyt, and R^ is aryl. In certain such embodiments, R6 is substituted Of ««substituted |dienyi, irs certain embodiments. L is €==©, Q is absent, and R6 is selected from hetercaryl and Cj^heiemaralkyl. In certain such embodiments, R’! is heteroaryi selected from pyrrole,, furan, thiophene,, imidazole, isoxazole, oxazole, oxadiazole, thtazole, ihiadiazoie, MaZole, pyratmle, gyfrdihe, pyrazine, pyridazme and pyrimidine. In certain alternative such embodiments, R6 is Gi^eteroaralkyl selected from pyrmiyimetbyl, feanylmethyl, thienylmethyl, i.mida20iyi*i^iy|, oxazoly!methyl, oxadiazolyiraethyl,

thiazdiyimetby!, tMadiazoiyimetbyl, tdazolylmefhyl, pyrazolylmethyl, pyridylmetkyl, pyfazinylmethyl pyridazinylmethy! and pyrimidinylmeikyL

In certain embodiments, LIsCK), Q is absent or Q,: and R4 is carbocyclylM-, wherein M is Co-folkyL In certain such embodiments, Rs is:cyefOprppyl or eyeldlfekyh

In certain;embodiments, L: and A are €-0, Q is absent, :Z is0, M is Cusalkyl', preferably methyiene,:and R6 is selected from R!!ZA~C5.¾¾Iky]-, RMZ~Cs.¾¾Ikyl*. R5}ZA-C^8alkyl-ZAZ«:Cpsaikyl>, (R;l A}}{E520}Pfr<})C><;,4aikyi~2AZ-e,,:8aIkyl·, (ItH0}{RliO)F(=€))©-Cr.:galkyI-Z-C^.8alkyl->: and heteroeyclyllylZAZ-Cusalkyfo wherein, each Occurrence of A is independently other than a covalent bond. In certain such embodiments, is IfetemeyclyiyZAZ-Coselkyl- where heterocyelyl is substituted or unsubstituted oxodiokolenyi or Ν{1!ίί}(11! % wherein Riis and Rn together arc Cs^alkyh y-C j.6alkyi, preferably Cioalkyi- Y-Croalkyl, thereby forming a ring.

In certain preferred embodiments, I, is OO, Q is absent, M is Cs^alkyl, and R's is selected ‘0)^^ {E0)3NCfosa!kyis {;RB}3N%|,sdlkyi-5 and heieroeyclyfrlvfe In eedain such embodiments, R&amp; is {R*5)2NCi4'?%l or galkyb, where R53 isjGt^aikyL: In certain other such embodiments*.·^ is heiefoeyelylM-> where heterocyelyl Is selectedij-pm morpholine, piperidine, plperazino, and pyrrolidine.

In certain embodiments:; b is: 0:==0,, R^iis Ci^alkyL Q is seiepted fromjO· and MB and R6Is selected from Ci^alkyl, eyeloatkyfrlyl, Gi^aralkyi and; C:j,feheteroara!kyl. In other embodiments, L is Cfo0, R* ;is,€?4ai,kyl, 0 is selected from: © and NMS and R* is 0¾.. ^aikyl, wbereC^6alkyl is selected from methyl, ethyl, and isopropyl. In further embodiments. L is C-0, R5 is Gf^alkyl, 0 is selected from 0 and NI I and R* is Cfo i,aralkyl, where aralkyl is phenylmethyl. In other embodiments, L ss C:::0, lG is Chalky!, ΐγΐη selected from O andMH, sod φ i$ Cifrieteroaralkyl, where heteroaralkyl is (4-pyridyl}methyL

In certain embodiments* L is ahsenborls C=Q. add R" and R6 together are€;. «aikybY-Cs-galkyl, Ci^alkylbEAXYsalkyl, dr Cpsaikyl-A, wherein each occurrence of Z and A is independently other than a covalent bond, thereby idrmjng: a ring. In certain preferred embodiments, L is O-Q, f| andY are absent,'dad Ik5 and R6 together ars Ci. iaikyl-Y-Cf^alkyl In another preferred embodiment, L and Q are absent, and Rs and R* together are ^^IkylYrCg^IkyL In another preferred embodiment L is €=0, Q is absent, Y is selectedfroth MM '.addfr~Cs.ga|kyi:> and R5 and R* together are CojalkyRY-Ci-jaikyL In another preferred embodiment, L is:€=0, Y Is absent, and R3 and 1R* together are Ciaalkyl-Y-Oi^aikyL In another preferred embodiment, L and Aiare €=0^, and R3 and It6 together are C^alkyl-ZAfr^alkyl In another preferred embodiment, L and A are C-O and R3 and R<J together are €;gy$fciyI~A·.

In certain embodiments, R2 is Cuiaralkyfrand R5 is selected from €>.«alkyl, €,. ghydroxyalkyl, €rt:;a!koxy alkyl, aryl, and Cs.«aralkyl, any Of which is optionally substituted with one or more of atnide, amine, carboxylic acid (or a salt thereof), ester (including Chalky! esteryCj.«alkyl ester, and aryl ester), thiol, or thioether suhstitnems.

In certain preferred embodiments, the chain of amino acids has a structure of formula (XV) R1 Rs q

JL

N Y Y X R4 0 R2 (XV) wherein: L is selected frorn C=0, C~S, and 30¾ preferably C=0; X is COOH or an activated form thereof, preferably K is COOH, COO, or C©M(Me)(Oye)f most preferably X is COOH or COCI; Z is ..absent* C^alkyl, €yx,alkoxyS:Or M E, e.g., absent, Og&amp;aIkyI> or Cs^alkoxy, preferably absent; R is selected from H and €i.«alkyl, preferably H or €H<;

Rs and R2 are each independently selected from hydrogen, C 3 ^salkyl,

Gj^alkynyl, Cr-shydroxyaikyl, Cj^alkoxyalkyl, aryl, dj^araikyi, heieroaryl, freierocyciyl, Gi^heterbeycloalkyL C}.«h«derbaralkyl, earboeyclyl, arid Gs.«earhdcydolalkyI; R4 is selected from hydrogen, C*^aralkyl, and Gosalkyi; iG is heteroaryl; and

Ri! is selected fre-m hydrogen, Chalky!,, and Ct^alkyL M certaot embodiments. R! and R'; are independently selected from hydrogen. €;. ^alkyl. G^hydroxyalky!, Ci.^Ikoxyalkyi, C^&amp;aralkyk Civsheiemeyetoaikp, Cj.. ^kderoaraikyk and Gr.searboeyein1aikyk In certain emlmdimenis, R5 andIG ate independendy Cj^alkyl selected fmm methyl, ethyl, propyl, isopropyl butyl, sec-butyl, and isobutyl. In certain embodiments. R* and R2 are independently C^^ydroxyalkyl. In certain preferred such embodiments, R1 ami R2 are independently seleeted from hydroxymethyl and hydraxyethyl, preferably hydroxymethyl- In certain embodiments, R' and R“ are independently Cj.mlkoxyalkyl. In certain such embodiments, ft* and R" are independently selected from methoxymethyl and methoxyethyl preferably methcxymethyl. In certain embodiments, lG and R" are independently C^dteieroaralkyl. In certain such embodiments,: R ‘ and R': are Igdependehtly seleeted from imidazolylmetiiyl, pyrazolylmetbyl, and tbiaaolylmethyb iand pyridylmethyl, preferably: imidazoi~4-ySmetbyI, tbiaxob4~yImethyl, 2'pyridylmethyl:, 3-pyridylmethy3s br4~ pyridyimethyl. In certain embodiments, R! and R2 are independently Cs.6aralkyi, In certain such embodiments, R* and R^ are are independently selected from phenyimethyl (behxyl) and phenylethyl preferably .phenylmethyl. In certain embodiments, iG and R2 are independently Ci.6carboeye!oalkyi In certain such embodiments Rs is eyciofrexyhneftiyL· In certain emhodiments Rs and Ryare dl Rerent. In certain embodiments, R* and R" are thesame. hi eertain embodiments, at least one of R* and R2 is selected from Go xhydroxyalkyl and Ch-galkoxyalkyL hr certain such embodiments, at least one of R* and Rf is alkoxyalkyl. in certain such embodiments, at least one of R! and R' is selected from meihoxyrnethyl and metboxyethyl.

In certam embodiments, R4 and |Gare independently selected from hydrogen and methyl, preferably hydrogen, 1« certain efebodimtirtts, R* is a 5- or 6~«>embered:heteroaryl, In certain such embodiments, Ra is selected from isoxazole, isothiazole, foran, thiophene, pxazole, thia^olej pytasiolej or: imidazole, preferably isoxazole, ihra% or thiazole.

In certain brnhodimetris, R5 is a bicydie heteroaryL In certain such embodiments fefeyclic lieteroaryi is selected from besizisoxazoie, benzoxazole, benzoihiazole, benxisothi azole.

In certain embodiments, L is C:::€>, Z is absent, and R* is a 1 ,34hiazoi»S~yl or 1,3» thiaxol-4-yl. In certain such emhndiments,%heft the tliiaxole is substituted, it is substituted at least at the 2~po$ifem In other embodiments, R^ is an unsuhstituted l,3-tMazoi~5»yl or l,3~thiazo!-4~yL

In certain embodiments, L is O0, 2 id abserh, pid R' is a substituted i,3»ihiaxol· 5-yL In certain such embodiments, R* is 1,3»ihiazol~5»yi substituted with a substituent selected from Cs^alkyL Ci^alkoky, Cj^alkoxyaikyl, Cpshydroxyalkyl, carboxylic add, ami nocatboxyiate» C j .gal kyl am moearhoxylate, * .saifcyilsami nocarffexy I ate, G u salkylcatboxylaie, Cpgheieroaratkyi, Cs.^sralkyi, Cn^heteroeycfeaikyl, and Ci. scarhocycloalkyl., In certain preferred such embodiments, Ri is 1 ,3~thiazol-S-yI substituted with a substituent Selected bom methyl. ethyl, isopropyl, and cyciopropylmethyi.

In certain embodiments, L Is CO, Z is absent, and is a substituted l,3»tlnazoI»

4-yl. Iri certain such embodiments, R"’ is l,3 -thiaxoh4»yl substituted with a substituent selected item Ci^lkyl, Cj-salfeoxy, C^alkoxyalkyl, Ci^hydroxyalkyl, carboxyhe acid, aminocathoxylate, Cii-plkylaminocarboxylate, {C^alkyJ^ammocaiboxylate, Cs. ^alkylcarboxylate, Cs„sheteroanj]kyl, Cj^atalkyl, Cj^beterocycloaikyi, and Cs.. ftcarbocydoaikyl, lit certain preferred. such embodiments, Rs is jj-tlbazoM-yi substituted with a substituent selected from methyl, ethyl, isopropyl, and oycfopropyfmeihyL

In certain embodiments, 1.. is C Ο, Z is absent, and R^ ismu isoxazOl-3-yI or Isoxaxol-5-yL hi certain preferred such embodiments, when the isoxazol-3-yl is substituted, it is subsiituted at least at the 3-position, In certain preferred embodiments, when the isoxaxoi-5-yl is substituted, it is substituted at least at the 3-position.

:Ίϊι. cert'ain^eni'fe^dirneiits, L is CO. Z. is absent, an<i R' is an unsubstituted isoxazol-S-yL

In certain embodiments, L is OO, 2 is absent, and R5 is a substituted isoxaxdl-d-yl. In certain sued embodiment^ Rs is !ioxt®ob3-yl substituted With a substituent selected imm Ct^alkyl, Cusalkoxy, Ci,saIkoxyalky]; Cj^hydroxyalkyl, carboxylic acid, aminoearhoxyiate,: C} ^alkyiananocaf hoxyl ate, (Cii ^al kyl)2aau:ttoearbox y I a te, € i, 6aikylearboxylate>: Ci^heteroaralkyl, Ci^aralkylj 'Ci^heterocycloaikyl, and €}. ftCarboeyeloalkyL In certain preferred sued embodiments R* is isoxazdle-3^yl substituted witb a subsiduent selected fom methyl, ethyl, isopropyl, and cyciopro py 1methy i.

In certain embodiments L is ^ is absent, and R* is isoxaxoI -3-yl substituted with a4- to b~mernbered idtrogemeoniaining Ci^gbeteroeycbalkyi In certain such embodiments, R* is isdxaxoioS-yl Substituted with azetidinyi methyl, preferably axetidln-1-ylmethyl. In eertein alternative such wnbodiments, I is C-O, Z is absent, and JR5 is w"’ vy I, .JL % . ...

isoxazob3-yl substituted with ’w*" >-, wherein W is 0, IsR, or CHa, and R is H or Ci-^lkyb In certain such embodiments, W is O.

In certain embodiments, L is OO, Z is absent, and R5 is IsoxaxolOyi substituted with Sunembered ditrogen~c0htaining Cb .<;betett3amikyl, such as pyraxolylpethyl, imidazoi yim eth yl, tel azoU-ylmethy 1, preferab ly 1,2,4~tri axo 1-5- yimethy I.

In certain embodiments, L is OO, 2 is absent, and Ra is isoxazoKVyisubstituted with Cr^altoxy or C^lkdxyalkyl* preferably methoxy, ethoxy, methoxymethy!, or methoxyetbyL

In certain embodiments, L is CO. Z is absent, and R' is isoxazol-3-yl substituted with Gu$hydroxyalkyL preferably hydroxymethyl or hydmxyelhvL

In certain embodiments, L as 2 is absent, and ?0 IsssoxaxoiOyi substituted with:a carboxylic acid. ajmnoearboxylaie, Ci^alkylnminocarboxylate,: fCi. iialkyllsamlnocarboxylate, pr Ci^alkylearboxylatei Jmcertain such embodiments, lO is substituted with metbyj eatboxykte or ethyl earbuxyiaie, preferably methyl carboxylate.

In certain embodiments, L is C O, 7.. is absent, and 10 is an unsubstituted isoxazol-S-yl

In certainembodiments, L is 0*Q>'Z is absent, and Es is a substituted Isoxazdt-S-yl In certain such embodiments, R* is lsoxazoi~5~yl substituted with a substituent selected from Ci,#lkyl, C^alkpxy, Ci^alkoxyafkyl, Cj^bydroxyaik>15 carboxylic acid, aminoearboxylate, C^alkylaminocarboxylate, {Gr4alkyl}3:ammocaiboxyl:ate, Go saikylearboxylafe, Cj,.gheteroatatky1, G^&amp;ralkyl, Gnsheteiocyoioalkylyand '€*„ searbheyeioaikyl In certain preferred such embodiments R3 Is isoxazole~5~yl subsisted with a substituent selected from methyl, ethyl, isopropyl,: and cyeloptopyinrethyk

In certain embodiments L is C-G, £ is absent^. and is isoxazol-S-yi substituted with a 4- to 6~memhered nitrogenmontamiug CixheferocycJoaifcyl. In certain such embodiments, R3 is isoxazoI-S-yl substituted with axetidinylmethyl, prefembly azetidin-i-ylmethyl. In certain alternative such enxbodiniehts, L is Z is absent, and Rs is Γ1 x

isoxazphfwyl: substituted with wherein W is O, NR, or €1¾ andE is M or Chalky!. In certain such einbodjincnts, W is O.

Itt certain embodiments, L Is G=0, Z is absent, and Rs is isoxazol-S-yl substituted with Son'embered nitrogen-containing £; ^hetcroaraikyf, snNi as pyrazolylm ethyl, imidaxolylmethyi, iriazol-:$~ylmethy!s preferably RZNrtriazoNS-ylmethyk

In certaip embodiments, t is CO. Z is absent, and R3 is isoxaxol-5-yl substituted with: Guplkoxy or C^alkpxyaikyl, preferably methoxy, ethoxy, rnethoxymeikyl, or mefhoxyethyl.

Id certain embodiments, L is Ο™Ο, Z is absent, and R" is isoxaxoI-S-y! substituted with Ci^bydroxyaikyk preferably hydroxymethyl or hydroxyethyk

In certain embodiments, L is O0, Z Is absent, and R* Is isoxazol-5-y! substituted with a carboxylic acid, aminpearhoxylate, Ci.pJkyl aminoearboxylate, (Cp §alkyi Jjaminocarhoxylate, or G; ^aikylcarboxylatei In certain such embodiments.,· R3 is substituted with methyl earboxylate or ethyl earboxylate, preferably methyl earboxylate.

In certain embodiments,Z is NR, preferably Nli.

Grderly protein degradmion Is crucial to the maintenance of normal cell InnCtions, and the proteasome is intepal to the protein degradation process. The-proteasonie controls fee levels of proteins feat are important for cell-cycle progression ami apoptosis in normal and malignant cells; for example, :eye!mss caspuses, BCL2 and nFfeB (Kfematod el ah, Free. Natl Acad. Sol. USA (i 990} 87:7071 -7075; AI monel et at. Leukemia (2002) 16: 433 -443). Thus, it is not surprising that ininbitingproteasoTne activity Can translate into therapies tptreat various disease states, such as malighaM, non-mafigriantand autoimmunediseases, dependingon ihecells involved.

Bothfe vitro and in· vim models have shown tlmt malignant cells, in general, are susceptible to proteasotne inhibition.. In fact, protOasome inhibition hails already been validated; asa therapeutic strategy for the treatment of multiple myeloma.. This could be due. in part, to the highly prolifertilye nndignant eelLs dependency on fee proteasome system to rapidly remove proteins (Mlfo etaL, I. Mol Med. (1!|97); 7|:5“Π; Adams, Uature {2004)4: 340-360). 'Therefore, certain cofoudifeents of the1 invention relate to a method of treating: cancers composing administering fo a:subject in need of such treatment an elective amount: of the proteasome inhibitor compound disclosed herein.

As used herein, the temi foaneer” Includes, but is not limited to, blood bom and solid tumors. Cancer refers to diseasuof blood, bone, organs, skin tissue and fee vascular system, including, but not limited to, cancers of the bladder, blood, bone; brain, breast, cervix, chest, colon, endremetrium, liver, luogj lymph nodes, mouth, renal, skin, stomach, testis, throat, and uterus. Sped Be cancers include, but are not limited to, leukemia (aetue lymphocytic leukemia (ALL), acute iyelogehous leukemia {AMI.}, chronic lymphocytic leukemia (CLL), chronic rayelogermas leukmma (CMIfo hairy celt leukemia), mature B cell neoplasms (small lymphocytic lymphoma, B cell profymphocytic leukemia, 1 ymphoplasmueytie Iyofohoma (such as Waldenstrom’s oracroglobuiioemia), splenic marginal zone lymphoma, plasma cell myeloma, plasmacytoma, monoclonal immunoglobulin deposition diseases, heavy chain diseases, exfranodal marginal zone IT cell lymphoma (MAUTJymphoma), nodal marginal zone B eel! lymphoma (NMZL), follicular lymphoma, mantle cell lymphoma,fei%seB cell lymphoma, mediastinal, (thymic) large B cell lymphoma, intravascular;large B cell lymphoma, priniary effusion lymphoma and Burkitt lyraphoma/feukenna), mature T cell and natural killer (NIC) cell neoplasms (T cell pro!ymphoeytie leukemia, T ceil large granular lymphocytic leukemia, aggressive HK cell fookemla, adult T ceil leukemia/lymphoma^extranodal NK/T cell lymphoma, ^t^patby-type Teell. lymphoma, hepatuspienie T cel. lymphoma, bkstie NK. cell lymphoma, mycosis fUPgoides (Seaary syndrome), primary cutaneous anaplastic large eel lymphoma, lymphomatoid papulosis, angioimmunohlastie T cell lymphoma, unspecified peripheral 1' coll lymphoma and anaplastic large cell lymphoma):, Hodgkin lymphoma:'(podukr sclerosis, mixed cdluarity, lymphocyte-rich, lymphocyte depleted or n©t dcpieted, noilplar Iymphoeyte-pmdoniinanlXmyeloma (multiple: myeloma:, indolent Myeloma,: spioldering myeloma), chronic myeloproilieratiyc disease, niydodysplastic^myoloprohiemtive disease, niyelodysplastie syndromes, 3mmunodefie;ency-associated lymphoptoliterative disorders, histiocytic and dendritic cell neoplasms, mastocytosis, chondrosareOma, Ewing sarcoma, fibrosarcoma, malignant giant cell tamer;, myeloma bonedisease, osteosarcoma, breast:cancer (hormone dependent, hormone indepatden% gyneeologiea! cancers (cervical, endometrial,:ihilopian tube, gestational trophemiastic disease^ ovarian, peritonea!, uterine, vaginal and vulvar),, basal cell carcinoma (BCC), squamous cell carcinoma (SCC), malignant melanoma, dermatofibroSarcOma protuberahs,: Merkel ceil carcinoma, Kaphssfis sareomaj astrocytoma, pilocytic astrocytoma, dysembryoplastie neuroepithelial tumor, oligodendrogliomas, ependymoma, gliofelastoma .multiforme, mixed gliomas, oiigoastrdeyiomas, msdulifdslastorha, ridinohlsstoma, heurpfelastoma, gemnndma, teratoma, mali^aut mesothelioma (peritoneal mesothelioma, pericardial mesothelioma, pleural mesothelioma), gasito-Ontero*pancrea tic or gastmenteropmicreaiic neuroendocrine tumor (GEP-NET), carcinoid, pancreatic endocrine tumerfPET), colorectal adenocarcinoma, colorectal carcinoma, aggressive neuroendocrine tumor, leiomyosarcosrurmuesnous adenocarcinoma, Signet Ring cell adenocarcinoma, hepatocellular carciiicma, cholangiocarcinoma, hepatoblastoma, hemangioma, hepatic adenoma, focal nodular hypetplasia (nodular regenerative hyperplasia, hamartoma), non-small cell lung carcinoma (NSCLC1) (squamous cell lung carcinoma, adenocarcinoma, large cell lung carcinoma), small cell lung carcinoma, thyroid carcinoma, prostate cancer (hormone refractory, androgen independent, androgen dependent hormone-insensitive), and soft tissue sarcomas (fibrosarcoma, malignant fibrous hystiocytoma, dermatofibrosarcoma, liposareoma, rhabdomyosarcoma leiomyosarcoma, hemangiosarcoma, synovial sarcoma, malignant peripheral nerve sheath tumoriheurofibrosarcoma, extraskeletal osteosarcoma). bfany fomors of the haematopoietic and lymphoid tissues are characterized by an increase i» cell pro liberation., or a particular type of cell. The chronic m y elopro I feral i ve diseases, {CMPDs)^e clonal haematopoietic :.$teui cell disorders characterized by proliferation in the bene niamow ofone or more of the myeloid lineages, resulting in increased numbers of granulocytes, red blood cells and /or platelets in the peripheral blood. As such, the use of proteasorne inhibitors for the treatment of such diseases is attractive and being examined fCilloni et al, Haematblogiea (200?) 92: 1124 1229). CMPB can include ehmme myelogenous leukaemia, clironie neutrophilic leukaemia, chronic eosinophilic leukaemia, polyeythaemia vera, chronic idiopathic myelofibrosis, essential ihrombocythainnia and unelasstflahleeln'bmc myelogfoliforative disease. An aspect of the invention is the method of treating BMPD comprising administering to a subject in need of such treatment an effective amount of the pfofeasomeanbibitor compound disclosed herein.

Myelbdisplasticdmyeloptplifofotive diseases, such as clironie myelpmonocytic Ieukaemia, atypical clironie myeloid leukemia, juvenile myelomonocytio leukaemia and Urielassifiable:myefod:yspIastic/myeloproliferaiive <b sease, are characterized by hypereelluiarity of die bone marrow due to proliferation in one or more of themyefoid lineages. Inhibiting the proteasome with the composition described berem, can serve to treat these myelodisplatie/myefopoliforative disease by providing a subject inmeed of such treatment an effective amount or the composition,

MyeSodysplastie syndromes (MBS) refer to a group of hematopoietic stem cell disorders characterized by dysplasia and ineffoetive haematopoiesis in one or more of the major myeloid ceil lines. Targeting NF-k'8 with a proteasorne inhibitor iri these hematologic malignancies induces apoptosis, thereby killing the malignant cell (Braun et aLGeli Peaib''St^Bifferentjiatibp;C20^) 13:?4S~?38), A further embodiment of the invention is a method to treat MDS comprising administering to a subject in need of such treatment an effective amount of the compound disclosed herein, MBS includes refractory anemia, refractory: anemia: with ringed siderpblasts, refractory' cytopenia with; muliilmeage dysplasia, .reffaetbry anemia; with excess blasts, unelassi liable: myelodysplastic syndrome and myefodysplastse syndrome associated with isolated del(:5t|) ehrpniosome abnormality.

Mastocytosis is a proliferation of mast ceils and their subsequent accumulation in one or more organ systems. -Mastocytosis includes, but is not limited to, cutaneous mastocytosis* indolent systemic mastocytosis {ISM}, systemic mastocytosis with associated clonal haematological nonmmst-eelEimeage disease (SM-AHNMD), aggressive systemic mastocytosis (ASM), mast cell leukemia (MCL), mast cell sarcoma (MCS) and exfreutaneous mastocytoma. Another embodiment of the invention is a method to treat mastocytosis comprising administering an effect amount of the compound disclosed herein to a subject diagnosed with mastocytosis.

The prbteasome regulates NF-κΒ, whiclt in fent r^ulates genes involved in the immune and: inflammatory response. For example, hlf-κΒ is required for the expression of the immunoglobulin light'chain, κ gene. the gene, the class 1 major histocompatibility complex gene, and a number of cytokine genes encoding, for example, 1L-2,11.-6, granulocyte colony-stimulating factor, and IFN-p |Palohibel!a et ah, Cell (1994) 7^:911-785). Thus, in certain ^ib:oid.tm^d^the^v^imn':''m!3fes.to..mcthods: of affecting the Ιρνρ! of expression oflL-l, MMC-S, iL-6yTi9fq, IFhl-ff Of any of the other previonsiy-mentioned proteins, each: method- comprising administering to a subject an effective amount of a proteasome inhibitor composition disclosed herein. Ip certain embodiments, the invention includes a method of treating an autoimmune disease in a mammal Comprising administering a therapeutically effeedve amount of the compound described herein. An “autoipsmuhe disease1* herein is a disease or disorder arising from and directed against an mdtvidnahs own tissues. Examples of autoimmune diseases or disorders include, hut are not limited to, inflammatory responses such as inflammatory skin diseases including psoriasis and dermatitis (e.g. atopic dermatitis); systemic scleroderma and sclerosis; responses associated with inflammatory bowel disease (such as Crohn's disease and ulcerative colitis); respiratory distress synd«>me fincluding adult respiratory distress syndrome; ARDS); dermatitis; meningitis; encephalitis; uveitis; colitis; gfememloncphritis; allergic conditions such as eczema and asthma and other conditions involving infiltration of T cells and chronic inflammatory responses; atherosclerosis; leukocyte adhesion deficiency; rheumatoid arthritis; systemic lupus erythematosus (SEE); diabetes mellitus (e.g. Type I diabetes mellitus dr insulin dependent diabetes mellitis); multiple sclerosis; Reynaud’s syndrome; autoimmune ihyroiditts; allergic encephalomyelitis; SjOrgen’s syndrome, juvenile onset diabetes; and immune responses associated with acute and delayed: hypersensitivity mediated by cytokines and T-Iymphoeytes typically found in tuberculosis,: sarcoidosis, polymyositis* granulomatosis: and vasculitis' pernicious anmmia (Addison's disease); diseases involving leukocyte diapedesis; central nervous system (CNS) inflammatory disorder; multiple organ injury syndrome; hemolytic anemia (including, but not limited to cryoglobinemta or Coombs positive anemia); myasthenia gravis; antigen-anti body complex mediated diseases; anti -glomerular basement membrane disease; antiphospholipid syndrome; allergic neuritis; Graves* disease; Lambert-Eaton myasthenic syndrome; pemphigoid bullous; pemphigus; autoimmune polyendocrinopatlnes; Reliefs disease; stiff-man syndrome; Beheet disease; giant cell arteritis; immune complex nephritis; IgA nephropathy; IgM imlyneufopatlues; immune thrombocytopenic purpura (1TF) or autoimmune thrombocytopenia.

The immune system screens for autologous cells that arevitally infected, have undergone oncogenic, transformation or present nnfami liar peptides on their surface. Intracellular proteolysis generate small peptides for presentation to T-lymphocyt.es to induce MHC class 1-mediatsd immune responses. Thus, in certain embodiments, the invention relates to a method of using the compound as aat immunomodulatory agent for inhibiting: or altering antigen presentation in a cell, comprising exposing the ceil (or administering to a subject) to the compound described herein. Specific embodiments include a method of treating graft or transplant-related diseases, such as graft-versus-host disease m host yersus-graft disease in a mammal, comprising administering a therapeutically effective amount of the compound described herein. The term "graft*1 as usedherein refers to biological material derived from a donor for transplantation into a recipient. Grafts include such diverse material as, for example, isolated cells such as islet cells; tissue such asThe amniotie membrane of a newborn, bone marrow, hematopoietic precursor cells, and ocul ar tissue, such as corneal tissue; and organs such as skin, heart, liver, spleen, pancreas, thyroid lobe, lung, kidney, tubular mgans (e.g,, intestine, blood vessels, m esophagus). The tubular organs can be used to replace damaged portions of esophagus, blood vessels, or bile duet. The skin grafts can be used not only for hums, but also as a dressing to damaged intestine dr to close certain defects such as diaphragmatic hernia. The graft is derived from any mammalian source, including human, whether from cadavers or living donors. In some eases, the donor and recipient is the same mammal.

Preferably the graft is bone marrow or an organ sudfeas· heaftbnd the donor of the graft and the host are matched for HLA class I I antigens.

Histiricytie and dendritic cell neoplasms are derived imm phagocytes and accessory celts, wbi eh have major roles in the processing and presentation of antigens to lymphocytes. Ifeplefing the proteasome content in dendritic ceils has been shown to alter their antigen-induced responses ^Chapaiic et at. Gaoeer Res. ¢006) :66:3461-5468).

Thus, another embodiment of the invention comprises administering an effective amount; of the composition disclosed herein to a subject with histiocytic or dendritic cell neoplasm. Histiocytic and dindritirc cell neoplasms include histiocytic sarcoma, Langerhans cell histiocytosis, Langsrhans eellsareoms, interdigitating dendrihecell sareorna/tnmor, feflicelar dendritic cell:sareoma/tumor and non-specified dendritic cell sarcoma.

Inhibition of the proteasome has been showri to be heheficial to treat diseases whereby a cell type isproliferating and immune disorders; thus, an embodiment of the invention includes die treatment of iymphoprolifefative diseases (LFiD) associated with primary immune disorders (PIP) comprising administering an effective anmnnf of the disclosed compound to a subject in need thereof The most common clinical settings of immunodeficiency associated with an increased incidence of Iymphoproliferati ve disotdersi including B-ce 11 andT-cell neoplasms and lymphomas, are primary Immunodeftmency syndromes and other primary immune disorders, infection with the human immunodefieieney vims: {H1V)? iatrogenicimmunosuppression in patients who have received solid organ or bone marrow allografts, and iatrogenis immunosuppression associated with methotrexate treatment. OtlferPIDs commonly associated with LFDs, hut not limited m, are ataxia telangiectasia fAT), Wiskott-Aldrich syndrome fWAS), common variable innhnnodefieieney {GV10), severe combined immunodeficiency (SC1D), X-linked lymphoproliferafive disorder (XU51), Nijmegen breakage syndrome (MBS), hypetvIgM syndrome, and autoimmune lymphoproliterative syndrome (ALPS),

Additional embodiments of the invention·relate to methods for affecting the prpteasome-depeudent regula tion of oncoproteins and methods of treating or inhibiting cancer growth, each method comprising exposing a cell (in vhej, e,g,,in a subject, or in viop) to the proteasome inhibitor composition disclosed herein, HFY-l 6 and BpM-fS-deriyed'Bb proteins stimulate ATF-and UbiqpjthPdependem cemUgafioftaod degradation of p53 is CfU#ietic«locyt<5 lysates. The recessive oncogene p53 has been shown to ac<^ttiatei8i^e:nonpemttss(ve.ientpe®@ilBt« in a cell, linewitha mutated thermolafsile EL Elevated levels of pS3 may lead to apoptosis. Exatopies of proto-oncoproteins, degraded by the ebiquitin system Include c-Mos, e-Eos, and e*Jnn, 1« certain embodiments, the invention relates to a method for treating p53-related apoptosis, comprising administering to a subject an effective amount of a proteasome inhibitor composition disclosed herein.

Another aspect ©C the invention relates to the use ©f groteasome inhibitor coinposidons disclosed herein tor the treatment of neurodegenerative diseases and conditions, including, but not limited to, stroke, ischemic damage to the nervous system, neural trauma (e.g., percussive brain damage, spinal cord injury, and traumatic damage to the nervous system), multiple sclerosis and other immane-mediated neuropathies (e.g.. Guillain-Barre s^mdrome and its variants, acute motor axonal nemxqaathy, acute inflammatory demyeimatihg polyneuropathy, arid Fisher Syndrome), BlVfAilElS dementia complex, axcnomy, diabetic neuropathy, Parkinson's disease, Huntingtorfs disease, multiple sclerosis, bacterial, parasitic, fungal, and viral meningitis, encephalitis, vascular dementia, multi-iniaret dementia, Lewy body dementia, frontal lobe dementia such as Pick's disease, subcortical dementias (such as Huntington or pmgressive supranuclear palsy), focal cortical atrophy syndromes (such as primary aphasia), nietahelie-ioxic dernentias (such as chronic hypothyroidism or B J 2 deficiency), and dementias caused by infections (such as syphilis or Chronic meningitis).

Alzheimer's disease is characterized by extracellular deposits of jLamyloid protein :(|)-AP) in senile plaques and cerebral vessels. β-ΑΡ is a peptide fiagment of 39 to 42 amino acids derived 'from an amyloid protein precursor (AFP), At least three isofomrs of APP are known (695, 751, and 770 amino acids). Alternative splicing of mRMA generates the isofomisf normal processing affects a portion of the· β-ΑΡ sequence, thereby preventing the generation of β-ΑΡ. It is believed that abnormal protein processing by the proteasome eontributes to the abundance of β- ΑΡ in the Alzheimer brain,- The APP-proeessing ^z^iejm^jcontdihsa^ptfon different subunits (22 kDa-32 kDa). The 25 kDa subunit has an 'M4-terminal sequence of X-Oln-Asn-Prp-Met-X-Thr'-Gly-Thn-Ser, which is identical to the ^subunit of human maeropam (Ko^ima, $:, et ah, Fed. Em*.

Biochem. Sac., (1992) 304:57-60). The APP~processing enzyme cleaves at the Gin —~

Lys16 bond; in the presence of calcium ion, the enzyme also cleaves at the Met——Asp5 * bond, and the Asp —Ala" bonds to release the extracellular domain of β-ΑΡ,

One aspect of the invention, therefore, relates to a method of treating Alzhei mer-s disease, comprising administering ip a subject an effective amount of the pfoteasonie inhibitor composition disclosed· herein, Sneh treatment includes reducing the rate of β-ΑΡ processing, reducing the rate of p~AP plaque formation, reducing the rate of β~ΑΡ generation, and reducing the ciinicai Signs of Alzheimer’s disease.

Fibrosis is the excessive and persistent formation of fibrous connective tissue resulting from the hypstprofiferative growth of fibroblasts and is associated with activation of the 'FGF-β signaling pathway, f ibrosis involves extensive deposition of extracellular matrix and can 'occur within virtually any tissue or across several different tissues, Normally, the level of intracellular signaling protein (Smad) that activate transcription of target genes upon TOΡ-β stimulation is regulated by protea some activity (Xu et ah, 2000). i However, accelerated degradation of the TGF-β signahng components has been observed in fibrotic conditions, such as cystic fibrosis, injection fibrosis, endomyocardial fibrosis, idiopathic pulmonary fibrosis, myelofibrosis, retroperitoneal tforosis, progressive massive fibrosis, nephrogenic systemic fibrosis. Other conditions that are often associated with fibrosis include cirrhosis, diffuse parenchymal lung disease, post-vaseetprny pain syndfome, iubcrculsis, sichle-ecll anemia and rheumatoid arthritis. An embodiment of the invention is the method of treating a fibrofie or fibrotic-associated condition comprising administering an effective amount of the composition described herein to a subject irr need of such treatment. rise treatment of bum victims is often hampered by fibrosis, thus, in certain embodiments, the invention relates to the topical or systemic administration of the inhibitors to treat bums. Wound closure following surgery is often associated with disfiguring scars, which may be prevented by inhibition of fibrosis. Thus, in certain embodiments, the invention relates to a method for the prevention or reduction of seaifmg.

Overproduction of fipppolysaecbaride (LPS)-^induced cytokines such as TNFu is considered to be central to the processes associated with septic shock. Furthermore, if is generally accepted that, the first step in the activation of cells by EPS is the binding of LPS to specific jpemhrane receptors, The «- ami β-submuls of the 2 OS proteasome complex have been identified as LPS-binbing proteins, suggesting that the LPS-Mdoeed signal tmnsduetion may he an important therapeutic target in the treatment or prevention ofsepsisfQumshi. N, et al,,/ Immun. (2003) 171; 1S15-1S25), Therefore, in certain embodiments, the proteasome inhibitor composition may he used i)r the inhibition of THFs to prevent and/or treat septic shock.

Ischemia and repertksion injury results in hypoxia, a condition in which there is a deficiency of oxygen reaching the tissues of the body, This condition causes increased degradation offcBa, thereby resulting in the;.^yatiph#MF-kBi(lC^ng et a!., 1994).

It has been demonstrated that the severity of injury resulting in hypoxia can be reduced with the administration of a proteasome inhibitor (Gao et ah, 2000; Bao et ah, 2001; Pye el:#., 2003). Therefore, certain anbodimeats of the invention relate to a method of treating an ischemic condition or teperfusion 'injury composing administering to a subject in peed of Such treatment an effective amount of the prpie&amp;Spfne inhibitor compound disclosed herein. Examples of such conditions or injuries include, hut are not limited to, acute coronary syndrome (vulnerable plaques), arterial occlusive disease (cardiac, Cerebral, peripheral; arteriafapd vascular: iicclusiens)^ sclerosis, coronary artery disease), infarctions, heart failure, pancreatitis, myocardial hypertrophy, stenosis, and restenosis. HF-κΒ also binds specifically to the HlV-enhancer/promOter, When compared to the Nef of mae239, the 111V regufatpry protein Kef of pbji.4; dlfifelfeyllwo'iamino acids in the region which controls protein kinase binding. It is believedthat the protein kinase signals the phosphorylation of l&amp;B, iriggeringlkB degradation^ thfdugh^the nbiquitin-proteasome pathway. After degradation, KF-kB is released into the nuclens, thus enhancing the transcription of IIIV (Cohen, J., Science^ (1995) 2d7;9dO). In certain embodiments, the in ventlpn relates to a method for inhibiting or reducing HIV infection in a subject, or a method for decreasing the level of viral gene expression, each method comprising administering to the subject an effective amount of the proteasome inhibitor composition disclosed herein.

Viral infections contribute to the pathology of many diseases. Heart conditions such as ongoingmyocarditis and dilated cardiomyopatby have been linked to the coxsackievirus BT. In a comparative whale-genome: microarray analyses of infected mouse hearts, specific proteasome subunits were uniformly up-regulated in hearts of mice which dev^^cfet^^ypeanjitia iSzalay ei aS, Am J Pathol 168: 1542--52, 2006). Some vimses utilize the ubipuitin-proteasome system in the viral entry step where the vims is released from the endosome into the cytosol. The mouse hepatitis virus (MHV) belongs ip tlii CmrpBirvik^e femily, which also includes the severe acute respiratory syndrome (SAKS) coronvirus. Yu and Lai (I Virol 79:644-648, 2005) demonstrated that treatment of eel Is ittiepted With MHV with a proteasome inhibitor resulted in a decrease in viral replication,, correlating with redueed viral titer as compared to that of untreated cells . The human hepa titis B virus (JdBV), a member of the Hepadnaviridcm virus family, likewise requires virally enpoded envelop proteins to propagate. Inhibiting the proteasom e degradation pathway causes a si gnificant reduction in tlfe amonjri of secreted envelope.pmtpifts^SsmsekfetialrJ Virol 79:12914-12920,2005). in addition to HBV, other hepatitis viruses t2, D and E) may also utilize the ubkjuitin-protcasome degradation pathway fer seeredon, morphogenesis and pathogenesis. Accordingly, in eepain emhodirnents, the invention relates to a method for treating viral infection, such as SARS or hepatitis A, B, C, D and E, comprising contacting a cell with tor administering to a subject) an effective amount of the compound disclosed herein.

In certain embodiments, the disclosed compositions may he useful for the treatment of a parasitic infection, such as infections caused by protozoan parasites. The proteasome of these parasites is considered to be involved primarily in cell differentiation and replication activities (Faugam et al,,. Trends Farasitof. 2003, 2 9(2): 65-39). Furthermore, entamoeba species have been: shown to lose encystati on capacity when exposed to proteasome inhibitors (Gonzales, ef ah, Arch. Med, Res, 1999,: 28, Spec Mo: 139-140). hi certain such embodiments, the administrative protocols Tor the proteasome Inhibitor compositions are useful for the treatment of parasitic infeetions in humans caused by a protozoan parasite selected fopm Plasmodium: sp$. (Including P, falciparum, F. .vivax, p, malariae, and F. ovale, which cause malaria), Trypanosoma sps, {mchiding T. cruzi, which causes Chagas' disease, and TVbrucei which causes African sleeping sickness), L.eishmania sps. fineinding L, amazonesis. 1,. ddndvanf L, infantum, L. mexicana, eted, Fneumoeystis earinii (a protozoan known to cause pneumonia in AIDS and other iromunosuppressed patients), Toxoplasma gondii, Entamoeba histolytica,

Entamoeba invadens, and Oiardsa Iambi ia, I» certain embodiments. the disclosed proteasome inhibitor compositions are useful for the-treatment of parasitic infections in animals and livestock caused by a pmtozpan parasite selected from Plasmodium hennanL Cryptosporidium sps., Echinococcus granulosus, Bimeria tenella, Sarcocystis ncurona, and Memx>spora crassa, Other compounds that act as proteasoirse inhibitors in the treatment of parasitic diseases are described in WO 98/10779, which is in<x>rpdrated herein in its entirety.

In certain embodiments, the proteasome inhibitor compositions inhibit proteasome activity in a parasite without recovery in red blood cells and white blood cells* In certain such embodiments, the long bait-life of Mood cells may provide prolonged protection with regard to therapy against recurring exposures to parasites. In certain embodiments* the proteasome inhibitor compositions may provide prolonged protection with regard to chcnmprophykms against future ihfection.

Prokaryotes have what is equivalent m the eukaryote 20S proteasome particle. &amp;!heih the subunit composition of the prokaryote 2 OS particleis simpler than that of eukaryotes, if has the ability to hydrolyze peptide bonds in a similar manner. For example^ the nucleophilic attack on the peptide bond occurs through the threonine residue on tlte hl-terminus of the p~suhunits. Thus, an embodirnent of thisviag^tipp j^MesTp a method of treating prokaryot ic infections, comprising administering to a subject an electi ve amo unt of the proteasome inhibitor composition disclosed herein. Prokaryotic infections may include diseases; caused by either mycobacteria (such as tuberculosis, leprosy or 8u:mh Ulcer) or arehaebacteria.

It has also been demonstrated that inhibitors that bind to the 2OS proteasome stimulate bone formation in hone organ cultures. Furthermore, when such inhibitors have been administered sysiernieuHy to mice, certainproteasome inhibitors increased bone volume and hone formation rates over 70% {Garrett, E lRu et ah, J. Clm (200¾ 111: 1771 -1782), therefore suggesting that the ubipustin-proicasome machinery regulates osteoblast differentiation and hone formation. Therefore, the disclosed proteasome inhibitor composition may be useful in the treatment and/or prevention of diseases associated with hone loss, such as osteoporosis.

TliiiSj ΐ» #itain embodiments, the invention relates to a method for treating a disease or condition selected from cancer, autoimmune disease, graft or transplant-related condition neimadegenerative disease, fibrotic-associated condition, isehemiwrelated conditions, iofesaion (viral, parasitic or prokaryotic) and diseases associated with bone loss, ccmprising administenng a crystalline compound of Formula (0).

Compounds prepared as described herein can be administered in various forms,: depending on the disorder to be treated and the age, condition, and body weight of the patient, as is well known in the art. For example, where the compounds are to he administered orally, they may be formulated as tablets, capsules, granules, powders, or syrups; or tor parenteral administration, they may he formulated as injections': (intravenous intramuscular, or subcutaneous), drop infusion preparations, or suppositories. For application by the ophthalmic mucous membrane route, they may he formulated as eye drops or eye ointments. These formulations can be prepared by conventional means, and if desired, the active ingredient may be mixed with any conventional additive or excipient, such as a binder, a disintegrating agent, a lubricant, a corrigent, a solubilizing agent, a suspension aid, ast emulsifying agent, a coating agent, a cydlodextfin, atid/bf a buffer. Although the dosage will vary depending on the symptoms, age and body weight of the patient, the nature and severity of the disorder to be treated or prevented, the route of administration and the form of the drug, in general, a daily dosage oftforn O.01: to 2000 mg of the compound is recommended for an adult-human patient, andfhis maybe administered in a single dose or in divided doses. The amount of active ingredient which can be combined with a carrier material toproduce a single dosage form· will generally be that amount of the compound which produces a therapeutic effect.

The precise time of administration aud/or amount of the composi tion that will yield the most effective results in terms of efficacy of treatment 1h a given patient wtii depend upon the activity, plmrmacokinetics, and blouvailahillty of a. particular compound, physiological condition: of the patient (including age, sex, disease type and stage, general physical: condition, responsiveness to a given dosnge1: and type of medication), route of administration, etc, However, the above guidelines:can be used as the basis for fine-tuning the: treatment, c.g., determining the optimum time and/or amount of administration, Which will rephire no more t ban routine experimentation consisting of monitoring the subject and adjusting the dosage and/or timing.

The phrase “pharmaceutically acceptable” is employed herein :to refer; tp those ligands, materials, compositions, and/or dosage forms which are, within the scope of sound medical judgment, suitable lor use in contact with the tissues of human beings and animals without excessive toxicity, irritafion,allergic response, or other problem or complication, commensurate with a reasonable beneiit/risk ratio.

The phrase “pharmaceutically acceptable carrier” as used herein means a pharmaceutically acceptable material, composition, or vehicle, such as a liquid or solid fi ller, diluent, excipient, sol ven t or encapsulating material Each carrier must be “acceptable*’ in the sense of being compatible with the other ingredients of the formulation and not injurious to the patient. Some examples of materials which can serve as pharmaceutically acceptable carriers include: (I) sugars, such as lactose, glucose, and sucrose; (2} starches, such as corn starch, potato starch, and substituted or unsubstituted ITcyelodexIrin; (3) cellulose, and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose, and cellulose acetate; (4) powdered foagaeanth; (5) malt; (6} gelatin; (?) talc; (8} excipients, such as cocoa butter and suppository waxes; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, com oil, and soybean oil; (If)) glycols, such as propylene glycol; (II) polyols, snob as glycerin, sorbitol, mannitol, and polyethylene glycol; (12) esters, such as ethyl oleate and ethyl laurate; (13} agar; (14) buffering agents, such as magnesium hydroxide and aluminhm hydroxide; (15) algipie acid; (! b) pyrogen-free water; (1?) isotonic saline; (IS) Ringer*$ solution; (19) ethyl alcohol; {20} phosphate buffor solutions; and (21) other non-toxic compatible substances employed in pharmaceutical fbripulatipns. In certain embodiments, pharmaceutical compositions of the present Invention arc non-pyrogemc, to,, do not induce significant temperalure elevafions when administered to a patient.

The term “pharmaceutically acceptable salt” refers to the relatively nomtoxie. Inorganic and organic acid addition salts of the inhibitors). These salts pan beprepared in situ during the final isolation and purification of the inhibitors}, or by separately reacting a purified inhibitor(s) in its foee base form with a suitable Organic or inorganic acid, and isolating the salt thus Tunned. Representative salts include the hydrobromidc, hydroehloride, sulfate, bisulfate, phosphate, nitrate, acetate, valerate, oleate, pahrritatoj Stearate, faurafo, benzoate, lactate, phosphate, tosylate, citrate, maleate, fomarate, succinate, tartrate, naphthylate, mesylate, glucoheptonate, iactohionate, laurylsulphonate salts, and amino acid salts, and the like. (See, Jar example, Bdrgeet at. (-1:977) “Pharmaceutical Salts'1, J. Pbarm, Sci. 66: 1-19.) 1« other cases, the inhibitors useful to the methods of the present in vention may contain one or more aeidie ftmctional groups and, thus, are capable of foffiilhg pharmaceutically acceptable salts with pharmaceutically acceptable bases. The term pharmaceutically acceptable salts” in these instances refers to the relatively non^toxic inorganic and organic base addition salts of an inhibitor(s). These salts can likewise be prepared in situ during the final isolation and purification of the inhibitors), or by separately reacting the purified inhibitors) ih Its tree acid form with a suitable base, such as the hydroxide, carbonate, or bicarbonate of a pharmaceutically acceptable metal cat ion, with ammonia, or with a pharmacenticaliy aeceptable organic primary, secondary, or tertiary amine. Representative alkali or alkaline earth salts include the lithium, sodium, potassium, calcium, magnesium, aird absminijm salts, and the like. Representative organic arnines useful for thb'^fhthtiobdfb^^diftoh salts include ethyl am ine, diethylamine, ethyienediamme, Mhanolamine, diethanolamine, piperazine, and the like f see, for example. Beige el ah, supra).

Wetting agents, emulsifiers, and lubricants, such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, release agents, coating agents, sweetening, Savoring, and perfuming agents, preservatives and antioxidants can also be present in the compositions.

Examples ©fpbannaceutieally acceptable antioxidants include: (I) water: soluble antioxidants, such as ascorbic acid, cysteine hydfoehlpridd, sodium blsu I fete, sodium metabisulfste, sodium solfite, and .the dike; (2) os hso ruble an t iox klan is. such as aseorbyj pahnitate, bUtylated hydrpXyanfsole (BBA), butyiated hydrexytojitene (BBT), lecithin, propyl galiate, alpha-tocopherol, and the like; and (3) metal chelating agents, sued as citric acid, ethylenediai’nine tetraac^ie acid f EE3TA), sorbitol, tartaric acid, phosphoric acid, and the like.

Formulations suitable for oral administration may he in the form of capsules, cachets, pills, tablets, lozenges («sing a flavored basis, usually sucrose and acacia or tragaeanth), powders, granules. or as a solution or a suspension irr an aqueous or uori-aqueous liquid, or as an oi!rin-water pr wateron-bil .liquid· emulsion, or as an elixir or syrup, or as pastilles (using an inert matrix, such as gelatin and glycerin, or snemse and acacia) and/or asmouthwashes, and the like, each containing a predetermined amount of aninhihitor^sf aa lh'aciixie ingredient. Λ composition play also be administered as a bolus, electuary, or paste.

In solid dosage: forms for ora! administration (capsules, tablets, pills, dragees, powders, granules, and the like), the acti ve ingredient is miked With one Or more pharmaceutically acceptable earners, such as sodium citrate or dicalciurn phosphate, arid/or any of the following: (!)fiilers or extender, such as starches, cyolodexinns, lactose, sucrose, glucose, mannitol, and/or silicic .acid; (2) hinders, such as. for example, earboxymethyloollhioso, alginates, gelatin, polyvinyl pyrrolidone,:sucrose, and/or acacia;: .(3) humectants, such as glycerol;,,(4) disintegrating: agents, Inch, as agar-agar, Calcium carbonate, potato or tapioca starch, alginic acid, certain :s*beaies, and sodium earhonate; (5) solution, retarding agents, such as paraffin; (6) absorption accelerators,, such as quaternary ammonium compounds; (?) Wetting agents, such as, for example, acetyl alcohol and glycerol monostearate; (§> absorbents, such as kaolin and bentonite clay; (9) lubricants, such a talc, calcium stearate; magnesium htearate, solid polyethyiene glycols, sodium fauryl sulfate, and mixtures thereof; and 0 0} coloring agents. in the case of capsules, tablets, and pills, the pharmaceutical compositions may also comprise huiforirsg agents. Solid compositions of a similar type may also be employed as fillers in soft and hard filled gelatin capsules usingsuch excipients as lactose or milk sugars, as well as high molecular weight polyethylene glycols, and the like. A tablet may he made by eompression or molding, optionally with one or more accessory ingredients. Compressed tablets may be prepared using binder (for example, gelatin or hydroxypropylmethyl cellulose), lubricant, inert diluent, preservative, disintepapt (for example, sodium starch glyeolate or cross»ltnked Sodium carhoxymctbyl Cellulose), surfoee-active or dispersing agent. Molded tablets may be made by molding in a suitable machine a mixture of the powdered inhibited» moistened with an mert liquid diluent.

Tablets, and other sol id dosage fornts, such as dragees, capsules, pills, and granules, may optionally beseored ©r prepared with coatings and shells, such as enteric coatings and other coatings well known in the phannaGeutlcai-fornmlating art. They may also be fonnulated so, as to· provide slow or controlled release of the active ingredient therein nslngj for example, hydraxypropylpi'e^l 'eellttloseip Vising pr^ppjjions to provide the desired release profile, other polymer matrices, liposomes, and/or raicrospheres. They may fee sterilized by, for example, filtFatioir tooui^t a bdeteria-retaining fi lter, or fey incorporating sterilizing agents in the form of sterile solid compositions which can be dissolved in sterile water, or medium immediately before use. These compositions may also optionally contain opacifying agents and may fee of a composition that they release the active ingredients} only, or i preferentially, in a certain portion of the gastrointestinal tract, optionally, in a delayed manner. Examples of embedding compositions which can be used include polymeric substances and waxes:. The active ingredient: ciin also be in miao-encapsplated form, if appropriate, with one or more of the aboverdescribed e*eipients.

Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, microemnlsions, solutions, suspensions, syrups, and elixirs, in addition to the activeingredient, the liquid dosage forms may contain inert diluents commonly nsed in the art, such as, for example, water or other sotvems, sotubiIking agents, and emulsifiers such as ethyl alcohol, isopropyl alcohol... ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glpol, oils (in particular, cottonseed, groundnut, corn, germ, olive, castor, and sesame oils), glycerol, teimhyimforyi alcohol, polyethylene glycols, and fatty acid esters of fofbitam and mixtures thereof

Besides inert diluents, the oral compositions can also include adjuvants such as wetting: agents, emulsifying and suspending agents, sweetening, flavoring, coloring, perfoming, and preservative agents.

Suspensions, In addlbonto the aciivelinhifeito^j may contain suspending agents: as, for example, ethoxylated sorbitol and sprbltan esters, microerystalline cellulose, aluminum metahydroxide, bentonite, agar-agar and fragacanth, and mixtures thereof.

Formulations for rectal or vaginal administration may be presented as a suppository, which may be prepared by mixing one or more inhibiiorCs) with one or more suitable nonirritating excipients or carriers comprising, for example, cocoa butter, polyethylene glycol, a suppository wax or a salicylate^ which is solid at room temperature, but liquid at body temperature aud, thereldre, wilt melt in th® rectum or vagina! cavity and release the active agent.

Formulations which are suitable for vaginal adnhmstmtion also include pessaries, tampons, creams, gels, pastes, foams, or spray formulations containing such carriers as are known in the art to he appropriate.

Bosage forms for the topical or tmusdepual administration of an inMhitor(s) include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches, and inhalants. The active component may he mixed under sterile conditions with a pharmaceutically acceptable carrier, and with any preservatives, buffers, orpropellants which may he required.

The ointments, pastes, creams, and gels may contain, in addition to inhibitorfs), excipients, such as animal and vegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicic acid, talc, and xine oxide, dr mixtures thereof.

Powders and sprays can contain, in addition to an inMbifdrfsf, excipients such as lactose, talCj silicic acid, ainminum hydroxidp, calcium silicates, and polyamide powder, or mixtures of these substances. Sprays can addi tionally contain customary propellants, such as ehlorofiuofphydroearbons and volatile unsubstituted hydrocarbons, such as butane and propane,

The InhibitorCs) can bo alte^abyelyradhitiBisfercxI.hy'^^spl.. This is accomplished by preparing an aqueous:: aerosol, liposomal preparation, nr solid particles containing the eompcsitiop, A honaque^us f e.g., fluorocarbon propel I ant) Suspension could he used, Sonic nehulixere are preferred because: they minimise.exposing the agent to shear, which can result in degradation of the compound..

Ordinarily,1 an aqueous: aerosol is made by formulating an aqueous isolation or Suspension of the agent together1 with convenfionafphannaceMtipaliy acceptable carriers and stabilizers. The carriers and stabilizers vary with the requirements of the particular composition, but typically include noniqnie surfactants (Tweens, Fluronlcs, sorbitan esters, lecithin, Cremophors), phaonaceuii cally acceptable co-sol vents such as polyethylene glycol, innocuous proteins like serum albumin, oleic acid, amino acids such as glycine, buffers, salts, sugars, or sugar alcohols, Aerosols generally are prepared front isotonic solutions,

TransdSrmal patches have the added advantage of providing controlled delivery of an inhibitor^) to the body, Such dosage forms cart be made by dissolving or dispersing the agent ip tife proper medium, ABso^tioo eulianeet's can also be used to increase the d ux of the inin bi toits) across the skin. The rate of such flux can be eontro ll ed by either providing a rate controlling membrane or dispersing the inhibitors) in a polymer matrix orgei.

Pharmaceutical compositions of this invention suitable for parenteral administration comprise one or more inhihiCor$(s) in combination with one or more pbannaeeuiically acceptable steriis aqueons of nonaqneons solutioris, dispersions, suspensions or emuisions, or sterile powders which may he reconstitnted into sterile injectable solutions Or dispersions just prior to use. which may contain antioxidants, buffers, baeteribstats, solutes which render the formulation isotonic with the blood of the intended recipient or suspending or thickening agents,

Exanm^Ps of snitabie aqueous and nonaq ueous carriers which may be employed in the pharmaceutical compositions of the in vention include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene gfyeol, and the like), and suitable mixtures thereof Vegetable oils, such as olive oil, and injectable organic esters, such as ethyl oleate, ErOper fluidity can be maintained, for example, by the use of coating materials, such as lecithin, by : the maintenance of the required particle size in the case of dispersions, and by the use of surfa ctants .

These compositions: may also contain ad?uvants::$oeh: as preservatives, wetting agents, emulsilyiug agents, and dispersing agents, Prevention of the action of microorganisms may be ensuredlby the;inclusion of various antibacterial and antifungal agents, for example, parahem ehldfobutanol, phenol sorbic acid, and the like. It may also be desirable to include tonieiiymdjusting agents, such as sugars, sodium chloride, and the like sum the compositions. In addition, prolonged absorption of the injectable pbarmacentical form may be brought about by the inclusion of agents which delay absorption such as al urainuirt monostearate and geiatin.

In some cases, in order ίο prolong the effect of a dmg, it is desirable to slow the absorption of the drug from subcutaneous or intramuscular injection. For example, delayed absorption of a parenteral ly administered drug &amp;fm is accomplished by dissolving or suspending the drug in an oil vehicle.

Injectable depot forms are made by forming tnieroeapsole matrices of inhihitor(s) in biodegradable polymers such as polylaciide-polyglycolide. Depending on the ratio of drug to polymer, andf he nature of the particular polymer employed, the rate of drug release can be controlled. Examples of other biodegradable polymers include polyforihoesterSf and po!y{anbydrides). Depot injectable formulations ard also prepared by entrapping the drug in liposomes or microenmlsions: which arc compatibleiwith body tissue,

The preparations of agents may be given·'.orally,:parenterall'yj topically, or rectal ly,: They are, of course, given by Torexample, they arc administered i n tablets or capsule form*. by injection, inhalation, eye lotion, Ointment, suppository, snfusion; topically hylotipti or ointment; and fecial ly by suppositories. Oral administration is preferred.

The phrases “parenteral administration" and '‘administered parenterally” as used herein means modes of administration other than enteral and topical administration, usually by injection, and ideludesf without limitation, intravenous, intramuscular, intraarterial, intrathecal, intraeapsular, intraorhital, intracardiac, mtiadenna!, intraperitoncal, transtracheal, subcutaneous, subcuticular, intraarticular, suhcapsular, subarachnoid, rntraspinal and intrastemal injection, and infusion.

The phrases “systemic administration^' “administered systernically/! “peripheral administration" and “administered peripherally5’ as used herein mean the administration of a ligandj drug, or other materia! Other than directly into the central nervous system, such that it enters the patient’s system and thus, Is subject to metabolism and other like processes, lor example, subcutaneous administration.

These inhibitorsfs) nmy be administered to burnans and other animals for therapy by any sin table route Of administration, including orally, nasally, as by, for example, a spray, rectaily, miravaginalSy, parenterally, iniracisiernally,and topically, as by powders, ointments or drops, including buccally and sublingually.

Regardless of the route of administration selected, the inhihiforfe), which maybe used in a suitable hydrated form, and/or the pharmaceutical compositions of the present invention, are formulated into pharmaceutically acceptable dosageforms by conventional methods known to those of skil l in the art.

Actual dosage levels of the active'iogredients in the pharmaceutical compositions of this invention may he varied so as to obtain an amount of the active ingredient which is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration, without being toxic to the patient.

The concentration of a disclosed compound in a pharmaceutically acceptable mixture will vary depending on several foetors, inefodihg the dosage of the compound to he ad ministered, the pharmacokinetic characteri stics: of the compound(s) employed,: and the route of administration. In general, the compositions of this invention may be provided in an,aqueous solution containing ahoutOJ-ICfekw/v of a compound disclosed herein, among: other substances, for parenteral administration. Typical dose ranges are from about 0.01 to about 5ffmg/kg of body weight per day, given in 1-4 divided doses. Each divided: dose may contain the same or dififereiit compounds of the invention. The dosage will he an effective amount depending on several factors including the overall health of a patient, and the foundation and route of adnunistrahou of the selected compmmd(s).

The term vcs.yaikyl” refers to substituted or nnsubstituied saturated hydrocarbon groups, inefodmg straight-chain alkyl and branched-cltain alkyl groups that contain from x to y carbons in the chain, including haloalkyi groups such as trifluoromethyl and 2,2.2· trifiuoroethvL etc. Coalkyl indicates a hydrogen where the gronp is in a terminal position, a bond ifmterhaL The 100118 ‘T^pailkeiiyP and Xfovalkynyr! refer to substituted or unsubstituted unsaiurated aliphatic groups analogous in length and possible substitution to the alky!» described above, but that contain at least one double Or triple bond respectively.

The term folkoxy^ mfers to an alkyl group having an oxygen attached thereto. Representative alkoxy groups include methoxy, ethoxy, propoxv, tert-butoxy and the like. An ‘"ether" is two hydrocarbons covalently linked by an oxygen. Accordingly, the substituent of m alky! that renders that alkyl an ether is or resembles an alkoxy.

The ten» refers to a Chalky! group substituted with an atkcxy group, thereby forming an ether.

The term 'TfosamlkyPy as used herein, refers to a C^lky! group substituted with an ary! group.

The terms “arhirie” and “arnin# are ari-reeognized and refer to doth tmsafestituied and substituted amines and salts thereof, e.gi} a moiety that can he represented by the general formulae:

Wherein R'y.Ri0 and Ria each independently represent a hydrogen, an alkyl, an alkenyl, -{CHi^-R8, or Ry hnd R,&amp;;taken together with the N atom to which they are attached complete a heterocycfo having from 4 to 8 atoms: in the ring structure; R* represents an aryl, a cycloalkyl, a eyelpalkenyl, a heferpeyelyf or a polycyclyl; and m is aero or an Integer from I to S,. I:p.pr^f^n^,fpibodia?ents, pply one of R? or Km can be a carbonyl, e.g,, .¾ RH!, and the siitrogen together do not form an amide, ίο even more preferred embodiments, R<- and R10 (and optional ly E!i!): each independently represent a hydrogen, an alkyl, an alkenyl, or ~(CH2)m-Rs. In certain embodiments, the amino group is basic, meaning the protemaied form has a pKa > ΊΜ. The terms "amide'5 and “amide” are art-reeognixed as an amino-suhstituted carbonyl and includes a moiety that can be represented by the general formula; wherein R\Ris are as defined above; Preferred emhodimentsoftlie amide will not include imsdes which may be unstable. The team “aryl” as used herein includes 5--, 6-, and 7-meffibered substituted or unsubstituted Single-ring aromatic groups in which each atom of the ring is carbon. The term %ryl“ also includes polycyclic ring systems having two or more cyclic rings in which two dr more carbons are common to two adjoining rings wherein at feast one pf the rings is aromatic, e.g., the other cyclic rings can he cycloatkyls, cyctoalkenyls, eydoaikynylSi aryls, heieroaryls, anchor heterocyciyls. Aryl groups /include benzene, naphthalene, phcnamfoene, phenol, aniline, and the like. ITie terms “carijoeyeie” and ^arbpeyetyr, as used herein, refer ίο a hon-arphtatie substituted or unsubditoted ring in which each atom of the ring, is carbon. The terms ^earboeye!#* and ^-carbocyclyF also include polycyclic ring systenia haying two or more cyclic rings in which two or snore carbons are cgmrmon to two adjoining rings wherein at least one of the rings is earbocyclic, e.g., the other cyclic rings can he cycloalkyis, cycloalkenyis, cyeloalkynyls, aryls, heteroarylsj and/of heterocyciyls.

The term “carbonyl” Is art-recognized and includes such moieties as can he represented by the general formula;

wherein % is a bond or represents an oxygen or a sulfur, and R represents a hydrogen, an alkyl, an alkenyl, TCH^-R8 or a pharmaceutically acceptable salt, Ru represents a hydrogen, ah alkyl, an alkenyl or where m and It* are as defined above*

Where X is an oxygen andJt* * or It·1 is not hydrogen, the formula represents an “ester55. Where: X is an oxygen, and ltsl is a hydrogen, the: formula represents a '‘carboxylic acid”.

The ternrs ‘foeteroaryl5' iriclodes substituted or unsubstitnted aromatic:5- to 7~ membered ring::Struetures,: more preferably 5- to 6-membered rings, whose ring structures include one to tour heteroaforns. The term ''heteroaryF also includes polycyclic ring systems having two or more cyclic rings in which two or more carbons are common to two adjoining rings wherein at least one of the rings is heterearoroatie, e,g., the other cyclic rings can he^cJoalk^.ieyciPiiifeenyls, cyeloalkyiiyls, aryls, heteroaryls, and/or heterocyciyls. Hctcroaryl groups include, for example, pyrrole, furan, thiophene, inndazofo, oxaxole, fhiazole, triazole, pyrazole, pyridine, pyraxsue, pyridaxine and pyrimidine*and the like.

The term “heteroatom5* as used herein means an atom of any element other than carbon or hydrogen. Preferred heieroafoms are nitrogen, oxygen, phosphorus, and sulfur.

The: terms; ‘foetetooyeiyP of:“heferoeyche group” referio substituted or umrdistituted UOft-arorastic 3- to Ifomenfoered ring structures,, more preferably 3» to: % memhered rings,: Whose ring structures include one to fonrhoteroatoms. The term terms Theteroeyclyl” or “heterocyclic group” also include polycyclic ring systems having two or more cyclic ripgs ln which iwo or more carbons are: common, to two adjoining nogs wherein at least ono of the rings is heterocyclic, <&amp;·$* the other cyclic rings can be eyc'baikyis, cyelpatkenyis, cydoalkyhyls, aryls, heteroaryls, and/or hetemeyelyls. Heterocyefyf groups include, for example, piperidine, plpemine, pyrrolidine, morpholine, lactones, lactams, and the like,

Tie term “C^hydmxyalky'r refers to a Chalky! group substituted with a hydroxy group.

The terms “polyeydyP or “ppiycydhc” tefisrTp two or more rings (e.g., eycloalkyls, eycloalkenyls, cycloalkynyts, aryls, heteroaryls, and/or hetemeyciyls) in which two or more carbons are common to two adjoining rings, e.g., foe rings are “fused rings:’. Each Of the rings of the polycycle can be substituted or uosuhsiituted.

The icon “protcasome” as used herein is Tneapt to include immune·· and constitutive proieasomes,

The temi “substantially pure” as used herein, refers to a crystalline polymorph that is greater than 90% pure, meaning that contains less than 10% of any other compound, including the Corresponding amorphous compound. Preferably, the crystallinepolymorph is greater than 935i pure, or even greater than 98% pure, lie term *lubshtuied” refers to moieties having substituents replacing « hydrogen on one or more carbons of the backbone. It will be Understood that “substitution” or “substituted with11 includes the implicit proviso that such substitution Is in accordance with permitted valence of the substituted atom and the substituent, and that foe substitution ^results' in a stable compound, e>g., which does not spontaneously undergo transformation such as by rearrangement, eyclization, elimination, etc. As used herein, the term ‘substituted” is contemplated to include all permissible substituents of organic compounds. In a broad aspect, the permissible subshtuenis include acyclic and cyclic, branched and unhr anehed. ca rhoeyc I ie and hei eroeycl ie, aromatic and non-aroma tic substituents of organic compounds. The pcntrissible substituents can be οπό or more and . AO _ the same orblffereni for For purposes of this invention, the heteroatoms such as nitrogen may have hydrogen substituents and/or any permissible substiteems of organic compounds described herein which satisfy the valences of the heleroatoms, Substituents can include, for maniple, a halogen, a hydroxyl , a carbon yl (sndi as a carboxyl, an alkoxycarhonyh osformyl, or an acyl), a iMocarbopyl (such as a thioesfer, a ihioacetate, or a ihiotormafe), an alkoxyl, aphosphoryi, a phosphate, a phosphorate, a phosphinste, an amino, an arnido., an arPidinc, an imine, a cyano, abitro, an azido, a sulfhydryl, an alkylthfo, a sulfate, a:sulfonate, a sulfamoyf, apnlfonamldo, a soifenyl, a heterocydyl, an aralkyl, or an aromatic Or heteroammatie moiety, It will be understood by those skilled in the art that the moieties substituted Oh the hydrocarbon ebaitt: can themselves he substituted., if appropriate. A:: “therapeutically effective arnounf’ ofa compound with respsef to the/subject' method of treatment, refers to an amount of the compoundfs) in a preparation which, when administered as part of a desired dosage regimen (Id a tnanimahipreferably a human) alleviates a symptom, ameliorates a condition, or slows the onset of disease conditions according to clinically acceptable standards for the disorder or condition to be treated or the cosmetic purpose, e.g,, at a reasonable beneht/nsk ratio applicable to any medical treatment.

The term “thioether” refers to an alkyl group, as defined above., having a sulfur moiety attached thereto. In preferred embodiments, the "hhioether5 is represented by -S~ alkyl. Representative thioether groups include methyithio, ethyhhio, and the like.

As used herein, the term 'Seating**#· includes reversing, reducing, or arresting the symptoms, clinical signs, and underlying; pathology of a eondition in manner to improve or stabilize a subject’s condition.

Example 1

Synthesis of Compound 1

Synthesis q/(B)

Mydmxybeir^iri:azole<l|0BT) tlOJl g, 80.0 mmol) and 01¾¾ (200.0 mmol 25 JS g. 31 ml.) was added to a solution of NBoc leucine (50.0 mmol,: 11.56 g) arid phenylalanine methyl ester (SOiG mmol, 10,78 g) in 500 ml, of DMP, The rniyinre was cooled to 0 °C: In an ice-water bath and benzotnaaol·1 -yloxyiris(dimetliylammo)r pbosplmnsPm hexaOuoropImsphate (Β0Ρ) (SO.O mmol, 35.38 g) was added in several pprtipns over five minutes. The reaction was placed under an atmosphere of argon and stirred overnight. The reaction was diluted with brine (1000 ml.) and extracted with EtOAe (5 X 200ml,), The organic layers were combined and washed with water (10 x 100 rot,} add brine (2 x 150 ml,} and dried over MgSCfi The MgSCfi was removed by filtration and the volatiles removed underreduced pressure to give (A) (18(17 g). To a 50 mL 0: “Cieodied solutiojv.of 80¾ TFA/DCM was added BocNITLeuPheOMe (45,80 anlieol,. 1|.0 g). Ifoe solution was stirred arid allowed to warm to room temperature over 2 hr. The volatiles were removed adder .reduced pressure to gi ve an oil, BdeNFlhPhe (45.80:mmoh 12,81 g), DMF (500 nVtl HOB I (73..37 mmol 9.91 g) and D!EA{ 183.44 mmol, 23,70: g>: 32,0 ml) were then added to the oil. The mixture was cooled to 0 °€ in an iee-waterijath and: BOF (73.37 mmol, 32,45 g) was added infive pmtttes, The reaction was placed under argon and allowed to warm to room temperature oyemight. The reaction was:diluted with MjO (1500 mL). and extracted with DCiVi (5 x 300 mL). The Orpnio layers wem combined and washed with Hj® (6 x: 300 mL) and brine ( I x 3Q0:mL) and dried over MgSCfi, The MgSO,? was removed by filtration arid the volatiles removed under reduced pressure to give a yellow solid. EtOH (200 mL 95%) was then added tp lbe:yelfow solid and the mixture was heated to 65 °G to dissolve all of the sol ids. The solution was then added to 1000 ml, of chilled H2Q and the resulting precipitate collected to give:(B) (21,5:9: g).

Synthesis (B) (1.80 mmol, 1 :.0 g) was mixed with TFA/DGM (80%) and was stirred at mom temperature for l hr, at which time the pixtufe was concentrated and placed under high vacuum for 2 hr giving foe TFA salt of the tri-peptide amine. To a 0 *C solution of the TFA salt (1,80 mmol) in DMF (10 ml^ Was added IT1EA (3¾ mmol, 0.7 ml) followed by chloroacetyl chloride (2.7 mmol, 0.215 mL). The reaction was allowed to warm to RT while sfirring ovemight under an atntosphere of nitrogen. The mix ture was then diluted with brine (15 mL) and extracted withElQAefo χ 15 pL), The organiclayers were combined» washed with HjO (2x15 mL) and brine (2 x 15 mL) and dried over NaaSCfo The NajSffowas removed hy filtration and the volatiles removed under reduced pressure, The crude material was suspended in EtOAe and filtered to give (€> (0.640 g)

Synihesis of (D) m (0.,019 mmol, 0:,0032: g)(and morpholine (0,1 10 mmol, 0,0096 g) were added to rsdlubou of (C) ¢0.094 mmolS: 0.050 g) in ΤΉΡ (10 mL) and the mixture was stirred O^emighf tinder an atmosphere ol’ditrogen. The volatiles were removed under reduced pressure and the erode material taken up in EtOAe (IS mL), washed with LLO (2x10 mL) and brine (2xl0mL) and dried over MgS04. "The filtration and the volatiles removed under reduced pressure to give (D).

Synihesis of {£}

Li OH (0.94 mmol, 0.023 g) was added to a slurry of (D) (0.094 mmol) in 4 mL of 3:1 MeOH/H^O cooled to 0 °C. After 12 hr at 5 °C the roaedon was quenched with 20 mL sal. MP4CI and diluted further with 10 mL LpD. The pH of the reaction mixture was adjusted to 3 with 1 hi BCI, extroeisd with DCM (3 x IS mL), and dried over MgSCV The MgS©4 was removed by filtmrion and the volatiles were removed under reduced pressure to gi ve (E).

SyniHesis ofCompmmtf / (E) (0,082 mmol, 0,046 g). DIEA (0.328 mmol 0,057 nit) and HOST (0,133 mmol, 0,018 g) were added to a stirred solution of (F) (0.082 mmol) in DMF (2 mL), The mixture:was cooled to 0 °C in an ice hath and 8DP (0,131 mmol 0,058 g) Was added in several portions. The mixture was stirred at 5 °C under an atmosphere of argon overnight. The reaction was. then diluted With HjO (15 mL) and extracted: With EtOAe. The organic layer:was1 Washed with water* sat. NaHCOj, and brine and dried over anhydrous MgSO«. The MgSCA was removed by filtration and the volatiles removed under reduced pressure to give compound I (0,034 g) (ICso 20S OTL<! OOnM:; IC50 Cell-Based CT-L<I OOnM).

Example 2

Compound I (LO g): Was dissolved in methanol (16 m t) heated to 80 HE. Water (4 mL) was then slowly added and the clear solution was allowed to cool to ambient temperature and the solution was brought to supersaturation by'evaporating:off i O rnL Of sol vent With compressed air. The restdli ng crystals were ft Itered. washed with f m L 1: | water-meihaK»!, asrd dried under vacuum for 12 hours to provide crystalline compound 1 (β<Ρ g) with a melting point of 212 °C,

The characteristic DSC curve of the sample is shown i&amp;Tigure 1 as recorded oh a TA Instruments Differentia! Scanning Calorimeter 2920 at a heating rate of 10 ^QrmnUte.

Example-3

Compound 1 (1,0 g) was dissolved in iacihonltrile (17 mL);'heated to 80 °C. Water (8 ®L) was then slowly added and the clear solution was allowed to eooHo ambient temperature and the solution was brought to snpersaturation by evaporating off 10 mL of solvent with compressed air. The respiting crystals were filtered, washed with 8. mi, 1:1 dsiomwsd water-acetonitrile, and dried under vacuum for 12 hours to provide:crystalline compound 1 (1),85 g) with a melting point of 212

Example 4

Compound 1 (CO g). Was dissolved m ethanol (1? mb) heated to 80 °C. Water (5 ml) was then slowly added and the clear solution was allowed to cool to ambient temperature and the solution was brought to supersaturatk® by evaporating off 15 mL of solvent With compressed air. The resulting crystals were filtered, washed With 8 rpL 1:1 deionised water-ethanol, and dried under vacuum for 12 hours to provide crystalline compound ! (0.82 g) With a melting point of 212 °C<

Exampie S

Compound I ( LI) g) was dissolved in ethyl acetate (30 mL) heated to 80 C<X Water (5 thL) was then slowly added and the clear solution was allowed to cool to ambient temperature and the solution was hn>ught to supersaturation by evaporating off 20. mL of solvent with compressed air. The resulting crystals were filtered, washed with 5 mL ethyl acetate, and dried under vacuum for 12 hours to provide crystalline compound ! (0.60 g) with a melting point of 212 ”€,

Example 6

Compound 1 (1.0 g) was dissolved in ethanol (15 mL) heated to 80 °C. Water (5 mb) was that slowly added and th# clear solution was allowed to cool to ambient temperature and the solution was brought to supersatumtion by evaporating off 10 mb of solvent with compressed air. The' resulting crystals were /filtered, washed with 10 mL 1:1 deionized water-ethanol, and dried under vacuum for 12 hours to provide crystalline compound 1 (0,54 g) with a melting point of 212 °C.

Example 7

Synthesis of(F)

Compound (0) (0.43 g) was prepared according to ICS, Pat. Application Mo, 2005-0256324 and was added to a flask along with Pd/E (10% wt, (5,10 gffbUowedby slow addition of TFA (35 mL). The flask was evacuated and back-flushed with hydrogen gas three times and then the reaction mixture was stirred under one atmosphere Of hydrogen at room temperature fhrtwo hours. The reaction mixinre was then filtered through Celite and the filtrate was concentrated under reduced pressure,

Diehloromethane (25 mL) was added and the volatiles removed under reduced pressure. The resultant thick yellow syrup was dried under high vacuum to a constant weigjbh The syrup was then transferred to 50-niL volumetric flask and rinsed with 8.5 ml, diethyl ether to yield .crystalline compound (F) (0.3 3 g).

Synthesis qfGampmmil I A 10 mb volumetnc flask was charged with I dtydmkybenxotnazoie (HOBT, 0.54 gland MjNjMf M'-tetrameihybQ-tlTi-benzmriaxol-l-yfJuronium hexailnorophosphate (HBTU, ! .54 g) arid diluted· to SO nrL with PMF, This stock solution of coupling reagents was 0.40 M for both HOST and HBTli, (£):(0.61 g), (F) (033g), and the coupling reagent stoeksolufion (2.7 mL), were added Ϊ6 a 1C mL volumetric flask and the mixture was cooled to 0 °C. DIE A (0.56 ml.) was then added dropwise to the cooled solution. The mixture was allowed to stir at 0 °C for t>0 minutes and was then gueoehed hy the addition of saturated sodium hiearhooat© {15 mL). lire mixture was diluted with ethyl acetate (35 mL) and thd layers staparated.

The organic layer was washed saturated sodium bicarbonate (3 x 15 mL), brine (2 x IS mL) and dried over sodium sulfate. The sodium sulfate Wasremoved by filtration and the volatiles removed under reduced pressure to give a thick syrup which was further dried under high vacuum to give a crude compound 1 as a team (0:59 g).

Example <$·

Crude compound 1 (0.390 g) was completely dissolved in methanol (11 mL) by stirring and heating in an oil bath (80 ”C) and deionized water (1 ? ml.) was added dropwise, The mixture was seeded with crystalline ccmpouod 1, stirred and allowed to slowly evaporate for 12 boom to approximaMy 20 mL to preeipitate compound f . The suspension was filter#, washed With(1:1 deionized water-methanol (4 mL). and dried under vacuum for 12 hours at room tmperatumio'^ddiwmpotmd I as a white solid (0.25 g). The:crystallisation -was repeated two additional times to yield crystailine compound I (0.13 g).

Crystalline compound 1 (0.3 g) was dissolved in isopropanol:(l5'UiL} by stirring and heating in an oil bath (SO t5C). The solution was potmentruted: under reduced pressure to reduce volume to· 5"mL, Deionized water (20 niL) was quickly added and the resultant suspension: was rigorously stiped for I hour. Thegiassy precipitate: was filtered, rinsed with deionized water fdS mL) and dried to yield amerphoiis Compound I (0.3 g).

The characteristic DSC curve of the amorphous sample is shown in Figure 7 Which was recorded: on a TA Instruments Di fferential Scanning Calorimeter 29:21) af a heating rate of 1, "Ormnute for the amorphous form .of Compound 1,,

Tlm:oh»mct«risiic.,^kay.di:ffm<dion pattern of tbe amorphous powder is shown In Figures and was recorded on the Shimadzu XRD-0G0D under Cu K.e radiation (voltage and current setat 40 k V and 40 mA| di vergcncc and scattering slits set at 'l ivand::reeei vins slit set at :0.15 mm; Nal scintillation detector used for diffracted radiation; a 8~20 continuous scan at 37min (0.4 sdc/O.02e step) from 2.:5 to 40° 20 was usedpsanmles were placed k an aluminum holder with silicon insert; and data collected and analyzed with lRD4ll/fOO0v.5.O].

Mcmipie9

Syfiikesis of (F) A flask was charged With I'd) and ethyl acetate (df)0 ml,) and thesolution was cooled in aft ice bathffor IS minutes with stirring, Trifieoroaeetic acid (2:001 mL) was added dmpwise, maintaining an internal temperature of less than 10 Pd/G (3,0 g) was added in one:portion and the flask: was purged under high vacuum and refilled with hydrogen three times.: After 2 hours, the reaction was filtered through Gel its and the filtrate evaporated under reduced pressure to a thick orange oil which wtetswirled gently with 170 rirtG dietllytl As the flask was smrled, fine crystals fonhed, The.:flask· was allowed to sit at mom temperature, and rapid etystellfeaiion occurred, After 1 hour at amhieht temperature, the flask was capped tightly and placed in the freezer .overnight {<~ 5 ?C), The resulting crystalline1 SdMd: was filtered and washed with ice: cold ethyl ether (MmLTand dried under high vacuum. Fine white crystals (14,1 g; melting point; 1.37 eC) dp :{F) were obtained.

Synthesis qfC&amp;mpounii l A flask was charged with (F){10g), {F}(i5.3 g), 11BTU (15.3 g), HOBt (5.5 g), aodDMF(30OmL), The mixture was stirred vigorously until dissolved and was placed in an MaCl/iee bath (¢-5 °C); After 15 minutes, DIE A (7.3 mL) Was added drapwise over <10 minutes, maintaining an internal temperature oftjess than -3 *G, After addition was complete, the reaction mixture was stirred in the hath for one hour and was quenched by addition of saturated NaBGOj faqd (200 rift)'. The slurry was extracted with ethyl acetate (1,3 L) and the organic layer was washed with sat. MaFtCOftaq.) (2: x 300 rat) and sat, NaCt (aq,) (200 nrL), and then dried oyer MgSCh,

The organic layer was concentrated to -50 mL under reduced pressure and methylethyi ketone (200 mL) was added, and the solution was again concentrated to ~50 mL. Methylethyl ketone (125 mL) was added again, and the solution was stirred in an oil bath (80 °C) until clear. The solution was then allowed to cool and was seeded with pure crystalline Compound I, The mixture was stirred for 2 hours at 25 °C and then o vernight at 0 °C, The white solid precipitate was filtered and washed with ice cold methylethyl ketone £300 mL) to give white solids. The solid was deed under high vacuum at ambient temperature to a constantweight to yield 13.5 g of pure compound 1 £mmpie.i&amp;

Synthesis of (F) flaslc was charged with (Η) (ISO g) [see: Bioorg. Med. Chero. Ifotter tSt§§, 9> 2283-8¾ dfid dihhti|jr0m^M^e'(3-|p0 the solotidh was edoled in an ice bath to 0*5 ®C. Tofluoroaeetic acid (136:.9 aiL) was added dropwise with stirring at 0-!0*G, after Which the reaction mixture was: removed Bora: the ice hath and stirred at room temperature for: 2 hours., Methyl tert-butyl ether (300 mL) was then added and 400 mL of solvent was evaporated under reduced pressure. iMTBE (206 m.L) was then added via addition funnel and the solution stirred for 20 minutes at 20 °C, then heptanes (1000 mL) were added within 10 minutes and the reaction mixture cooled to 0--5 °<%... The reaction mixture was stirred for 30 minutes and then the solids were filtered,: rinsed with cold heptanes (0-5 °G, 3 x 100 mL) and dried under high: vacuum to the constant weight' to yield 90.69 g of (F) as a white solid.

Synthesis qfXiompmmd I A solution of (F) (117,33 g) in DMf (900 mL) was cooled in a NaCI/ice hath to :»2 °C. IIBTU (138.06 g)sBG)BT (55.90 g}; (F) (90.00 g) and ice cold DMF (180 mL) were then added to the solution followed by addition of neat D1£A (67.19 g, 509.66 mmol) via a dropping funnel at a rate such that the internal temperature remained at --0 °C, After two hours, neat isopropyl ethyl amine (24.0 g) was added via a dropping fenei, The mixture was stirred at 0 °:C until conversion >99%. The reaction mixture was then transferred portionwise into a dropping funnel and slowly added to an ice cold half-saturated NaHCOj solution (3.6 L) (internal temperature maintained at 20 *€). The resulting slurry was sited with a mechanical stirrer tor 30 minutes and the solids were then filtered and the filter cake washed with ice cold water (2 x 1350 mL):, The solids were then dissolved in diddofomethane (2 ? L) and the organic phase vvas extracted with water {portions of2700 mL) untll^relative pettsenL&amp;jeh:fpr-ddOBt/MBTU was <15% hy HPLC (200 pL solution for HP LG sample). The organic phase was filtered through a plug of SCditno sulfate and subsequently inline '.filtered 'through a pad of active charcoal.

The organic phase was concentrated under reducedipressure arid methylethyl ketone'.(;|:330 hiLJ was added and thesolutlonconcentrated again under reduced pressure,; Methylethyl ketone:(13 50 mL) wasthen added and the solution concentrated again under reduced pressufe. The resulting concentrated solution was cooled to 0 °C until solids were formed; then the '.mixture was heated to 73 °C as orere methylethyl ketone was added fea. 730 mLf until complete dissolution. The solution was cooled to 65 °C and Seeded and the resulting shlutfon/slprfy was cooled at a rate of 0.5 "C/ininute to 20 °€ (Stir rate of 60-70 rpm), The slurry was stirred for a minimum of 5 hours at 20 -€ to allow lor completeerystallkailon. The solids were filtered Off and washed with ice cold methylethyl ketone (720 ml..) and the filter cake was dried under a stream of nitrogen for 1 hour. The solids were transferred into a round bottom flask and dried under reduced pressure to constant weight to yield IT 6 g of crystalline compound I.

Example / /

Methanol (200 mL) was added to crude Compound 1 and the mixture was concentrated to 100 rnL. Additional methanol (275 ml,) was added, along with del on i red water .(7:S''®L)cSnd:4he':mixi6re. concentrated to 400 mL. The clear solution was then seeded with pure crystalline Compound I . stirred and allowed to slowly evaporate under a stream of compressed air to 200 mL. The resulting yellowish solid was washed with deionked water (400 rnL) and !: 1 deionized water-methanol (300 mL.) until it turned white and filtrate turned clear. Compound I was then dried under vacuum for 12 hours.

The resulting compound 1 (1 ?.T'g) was: completely dissolved in methanol {275 mL) by stirring and heating in oil bath (bath set ah§5 ^ mixture temperature less than 65 ?C). Deionized water (75 mL) was added dropwise over 15 minutes, and was allowed to cool to room temperature. Seed crystals of compound '1 were added to the stirred solution, and the mixture was allowed to: slowly concentrate under a stream of compressed air to approximately 2:50 mL overO hoars. The crystals were then iltered and washed with 1:-1 deionized water-methanol pOOCmL), The1 white solid was dried under vacuum for ft hours at 22 *Citd yfols! -crystalline compound 1 (14,0 g).

Example 12

Cmdo compophd 1 (12.1 -g) was completely dissolved in methanol (50 mL) by :stirring::Md;:k^tirig' ia at 85 *€; mixture temperature less than 65 Λ0),

The clear solution was allowed to cool to room temperature and seed crystals of cOmpoundT were added to the solution. The mixture was allowed to crystal lice over three hours at room tempomiofo. The resulting solid was washed with tit deionized water-methanol (500 mL), filleted^ and dried under vacuum for 12 hours to yield crystalline compound tfig),

Example 13

Synthesis: of (F) A flask was charged With (Η) (1 g) and ethyl acetate (20 mL) and the solution was Cooled in an ice hath for 15 mi nutes with: stirring. Trihuoroacetic acid(H) mL) Was then added dropwise, while maintaitring art: Internal temperature of less than 3 °L. After stirring at 0 ?e for 2 hours, the reaction was allowed to warm to ambient temperature and was stirred for two additional hours. The solution was then evaporated under reduced pressure io n thick colorless ml. This crude mixture was swirled gently with·· 10 mL of diethyl ether and as the solution was swirled, fme crystals fomted. After 30 minutes si ambient temperature; the flask was capped tightly and plaoed in the feeder overnight The resulting crystalline solid was filtered and washed with ice cold diethyl ether, and then dried on high vacuum to a constant weight to give tine white crystals of (ft) (670 mg).

Svnih&amp;sis efCompotmd I

Compound (E}(!4.2 g), HBTU (14.3 g), HOST (5.1 g) and DMF (300 mL), were added ίο (F) and the mixture was stirred at roo.ro temperature to complete dissolution.

The reaction was cooled in ice bath for 15 minutes, and DIE A (32 mL) was added over 15 minutes while maintaining an internal teHipcrainm of less dran 10*C, The reaction mixture was then stirred at 0 °C for one hour before it was quenched with saturated sodium 'bicarbonate ¢2()0 mL). The mixture was extracted with ethyl acetate (1.5 L), and the organic layer was washed with saturated sodium bicarbonate (2 x300 mL) and deiooiaed water:(.I x :2()0 mL)- The combined aqueous wash waa esiracted with ethyl acetate (200 mL) and the organic layers were combined (1.2 L)i

The combined organic layers (1,? L) were concentrated under reduced pressure to 100 .ns 1., followed'by addition of methanol (200 mL), and the mixture was again concentrated to lOO ntL Additional methanol (200;mL) wasiddedi deionized water (75 mL) was slowly added with stirring, and the mixture concentrated to dOO snL. The dear solution was seeded with crystalline compound 1, stirred and allowed to slowly concentrate under a stream of compressed air to about 200 mi* The off-white solid was washed until solid turned white and filtrate turned clear With a 4:1 deionized watem methanol (2 1.) and 1:1 deionized water-methanol (500 mL). The resulting solid was dried under vacuum for 12 hours at 22 WC to provide compound 1 (16,8 g).

Compound I waa completely dissolved in ethanol (200 mL) by stirring and heating in oil bath (bath set at 85 mixture temperature less than 65 "43)- The clear solution was allowed to cool to room tempemtore and seed crystals of compound 1 were added to the stirred solution, and the mixture was flushed with air and allowed to crystalline. The mixture was then filtered, washed with 1:1 deionized WaterWthaool (200 mb),and dried under vacuum for 12: hours at room temperature to yield ,10.2 g of crystalline compound 1,

Example 15 SyMimste offfi) A: SOf) ml, flask was equipped with &amp; tTmehanical stirrer thermocouple, cooling hath. :{<3) (12.5 g) was dissolved ίο ethyl acetate (125 mL) anil the clear solution was cooled to 0-5 followed by slo w addition of trifluoroaoetic acid (375 mL) roeh that the mtomaLtomperature was maintaines! below 10 °C. After 5% Pd/C (L2S g) was added and the reaction mixture under an atsfto^here of hydroger) for 2 hours, The reaction mixture was filtered through a glass fiber and nosed with ethyl acetate (50 mL), Thefiltrate was then concentrated under reduced pressure to yield a yellow oil. MTBE (50 mL) was added to the oil and eb-evaporated to yeltewoilat 25 °C, MTBE (60 mL) was again added and the mixture was cooled to -10 °C and stirred ft»' 00 minutes. Heptanes (120 mL) were then slowly added to the stirred mixture and stirring was continued at-10 °C for an additional 15 minutes. The solids were collected by filtration and die crystals were rinsed with heptanes (2 x 40mL) and dried under high vacuum at room ternperaihre (22 *€} to a constant wei^it (10,1 g), %mkesis of €{Mpimnd 1 A flask equipped with a mechanical;sti'rfervlherrpdpouple,·'cooling 'bath;nitrogen irilei and drying tube was charged ’with OMF, (T) (133.9 g), (E) (241.8 g), HBTU (242.8 ;g), and l:I0BT (86.5 g) and themuxture was stirred and cooled to 0-5 °C, DIEA (:156 mL) was then: added slowly over at least 30 minutes, while hiaintaining temperature: between 0-5 °C. Ilie reaetion mixture was stirred at· 0-5 *€ for one hour and was then poured Into a vigorously stirred saturated Sjdlftfift«Ofi^diaiEh;feicarbonaie (3630 mL) and: ethyl acetate (900 mi). Additional ethyl acetate (2000 mL) was added to extract the product and the organic, layer was separated. The aqueous layer was then extracted with ethyl acetate (1930 mL), The organic phases were combined and washed with saturated solution of sodium bicarbonate (2420 mL) and brine: (2420 mL):, dried over magnesium sulfate (360 g), hltered through glass fiber filter and rinsed with ethyl acetate (2 x 36(1 mL),

The resulting solution was concentrated to a sent iso lid under reduced pressure and methartol (725 mL) was added and co-evaporated under reduced pressure to yield semi-solid compound L The erode product was dissolved in methanol (5320 mL) and the solution was stirred while water (2130 mL) was added over twenty minutes. When addition of water was complete, approximately 0.3 g of pure crystalline seeds were added and the methanol/water solution was stirred for tliree hours. The resulting: cs^stalline white solid was isolated by Bltmtidn and the foie white efyftalfom product was pinaed. with a umlhanol/waier solution (I; 1, 1200 mL). The resulting solid was rinsed with rnethadol/water solution (1:1, 1200 mL) and the crystalline product was poured onto drying tray and dried to a constant weight under high vacuum at 27 °G under nitrogen bleed to yield crystalline compound I (230 g).

Example /b

Synthesis of (F) A 100 mL three-neck round bottom flask was charged with (G) (5 g) and dichfomrhethane (IS mL), The mixture was stirred until foe solids had dissolved, and then placed in an ice hath . After 20 minutes, the internal temperature had reached0.0 °C and trifluoroacetic acid added dropwise over 3 tnin. After the addition was complete, the flask was allowed to warm to room temperature. After 2 hours, MIBB was added to the flask (35 mL) and theouxfure was cooled in an ice bath, wherein (ft) began to crystalline during cool ing. Heptanes (65 mL) wefo iften added to the Hash dropwise over 15 min and the: flask was placed in the freexer (~5 ®C). After 1 hour, the solid white product was collected and washed with heptanes (ID mL) to provide 4,57 g of (F),

Bmmpte 1 ?

Synthesis o f Compound / Citrate Sail

Compound 1 (10 g| and citric acid (2,7 g) were dissolved in THF (75 mL) and acetonitrile (SO mL). The solution was then stirred for 2 hours at room temperature, at which time a white precipitate formed, The flask was then cooled' to *10 °C andstirred overnight. Tbe-sblMs -washed with 100 nrt, acetonitrile to give 1:1.52 g of the citrate salt pi compound I >

Example JS

Synthesis {$(&amp;) and (Q)

Synthesis of (I) A suspension of diraeAytliydmxylamine hydmc^feride {10.53 g} 108 mmolf in PCM (270 m.L) under an aimosphefe of argon was stirred vigorously ίθ*:ίΜ>.: hours'; followed·, by .addition' of TEA (10,02 §, 14,75 rob. 1 OS ϊβϊβοΙ) via addition funoel. A solution of Bpe~ bones ne-OB (25.0 :gS:10S rpmo!) in PCM (270 nst) was cooled to 0-°C: followed by dmpwise addition pf isobutyldTiorotonmte (!4.73,g, 13.98 rnL, I OS rnmoS) via addition tonne!, The mixture was further cooled to -20 °C and NMM (10.92 g, 107 rnL, 108 mmol) wps added via addition tonnel at such a rate to maintain the internal temperature below ~10 °€„ After stirring for 5 minutes at -20 *£> the previously prepared dimethylhydroxyiarnine solution was added via1 a wide bore Teflon, cannula. The reaction mixture was removed from the eooling'hath and allowed to warm: to room temperature pyerrdglrt The mlxture was then diluted with water ¢100 ml.0 and stirred for 15 minutes.

The layers were separated and th^afpeoss ;liyear-extracted'':Wi0i OiCM (2 x'JO'mt). Tie organic layers were combined:, washed with I N HO (4 x 150 ai), water (1 x 1 50 mL·);, sat, BaHCCfi (2 x 100 ml), hole (1 x 250 mL) and dried Oyer NajSCL, The !%2SG4 was removed by filtration and thevofatiles removed under reduced pressure to give (1) {28.05 g, 102 mmol).

Synthesis &amp;f (J)

To a 0 °C solution of 0} (10.0 g, M.4 mmol,) in i 00 roL of dry THF, under an atmosphere of argon was added isopropenyl magnesium bromi de {364 mL, f82 mmoL 5,0 eq, 0.5 M solution in THF)dtsm wise using an addition funneL The rate of addition was adjusted suedThat the internal reaction temperature was maintained below 5 aC.

Aler six lonrs tbe reaction mixture was poured into 250 mL of sat. Nl-^CE and 500 mL wet ice. Afterstirring Hr 30 minutes the mixture became clear and the volatiles were removed under reduced pressure and the crude material diluted with BtOAe {200 mL). The layers were separated: and the aqueous layer extracted with BtQAe (3 x 150 mL), the organic layers were combined, washed with water {2 x 150 mL), brine {2 x 150 ml.) and dried pverMgSCL, The MgSQs was removed by filtration and the volatiles removed under reduced pressure. Purihcation by flash chromatography {15:1 hexanes/ EiGAc) gave (j) as a soiid {2 J g, 29.3? mmol).

SyntHmfs qf(K) mtrf{L)

To a 0 °C solution of (J) (S3) g, 19.58 mmol) In 2(i0 ml.of MeOH was added CeClrTBjO (8.75:¾ 23.50 mmol). The solution was stirred under an atmosphere of argon unfiithe CeGL-THaO was completely dissolved. To this solution Was added hlaBH4 (0.88 g, 23,50 mmol) in 10 portions over 2 minutes. The reaction was then stirred under an atmosphere of argon at 0 °C for 6 hour's. The reaction, was quenched at 0TG: with approximately 2.5 PiL of glacial BGAcaod afier 30 minutes of additional stirrinl nt 0 aC the mixture become clear. The volatiles wereremoved under reduced pressure and the remaining oil taken up in EtGAc (200 m:L). The organic layer was washed with water (2 x 100 mL), bride (2 x 100 mL) and dried over MgSCL. The SjlgSO* wasremoved by filtration and the volatiles removed under reduced pressure giving (K) and (L) as a waxy, white solid (4,75 g, 18,5 mmol). Ratio of diastereomers 4.5:1 as detemuned hy/HELCL

Synthesis of (Mf; (19), (Q) ani (Jf*)

To a solution of (K) and (L) (0.025 g, 02)9? mmol) in PCM (1 mL) was added mCFBA (02)1¾ g} 0,10? mmol), The mixture was stirred airDom/timiperatufe^ftir one hour at which time the mixture was diluted with sat NaBCOs (5 mhf> The layers were separated and the aqueous layer extracted with DCM (2x2 ml.}. The organic layers were combined and washed with water (2 x S mL), bride:£2 x S mL):a;nd dried over MgSOa; The MgSTA was removed by filtratioh and the Volatiles- removed:udder reduced pressure to give an oil,

SpalMsisoffif) and (Q)

To a solution of Dess-Martiu Periodinane ((>.023 g, 0,055 mmol) in 1 mL iVfeChf at 5 was added a mixture of(M), (N), (O), and (P) (0,010 g; 0.03? mmol) as a solution in MeClS5 (1 mL). The mixture was placed under an atmosphere of argon and allowed to warm to room temperature while stirring overnight. When complete, a white precipitate had ferined and the reaction was cooled in an ice-bath-and diluted with 2 mL sat. hiaHCOs. The mixture was diluted with 10 roL ofBtCtAe and the solids removed by; iiltering througli a plug of Cellte. The mixture was transferred to a separatory fennel and the layers .separated. Tile aqueous layer was extracted with EtOAe(2 x 5 mL) and the organic layers combined, washed with water ¢3 x 5 mL) and brine (1 x 10 mL) and then dried over NafSCfe The Na^SD* was removed by filtration and Che volatiles removed uhderfedaced pressure: to give a mixture of (Li) and (Q) as a li ght, yellow oil.

Example 19

Aftemiiie Synthesis of f?/; ami (Q)

Altmmpe S^mikesis of:(0)

To a -5 °G solution of (R) (0-200: g, 0)78 mmo!) in pyridine (3 mL) was added 10% aqueous MaGCI (1.5 tnL}dropwi$e at a rate such that the internal reaction temperature remained heiovv »4 *€. After the addition of BaOCl was complete, the reaction flash was placed in a 0 °C hath and stirfed fer two hours.. The mixture was then diluted with .EtQAc (10-nit), washed with:water· (2 x 10 mL), hrine(2 x 10 rnLfand dried over Na^SO*. The Na^CL was removed fey filtration and the volatiles removed under reduced pressure to give the crude mixture of ill) and (Q). Purification by flash chromatography (20:1 hexanes/ BtOAe) gave 0¾} as an oil (Bd)S9 g, 0.216 mmol) and (¾) as a solid (0.023 g, 0,085 mmol).

Exampie 20

Smthesis {>f €ompannE l Synthesis qf {F)

To a tOmL round bottomed flask was added (H) (0.050 g5 4.18 mmol) and DOM (0.80 mL). The mixture was cooled id 0 eC and neat TEA (0.20 mL) was added dropwise. After the addition of TFA was complete the Bask was allowed to warm to room temperature while stirring for one hour. The volatiles were then removed under reduced pressure and the respiting oil was chased with PGM (2 ml. x 2) and the volatiles removed under reduced pressure.

Synthesis ofCompound I

To a 10 mL round bottomed flask containing (F) was added (E) (0.08$ g> O.tS mmolf MeCN (2.0mL), HOBT (0.031 g, 0.23 mmol), and HBTU (0.087 g, 0.23 mmol) and the mixture was cooled to 0 °C- To this mixture was slowly added D1EA (0.077 g, 0 104 mL. 0.6 mmol) and the mixture was allowed to stir at 0 °C for one hour before quenching with saturated NallCXT (5 ml..). The mixture was diluted with EtOAc (15 ml.) and the layers were separated. The organic layer was washed with saturated RaHCOj (3 x Sfflii), brine (2 x 5 mT) and dried over Na^SO^ The NasSO* was removed by Oltratioja and the volatiles removed under reduced pressure: to give a thick oil. To the flask containing· the oil was added DCM (1 ml) and the placed: under high vacuum while swiriing giving €»tnpound 1 {O.IOOg, 0:.14 mmol) as a foam.

Example Jl

Synth esis e>f Compound I

Alternate Synthesis of(S)

To a 10 mL round bottomed flask was added (G) fO.OSSg, 0.18 mmol), formic acid :(2 ml.), and Fd/G (S% wt, 0.05 g). Once the deproteehon was deemed complete by TLC and LCM% the volatiles were tenioved under reduced pressure. The oil was chased with DCM/(2 mL x 2) and the volatiles removed under reduced pressure.

Synthesis o/Compmnui 1

To a lO ntL round bottomed flask containing (S) was added (E) {(3.085 g, 0.15 mmof). MeON (2.0 mh| BOB'! (0.03! g,0.23 mmol), SHBTU (0.087 ¾ 0:23 mmol) and the mixture was cooled to 0 ®C. To this mixture was slowly added DlBA (0.077 g, 0.104 ml, 0.6 mmol). The mixture was then allowed to stir at 0 “C fbr 60 minutes and was quenched by the addition of saturated NaBCQjs (S mL). The mixture was diluted with EtOAc (15 mL) and the layers separated. The organic layer was washed with saturated NaBCQj (3 x 5 mL). brine (2x5 mi) and dried over KajSCh· The NaaStT? was removed by filtration and the volatiles removed under reduced pressure to give a thick oil. To the flask containing tlie oil was added DGM (I mL) and the mixture placed under high vacunni while swirling giving Compound 1 as a foam. "'$%ampie 22

Synthesis qf(M}

Water (214 ml.) was added ίο a three neck flask -equipped with a mechanical stirrer, an addition funnel, and a thermocouple with display and cooled to an internal temperature of-5 to 0 °C. Solid calcium hypochlorite (107 g, 748 motel) was then added over approximately 5 minutes, while the temperature of the mixture is-maintained at approximately -3¾ to 0°C> The mixture was then further cooled to -10¾ to ~5°C and stirred for 10: minutes followed hy addition of NMF (1000 mL) via addition funnel at a rate to maintain internal temperature between Ί Ο °C to ~5 °C. The reaction slurry was then stirred at -10 °C for Ifommufes. (11) (47,8 g, 187 mmol) was dissolved in NMP (40 OmL) and added dropwiseto the reaction mixture while maintaining the internal: temperature between »15 0C and -10 ^C. The reaction mixture was then stirred at ~5 ®C td 0 °C until the reaction was complete: hy TLC. Upon reaction completion, the mixture was quenched by slow addition of 1.0 0 sodium thiosulfate solution (300 mL), maintaining: M:M^fta1:temfSeratureof-l(J1C to ~S °€L Ethyl Acetate (I G00 mL) was then added, the layers were separated and the aqueous layer was extracted twice more. The combined organic layers were; washed with water (500 mL) and brine (500 mL), dried over mapresi urn sulfide* filtered and concentrated under mdueed pressure to a to yellow oil which was dissol ved in hexanes (000 mL) and filtered through a plug of silica to provide (H.) as a pale yellow oil (20.8 g),

Euuivalcnts

Those skilled in the art will .recognize, or be able to ascertain using no more than routine experimentation, numerous equivalents to the compounds and methods of use thereof described herein, Such equivalents are considered to be within the scope of this invention and are covered hv the following claims. AH of the above-cited references and publications are hereby incorporated by reference.

Claims (9)

1. A pharmaceutical formulation comprising a pharmaceutically acceptable carrier and a crystalline compound of Formula (II)

wherein the crystalline compound has 2Θ x-ray powder diffraction values 6.10; 8.10; 9.32; 10.10; 11.00; 12.14; 12.50; 13.64; 13.94; 17.14; 17.52; 18.44; 20.38; 21.00; 22.26; 23.30; 24.66; 25.98; 26.02; 27.84; 28.00; 28.16; 29.98; 30.46; 32.98; 33.22; 34.52; and 39.46.

2. The formulation of claim 1, wherein the crystalline compound has a DSC thermogram in accordance with Figure 1.

3. The formulation of claim 1 or claim 2, wherein the crystalline compound has a melting point of 205 to 215 °C.

4. The formulation of claim 1 or claim 2, wherein the crystalline compound has a melting point of 211 to 213 °C.

5. The formulation of any one of claims 1 to 4, wherein the crystalline compound has an XRPD pattern in accordance with Figure 2.

6. Use of the formulation of any one of claims 1 to 5 for the manufacture of a medicament for treating a disease or condition selected from multiple myeloma, Waldenstrom’s macroglobulinemia, mantle cell lymphoma, diffuse B cell lymphoma, mediastinal (thymic) large B cell lymphoma, intravascular large B cell lymphoma, cancer of the lung, and cancer of the pancreas.

7. The use of claim 6, wherein the disease or condition is multiple myeloma.

8. A method for treating a disease or condition selected from multiple myeloma, Waldenstrom’s macroglobulinemia, mantle cell lymphoma, diffuse B cell lymphoma, mediastinal (thymic) large B cell lymphoma, intravascular large B cell lymphoma, cancer of the lung, and cancer of the pancreas , comprising administering to the subject in need thereof the formulation of any one of claims 1 to 5.

9. The method of claim 8, wherein the disease or condition is multiple myeloma.