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(11) Patent Number: KE 587 (45) Date of grant: 1 0/07/20 1 3
(12) PATENT
• (51) Int.C1.8:
A 61P 19/08, 25/28, 31/00, 33/00, 35/00, 37/00, 37/06, C 07D 303/36, C 07K 5/10, 5/107
(21)Application Number: KE/P/ 2010/ 001070 (22) Filing Date: 03/10/2008
(30) Priority data:
60/997,613 04/10/2007 US and 61/008,987 20/12/2007 US
(86) PCT data
PCT/US08/011443 03/10/2008
WO 2009/045497    09/04/2009
 
(73)    Owner:
ONYX THERAPEUTICS, INC of 333 Allerton Avenue, South San Francisco, CA 94080, U.S.A.
(72) Inventors:
PHIASIVONGSA, Pasit, 2712 Saint Regis Avenue, Brentwood, CA 94513, USA; SEHL, Louis, C., 403 Alameda de la Pulgas, Redwood City, CA 94062, . USA; FULLER, William, Dean, 3085 Driscoll
Drive, San Diego, CA 92117, USA and LAIDIC, Guy, J., 329 Waverley Street, Menlo Park, CA 94025, USA
(74)    Agent/address for correspondence:
MURIU, MUNGAI & COMPANY ADVOCATES, P.O. BOX 75362-00200, NAIROBI
 
(54) Title:
CRYSTALLINE PEPTIDE EPDXY KETONE PROTEASE INHIBITORS AND THE SYNTHESIS OF AMINO ACID KETO-EPDXIDES.
(57) Abstract:
The present 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 are stereoselectively converted to the desired keto epoxides.
 
WO 2009/045497    PCT/US2008/011443
CRYSTALLINE:PEPTIDE, EPDXY KETONE.PROTEASE .ThalIPITORSAND . •
•    - THE SYNTHESIS.OF AMINO ACID KETO-EPDXIDES
Related Applications
•    This .application:Claims 'the benefit of priority of U.S. Provisional Pattat.- - 5    Applidati on, S    60/997,611 filed-"on,October;4,.2007 and -U.S..- ProVisioriaLPaterit
Applidation1Serial.Nw...61/00g,987-filed-onbecerhber.20; 2007 ;.;.th tedchings -of
. are incorliorated by reference in their entirety.
. Background of theInvention
In 'eukaryotes, 'protein .degradation is predominately .mediated througl, the
. tibiquitin.pathway in:which proteins targeted. for destruction are •igated tO.tric '76 amino: ...
    't7 • 2 racid.polypeptideubiquitin.:-Once targeted,: ubignititiated.proteins,thenserkeaS •substrates,    ;
- • • krthe .26S proteasOme;-a niulti catalytic protease; which el eaves•proteins into short
-peptides through the action-of its three major proteol:,,nc activities. While having a
:general-function in intracellular protein turnover-, protcasonie-mediated degradation alz.k • 5    plays a key role•in many processes such-as major i-iiStocornpatibility
: class I antigen presentation, apoptosis,. cell growth    s oo, NF-rd-Li ac-,
. processing, :and transduction of pro-inflammatory signeis.. •    .:.. •
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.Outor rings and 7 different •P subunits comprise - -the inner rings. The a subunits serve as binding sites for the 19S (PA700) ann 118 (PA23)-regulatorY complexes, as .wel• as a physical-barrier ibr the inner protc:ol ylic chamber•formed. by, the two .• subunit'rings... Thus-,' in vivo, the proteasome is believed .to.
•    exist as:a 26S.particle ('the 26S' proteasome"),.- In kivo :experiments have showir.that iral bi ti on of the 20S..fbrm :of the proteasome Cali be readily. correlated to int!ibi ion, of 268 pcoteaso me:. :Cleavage of amino.-terminal•prosequences off3.subun its -du fin g particle: .- _formation expose amino terminal threoni•ne:residuesy which serve as the catalytic .. •    •
•    nucleophiles, . The:subunits resp.onsible,for catalytic-aetivity in proteasomes Anus: possess¬. . an,arnino- terminaLnucleophilic resid•ue;.and-t•lieSesubunits belong,to the' familyof
30.. - - terminal nueleoPhilefNtn)..hydrolases; (where thenueleophilic.N-terminal residue' is; for
..exarriple; Cys;•Ser;.Thr,..and other .nucleophilic -Moieties),    ineludes,..for
-1-
 
WO 2009/045497    PCT/US2008/011443
• example, penicillin Gacylase (PGA), penicillin V aeylase (PVA) glutamine PRPP.. amidotransferase (GAT); and bacterial glycosylasparaginase. In addition to the
ubignitously -expressed .subunits, higher vertebrates' al so possess three interfercni-77 inducible p. subunits (LMP7,.LMP2- and-MECLI), which replace. their normal • counterparts, X, Y. and- Z respectively,- thus aliering the -catalytic activitiOs'a he • • proteasome:.-Thro ugh ...the .nse. of different epti•de -substrates, three ma jorproteol ytie,;
What is needed are improved compositicus, and IT etliods for preparing ;.1.-:;c1 . fOrmulating proteasome inhibitor(s).
.Summary of the invention    •

The invention generally relates to the synthesis of proteasome inhibitors and the •
•    20    preparation and purification of intermediates useful therefor.
One aspect. of the invention relates to crystalline compounds having a structure of Formula (I) or a pharmaceutically acceptable salt thereof,
 
. 0 R6 P    0 R8
N
1;1Y N R5 R2 C7    R7 R4
 
25    wherein
X is 0, NH, or N-alkyl, preferably 0;
''Y is -N-H, N-alkyl, 0, or C(R9)-2, preferablyN-alkyl, 0, or C(R9)-i;
..Z is O or C(R9)2, preferably C(R9)2;
-2-
 
WO 2009/045497    PCT/US2008/011443
RI R2, R3, and R4 are hydrogen;
each of R5, R6, le, R8, and R9 is independently selected from hydrogen, Ci.6alkYl, Ci..jiydroxyalkyl, Ci_6alkoxyalkyl, aryl, and C1_6aralkyi, each of which is optionally
/    substituted with a group selected from alkyl,-amide, amine, carboxylic acid or a
5    •harrnaceutically acceptable salt thereot.earboxyl ester, third, and thioether, preferably .
RS;    and R8 areindependently 'Selected:from    Ci:.6hlydroxyalkyl,. and    .
6aralkyl 'and • eaCh R9.. is'. hYdrOgen; more pleferably, R6.. and R8'are indePendently
R5 and R7 are iridep'eridently 'CI.6aralkyl and each R9 is 13;
-rri is an integer from 0 to 2; and
10    :n-is 'an integer from C to .„-preferably 0 or 1.
.-Anothefiaspec• of tlieinvention- relates to.acrystalline compound of FOrmixla,(III)
 
(III) wherein X is any suitable counterion.
_ 15    Another aspect of this invention relates to methods for the synthesis of amino acid
keto-epoxides according to scheme (I)
         R3
0
R1- N    11
R2
wherein    (I)   

     20 .    selected from a.protecting group or a further.chainof.amino acids, which :itself may
.beOptionallysubstituted„ preferably-a protecting group, most preferably an electron withdrawing protecting group;
R2 is selected from hydrogen and C2_6alkyl;
- 3 -
 
WO 2009/045497    PCT/US2008/011443
R3 is selected from hydrogen, C1_6alkyl, Ci-salkoxyalkyl, hoterocycly1„ aryl, heteroaryi,..: Ci.6neteroaralkyl, and CI.6aralkyl; and
wherein the method comprises a Vereoselective-epoxidation under .epoxidiiing conditions; preferably an aqueous sodium hypochlorite (bleach) or. calcium : hypochldritesolution in the presence of a cosolvent selected from pyridine, icetoritiile,-dirnethylforrriamide,(13MF);:dimethylsulfcixide (DMSO);N: . .
:t.:-.Inetlisdpyrrolidirie.-(NMP),.-dirnethylacetariiide:(DNIA), tetrahydiofuran (TFIF); - 7"1- - and nitromethane.
Brief Description of the Figures
10    Figure .1 shows a DSC .(differential    calerimctry)
crystalline compound 1.
'Figure -2 ShoWS an -XRPD• (X-ray powderdiffraction) pattern of crystalline- • compound 1.
Figure 3 shows a TG thermogram of crystalline compothld 7.
)5    . Figure 4 shows a DSC thermogram cif !.1:norphous compound
DSC thermogram of crystalline compound 1..
Figure 5 shOwS an XR•.D. pattern. ofamorphOus. compound cximpare:"..tb'th;,. ..XRPD pattern of crystalline compound 1.
Figure 6 shows a TG thermogram of amorphous compound 1 compared to the TG 20    pattern of crystalline compound 1.
. -Figure 7-shows a DSC curve of an amorphous si,,,rnple of compound 1. • Figure 8 shows the.XRPD pattern of an-;crphous compound 1.
Figure 9 shows a DSC. curie of a crystalline Compound F.
gure 10 shows an XRPD. pattern of a 4.wystallinc.-cOmpound F.
.25    Figure, 11 shows.a.DSC curve of a. crystalline citrate salt. of compound :1.1.-.
Figure 12 shows an XR PD. pattem of a,2rystalline citrate salt of compound .1.-
- 4 -
 

WO 2009/045497    PCT/US2008/011443

Detailed Description of the Invention
In certain embodiments, the invention relates to crystalline compounds having a structure cifortrtula (1)--or a. pharmaceutically acceptable salt thereof,
 
5
wherein
Xis 0, NH, or N-alkyl, preferably 0;
Y:is.NH, N-alkyl, 0, or C(R9)2, preferably N-alkyl, 0,or C(R9)2;
:Z is.0 or C(R9)2, preferably C(R9)2; ' R', R2, R3, and R4 .are hydrogen; -
each of R5; R6, R7, R8, and R9 is inc=e-:,mdently selected from hydrageui
Cikhydroxyalkyl, C1...6alkoxyaIkyl, aryl, and C    cacirof which is oplOgal
substituted. with a group selected -from alkyl,    arrate, carboxylic acid
.. ...pharmaceutically acceptable salt,thereot_Carbc,IyLeiter, thiol, and thioerne:,
15    -    R.7,. and. R8 are-independently selected.frOm Ci. a1Icyl, C1.6hydroxyalkyl, and
6aralkyl. and each R9 is hydro.gen more.preferably„R and. R8are independently Ci_6alkyl,-; R3 and R7 are independently Ca..6aralkyl and each R9 is H;
m is an integer from 0 to 2; and
it is an integer from 0 to 2, preferably 0 or 1.
20 •    In certain embodiments, .X is 0 •and_RI, R2, R5, and R4 are all the same, preferably
R1',,R2, R3, and.R.4, are all hydrogen. In certain such embodiments, Rs, R6, R7,-and R8 are
. 'independently selected from CE,6alkyl., Ci.6hydroxyalkyl, and ;Ci4aralkyl, more preferably," and re are independent] yCi,..6alkyl arldR5 and R7are independently. C1 6aralkyl.
25.    In certain preferred.embodiments, X is 0, Ri,-R2, R3, and R4 are all hydrogen, R6
and R8 are both isObutyl, R5 is phenylethyl, and R7 is phenylmethyl.
-5-
 
WO 2009/045497    PCT/US2008/011443
.In•certaimembodirnents„ the inventiortrelates to a method .for the preparation.of a
•    crystalline compound of formula (I)-or (II) comprising oneor more of: (i) :preparing a -... solution: of the _amorphous compound; which compound may be prepared.acoording tp,. for example, U.S.. Patent No,: 7,232,818, in an organic-solvent; (ii) bringing the -solution :4;c
5    supersaturation' to. cause fonnation of Crystals; and (iii), isol    e.g.,. by
:-:filtering:the.crystals,-by decanting _fluid:front:the crystals-;•-or by any other.Suitable;:.': separationtechniquel.-Ancertairrembodiments; preparation farther comprises inducing..::.-, :crytallizatio.n:Ara:c6rtain-embo.diments,. preparation further •comprisc.:s washing •-the',)
filtered crystals:; e.g.:, With a §oiv.erA pr.110U-solverst    certain .igiaboc.i.imentS, •
•    1.0 - --: preparation' fUrtlier coinprises drying,,preferably under reduced pressure; such-as under vacuum pressure.
certain embodiments, the-amorphous compound may be 'dissolved Man organic-
:- sblvent -selected, from.acetoriitrile, methanol;: ethanol; ethyl 'acetate.,• isopropanol;.isopropylt: '. acetate; isobutyl acetate, butyl acetate,•ropyl acetate; methylethyl ketone, methylisobutyl,
15    ketone, and acetone, or any combination thereof. In certain embodiments, the arnc.' $,7pus-
compound may be dissolved in an organic solvent selected from acetonitrile,-
ethanol; ethyl acetate, -isopropyl acetate, methyletly,-71 ketone, and recto= e,    at y
combination thereof: in certain embodiments, tilt: Amc,:pitous compound
dissolved in an organic solvent selected. from acetonitrile, •methanol, ethanol, ethyl
•    20-=        :acetate;Triethylethylketone, or any combination thereof: In -certain-embodiments;,-the! • -
organic solvent or solvents may be combined with water,

 
In certain embodiments, hringing'the solution to supersaturation comprises the .additiOT.1;.of an!anti-solvent;:such as water or another polar liquid M SC i';.1 With tale organ: solvent, allowing the solution tO.,cool, reducing the-volume of the sol'utien.; of any.
25    combinationdiereof•    ertain embodiments; bringing the solu'tion:to supersaturation-

'comprises adding ananti-solventxdoling the:solution to arribientremperature Orlow.er, -atidrednc.:i rig the volume of.th,D Solution;•e:g:, by eVaporatirn     th-e.sOution.
certainembOdiments„ allowing thesolutiontb • cool' may be passive
,_Solution to: stand at ambient tetnp erattire). or acti ve: (e.g.:, cooling theuti on 30    bath or freezer).
In certain.embodiments, the method further comprises inducing precipitation or -crystallization. in certain embodiments inducingprecipitation or crystallization
- 7 -
 
WO 2009/045497    PCT/US2008/011443
:coinprises-secondary nucleation, wherein nucleation occurs:in the presence of seed
erystalsTor interactions with th.e-environment (crystallizer walls,-stirring impellers
•    sonication, etc.).
•    In.certain-embodiments; washing the crystals comprises washing with a-liquid selected from: anti•Solvent, acetonitrile,.methanol, ethanol, ethyl acetate,- methYlediyi ketone; acelorie, or. a -combination thereof : Preferably the crystals are washed with- a  -1.,cornbination of.antizsoivent and•the•organie solvent: In certain ernbodiments; theanfi¬solvent is water.
In certain embodiments; washing- thecrystals•eomprises washing the crystalline : 10    -•-con ound of Formula (II) with methanol and water..
•    ,    in:certain embodiments, a crystalline compound of Formula (II) is substantially
pure.: In certain •embodiments,: the melting-point of the crystalline compound' of Formula : • • '(II) is in the range of about .200 to about 220 °C; about 205 to about 215 °C, about 2 Lt. to '
r.
about 213 °C, or even about 212 °C.
15    In certain embodiments, the DSC of a crystalline compound of formula (1i).
sharp -endothermic maximum at about 212 °C, e.g.. r-.7-niitiryz frcm melting F-nd decomposition of the crystalline form as shown in
''In :certain embodiments, the X-faypowderpattern of a crystalline compound-of.'
is•(0-20°)::-6.10; 8:10; 9:32.; .10.1•0;. T1.0.0; 1.2'.1.4; 122.50; 13:64;    :
20        17.52; 13A4;•20.38; 21.00; 22.26; 23.30; 24.66; 25.98; 26.02; 27.84;'28.00; 28.l 6; 29,9k-4:-
30.46; 32.9'8; 33.22; 34.52; 39.46 as shown in Figure 2.
. n certain embodiments, the    therinogram of a cryStallinecompouri of
•    Fon-hula (II) exhibits from 0.0 to 0.1% weight loss in the -temperature- range of 2.5 •to:200 °C as shown in Figure 3.
in certain embodiments,-a 'crystalline col-11pound of Formula (11) is.not solvated: -.._(e.g.,.the•crystal lattice does. not-comprise molecules of a:solvent): In-certain-.aiternative- -• embodirnents; a crystalline compound of .FonnulaJII) is solvated.
•    In zertannerribodiments, the invention relates to a method for the preparation of.an amorphous:conipoilnd of Formula.(II)- comprising one or more of (i) dissolving the , . 30    crystalline compound in-an organic solvent; (ii) bringing the solution to sUpersaturationto- •
-8-
 
WO 2009/045497    PCTJUS2008/011443
causeformation .Of.crystals; and (iii) isolating the.crystals, e.g., by -filtering. the crystals, by decanting fluid.frornthe.erystals, or•by.any other suitable separation technique.: hi • ;certain 'embodiments, preparation furtheroompris.esindttcing certain traddirnents;preparatiOn fiirthercomprises washing the amorphous compound—In
:.5 •    certain 'embodiments the method further co .mprises.drying, preferably under-reduced.- ,
.,,:.pressure,• stich':as.-iitide.r vacuum pressure: In-:certain -embodiments, the in=
-:-...:a.-ci-ystallinesaltOfraleompoundof.Forrnula.(I).or.04;..:wherein the .s41t.coUnterion is
. ...Seleeted frotri.brornidoehloride;.sulfate-,•phosphafe,,nitrate;.acetate,-trithloroacetate2;.:..,. ;.
citrate.; merharresulfonate,•.valera.te, oleate; palmitate, .stearate,laurate,.benzoato„
.10        • .suctiriate„, toSylate, malonate,-maleate, furnatate;'succinate, tartrate, mosylatc, 2-,-
hyd'mxyetansuifoPete; end the.like. In certain-.sni.th.emboclim,.,..--;-s, the salt poi int,an-ion- it:.
;selected from-citrat4tartrate,,trifluoroacetate,'Inethanesulfonate, toluenesulfonate,.:: 'chloride; :and btornidei...preferably citrate:- .    •:,
• In certain embodiments,.the invention.relates.to a method for the preparation,' uf .n- • •
15    - crystalline salt ofa compound of Formula (II); comprising one or more of: (i) preparing
the amorphous compound e.g., according to U.S. Patent No. 7,232,818.; (ii) dissolving tr . amorphous- compound in an organic solvent; -(iii;.brir4nrir the se_stlut-on to supersaturatior
to cause formation of crystals; and (iv) isolating    -crystal    by (raring
....by decanting fluid ,from the crystals,. orby any:other-suitable separationteehnique,.:.:',In.:;:.
a    '.".certaircernbodirrients,-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 under vacuum pressure.

In certainernbbciirnentS, the invention relates to a method for the preparation of a . 25 -*. - crystalline-salt of a.compound of Formula (II), comprising one or more of (i) preparing a.: :Srbiti on of a,cornponnd of FonTrula.Min an Organic _solvent; (ii) adding a.-suitable.7acii-11 ...(iii) brine-,ingthesolution to supersaturation to causeferrnation ofcrystals; and (iv)
rig the crystals; e.g:, bylilteringthe cryitals,-by decanting fluid from the.crysIals,.or • • . by any other suitable separation technique:- In 'Certain embodiments, preparation further .30- •    -comprises induCing.crystalli.zation..-...lneertain embbdiments;.preparation further
comprises4aShing'thetrystals;    with a solvent, ormon-solvent.fluid.. In certain

embodiments, preparation further:coinprises. drying..,. preferably under reduced .pressure,
-9-
 
6/1 2
 
0
0
erl
0
VI
rs1
0
Lf1
0
O
r4
0
0
,'.
0
LO
0
O
31VDS A A8V811811V
 
SUBSTITUTE SHEET (RULE 26)
 

 

 
WO 2009/045497    PCT/1TS2008/011443
9/12
SUBSTITUTE SHEET (RULE 26)
 

WO 2009/045497    PCT/US2008/011443

10/12
_o
 
O0
0
(ScD)
SUBSTITUTE SHEET (RULE 26)
 
12/12
 
(SdD) AlISNUNI
SUBSTITUTE SHEET (RULE 26)
 
WO 2009/045497    PCT/US2008/011443
such:as under vacuum' pressure: In certain embodiments where the salt is less soluble•in a. solvent than the-free .base, adding the acid to a solution may itself be sufficient to bring. . •  the solution to supersaturation:
 

•    In certain embodiments, thesalt counterion is selected from selected from
. 5    . bromide, chloride, sulfate, phosphate, nitrate,. acetate; trifluoroacetate,, Citrate,.    .
•    methanestilfonate; yalerate, oleate-;pahnitate,:stearate,, laurate; benzoate,.lactate,

: . sUceinate,:loSylate,rnalonate, inaleate; funiarate;:succinate;'-tartrate, mesYlate,
    -    hydroxyethansulfonate; arid-the like: In certain subh'embodiinents, the salt counterk)n
selected .fronicitrate,‘ tartrate, trifluoroacetate,". methanesulfonate., teluonesulfonate,-
    10    chloride, and bromide, preferably citrate.
•    ..    In..certain embodirnents,. the -organic solvent 'is selected from 171-1F; •acetonitrile, .
•••-    etherv.and.M.TBE:;.-otany [cornbination-thereotpreferably THF or acetonitrile5or:.-a    •    .
combination thereof •    . •
In- certain embodiments, a crystalline eitrate'salt of a compound of Formula (II.) is. . •
    15 .    substantially pure. • In certain embodiments, the melting point of the crystalline citrat-sa1t• . • .•
of a-compound of Formula (II) is-in the range of about 180 to a.;)ont 190 °C or eveni,'•.5.0;c4':''.-• • 184 to about 188 C.
.    •    embOdiments,.the DSC.of,a crystalline eitrate.salt of a compound _of
_Formula: (1.1)-has a sharp-endothermic maxirnum,at.about 187 °C„    resulting 
20 • 'melting-and decomposition of the crystalline form as shown in Figure 11.
•    •• • • In :certain •embodiments-,•the X-ray powder pattern of.a crystalline citrate salt of a
    _    uoniPound'ofFornihla .(II)is (0-20°).: 4.40; 7.22; 9.12; 12.36; 13.35; 14:34;1554; 1614;:.    ".    •    1--••••••'.:-.
    .f6,54;.17.00; 18.24;1.8..,_58;- 19,70; 19.9Q; 20.3_0;20.42; 21.84; 22.02; 23.34;    23.84; 24.04.; .-   
    ':,9-4.0824.48;.:24,76; 2548.;,26.18; 28•,1:428.20;• 28..64; 29.64;.31.04;:3.1.84;    33.00;    •    .    t
.25- -.    ••. • 34:06;-.3A.3-0; 34:50;15..18; 37...48;,-37.:90;-.3.9.48. as shown-in Figkire' 12:    .    •   

•    - ..• •    hi.certain.embodiments; the:invention •rthktes to a 'cr•ystatline.eOmpound of.    •    .• ,
Formula (ill)
 
WO 2009/045497    PCMS2008/011443
 
wherein )..c: is any suitable counterion.
COUriterion.Selected .frOm bromide chloride, .• sulfate;.Phosphate., nitrate, acetate,.trifItioroacetatei.citrate, methaneSulfonate,.:valerate,,
Staiate,,iautatei benzoate, ••ctate-;:sticcinate, tosylate,:mainnatei.-. maleate,.thniarate,.:succinate; tartrat;.rnesylatei:2:hydroxyethansulfonate, and the like.
(See•or .eaniplei.Berge et al, (.197.7)-"?harmaceunoal: Salts", J. Pham-r. Sci.    • ...
.4n,certain'embodinients...X,js.selectedfroin trifluorOacetateilnethanesulfonate,.,
10 :    *.:toluenesUlfonateacetate,:Chloride, -and bromide, 'preferably: trifluoroacetate. -
-In certain embodiments, the invention relates-to a method for the preparatio::?:01.:.a :crystalline compound of Formula (III) comprising one or more of (i) preparing d compound of Formula (IV), e.g.;- according to Bioorg. Med. Chem. Letter i99, 9, 2'2
88 Or US, Patent Application 2005-0256324, wherein PG is a suitably: protecNi?.:
15    (e.g., Boc or Cbz)
(IV)
(ii). dissoiVingthecompOurid•of Formula (IV) in an organic solvent; (iii) adding a .i.acid;,(iv) bringing the solution- to.supersaturation.to, cause formatioh.of crystals; -and:M.
20    isolating the crystalSi.e:gli-by filtering the crystals, by decanting fluid- froni.the:crystals,. or:•.
•    ' hyany .other.suifable.sep,arationtechnique2ln certairrernbodiments,-•preparation 'further.,    • •t
comprises. inducingcrystallization• In •certain embodiments, preparation further,:
•    comprises washing thecrystals,    with a solvent.or non-solvent 'fluid:- In certain- :
embodiments;. preparation-further comprises drying,.:preferably under reducedpressurei• 25 .    such as under vacuum pressure.
 
WO 2009/045497    PCT/US2008/011443
-In certain:embodiments„the invention relates to a method for the preparation of a: . crystalline compound-of Formula OD, comprising-one-or more of: (i) preparing a
•    5:01:Ution of ar•arnorphous compound' of Formula (I;V); e.g., according to :13iaorg, Med.. e Ghent Letter 19.99; :9, .228-3.-88 'Or U.S: Patent Application 2005-0256324,.. in an organic . . ..;
solvent;, wherein PG is a suitable protecting gaup -(e.g.,..Boc-or Cbz).:
 
0
(ry)
•    •    (ii) bringing•the solution to-superSaturation]to.causelorniation of crystals; and

:    '    :-    • iSolating thecrYStals;:e.g.,;;by-filtering the crystals,i. by decanting fluid froni the crystals,--,,ot
.s:,..lay• any .othet-suitableSeparation-technicine.:,:in.Certaiwemboditnents;-.preparation.furthev,,.:. comprises inducing crystallization. In certain embodiments, preparation farther,
comprises washing the crystals, e.g., with. a solvent or no•solvent fluid. In certain
...embodiments, preparation.fiirther comprises drying-, preferably under reduced. nre.ssUI.6;
•    such as under vacuum pressure.
•    In Certainernbodiments-thee-acidis selected from hydrobromic; hydrochloric,I• sulfuric, phosphoric, nitric, acetic, trifluoroacetic, citric, methanesulfonic, valeric, oleaic,. palmitic„ stearic, laurit; benzoic, lactic,. sticcinici.p-toluenesulforiic, citric, rrtalonic,• .    fumaric; ;311Ceiri c.,.tartatic;,.rnethanesuifonic,.24-1-ydroxyethanesulfonie, and the
like. Preferably the acid is trifluoroacetic acid.
20    In certain embodiments, X•is a counterion selected from .hydronvomide., .
.--,-.:.•ydrochloride,..sulfate,,Phosphate,.nitrate; acetate, trifluoroacetate, citrate,
.methandsulfonate;.valerate,..oleate,epalinitate, stearate,:la.urate,.benzoate, lacta.te,,
succ-inate.,,tosyleate,-malonate,:maleate fumarate,..succinate; tartrate, .rues
......-ydrox.yethanSulfonate, and. thelike: .(See„. f9r example, Berge et .al, -(19:77) • -.
•    25    f•TharrnaCentical,SaltS7%-k.:Pharm.....SCi.. 66:: .1-19.-).-Incertain embodiinents:. X. is selected
from trifluoroacetate;.nethanesulfonate,-toluenesulfonate,e acetate, chloride; and bromide,. preferably trifluoroacetate.
-12-
 

WO 2009/045497    PCT/US2008/011443

In certain enribodimetits, the-.compound of-Formula (IV) may be dissolved in an    Y•,
-7- ,ore;anic..'solvent selected from dichlorornethane, ethyl•acetate, isopropyl acetate,- is.obutyl: a.c..'etate,:butyl acetate, propyl- acetate,•:diethyl ether;:methyttert-butyl.ether.(MTBE),,or. any combination..thereof. certain ernhodiments;the -organic. solvent is s.tlecte.d-frOrn. • dichloromethane,-ethyLac.etate,..IVITBE,;07 .any combination thereof,.prefevably either.:• • •. dichlorometl ane. arid r.4'1'BE.or.ethyl.acetate and MTBE. . ;. .
In certaiiV,enitiodiments;,bringing:th-e solution.•to-supersaturation ecrnprises the    ,:
-anti--;sblvent; •such- as -hexanes. or-heptanes or another liquid 'rniscibleWith. • - • -the organic solvent;tallowing•thesoltation to .cool,:reducing.the'votume-oLhe
1 CI • any:combination thereof .: certain embodiments, bringing the solution
..supersaturation .corripriseE; Fiddingian.    copillig the solut:ori.tt.7.: anrIbient

:temperature9rlower,and.roducipg-the volume of the:solution, e.g.,..by.evaporating..,i.,=;•
:solventfrent•thesolution:„In certain embodiments; the anti-solvent.is.•exanes,or-.,:::,;..,.    =
heptanes, preferably heptanes.
    15    . • -In certain embodiments, washing the crystals comprises washing with a    •
. selected .ftorn anti: solvent, ethyl acetate, .dichloromethane, or,a combination thereof*,: .Preferably the crystalS.are: waShed.vi.fith ',anti-solvent, preferably hep:anes.
•    •    diicertai• embOditnents, theDSC•of a crystalline compound of Formula (III)liasa
sharp endothermic maximum at about 137 °C, e.g., resulting from melting and
    . 20    &Composition of thecrystalline form as shown in Figure 9.
- In certain embodiments, the X:-ray powder pattern of-a crystalline. c,..6npounc: of•
Formula (II) is (0-20°):-.8.84; 15.18; l5:32;-16;20; 16.82; 17:66; 18.26; 19.10; 2120;.
22.58; 23:06;.23,52;.25.32; 26,58; 28.60;.30.08; 30.48; 30.84; 32.20; 36.14, ;37.19.as shown in Figure .10.-
•    -    in 'Certain enibodiments,:sa. Crystailinecompound Of Formula    not-solvated • -
the .crystal. lattice:does not cOMprise moitCtfies of a-solvent), In t-;.e.-rtainalterriativer. embodimentS,...a crystalline ccimponnd. of Fbrmula.(111). is solvated.
'In certain embodiments; the invention relates to a method for the preparation-off-a •
--) crystalline compound of Formula:(II), comprising one or more of (i) preparing- asolution
    3.0    • of compound of Formula .(1V) wherein PG, is a suitable protecting group-('e.g_, Boo or .
Cbz), in a first organic solvent
- I 3 -
 
WO 2009/045497    PCT/US2008/011443
palmitic, stearic;.lauric; b.enzoic, lactic; succinic,.p,toluenesulfonic, citric, .malonic,... • maleic;fumaric; succinic, tartaric,. Triethanesulfonio- 2-hydroxyethansulfonic, and-the    .
•    .- Preferably the acid is tifluoroacetic acid.
• in certain embodiments; -X is .a-counterion selected fromhydrobrc.--irnide, -hydrochloride;.sulfate,...phOsphate,:nitrate; acetate, trifluoroacetate, citrate,' • ,••• methanesulfOnate,• .valerate;-oieate;•almitate; stearate, laurate,benzoate; lactate,:
-.-.hydroxyethanesulfonate,-and-thelikel (-S for example; Berge et al (1.977)
cPhatmaceutical-Salts    Pharm.. Sci. 66:. I    .certain ernbodiinents,. X;is selected    •...•
•    10        .from.trifluoroacetate;.methan.esulfonate,-toluenesul-fonate, acetate, chloride, and bromide;..
pieferably trifi'uoroacetate.
Irceertain •embodirtientS';-theilistorganic solvent is:selected from. . dichlorornethane;ethylacetate; isopropyl acetate,..isobutY1-acetate.„•butyl-*acetate,..propyl.    .
acetate; diethyl ether,-methyl tert-butyl• ether (MTBE); or any combination thereof. Ia.,.
1.5 -        certain embodiments; the organic soIvent.is.selcctalfrorn di•chloroniethane, ethyl •acetate,
.MTBE, or any combination thereof, -preferably. either dichloroMethane and
ethyl acetate and MTBE.
certain- •embodiments,..the.second..organic solvent. is Selected from •acetoilitrile,..-•
methanol, ethanol, ethyl acetate, isopropanol, isopropyl acetate, isobutyl acetate, butyl
'20 • • acetate;•propyl acetate, methylethyf ketone, methylisobutyl ketone,- and acetone, 'or any .• combination thereof In certain embodiments, the amorphous compound taay 1-ye • 'dissolved in an organic solvent selected from acetonitrile, methanol; ethanol, ethyl
..acetate, acetone, or any:combination -thereof - In certain embodiments, theorganics.olvent • . orsolvents may be combined with water.
.25    In certain ..embod iments,•preparati Orr further .comprises.Washing -the crystals. of
either.orboth•of Formula (H) or .(III).- In certain embodiments;. Washing-the- crystals: of a -
 •ornpourict.of Formula (I1) cornprises•-washing with:a liquid seleCtedfromanti-Solvent,: .• -.1.-acetonitrile,..methanol;•ethanol,.-ethyl.acetate, acetone, or a--combination-,thereof. .."..Preferably the crystals.of:a-compound of Form-ula-(II), are washed with a cOn-ibiriation
- 30, ."- 7.7anti‘sdiventand.the..Organic .solvent..71n. certain embodiments,' washing the-crystals.
comprises-washing thecrystalline compound:of Formula (II) with methanol and Water:.i.ln-
- 15 -
 
     WO 2009/045497    PCT/US2008/011443
certain embodiin ents;•washing the crystalsof a .compound of Formula MI). comprises •.
•    -. washing with a•liquid•selected from .anti-solvent,. ethyl acetate, dichloromethane, ,•combination thereof: ;Preferably the crystals., of a-compound of Formula    •are. washed
with anti-solvent, preferably heptanes.

,Irr certain embodiments, preparation further cornprises drying thedry-scats of either
. onboth otTormula: (14: o (III);.preferabl und erreduced-press ure,• su eh, as -under vacuum:. pressure.    r., .
, • In:Certain. embodiments; the invention relates:to a phanna eel)    co; posi Lion
,comPrisifig•a.erystalline,Cerripound.ofForrntils (I):•or (II) .arid-a• phannacciitically, acceptable. carrier,:In, certain embodiments,. -the.,:pharmaceuti cal 'composition is s elected •
•    from-tablets, capsules, and injections..
:This invention also•relates to.•:methods,:for the Synthesis of epOxyketones; such as fermidae and-(IV) above.• Thus, in• another aspect, the invention provides a ibethe'd for preparing amino acid keto-epoxides according to scheme (I)
15         R3 I
2 0
    (I)   

wherein
     R1 is selected from a.protecting. group or a further chain of amino acids, hich itself may . •
be optionally substituted, preferably. a.protecting.group„ Most-preferably an .
    2.0    electron .withdrawing protecting group;•.
is selected from hydrogen and C1..6alkyl; and
13.• is,Selected from hydrogen, C1.6alkyl,    hoterocyclyl, aryl, heterbaryl;
1• ...:.:01:6'neteroaralkyl, and 1.6aralkyl; and -
:.•wherein.the:method .compris es. a- stereoselecti veep° xidation und er.epc x idizing: .!; :    . •
    .• 2-5 . •    •• ctindi•tions,: preferably an.-aqueous Seditim.bypochlorite(b1 ea ph) or calcium -    •
...hypochlorite.solution: in the presence.of a, cosol vent selected from pyridine;
•    • acetonitrile; DM E,    N MP,. DMA; THF, and,nitromethane.

- 16-
 
WO 2009/045497    PCT/US2008/011443
:In certain embodiments; theCosolvent is selected from NMP and pyridine, .. preferably pyridine.
•    in oertainembodiments,- the iepoxidation •s.Performed using aqueous sodium .. ,:hypodhlorite in: the .presence of-a-cosolVent .selected from pyridine,,aceionitrile„DI,v1F, - • ,DMS0,-.NMP; DMA., THF; antFnitrOmethane; preferably NMP or pyridine;_more,. preferably.pyridinert:certaimerribodiments;,the-.,epoxidatiottis,,performed using aqueous•sodium^hypochloriteisolution.. in certain .embodiments, the epoxidation;is performed-Using -IOW aqueous 'sodium hypochlorite .soluti in'the.presencebf pyridine:. yin certain embodiments, , the -,e.poxidationi& peifortned -using .7..calcium hypo Chi
•    0    solution in the presence of NMP.
. In:certain embodiments„.W.is'selected from a protecting group or..a further .chain:.    amine-acids,.whichitselfrnay he optionally. slabstituteth •In certain such .embodintents,    : ,
preferably.an el ectron. withdraw ing protecting group.:    ‘,    „
•    In 'certain embodiments, R1 is selected from t-butoxy carbonyl (Boo); benZoyl • •
15    (Bz), fltioren-9-ylmethokycarbonyl. (Fmoc), trichloroethoxycarbonyl -(Trot), and
benzyloxy carbdnyl (Cbz). In certain such embodiments, Rt is selected from t-but6xy, carbonyl.(Boc),•benzoyl (Bz), trichloroethokycarbOnyl (Trot), and benzyloxy-da.-140':il
,(Cbz); preferably CbZ..or Roc,- -cerfain"'preferred embodiments;    •
In. certain..embodiments, eis.selected..from hydrogen,
20    • heterocyclyl, aryl, heteroaryl, C1_611eteroaralkyl, and C1.6aralkyl: In preferred .
embodiments, .R.; 15 C.r...6alkyl, preferably isobutyl.- in certain preferred embodimf.Ints,    -
. is CI aralkyl, preferably phenylmethyl, 4-hydroxyphenylmethyl, or.2.-phenyl ethyl .
In certain embodiments, the stereoselectiw epoxidation is .performed under.
conditions,that.do 'not:result:in signifi cant _epimerization•of the carbon .bearing R3; :such:    :
•    2';!' . • • thatthere-is-,less.-th-an -1.0%;' less than.5%.,.- less, than'2%,•or.-even lest •than.1% epirricrizatioir •
.of the.carbon bearing-R3• in certain .embodiments,rthe-stere0selective 'epoi-cidation is,
•    .,perforrned.such,that.the.product is,.-g.,reater than about 90%„greater than. 95%,
..98%;"or even, greater than .99% diastereanierically. pure.
In certain. embodiments; the epoxidation is performed at a temperature in the
. 30' _ range of about -1.5.°C to about 10 °C; about 0,°C to about 5 00, or even about-45,PC to about 0 °C.
-17-
 
WO 2009/045497    PCT/US2008/011443
•    In certain embodiments; the compounds in scheme I have the following stereochemistry
R3    R3    h.
-k°
•    R1-1,1 R U    R2 0
L.:•In_certahrembodirnents•..the stereoseleetive-epoxidationtis•perfortned-,suchthat,tbe.,:;,.....-
zreaterthan about.90%.;,greater,,than:9594s, igreater -than 98%, !or.:•evengteater.i,-:.•
than 99%-diastereomerically pure,
•    ..The:usesof various 1\1•Aprotectirig,groups;',the benzyloxy.carbonyi group:or the.,
•    1,•hutyloxycarbonyl group -(Boc:)., various coupling reagents,
...1;:dicyClOhekylcarbodiirnide (DCC), 1;3-diisoPropylcarbodinnide (D.IC), I -(3,
,1.0 ;f7.; =dimethylanaitopro pyl).:3 ethylc arbodiimide ,(ED.C)*;',.1-hydrokyazaberizotriazoletHAX14.,: ,
.;.,:_if,,carbOnyldiimidazoleor,1•.,thYdroxibetiiotriazole„rtiortohydratQ' WORD); and.various..,-,:..,
cleavage .conditions: .forekample, trifluoracetic acid (TFA), HCI in dioxane .
. hydrogenation on Pd/C inorganic solvents (sucl-ias methanol orethyl -acetate), boron ...
-.--tris(trifluoroacetate), and•cyanogen bromide, and reaction in solution with isolation-and. -15 .•    -purification of intermediates. are well-known in the art of peptide.synthesis,and.-aiTL.7,_ •
..equall.y.aPplica.ble to the preparation of the subject-compounds (Greene,.T.V.;
•    • PG.:1\4...Fratectiv..e.- 6i-orips in Organic. Synthesis; aYo. ed.; •Wiley:N ew York; I999):':    •
In certain erribodiments,..the amino. acid ketoiepoxide maybe further modified by    ••. •
•    • -
deprotection theT amine, if applicable, and coupling with a chain of artl3pc,    •
20    Methods-for the ,coupling of such fragments are well known in the art (Elofsson, M.,- et al.:    ••
999).Chemistty. Biology, 6:81 / -822;- Elofsson;    al, (1999) Chernisoy• Biedo_
-6:8-11422); n. a.preferred embodiment, the chain. of amino acids comprises one. to three- , amino acids.
In.certnin embodiments; thechainof arnino:„ecids    structure of formula (V.I). ,
25 ..    Orapham-raceutically acceptable salt thereof
Rs R13
li NYC

5 R12 0    R7
(VI)
-18-
 
WO 2009/045497    PCT/US2008/011443
.•-:-Wherein each kis:independently selected: from e-..0;,C=.:-S,•and SO2, .preferably.e---.0; or Kis optionally a covalent bond when adjacent to an occurrence of Z; .
L is absent or. is-selected from.    C•7--S, and SO2; preferably L is absent or e.0; •
•    M is absent or is ei_12alky1,preferably ei_salkyl;- •
Q-1S,absent'or, is selected from. 0fINH-•,. arid,:N-,01.f6a1kM.Keferably.Q• is. abSerit3 Most preferably•Qis..absent or 0;
`X COOH r r an activated fOritifferedf, prefetablSi X is-e0OH, ::,.:-:.::•....::....ePN(Me)(01■4e),.-most preferably;) is.00011 or e,Qe1;
YfiS all-Sent    selected 'frorri 0, NH, N-c1.04:34., S.; SD, SO CHOR.1.7 t, d r•FiC0i5f17.A.:
    . _l0.    each-Z „i ndepend ently. s elected from .0;, S ; NH, and N-Ci_olkyl, preferably 0; or -
..--:Dis:optiOnallyz, covalent bond.when adjacent to an pccurrence:of A;
R5, R6, and R7 are each independently selected from ej_6alkyl, _6hydroxyalkyl,
'6alkoxyalkyl; aryl, and ei_oralkyl, any of:which:is- optionally substituted with one
or more of amide,-amine, carboxylic acid (or.a salt thereof), ester (including,el...
    15.    .6alkyl•and 01.15alkyl ester and aryl ester), thiol;or thioether substituents; ,
R9 is- N(R'')LQR''' ;
and 12.1..are.independently.selected from hydrogen, OH, and
preferably, RIP:is selected fromhydrOgen, OH; and el..6alkyl, and R.'2` and Ru are independently selected froinhYdrogeri and    preferably hydrogen;
    20    • R'1 is selected from hydrogen; C1.6alkyl, C, 6alkenyl, C1,6alkynyl, aryl, C1:.6aralkyl.,.
_heteroaryl, ei,shete,roaralkyl, R'5ZAZ-Ci,.8alk.y1-, R ''Z-e I _8 alkyl-, ...(R.'5:0)(W.60)P.(=0)0-C14;alkyla-ZAZ-.01_8alkyl- RisZAZ,e1_8alkyl-ZAZ-e1_.
•    8alkyl,.. h.etereocYcl ylivIZAZ-,C1,8 alkyl-, (R113)(1'.Z. 60)P (.-0)0-ei _8 a lkyl-; (R 7)2N-,
•    -01.:12alkyl.-.,-(R1.7)3V,C1,6alkyl;=,,hetefoc:;)icl,Y1W,.carbocytiy1M-, R'8S-02 C -8 alkyl,
    7.5    :    and    5-.S02.41;: preferably ei6alkyl,-e.[..6alkenyl, ei.:Galkyny1;••aryl;•Ci .6 aralkyl,
•    ,neteroary1;;Ck.6heteroaralkyl,•1115ZA-ei_salkyl-;
•    (R. sox.R1.60)p    (11'50)(R160)P(----0)0-e1_3alkyl
;•    heterocycly1MZAZ-elL8alkyl!,.; •
-19-
 
WO 2009/045497    PCT/US2008/011443
heterocyclyIM-,- carbocycIY1M-.; RI.8S02C1Lsalkyl-, and RI 8S011\1H, wherein each . `.:occurrence ofZ- and A is. independently other than a covalent bond; or
R."9- drid.Ri': together are CI -.6alkyl-Y.-Ci_6alkyl, CI _6 alkyl-ZAZ-C1.6alkyli .ZAZ4C r.i6alkyl-: • •
Ci,6alk-yl-A,- thereby forrni ug .a
preferably . CI -..2alky17.Yei_2allcyl, 61::2alkyl•-.ZA-C1_2a•lky4.
,alkyl;=A C1    or    wherein•each pccurrence,of Z :and• A..its
•    independently other than a covalent-bond;    •    ;    •
R.'5'and R16 •are independently selected from hydrOgen, metal cation alkenyl,:.Cr_6alkYriy1; aryt.hetetaafyI,, C 1 bar-alkyl, and -Ci1:6heteroaralky4,
• I:0    preferablyifrOrrithydrogen, metal 'caticiii :and.C)•_•6 alkyl,- or R15 and R ' 6. together
C'1-6,alkyl, thereby forming. a ring; •
each°R'7 is independently Selected.froinhydr6gen'-and
8. is: independently selected from hydrogen, 01-I, Ci_6alkyl,-C1•_6alkenyl, C.1.6alkyny4.• carbocyclyl, heterooyclyl, aryl, heteroaryl,'C1_6aralkyl, and CI.6heteroaralkyl;:
15    , provided that .in any occurrence. of the.sequence ZAZ, at least one member o.fthe.- -
sequence must be other than a .coyalent bond.
and :R.:7.are..selected from .Ci•._6allcyl'orrictaralkyl..,-,,
•    In Preferred ernbOdiments, R.6 IS C1_6alkyl and R5 'and R7 are C1_6aralkyl. In the most .    •
•    -preferred •embodiment; R6 -is isolitityk •R-5, is 2-phenYlethyl; • and R7 i•phenylmethyl.    .    .
20 .    .    cert44-1 tipbod .1311 ent8;    ).tad Q    ;J,r,d .R1.1 iS sele..cte&from.C1:.6alk:.y.1;
6alkenyl, C1.6alkynyl, C1_6aralkyl, and C1_6heteroaralkyl. In certain such embodiments,.
•    RIP is Cf.6alkyl•and RI "is -Selected:from butyl, ally•, prbpargyl, phenYitnethyl, -2-pyfid;.• 3-pyridyl, and 4-pyridyl.
•    •    bther-embodiments,, L. is. S0.„ is. absent,.•and ,R '. is selected: frorn.0 i_oalk.y1    ,
    25,    and:aryl. In:certain such.,..embodiments,•R'.."'is.selected from ineth and :ph enyl;
In:certain:reinbodirnenfs: L is    4S:selected from .C.1._aikyl;
•    •Ci.alkYnyl: aryl, Cri.:6aralkyl,Theteroaryl;    ...iheteroaral•kyl,


    30:    • heteilocYcly1MZAZ,Cialkyl-,.(RI7-)?N-Gialkyl-;-
-20
 
WO 2009/045497    PCT/US2008/011443
darbocycayl.M,-,-W8SGiCr_aikyl-.and.RI.S021\1I1,,'.wherein. each. oceurrence•of Z andA: is •-•
independently -other than a covalent bond. In certain embodiments, •is.C=0, •Q is    .
•    absent, and R11 is H.
In. certain    R•is •Ci_6alkyl, Q. is. absent, and :I., is
In certain such trribodithents..-R1• is-ethyl, isopropyl, 2,2,2-trifinproethy4 or 2 - (methylgulfonyl)ethyl.. • •
•    -In othereintiodinierifs, L is •C=0,.q:is• absent; and R!!-is Ci..6atalkyl: In certain - -siich:erribodimentS;    ' is:Selected from 2-phenylethyl, pheriylinethyl,
methox,YphenY1)triethyl,- (4=chloropheriY1)rnethyi; andv.(4-fluoropheriy1)methyl:-
10 1- '    -It-other elilbodiments, L is &-i0, Q is absent, RI° is C-;:alkyl, and R:1'is aryl ..In
certain such- einbodirnents.,:W1 is substituted or unsubstituted phenyl.
.:•Iiidertain:embodiments, L. is C=0,.Q is absent or 0„ n is.0:0n1.; and :e is
(CH2),.,carbocyclyl. In certain such embodiments, R1.,' is cyclopropyl or cyclohexyl,. .
.In certain embodiments, L and A are    Q is absent, Z is 0, n is an integer._ .
15    frorn.l to•8 (preferably. I); and .R I 1 is .selected from R15ZA-C1_salkyl-,
(R1:10)(R_IC.R(-t-0)0-,C1_8alkyl-Z-C1_galkyl-, and heterocycl ylMZAZ-C1.8alkyk. each: ecurrence of•A, is independently otherThan.a:covalent bond. An. certain such .V •
. • .-emboditherits;R7.-isTheterocyclylIVIZAZ-Ci_galkyl- wh-ere.hetc.Tocyclyi is.sub5tituhed or'.
20 .    unsubstituted oxodioxelenyl or N(R12)(RI3), wherein R'2 and R13 together are Ci_olkyl-
Y-Ci.:6alkyl, preferably CI-3alkyi-Y-C1_3a1ky1, thereby forming a Ting.
•    In.certain prefen-ed embodiments; L is C=0, Q is absent, n is an integer from
8, and R''-is selected from (e0)(R160)P('--0)0-Ci_salkyl-, (R17),NCI_sa11cyl,
and.heterocyClyl-M-. In certain snob embodiments, R' ' is -CI_ - • . • . 25:    3alkYIN(R-12).5 or-C1_8alkylN+(k'7),' where R17 is C1.5alkyL_ in -certain other such
. embodiments, k3 isheterocyclyllv1-, where heterocyclyl is selected froth morphalizo,-
piperazino and pyr.olidino.
,In'certai•ernbodiments, Lis    RI°    Q is selected.from•0 and-NH    •
.• •    and R'.1 iS -selected frOrnCi_6alkyl, cS/oloalky1-1V1;-CIL6aralkyl, and Ci_61ieteroaralkyl-. In-
30    • other ernbodimentS.,:.bis' C=0, •Ri..-Q is Ci_olkyl„ Q is selected from 0 and: NH; and R•'.-1 is
 
WO 2009/045497    PCT/US2008/011443
:C.i.:6alkyl„ -where ic 11-..0.1kyl- is selected from methyl;••ethyl, and isopropyl. -.In further
•    :• embodiments, Lis:C•0,    is    is selected frOrn.-..Q and.NH• and R11 • is ,C•1„,
baralkyl; 'where aralkyl is phenyhriethyl.. In•other.ernbodiments, L is.C=0, R10. is.    •
76hikyi, Q is-selected from .0 and NH; ..and l• is -ei_Jieteroaralkyl.; Aki-refe.heteroaralkyl is.    •
•    (4-pyridyl)rnethyl. .
..In-certain embodiments, .L:isabsent or is-C=0; and-e-.and    togethef-aTe.er--...'-',-•
•    -j6allt5)1,Y,.C16alkyl,. i4alkyl-TA=.01-alkyl;    eaOh-occUrrenCe.-OfZ:';',,
-and A.-is•independently-other than.a covalent bond; therebyforrn ing Ting: .•
preferred_ernbodiments, L iS•-c-----0;14 and    .
. • ID:. .: 3 al kyl-Y    In. another preferred embodim ent; :L-and Q are absent, and• R and . .
TO). to 2-,atiet• are C-i_-3:alkyl-Y-C1..3alkyl.::lit antjtheif    f..f.1.-1-red • erribodiiireilL    .
absent;-•Y is selected:froin:NH .and•N•,:eialkyl;and:R-1:°•and R`Ii.together-are :C i..:3,alkyl4Y,; • ei.3alkYL..Ineriother'preferrecl.emboclinient;t.iskYis..absent,,,andR1.9.•arid together•are.C1.:3alky14Y-Ci..3alkyl. • In another preferred embodiment,. L and Aoare,,-C=0,. •
15        and -R1°• and R' ' together are C1_2alkyl-ZA-C1_2alkyl. • In another preferred' embodiment, L
and A are C=0 and R10 and RH. together are C2_3a1ky1-A.
•    • - In certain embodiments, the' chain of amino acids has a- structure of for2.01a..0,711).- .
 
(-'111)
20    wherein
-.each    independently.selected from C7,0, C=S, and S0a, preferably 0=0; or
A:is:Optionally:a covalent•bond wheri adjacent to. an occurrence of Z; •
eachB..is independently selected frorri 0=0,-.0=5,. and SG), preferably    •
D is absent or is C)..salkyl,. •
25 • .    selected. from 0, NH, and N-Ckbalkyl;
•    yK,:is• absent or' Seledted :from•-:C=0,    SOiipreferably.K is abs ent.or. is•C-,=0; :
L.is.absent or is selected from C-4.0;.0=--S;.and S02; preferably L is absent or C=.0.;    . •
 
WO 2009/045497    PCT/US2008/011443
M is absent or is Ci.salkyl;
Q‘is absent or is:Selected from 0, NH, andNICI_oalkyl, preferably-0 is absent, 0, o•NITI, most preferably Q is absent;
X is. COON or an activated font- theieof,preferably X is COOH, COCI, or 5    CC.N(Me)(0MO, inost preferably X•is COOH. OrCOC.1;
. ; each.V-is independently absent OP is selected from '0',...S;...-NH5.andN-.0 1 _6alkyl, V is absent or 0;
: -.-W•is .absent or is. independently selected. from    -NH, and N•C1.6alk41, nrr-fera4bly
.Y is    or is 'selected from 0,    S',;`40,    CHORI7., and CI-1(.202R17;
10    each Zisindependently selected from 0.;.S; NH; and N,C:i16alkyi, preferably. 0;
...-ZiS:optionally_a.'covalent,bond-when-adjacent    o.ccurrerice.of A; • .
•    R5,-R6,-an-d.R7.are each* independently selected:from
6alkoxyalkyl,.aryl, C1:6aralkyl, and RI6DVKOCI_3alkyl-, wherein at least one of R5 and R' is R16DVKOCI.3alkyl--,
15 • R9 is N(RI°)LQRI I;
R10 is selectedfrom.hydrogen, OH, and-C16alkyl, preferably hydrogen or Cf.6alky1;. ;.•
a. further. chain-of amino acids, hydrogen,. a. protecting .group, ,aryl,. or. hetero aryl, . any of which is optionally substituted with halogen, carbonyl, nitro, hydroxy, aryl, -C1.5aikyl; or 1E.11-1s-seiected from
20    C1,6heteroaralkyl, 111.2Z,AZ,C1.8alkyl-, R'5ZAZ-C1_5alkyl-, (1120)(R130)P(=0)0-
heterocyol-yliviZAZ--
: ,•        (R.120)(R1.3'0)P.(F-0)O-Ci_salkyl-JR14)2N-C1.8alkyl-, (R14)3N+-e1-
heterOcYcly1M-•,.earbocycly1M-.,.RI5S02C14alkyl-, and P,_15.SO2NH; or: .
R.'° and Ril;togethser: ate C1_6alkyl-cYCIL64lkyl, Ci..0Wyl-ZAZ-Cz_6alkyr.; ZAZ-Cji60*1,-
.Z.A2-C-c.:6alkyl-2AZ, or Ci_6alkyl-ZAZ; ,
•    '• 11.1-?'and-Riare.independentl y selected frorn.hydre p.-en;. metal cation,r1alkyl; C,
.6alkenyl,'.C1,6alkyrty•; aryl heteroaryl,.C]:_6aralkyl, and:01,6heteroaralky,1,-...
preferably froin.hydrogen,rrietal cation,:and.C, alkyl; 'or R12--and R13 together are..-•
thereby forming a ring;
-23-
 
WO 2009/045497    PCT/US2008/011443
each R10 is independently selected •-from h.ydrogen.and. C1.6alkyl, preferably Ci.6alkyl;'...-
•    -each. R5• is independently selected from -hydrogen,. -N    Ci_Galkyl; C1..6a1k6I1Y1.;
5alkynyl, carbocycly1; :heterocyelyl, aryl, heteroaryl,:earalkyl, and C. - • 6heteroaralkyl;
5 .    seleotedirom hydrogen;-..(R170)(R180)P(7,--0)W-,R-7.:9B7, heterocyclyl-,. (R.19 )2N-•.;
GB&,.arid,    .where the Ci4alkyl moiety-is. • •
:'-optionally substituted,With-01-1,,CiiialkY1W.(optionally:stibstituted-with
•    •.• • preferably fluorine),.aryl, bete:roar:A; carbocYclyl,    and Cl_62.1.11Cyl;
:... :prefeiabltIeast one-occurrence-of R;•(1.7is otherthan-hydrOgeri; • -
•    .1O .    R.1:7 and .W8 tre•independently selebted '-fr:.'an hydrogen, metal • cation,-    Cl•:.
1,6a1kynyl, aryl.,,Iiteroar.Y1; 7C•ii6aralkyl;. and- c 1.6hetero aralkyl;:' ••, . ,
LL' preferably+ from hy.drogen;•metal,.catitifOand..01.::6alky.1„,•or.R17: and_ R!8.. together,. ate•.7

.C1.6alkyl, thereby forming a ring; and
•    each R19 is independently selected from hydrogen, ORM, C1_6alkyl, C1_6alkerryi; CI_ 15    6alkynyl, carbocyclyl, heterocyclyl, aryl, heteroaryl, C1_6aralkyl, and C•
6heteroaralkyl; and
D, C, V, K, and W are selected Such that there are_no 0-0, N--O, S-N, or S-C bOtids. •
In ..certinrembodiinents,-Rs; .1e, and R7 are' each independently selected- froth. C12
..6alkokyalkyl, 'aryl, C-1.6aralkyl, and.R-16DVKOC11.3alkyl--• •
20 - wherein at least one-.61R5 and R7 is R.16DYKOC1_3alkyl-. In preferred ern.bodinients„one..
of R' and R7 is C1_6aralkyl and the other is R16DVKO.C.13alkyl-,.and    is independeritly..,
•    Cr4;alkyl..7 In. the -most preferred embodiment,. one of R.5 and R7 is .2.-pheriyIethyl.or plienylmethY1 arid the•other.is R 6.p VKOCH::- or RI.3,VKO(C:113)C17.1-, and R6.•is,i!:;obotyl.
. - • 7 - Ineertain.7.embo•dimentS;:7eachkt.57 is independently selected from. hydrogen,:Ci..•
•    •    heter.b•cYclyi,,-,ary).;- hetevoarA
•    and C-'1.6heteroaralkyl.
•    certain•embodiments;. each RI9...is independently. .selected from hydrogen; •C-1.•,_ ,..r s...6alkyl;•.C1_64lkenyl,,ef_'6alkynyl,•7carbocyclyl, heterocyclyl, aryl; heterOaryl    f.6aralkyke. • •    -; •
and Ci_oheteroaralkyl.
•    - 24 -
 
WO 2009/045497    PCT/US2008/011443
in certain embodiments. t.and.Q are: absent and    is selected from hydrogen, a... •
•    . .:further. chain of amino acids; Ci4acyl,•a-protectingigroup,•aryl, heteroaryl,    ;
•    Cr.,6heteroaralkyl.. In certain such.embodiments,.::

p10 is:ei.6alkyl. and. RI I•is. seledted-froinbuty1,-allyt; propaTayl, phenylmethyl; 2-pyridyl;
•    5    . 3,pr-idyl, and 4-pyridyi.
•    .• .    • •    other embodiments, L    I, is ...s.elected-from
and aryl in certain such embodiments R1' is selected from methil ond pheny
•    -;    , In certain embodiments,•L CF-0,and    is selected from    C3 _Galkenyl;
    .ti•    - Oalkyl,,!ary1;,:€116aralkYl;:heteroaiy15-.C1...eieteroaralkyl; RZAC _galkyl-:    •
-    •:    • galkyl,(RI20)(R:' 30)p (--.,=o)O'-c: -g    • • •
    ,.•    . / • (RI0)(e.b)P(:-.70)0-Ci_8alkyl.,Z-C.14alkyl,, RI4ZA-_'Ci...salkyl-ZAZ,c.r.gaik.y1,,,.. •
,carlioeSTaly1M-5:RSOiCi:_8allcylA;7andRISO:y1s1H,. !.Inicertairi- embodiments, -Dis..C=.0,"Q,    •
is absent, and RH is H.
15    In certain embodifrients; RI° is C1_6alkylR.1 I, is C1.6alkyl, Q is absent, and- L is
C=0: in certain such embodiments; R is ethyl., isopropyl,.2;2,2-trifluoroethylior 2- (rnethYlsulfonypethyl.
. In other emb.odiments;'L is C=0, Q is absent; and RI' 'is CI 6aralkyl. In certain : . suchernbodiments.,1111 is selected.from.2-phenylethyl, phenylethyl, (4, . _ . . . , 20 ' • methoxyphenyl)rnethyli (4-chlorophenyOmethyl, and (4-fluorophenyl)methyl...
In other embodiments, L is'    Q is absent, RI° is Calkyi, and R1 is aryl. In..
•    -.certain 'such embodiments, R' is subStituted or unsubstituted phenyl..
•    :. In-certain ernbodiments,..1.5 is C=O, Q is absent .or 0, .and .R .-•-is (CH2LcarbocyClY1.. ;In certain such.. embodiments; .R is :c.yclopropyi .01,cycloheyl:    •    • :•-• .
--•••'.E.?•••    .. • -    • 25 .    •    • -incerndn' emhodinients„    A' are-    Q-is.absent,,Z.-is :0., and RY.is selected...
_r• Trorit'13.12.7,A,Ci _gal kyl-,-. R    I2Z/1/2.,C    _gal kyl    •    .; -
(RI.-z0)(R13:0)P(=-0)(3-0-1_8'alkyr,ZAZ,C14allsyl,-„    1
alkyh,yand heterocyclytMZAZ--Ci..:salkyt, In. certain such embodiments, R' .is    •
•    heterocyClylkIZAZ-Cf..galkyk-where heterocyclyl is:substituted.or-unsubstituted.•:
- 25 -
 
WO 2009/045497    PCT/1JS2008/011443
oxodioxolenyl.or N(R39)(R:1), wherein•R2° and re! -together. are C,_ealky1-Y-C1_6a1kyl,: Preferably Ci:.3a1ky1,Y-.C1_3a11<y1, thereby forrning a.•ring,
,,In•Certain preferredembodithents,'L is-C=0,;:Q is absent, and R11,• i's Selected; from-.
-• *326)Cki    (le'l•INC;i8illkYk.•:(14,4)31‘417(CI-12);-,‘-,,andineter6cycly-1-1.Vi-.
Odim eats, R    .;C ■,.,8alkyiN±(R!4)3, .wheteR'4=is
othe• such ernbodirnents,,le.1,is hetcrocyclylM•t,.wh.ere,•-heterocyely1
•    -    is.-seleeted fromznorphOlino; piperidino,    •    '
. • In'',";:ertain einbodinients, L    RI°. is :CI..6alkyl,.Q is-se,lected.fronr 0.and
...and.Rklis,Selected•.frorn •Ct.2alkyl?,scyeloalkyl-IVI,,C11:oraalkYl.;,and.C.,`..ateteroaraalkyh'I•n
otherenabodit41ents;-1.,    0 Is selected frorn 0    is •
is:selected-frOin methyl, ethyl, .and.-isopropyl:,,In:ardher .-tinbodinieritS;:LiS•C•=0,    iSselected:frbny 0 and NH. end:R1,:14s,C1.,.„,,-
isPhenylrnefhyLin other.emhodinients;    ,
'6 alkyl, Qis selected from. 0 and NH; and R '1- is -Cf.L.6heteroaralkyl, where hetept:majIzAis•
    15    (4-pyridyl)methyl.
.in certain embodiments; L is absent or is. C=0, .and-R'9 and R'' togethetar":.-C,i,i
e1-6alkyl-ZA-01.6alkyr,. or C 6ally1,-A., 'thereby formirTgL.,
certainpreferred.embodiments; L is .C=0;•.-Q and Yare'•absent,.and R19and•Rtogetber • • are Ci...3alkyl-Y-C1_3alkYl. In another preferred embodiment, L and Q are absent, and R'°
    -720-    •
and R''    are-C    In:another preferred embodiment, -L    .•
all-stilt, Y-    ected f•om• NI-Land    ar.,,d    R'.'    •
3alk-Y1-Y-C1alkyl. In another preferred embodiment, L is C=0, Y is absent; and -RJ5-and.
R''...togetherare    •In another preferred- .:,'rrkt.l.odi-3-11<7.';;2"    ;sid A
•    C=.0:"and.-R!° and R' together are CI-2dikyl-ZA-C1_2a1kyl.• •• In anotlaer'preferred •
    L and Aare•    C=0and.R1• and R'''. together. are C2-3aikylA.`
Certairt.ertihodink.-nts,.Ris    'fn certain suCh..
•    ernbodirrient,.:D,;    K, and:.W 'afevbsentTither`sitach:embodiments;    -Live
•    ahsent,:Dis    is. 0: ; In yet other such c..embodiments, D. is CI _salkyl,• •K.is

•    C=0, and V and W are 0.
•    .    1.- -In certain embodiments,    is-Rt7GR-. In preferred.    B is C=0, G is•
0, D is Cl.salkyl, V is 0, and K is C=0.
-26-
 
WO 2009/045497    PCT/US2008/011443
in certain.ernbodiurents; Rn is.heterocycly1-. In preferred. such embodiments, D-is : •    _galkyl,- in:certain:such embodiments, -V-is 0, K is 0=0; and heteroc.yotyl. is
oxoinxolenyl r Other sUch.--.embodiments, V is absent, is absent or 1. s C=0,    •
' • ...'-heterocyci yl 1\1(R2.°)(R2); where P22°.and.R21 together are 317:-.1 • 3-WB-1,,•.orB-,a-..-T-4    ,
Is:absent or is ..selected .from 0, NR.i.7,c .SO;S02,.CHO:R.39 CHCO7W.7., 00 ni72,    • -
- • '    lis,,absent or is Cj_3alkyl.
.    : .    r. .7 •    certain:embodirbents;,R16-is,(R    or EIV?-)3N--,-, and preferably y
such-enibodiments,-D is-C1.8alkyl and,K-is-absent-or-C=.0. In certain    ,
• ..ernbOdifrients-Where_V-is. absent and    is (W.,9:)21'4-:,,D .absent K is absent    -
1,0    preferably K is 0=0.
- • .    certaimrimbodiments; R'' is R'9S0-5,:3BG-:! inpreferreci sucherrbodiments, ;5,    ;
..,.,,,. .•.    is C=0, D, V, and K are.abSent, and G is NH or:NC1,6alky1.1
In Certain erribodirrients,    R..(7GB.Ci_galkY1=. In preferred ernbodimentk B is
. C=0, G is 0, and. the Ci..8alkyl moiety is optionally substituted with OH, Ci_0. 15    (optionally substituted with halogen, preferably fluorine), Ci...8alkylW, aryl, heterOaryl,
carbocyclyl, heterc cyclyl, and C:6aralkyL in certaiii such. embodiments, the    nW4. •
moiety is an urisubstituted, mono-, or disubstitured C. alkyl.    .    .■;

. In certain embodiments, the chain-of amino aCidS has a structure of formula (VIII) . : - - or (IX).or a pharmaceutically acceptable salt.thereof ,
11
X
20    .R6    R6
(VIII)    (IX)
wherein
fdepend tRitly an, aromatic or. heteroaromutic group optionally substituted- with . to 4. substituents;
.25    -I is abent or.is selected-korrr C=0, C=S; .and S02, preferably SO, or 0=0;    •,
'X is COOl-i-or an activated-form thereof, preferably X is COOH, COC1, or CON(Me)(0Me), most preferably X is COOH or COO-,
Y is absent or is selected from C=0 and S02;
- ?7
 
WO 2009/045497    PCT/US2008/011443
Z is absent or is Cl .6allcyl;
R5'and'R6 are each independentfy selectedfrorn CI _6alkyi, c1,6hydroxyalkyl, C1_6alkoxyalkyl; •aryl; and Ci -6aralkyl,..any-of which is optionally:substituted with.. one or inoiecif amide; amine, carboxylic acid-(or a salt thereof), ester (including
Ci_6alkyl ester, Ci.salkyl ester, and aryl ester); -thiol„ or thioether stbstituents;
R9 is N(RI°)L-Z-R";
-ARIPiS selected from hydrogen; OH; C1.6aralkyl-Y-; and C16alkyl-Y-, preferably: hydrogen;
R" is selected from hydtogerc.OR12,    carbocyclyl, and heterocyclyl:
and
I? is:selected, from hydrogen, Or_6alkyl;, and: C•_6aralkyl, preferably hydrogen.    4: •    :•
•    •    certaiii•embOdirrientS, I:: is- 'selected from     C=S,-:and SO2,. preferably 02', Lir . : • -
C=0 .
 
In 'certain embodiments,.R1° is selected from hydrogen, OH, C1.6aralkylOnd CI_ 15 •    6alkyl, preferably hydrogen.
Tn. certain embodiments, R" is seleCted from hydrogen, Cs,.6alkeny:: carbocyclyl, and heterocyclyl.
1-    hi:certain ettibodiments,:12.5.and le are each independentlyselected frora..C1:.6alkM,
C1_6hydroxyalkyl, and Ci_6ara1kyl. In preferred such embodiments, R5 is C    and .R6
20        is C1:6aralkyl: In more preferred such embodiments, R5 is isobutyl and 'R!'. is
phenylmethyl.
-Incertain.embodirn&xts, RI° is hydrogen; L is-C=Oor SO2, RI    Ar-Y-, and each •..
•    ' Ards' independently selected from. phenyl;    b,enzofuranyl; naphthyl,. quinoliny1;-.:.
pyrwzyl;    ;In:certain such embodiments, 4s;,,r,:may.:,.
.be.substituted withAr-Q- wl:iere'Q is sel ected from 4,ditEe.c;tbqad,_'07, and C    4R-
certairt•other such etabo'diments- where Z•is.Ci...6alky14,Zniay be. substituted, preferably, - v,ith Ar, e.g., phenyl.
In certain embodiments, R!°;is-hydrOgen, Z is absent; Lis C=O or SO2.,- and-R11 is. Selected from Ar-Y-.aridheterocyelyl. In certain-preferred such embodiments,
-28-
 
WO 2009/045497    PCT/US2008/011443
heterocycly1Y is..selectedfrom.chromonyl,- chromanyl, morpholino, and piperidinyh In certain other preferred such embodiments, Aris selected from phenyl, indoly1,. benzofuranyl, naphthyl;-quinolinyl,:quinolonyl,- thienyl, pyridyl,-pyrazyl; and-the
:..ln,ceritlinernbcdinients,le hydrogen,.L -is-    SO2,- Zi absent,. and IV Is
V.ihere.C4:60-kenyl- is a substituted vinyl group; where the substituent is
•    ,preferably:anarytOrbeteroaryl group; morelpreferably -a phenyl group -optionally.: ,-
•    : substituted with meth four substituents.
'    certainernbodiments, 111.2 is selected from hydrogen and Ci..6a11,..j.. In:certain.
preferred.:sud•ernbOdiments, 1Z.1..i.s:s•eleCted:from4iydrogen.and- methyl.- ,-In more •-
•    preferred such embodiments., -R12 is hydrogen..
-..In:certain:preferred embodiments; the.chain of amino acids has a structure of  formula (X)
 
(X)
15    Xis COOli or. am activated form thereof, preferably X is. COOH, COCZkir •
•    CON(Me)(0M-0), Most preferably X is COON -or COC1; •
R6, and.R7-..areindependently selected-from    01_6hydroxyalkyl, •
•    AL-,alkox.yalkyl, aryl; and C 1..6arallcyl,. each of which. is optionally substituted with' a grom. :
•    selected. from amide; amine, carboxylic acid or a pharmaceutically  acceptable. salt thereof, 20    carbox;.1 ,ester, thibl,•and thioether,•preferablY.R.,C)is..Ci_cialkyl and    andie 4t-t:
6atallkYL-most preferably, .K6 is isobutyl,.is.2-.0benyl ethyl, .ancl
iS 'afurther-chain::of amino acidS;liydrogen.,••ClaCyl,ia protecting group; -aryl,. rN h eteroat91; :wherestibstittients-inCludeThalogen; carbonyl„ nitro,thy:droxy,    Ci;
preferably'R9-is7Cf:6acyl, _niost.preferably-R9is acetyl.    .    ••
25.•    3n-certain.prefeiTedembo_diments, the chain. of amino acids has -a 'structure 'of • •
-formula (X1) .or apharinaceutically acceptable salt thereof,
 
- 29
 
WO 2009/045497    PCT/US2008/011443
 

 
ort,
 
wherein
s :absentbri§:selected from—CO, or .-C(=-7S)0;
5    .or an activated forni'theieof,.preferably X is COOH, COCI, Or.
CON(Me,,,(0Me), MOS t preferably `Xis  COOH or COQ
Y is NH,.N-alkyl, •0, or C(' 9)•j,. preferably N•-alkyl, n, or r (R9)2;
Z is Q or C(R)2, preferably 0(R9)2;
-and Ware-independently selectedfrom hydrogen and a•group of formirla .10    (XII), preferably; R'TR2-,- and R3 are all the same, more preferably-RI; R2, and R3 are all •
hydrogen;
0
11,0R1-,
,L.
OR"
R1° R11
(XII)
•    • each    ,.    is independent] yselected from..hydrogen,    CI- • •
15    6hydrOxyalkyl, Ci.6alkoxyalkyl, aryl, and ci_oralkyl, each of which is optionally    •
substitutedwith.a.group selected from alkyl, amide, amine, carboxylic acid .or a. .
-pharmaceutically acceptable salt thereof, carboxyl ester, third, and thioeth,er, preferably,
R6„ .and R7 are independently selected from C1_6alkyl, C1_61-iydroxyalkyi,. and CI-
6aralkyl:and•.eachle is hydrogen, more preferably, R6 is C1.6alkyl, Rs and R7 are ..! 20    independently Ci_oralkyl and each R9 is H;
and R'' are indepe'nderitlY Sele.cted.from hydrogen and CL...6alkyl,,Or RII) and • '!.together-form a 3-, tof6-membered ,carbocyclic or heterocyclic ring;
;    andR1:3. are independently. selected from hydrogen; a metal cation,
•    , or-R:.!:;:and R13 together represent Ci4alkyl, thereby forming a fin ;
25    m is an integer from 0 to 2; and
-30-
 

WO 2009/045497    PCT/US2008/011443

nis an integer from 0 to 2, preferably 0 or 1.
•    In certain embodiments, X is.0 and RI, R2, and R3 are all the same, preferably RI,
and •R.3 are all-hydrogen.. In Certain such .embodimentS; Rs-, ie; and. R7 are
.... independently, selected froin CI _6alkyl,. ci..6hydroxyallcyl, and .C1 _6aralkyl, .more: 5ti:.-.preferablY„.R6 is Ci..isalkyl and R5 and R.7. are independent-lye l'::6aralky1.-
In certain.preferred.embodiments;;R~;'_R2;    -are..all hydrogen,  R6 and. e.    ...-. •
' - both •isobutyl,RS is-ph'enylethyl, and R7 is phenylmethyl..
In. certain.embodiments; 'IV, R:6, and .R7..areindependently selected from hydrogen, .;. .G1.:6hydrOkyalkY3;CI:6:alkokyalkyl, ary1;.andEl_6aralicy13. each of.Whieh_is.;,-...
•    10- - : ....optiona.11y.substituted.with.a Rroup. selected 'frorn.alkY1,-ainide, arnine Carboxylic.,acid . apharmaceutically:acc.eptablesalt•thereof,„ carboxyl ester; thiol,..and thioether:". In pertain ...-f.:ernbodirrientS; at.-leasfone-of:W.:and le; is...Ci.6atalkyl-substituted. with:

•    referably shbstitined withperhalbalkyh fn certain-slieheriabodirrients,IR7,is Gi.;6aralky1.- • -substituted with trifluoromethyl.
15    In certain embodiments, Y is selected from N-alkyl, 0, and CH2. In certain such
embodiments; Z is CH2; and m and n are both G. In certain alternative such.erntibtliments,
•    Z is CH2, rn.is 0, and-n is 2•or 3. -In yet another alternative such embodirriem.,Atis 0, in is 1, and n is 2.
—hi-certain preferred- embodiments, the. chain. of- amino. acids has a' strutture- of . 20    formula (X111)    •

 
-wherein
.eaohAr is independently. an aromatic -or hetero,aromatic group Optionally: 25. substituted with 1 to 4 substituents;
each A. iS independently Selected froth    and SO2, preferably C=0; or
A. is: optionally a covalent bond when adjacent to ail occurrence of Z;
- 31 -
 
WO 2009/045497    PCT/US2008/011443 -
B is absent or is N(R9)Rl°, preferably absent;
-    I< is:absent or is selected from CO,=.    SO2, preferably SO2 or. CO;F--
•    M. is absent or is Ci_valkyl, preferably Ci_salkyl;
Q.is absentor is selected from 0, NH, and N-C1_6aBly1;
.5    'I.% X isCOOH or an activated form thereof, preferably X is COOH COCA, or..
;'CON(Me)(0.Me),-rribst Preferably X is COOH or.CO•C1;-    .
Y iS.absent or is selected from C=0 and SO2;
each:Z iridependentlY,sete,cted frorri'0,.S;NH, and:N-C1_6alkyl,, preferably 0;
-Z optiOna?ly a covalent bond •..yhen adjacent to an o,-;:curreiice cf A;
10    .,-,R1.-is.Selected:frorri:H;,:-Ci_6alkYl-B;'e.f--6bydroxyalkyl, CI _6alkoxyalkyl, aryl,
C1_6aralkyl;
. R2 iS selected from aryl, Ci.6aralkyl, heteroaryl, and Ci_6heteroaralkyl;
R4 is N(R5)L-Q-R6;
R5 is selected from -hydrogen, OH, ci_6aralkyl, and Ci_6alkyl, prefera1 l4bidrogen; 15    FAG is selected from hydrogen,- C1-6alkyl, C1.6alkenyi.    Ar- Y
,carbocycl•ly.lieterocyclyl, an NAerminal protecting group, aryl, C1_6aralkyl, heteroary1;.-...
'C'r,61heteraiialkyl; RJ•EA:1:•C:1•.:§alk"5/1-=-;:W 4ICI•ialkyi4AR 0)(R '20)P
ZAP',-c,..salkyl-, RI 1•ZAZ-Ci_.8alkyl-ZAZ-C    heterocycly1MZAZ-C1_
(R110)(R120)P(=0)0-01_8alkyi.-; (R0)2N-C1_12alkyl-,
20..111Cterocy0y1W carbocycly114-, R1:t02C    and P.)48021\114; preferably an1:1'-.-
. caOping group, more preferably t-butoxycarbonyl or benzylexycarbony1;• or
:R5 and .e.together :are:Cf..6*alkY1-.YCI._6alkY1,-, CI _6alkyl-ZAZ,Ci.6alkyli,
6alky1,ZAZr.c1.6alkyl,:ZAZ7C1:6alkyl.-ZAZ,.or C .6alkyl-A,..thereb y forming a ring;•:
R7. -lif,Selected.from. h?./drog,cn.;-C.1_60kyl;:and:C; -6 aralkyl, preferably hydrc)gcn;_.. • 25'; le..is'selected frOm %hydrogen:OH; C1. alkyl, preferably C1:_6alkyl;
N-terminal protecting group;
R-1-1- and R12.are independently selected from .hydrogen; metal cation, Cr.6alky.1, cr.    .
Ci_olkynyl, aryl, heteroaryl; CI_6aralkyl,•and. C1.6heteroaralkyl, preferably from
- 32 -
 
WO 2009/045497    PCT/US20081011443
hydrogen, metal .cation, and Ct.:6alkyl,.or R.' I" and -RI. together are Ci_6alkyl, thereby forming a ring;
each R13-is independently selected from hydrogen and Ci_falkyl; preferably C. • 6alkyl; and
5 •    R11.is Independently selected fitrn 'hydrogen; Ci_fAlkyl,
oarbocyclylrheterocyclA.-aryl,,heteioanyt;:eir.:6aralkY1-,-and-C-1_6heteroaralkyl;--: .„
•    .--.---procHdedf-thatirr-any occurrence of the .sequence ZAZ.,:atleast one member of.the sequente:must- be other than a covalent bond.
•    In Certain. embbdiments, R' .selected from -.•C I.6a1ky1..-13 and Ci_oaralkyl.. :In. 1,0- •    6c-;,--tah..s.i, c;h:i-j-:q.:6(iiit.erits:, P. is -substitO.Lt:d. with. one th-.    fl-orn
hydroxy;:b alogen,, amid e.;: am ine,- carbox,yli e. acid.,(or la:Salt: thereof) ester.. (including.C.r..„
••--:.6alkY1' ester, Ci,ialkyl ester, And. aryl. ester),Jhiol, 'or_thioether. In .certain Prefen-ed.:Such-.••-: ,embodiments, .R1 is substituted with one or more Tsubstituents selected from c4boxylic
acid and- ester. In certain embodiments,.R1 is selected from methyl, ethyl, isopOpyl,
15    carboxymethyl, and benzyl. In certain embodiments R' is -CI -6alkyl-B and CI _6aralkyl..•
In certain preferred such embodiments, B is absent:
in certain embOclimerits, R2 is selected. from C1_6aralkyl and C1..6heterci,i•Okyl. • In
certainSueltembOdiments-;_R2'is- selected-from .Cf...6alkyl-phenyl,,C.,:6alkyWndoly1;:01:...
OlkY14-thierbYt;'Cf.-6•alkYl=thiazoly1,,.and:Cr-6alkylis,othiazoly1,-wheteinthe.alkyl Moiety. 20 • • I may contain' six, five, -font, three, Iwo, or one carbon atoms, preferably one or two. • In •
. • certain Such embodiments, l•e.is substituted with one or more substituents. selected.from hydroXy; -halogen,.    amine,- carboxylic:: acid (Or a salt thereof, ester (including CI...
6alkyl ester; Ci..5alkyl ester, and aryl ester), thiol, orthioether. In certain such
:-..embediments,--R-2...is.substitutedwith:a.substituent selected. from alkyl., trihaloalkyl,1•• • .-    •
2$-alkoxyi hydroxy,. or: cyario.. In certain such embodiments, R2.is selected from    -
phenyl. and.    In certain preferred. such ernbod inients; .R2 is selected from,
 
WO 2009/045497    PCT/US2008/011443
 
D
DNir
 
R
•    • R = H or any suitable protecting group
•    wherein...D.:is selected from H,•0.1v1e, Q.134%.0q: CN, .C.F3 cmd    in certain .i.
erribodiments.,D. is selected :from H, 0114.e, OH, CN.„CF3 and CH-3...
•••••, :::...in:certainpreferred.sticirembodirneritS- where. Di s..attached to •.a..si x,:memb ered
.5    • : ring, D is attached. at the 4-position telative.to'thejpoint of.attachrnent; preferably
exchiding.embodiments where the 4-position of the ring is occupied by the nitrogen of.a pyridine ring.
_ in certain embodiments; R5 is hydrogen, Lis C=O or SO2, R6 is Ar-Y-pand each :.Ar.is independently .•Selected from ph.enyl,indoly1,-benzofuranyl,liaplith.yl,
quinolonyli-thienyh -.p,yridy,1;:p.yrazyl; and. thel ike.:..drrcertain such: embo.diments,-Ar may -.7,    •••
•    -• r -be: sub stituten,    where.F.:- is-selected frorita direct bond,    and, .0    r•-7:    • !'•-
. certain, other such..ernbo•diments where .O.is C1:6alkyl; Q may be substituted, preferably
•    with Ar,. e.g., phenyl.
In Certain eMbo.dime,niS,k5 is hydrogen, Q:is absent, Lis C=O• 1,.•;03,Elrit.1 15 • . ,s,elected:from Ar-Y and. heterocyclyl:• In certain preferred such embodiments, •
•    . heterOcyclyt is: seleCted from' chrornonyl, chtorrianY1, -i-norpholino; and -piperidinyl.. In .•• ...:eertainotherpreferred such embodiments.,-Axis. seleCted from phenyl,. indoly1 - • benz'ofuranyl,.naphthyl,..cillinOtinyl,•quitolonyi,.thienyl,.pyridyi, pyrazyl, and-theIke... •
In. certain embodiments, R5 ishydrogen,..L is G=0 or .S01, Q is.absent, and R5 is •1
•    20 . • 0•11.6alkeny1;4here•Ci_6alkenyl is-a s'ubstituted.vinyl group where the Substituent 'is • preferably, arraryl•orheteroaryl group, more•pre.ferably- a phenyl group:optio.nallY,-.:;.•..• substituted with one to-four substituents.
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WO 2009/045497    PCT/US2008/011443
In certain embodifnents, L and •are absent and R6 is selected from Ci_6alkyl,
•    -Ci_oalkenyl, C.1    c I -6 aralkyli and C1..6.heteroaralkyl. In certain such embodiments,
R5.is,C1.:,alkyl and 16 is. selected from ,butyl, ally1,.propargyl; phenylmethyl, 2-pyridy1,- 3•pyridyl, and 4•pyridyl.
5    in other embOdinients, L is SO9, Q is absent, and R6 is selected from Ci..6alkyl and
aryl: Its certain:_such,embodiments; R6 is selectelifi-orn Methyl and-phenyl.. -.    . • •-•
In certain embodiments, iS .C=0- and_116
:01:6alkyrtyl, aryl, CI    heteroaryl, Ci.6heteroaralkyl, RI IZA-C1..8alkyi-,
(R    (-_))P,(---20)&C    (R1 I    I 0)(R 2'0)P    . -
CI    (1 110)(1 !2:0)P(L-0)042.1-8a1ky1-Z7C178alkyl-; RI 1 A.. t.8a1_kyl--ZA-7-
heteroOyclyfiAZ4L-C1.8alkyl-, (RP.)21C1-C1_8alkyl-',:.(11,1: t!)31.,,,+- ,8 alkyl-, heterocycly1M7,, - •
carboCyClY1M--.; R:1,4.S.0`,Ci.::8alkil and leS02NIIL.,..*Iierein each occurrence .6f Z    AdS• •
•    .indeperidendy other. than 'a,cavalent bond., Iii certainemb.odiments, L
absent, and R6 is H.
15 •    In certain embodiment, R5 is C1_6alkyl, R6 is Ci_6alkyl, Q is absent, and L is C=0..
In certain such embodiments; R6 is ethyl, isopropyl, 2,2,2-trifluoroethyl, or (methylsulfonypethyl.
In other emboPTIPntS,.L is 0-70;.Q is..abseht): and R6 is Cl.6aralkyl. In certain...-such embodiments,..R6_is.selepted.from, Z-phenyletbyl, phenylinethyl,..(4:-
20    methoxyphenyl)rnethyl, (4-chlorophenyl)methyl, and (4-fluorophenyl)methyl.
In other embodiments,.L is C=0, Q iS.absent, R5 is Ci_6alkyl, and R6 is aryl. In certain such ernbodiments',R6.is substituted or un,substituted phenyl.
- In certain .embodiments,. L is    Q•is absent,• and-R6 is selected from heteroaryi.
,
and-Cii6heteroaralkyl:    certain,such embodimentsjeis heteroaryl selected from    :
•    25 ,    'fur* tbiophene,..imidazole; isoxazole; -oxazole,'cixadiazole,
pyrazine, pyridazirkt and pyrimidine.,.In.ti,,ertain alternatiVe    • ‘1
ts;_.R6    4,h Qt0oaralicyl sclected.-ftorn pyrtolyirnethyljuranylmethy14.,,:,,..-
'thienylmethyl;-imidazolylmethyl, iscxazolylmetbY1,.dxazolylmethyl,.0xacliazolylmethyl,,,..
..:thiaZolylmethyl; thi ad iazol yltrietbyl;.tri azolylrnethyl ;-,p,yrazolylniethyl 5 pyri dylin ethyl    - •
pyrazinylmethyl,:Pyridazinylmethyl and pyrimidinylinethyl..
- 35-
 
WO 2009/045497    PCT/US2008/011443
Imcertainembodiments,'L is C=0, Q is:absent or 0, and.R6 is earbocyciy1M-, wherein M-is- Co_i alkyl: In certain'such embodiments, .R6-is cyclOpropyl or cyclohexyl,:
In certain embpdimentS,. L and -A are C----0;-Q is absent, 2 is 0, M is C1,8alkyli preferably methylene, arid R6 is selected from RI 'ZA-C1..3alkyl-, R142-C 1:2    R! 1,2:A7
Ci zgalkyl-ZAZ-C g alkyl-, (R! ' 0)(R120)1)(=0)0- C1_8 aik)1.-.ZAL-C 1.8alkyl.=;
-
    10)(12 '20)1. (=-0)0-Cy_8alkyl-Z,C _g al ky.lc and- heterocycly1MZAZ-Cf,salkyl-;:wherein:-    -,
eaCh :oecurrence olAY it,independeritly other than ,a..'covalent -bond. In ‘certain such
••
embodiments le: is' hetero-Cycly1M-ZAZ-Cf.gal    where heterocyclyl is substituted- ot, .•    .
,unsubStildted okodioxolenyl N(R.!)(1V7), vy.hereiriR'' attd,R. 17 tCigetile:C.
0 • • y-c1,6alkyl; preferably C13alkyITY-Ci_3alkyl, thereby forming a ring.
-In-certain preferred.embbdiments,:L    Q is absent: IA, is Ci4alkyl,- and R' is
' seleCtedfrOm (R!;10)(R    k-F--".0)0 e.0    (R1b3Nt C0.8alkylarid.
beterocy.cly14-W. •lri certaiti,sn6frernbodimerit's,R6 is (R13)2NCI'_-8alkyl or
3alkyl-, where R'3 is CI _6alkyl. In certainother such embodiments, R6 is heteroCyclylM-, 15    where heterocycly1 is selected from morpholino, piperidino, piperazino, and pyrrolidino .
In Certain .embocliments, Lis    R5 is CI-.6alkyl, Q is selected from.0, and NH
and R6 is selected from C1•6alkyl, cycloalkyl-M, Ci_6aralkyl, and C1.6heteroal*lkyl; in .
•    other ethbodiments;L.is    C1_6alkyl, Q is selected from 0:and NI-1,:arid .12-6 is•
salkyl, where C1..6alkyl is selected from methyl, ethyl, and isopropyl. In further
-20 • • embodiments; L is C=0,'R5 is C1:6alkyl,Qis.selected from 0 and NH and R6 is •3..
,saralkyl,,whie.re aralkyl is pheilyhnetItyl. In other embodiments, L -is C=0, R5 it Ci alkyl,• Q is selected from 0 and 'NH, and R is Ci.dieteroaralkyl, where heteroaralkyl is (4:- pyi.idyl)methyl.
-In Certain embodiments, •Lis absent Deis C=0, and R5 and R6 together    .    •
25 6alkyl-Y.--c1.6alkyl, Ci,..-6alkl-ZA,C1.:6alkyi, nr Ci_6alkyl-A, wherein each, occurrence cif Z and A is independently other than a: covalent bond-, thereby forming a ring. in certain Toeferrt.:7d ethbodiments,,L .Q .and 1' ate absent, and Rs. and R6 together are-C1...: 3a11(0-61'..3-alkYl., In another-Preferred embociirnent, L and Q are absent, and.R..5,and
logether.are-Cl3alkyr,Y"-Ci.:3alkyl. 'In another' prefeired embodiment L is
30 ---absentrY is s'elected'frorri NH i-ItiN-CL6alkyl,, and •5 and R6 together are
C1_ alkyl f.ln:ariother -preferted'emboditnent', L is D--0,'Y is absent, and R5 and R6,
36-
 
WO 2009/045497    PCT/US2008/011443
 
.-together    -lit-another preferred embodiment,1 and A are.C=0,.
.    land R5. and R6. :together:are CIL2alkyl-ZA-Ci_2allsyl In•:anotherpreferrett embodiment,-.L.
•    and A ,:are..C=0 and RS and R6 tp2ether are    .
•    In:certain: einhodiments,•R7-    fromhydrogen    In.certain.
- • :*Prefen-ed.:Such .emboditnents,-R7 is selected Bona hydrogenandniethyl. .1nmore-preferred... such:embodiments, 117, is hydrogen
In    ernbOdirnents,    11.3 ate•eaCh.independently C1 6aralicyl; -and. R
selected 'from Ci -6alk y1; C 1..6-hydro x    'C 176 a licox. yalk yl ryl, and CI _Garai    of
which'i sOPti onallY.-sUbstifuted ;with one of-. in ore    n e,, earbox yl c acid '(or.av4',-- I .
thereof)-,..,:ester. tinc.Auclingef_Gq1lcy? estc:r...Cf_s3alkyl. enter,.and aryl ester);thioi zr    . .-
thioether. substituents.,-
•    pieferred•embodiments,:thethain of 'aniino acids has:a.' structure of    J:-
formula (XIV)    •
 
•    ndependently.anaromatic orheteroaromatic grOtip optionally:. sOstituted with 1 to 4 eubstituents;
each-A is independently selected from C=0, C=S, and 502, preferably C=O; or
 
WO 2009/045497    PCT/US2008/011443
is optionally a covalent bond when adjacent:to an occurrence of A; ..IV- is:selected •from. aryl, Ci_6aralkyl,-heteroaryl, and C1.6heteroaralkyl; R4 is N(R5)I.,-Q-R6;
is.selected.from hydn)gen, pH:, C1_6aralkyl, and Ci_6alkyl, preferaL2,4
R6 ..is:Sefected, from hydrogen,.C1:.6all,cyi, earbbey.clv,I;heterocyclyt,arr,N-tei-rninal.protecting,gronp, ary1;:Ci..:6aralkyl; heteroatyl; C_I'..:6heterdaralky4:1V ' ZAZ-C
t?.
C,_    (R13)3.14+-Ci.i 2 kyl-:,
.    carboc!iclylltil,T.:1'4.510fiC alkyl -; ?-nd.
capping group; more preferably t-butoxycarbonyl or, benzyloxycarbony1;. or,
ard-R,..together
6alkylZAZ-C1_6alkyl,.ZAZ-C1 _6 al kyl-ZAZ,. or Ci_salkyl -A. thereby forining.a ring;
R9 is selected from hydrogen, OH,. and CI_balkyl, preferably.C1,6alkyl; and • is an N-terminal protecting group;
RI' .and R12'rare independently selected from 2ydrogen, F fetal o..;nork,
C1,6alkynyl; aryl, heteroaryl,,,    preferably: frqrn.
•    hydrogen, metal catiom..and,Ci_olkyl,priR'!.and:R!.2, together are ,c14alkyl,. thereby lorming a ring;
20 •    • :each R13 is independently selected from hydrogen and Cr_oalkyl,.preferably
,alkyl; and
RED is independently.selecf.ed from hydrogeri,.'C1_6alkyl, C1:6alkeriy1,- C carb.ocyelyl, heteroCyclyl, aryl, heteroary1;:CI:6aralkyl,. and C1..6heteroaralkyli.
R ]:.; selected from hydrogen, Ci_oalkyl., C1,:6hydroxyalkyl:.C116alkoxY, -C(0)1411.0).6a)kyl, and Ci,baralkA:Prefera-bly
    hyill-cnyalkylonore preferablyrnethyl;setllyl,.hydrOxymethyt-,.and.2.-hydrolx.yethyli    t
•    provided-thatin any occurrence•of the sequence ZAZ; at:least one:mernber.of the sequence must be other than a covalent bond.
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WO 2009/045497    PCT/1JS2008/011443
In certain einbOdimenfs„ le-is-selected from Cj_6aralkyl and C1,6heteroaralkyl.
•    certain. stich embodinients, R2 .is selected from C    Cke,alkyl-indolyl,
-.011.cyltthierly1;.C.,i_6alkyl-thiazoly1,,: and C _6alkyl-isothiazol yI, wherein the alkyl -,N,toiety as y .contF4in,six,iive.;•four, three; two., pr one,sarbori atoms, preferably one or two,--; In; ,
•    • certain; .such...embodisnents;.R. is .E;ilbstitlIted •with :orie.•or more substituents.selec. ,hydrox7j;..hatogen,..arnide,,amine; carboxylic,acid-(or salt :there 0),- ester_ (;".'10Vrii'41g:0.-'' • , ,-...,..alkylestof-,..CoalkyLester;.■and aryl :ester),:thiol,:or-dno ether.. In certain    •
•    r embOdiinents3:R2.is.!substittited:witIra'subStituent.:selected from alkyl,-
•    alko:xy,.1r.ydroxy,.or cya4o, Iv certain.such-ernbodirrents,    is.select.O.fm.21,c
•    10    • • 'phenylrand-CT,6-alkyl=indolyl: -In certain ;preferred such 'embodim ents;    s-•sel ect e,c1. from,— ,.
pa    •
D
•    "\N"' N
I    j    stf
and
R
R = H or any suitable protecting group     

 
wherein D is selected from 14, OMe, °But, OH,' CN, •CI-73. and CH3. In certain- •
•    embodiments D selected..from.H, Oivie, 01-i, C.11,1,.(.7:3 and C.H3.
-preferred  such embodiments where TY is attached to a .71;-;.-inerribereri ring, D is ati.e.,'..hed at the 4-position relative to the point of attachment, preferably
. •:ex eluding embodiffients :where.the.4-position.of the:ring, is occupiedbrthe nit.rogen,of pyridinf: ring
t.p.cert4k; embodirnents•e is hydrogen,. L is .C=0 or SO R5 is Ar-"Y-;,;
•    is iri-derif.1ele0y. selected. fr.c.,srn phenyl; indolylobenzofUranyl,r naphthyl; •
70    q•inalonyl;'thieriy1; pyridyl, pyrazyl, and ,the    In certain such embodirnerits,.:.Ar:may • :-.•
be •substifuted    .E. is. selected from 4• direct , b ond,    and C :6allcyh • JP. .
•    certain -otheruchembodiments•where Q is. Ci.:6alkyl;-Q may be substituted, preferably- • with Ar, e.g., phenyl.
 
- 39 -
 
WO 2009/045497    PCT/US2008/011443
In certain einbodimentS, 1.?,.5, is hydrogen, Q•is absent, L is C=O•or SO2,•    .and R6 is :
•    ..selected from Ar-Y. and heterocyclyl. In ,pertain preferred such. embodiments, • -
•    heterocyclyl is-selected-from •chrorrionyl, ehrornanyl, rnorpholino, and piperidinyl,..• In. :certain other preferred such. embodiments, Ar is selected from phenyl, hildoly1,.- •-••• •.• .•
•    :benzofuranyt., naphtliyi; quinolinyl, quiTiolorryl,thienyl, pyridYI, pyrazyl, ?;i.nd In certain, embddiments;    hydrogen, -L is•C=0 or S02,. Q iS :absent, and    is':'
:•-•Gi_6alkenY1;:viliere'Cl_Olkeriy1•.is:a substituted vinyl group where the substituenuis„
•    -preferably. an aryl'br-freteroaryl' gr. oup-,•more•preferably 'phenyl. grOup optionally substituted with one to fbtir Substituents.
10    An:certain einbbdirnents,..t%and Qare. absent and R5 is selected from C.'1..6alkyl-; .
Ci_6alkynyl,,L).45aralkyl, !and.C•1_6heteroaraikyi: In bertainsuch.embodiments,'
•    RS is C1-L6alkyl, and:R6.1syselectedlfrorn -butyl„!all yl;prop.argy1', phenyhnethyl, 2.7.pridy1;;;:!-.:.. , 7 : ; 3-pyridyl, and 4-pyridyl.• •.

In other-embodiments, L is SO2, Q.is absent; and R6 is selected from C-;;:.Aikyl and
15    aryl. In certain ouch embodiments, R6 is. selected from methyl and phenyl.
in certain .embodimentsi, L is C=0 and R is selected from Ci.salk-yl,
aryli .c..i7sarallcyl,•heteroaryl, C1_6h&eroaralkyi, RI IZA-Ci.galkyl-,
-RI-"Z•-c:, _8    , (R.! .0-)(K120)7(-7-.0)0-.Ci_ialkyl--...,..(R110)(R120)P(=0)0-C1_0.1164.,,ZAZ;
,(R I 0) (RI!: 0)P (=0)0-Ci_salkyl-L-Ci_salky,1-, RI 12A-Ci_salkyl-ZAZ,C.i..salkyl,, 20.    leterocyclyINAZAZ-C1 -8 k34-, (R1)214-C1 _8 alkyl-, (R1.3)3N÷-C14    heterecycly1M-, •
•    :    R1:-SQ2e1..olkyl-; and R.1S02N1-1•,,.wherein each occurrence •of
...independently.other. than a,covalent bond. In certain embodiments, .L is-C=O, Q. is.. absent, and le is H.
In certain ernbodiments,iR5-is .C1...6alkyl•,    Q is: absent, and L
certain-such embodiments,. R6 is ethyl, isopropyl, 2,2„2-trithioroethyl,. or. 24
•    -(nethylsulfonyi)ethyl.
•    -In • other „embed iments; -L    Q- .abs.ent,_an d .R6.. is 12    alkyl in. certain
'Such:. errib o dirrients,•R6.. i selected.frorti /2-phen yl    phen Om ethyl, (4.    • -•    _
:•::metlfoxyphenyl)methYl," (4.--chlorophenyl)methyl, and (4-..11uOrophenyl)rnethyL
- 40 -
 
WO 20091045497    PCMS2008/011443
in-other embodirnents,L is C=0:,...Q is absent,-R5 is Ci..6alkyl, and R6 is aryl: In certairrsuch ernbodiments,,R.6 is substituted or.unsubstituted phenyl,:
-In. certain embodiments,.1.;is C=0,':Q is .absent;7.and-R6- is selected from hetei-oaryl nd.C.:1:.:61ieteroaralkyl.- In Ccrtai'n 'such ernboclitarits, -RP is het,--roar'yi‘selected from. ::pyrFole, fiaran-; thiophene, irszidzzole,jsbxazole, oxazale,
.`; tiiazoles pyrazole; pyridine; pytain'e,-pyri-dazine':and-pyrirriidine; In Certain, alfetriative-
 
thiertylmethyl,irriidazatylmethyl;'iioxazolylmethyl,!c}iazolylmethyL;bxadia,7-Oly.linethY1,,,:-.. 10 • - .pyrazipykrie,thyl,,t)yrjdazinyineihyl andpyrimidinylrnetbyl,,
In.certain ernbadirnents;Lis C,,-=0;Q As absent or 0; and.R`-,..;is„carbOcyclyliVi-,.
In certaiii•embodinients, L and•A are C=--0,-(2 is absent, .Z .is 0, M is CI ..g alkY1,2 • • .  preferably methylene,' and R6 is selected 'from R11ZA-C _salkyl-,
15.    CI _ialkyl-ZAZ-C1,8alkyl-, (R11.0)(R120)P(=0)0-Ci..8alkyl-Z.AZ-Cl_8alkyl-,,
..(R110)(1 .1,2.op(=o)o-el_galkyl-Z-Ci_salkyl-, and beterocyclyIMZAZ-CI _8alkyl-, w ere‘1.: . each occurrence_of independe:atly other than a .covalerkt bond. In certain suci% -eniboditnentsR6. is heterocycly1MZAZ-Cssalkyl-:•whereheterocycly1 is-substituted:or uriSUbSiitUted oxodioxoleriyI or N(itiE)(K- 17,), wherein R16 and R17 together are C1_6alkyl: •
20.    Y-Ci..6alkyl; preferably Ci .,3alkyl-Y-C1_3alkyl, thereby forming a ring..
In -certainprk--;-fel'red embodiments,iS'er-70,- Q. is absent, M. is.-5-2:)41alky-'..,-a')d-R
- selected from .(R-11.0)(R1.40)P(7-=-0)0-Calk.Y1,, .(R.1„3)2.1\ICI„salkyl, (R13)3-N-71-C    .
heterocyclyl-M-.. In. certain Such. embodirnentS, R6 is (R13)2NC),jalkyi or-(i413)31 ei-I
gallcyl-,7,where--W3.is..Cp,6alkyl:    certain-otb.er•suCh embodiments-,R6.is iseterocy.cly1W:
•    • 5    •_vthere.heteroeycl.yl.fssetected from morphOlino,..piperidirio,; piperainc, and    •
*In Certain'enibodirnents;LiS    " is -Selected Emirs    d    .7
acid R6..iS;.selected,fromC1
.,other:ernbodiments Lis. c-,20,:R.,5•is.C.1,6alkyl;.Q is :Selected.ifrOrn
ernbOdirrients;    C.r:6alkyl Q select-ed.-from .0 and:NI-I.:arid R6-is:
....::.,6aralkAwhere:aralkyLisphenylinethyl::10:other.embodiments,: L is    is Q...t6alk..yl,
- 41 -
 
WO 2009/045497    PCT/US2008/011443
Q is selected•froth:0    ,NH; and ..1Z.6is.Ci-heteroaralkyl, .where -heteroaralkyl
pyridyl)methyl.
-In certaiti7enibOdimentS,--L iSabsent- or- is -C=0; :and-Rand-12.6 together•are    -- •    7 ‘..,
or:0126alkylA,:wherein..each occurrence of:Z.
5.• •,• :".and A:is iridependentlYother than:a. covalent bond;,-thereb.y:forming-aring: . In. certain...
;.preferred,erribe:diments;•-L-is;C=0;-.Q.,andY areahsent,..a-nd.R.and,g.tOther,
.3alkyl=Y C.1 3alkyl ::In .ariother.'preferted..'enabodirrient;.L'and.Qateabsent;'and
drranother'preferred.embodimerit,.:Lis-&--0;.Qis
.    , 1:J.serit::-.Y,.:.is-SelectecarOm--NFI.:ancl..N-..Ci26alkyl; and.1i5, arid R6s.together: are    :    „
-10.    CI _3 alkyl.- :hr another preferred embodiment, Lis'e.;---0,'Y. is absent; .and Rs and
.togetherarec:r..alkyl7y:-C-1 _3 alkYl.. In another preferred•embodirnent, L and A are-C-=0,
- •:R6alkyl-,ZA-7q.:.,50k94..,Iii :another preferred.embOdimentf;L.-7,-.
•    - !1.:' and:-.A.-are,C=.0 and ie•Jand R6 together are C2..3alky1-7X
In_certain embodiments;    C1..6aralkyl; and 131' is selected from Ci_oalkyl, CI_
. 15    6hydroxyalkyl, C1.16alkoxyalkyl, aryl, and .C.I_Garalkyl, any of which is.optionally
..Substituted with one or more ofarnide,.amine; carboxylic acid (or a salt thereof),: ester.:. (including C1_6alkyl ester; Ci_salkyl ester, and aryl ester), thiol, or thioether substituents:
fl:-.:.:;,;,...=-In:certairrpreferred..embodiments;:the:chain-o.f.anaino.acids-has.a structure -of formula (XV)
20     

(XV)
wherein
L is Selected from C=0; C=S, and SO2, preferably C=O;
•    or an :aeWated. font thereof,. preferably X. is -COON, COCI,.. or .    C01■1(Me)(0Me); mast preferably. X is COOH or LOCI;
)-4Z iskabsenti.C1-61kyl;-Ci_6alkoxy., or NII.,4e;g::: absent, C _6alkyl, or Ci_oalkoxY;- preferably absent;
R is selected froin 'Hand Ci_6alkyl, preferably H or C1-13;:
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WO 2009/045497    PCT/US2008/011443
R1 and-le:axe-each independently selected :from. hydrogen; C1_6alk-yl, C2.6alkenyli• C2:6a1kynyl; Ci_jiydroxyalkyl,• C1.6alkoxyalkyl; heteroaryl, • heterocyclA. C 11.6heterocycloalkyl-,. C.I.:6heteroaralkyl,„carbodyclyl, and C _6carb oc:yelplalkyl;
selectedfron3 hydrogen, Ci_6aralkyl, and C1_6alkyl; 5..    R5* is heteroaryl; and
: R6.is selected fromhy.drogen,.CI_6alkyl, and e3..6aralky.1:". •
;:    4
.0 ..:611y0..roxYal IcYL:. _6alkcxyalkyl; CI ..6aralky4. ,6heterocycjoaikyl:.
,61-teteroa'ralkyl,, and C1...6carbecyclolalkyl, .. In certain embodiments, R1. and R2..are ;
l 1        iridependwtIY:Ci...6alky17selected fron.k.methyL ethyl; propyl,..isc-fpropA,-
and.i0btityl.:_,Iri.:certain;embodiments,•R!.:.:and.R2: areind6pendentlyC1_6hydroxyalkyk, In .•
-:.,:cett.ainiireferreds.uchernbodiments,.R.I. and,2:are.independently.selected•from hydroxymethyl and hydroxyethyl, preferably hydroxymethyl. In certain embodto„ents; R1 .. and R:2 are independently C1_6alkoxyalkyl. In certain such embodiments, R1 andR,.2 are
1•5    independently selected-from methoxymethyl and methoxyethyl, preferably
methoxymethyl. In certain embodimentS, R1' and R2 are independently C1_6hetergaralky14. In certain such .embodiments,-R.1 and R2•are independently selected from
•    iinidazolYhriethyL'pYrai.OlYtmethY1,:and thiazolylinethyl,..arid pyridylniethyl, preferably:. •: inaidazol-4-ylmethyl, thiazol-4-ylmethyl,..2-pyridylmethyl, 3-pyridylmethyl,. or 4.1..
20    •pyridylmethyl: .1n-certain embodiments, R1 and. R2 are independently Ci.:6aralkyl... In .
certain such embodiments, .R.1 and R2 are:areindependently selected from :74.ietaylmethyl..-, (benzyl)-and-phenylethyl, preferably phenylrnethyl....In certain embodiments, R1 an.d.R2. are independently C1.6carhocycloalkyll. -In certain such embodiments R.1 is
tIti•certain embodiments •R!. and -R2-are.different. In•certain. •  25 • • .embodiments, R1 and R2.are the.same.
-. In- certain erribodiMents-at least•one of R1 and R2 is selected from-Ci...
•    y,liy'clroxyalkyl-and .C.i.,01.koxya I kyl ,    certain such embodiments, at• least: one of,R1: and .
R.2,' is alleox•yalkyl-:    certain-sucherribadiinents.,•atieast. One of R1.-and R:2, is selected
methoxymethyl and methoxyethyl.
.30    • • :In certain embodiments, R4._and R.6-are independentlyselected frorn hydrogen and    ...
- methyl, preferably hydrogen.
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WO 2009/045497    PCT/US2008/011443
ineertain.embo'diments;:Rs•-•is a 5 or 6-membered beteroaryl.. In certain such. '.'embodiments., W.is.sele,ctedfrom.isoxazole;isothiazole,.furan,.thiophene,
pyrazole; or imidazole; preferably isoxazole,.furan.,. or•thiazole..
•• In certain_embodiments.,.W is .a-bic3;•clicheteroaryl. In certain-such embodiments,. bicyclic heteroarytis selected frorn benzi soXazole; benzoxazole,.benzothiazole, ,
•    benzisothiazole.
.    •.I•ceffain-ernbodirnerit, I:, is E.=0,-..Z is absent; andW is a 1,3-thiazol-5-yl
-thiazolL4-Y1. fri.certain.SUCh embodiments; when the.thiazOle is Substituted, it is subsiitirtedrafleas't af-the    Iri;•ather,csuch:einbodimerits;    UriSubStittited,;...,...
10    1 ,3-thi az.61-5 :::y1 or 1 ;3:41ii azol-4-yl.
i
in.certain embodiments, I., s.04), 'Z is. absent,• and Rs is a substituted 1,37thiazol7
 .    •
Certain.'stich:thbodiniehtsi leis,-,1,3,thiazOl-yl.substitutedwith a substituent.
•    'selected from Ci.:6alkyl.,*.C11.6alkOXY;" C1 .6alkoXyalkyl, C1_6hydroxyalkyl, carboxylic acid;. • aminocarboxYlate, CI .6 alkylaminocarboxylate, (C1_6alky1)2aminocarboxylate,
15    6alkylOarboxYlate, C1.6heteroaralkyl, CI _6 aralkyl, C1_6heterocycloalkyl, and Ci.
6carbocycloalkyl. In certain preferred 'such embodiments, R5 is 1,3-thiazol-5-y1,. . substituted with a substituent selected from methyl, ethyl, isopropyl, and
cyclopropylmethyl.
In certain_embodiments, Lis C=O,.Z is absent, and R..5 is.a substituted 1,3-thiazol¬20    •    In certain such embodiments, Rs is 1-,3-thiazo1,4-yl substituted with a substituent
.selected. from ,C1_6alkyl-,;C 1.L6alkox C    kokyal    C _6h y dro-xyalkyl, carboxylic ,aci    :    •,
.._..aminocarboxylate, Ci...6alkylaminocarboxylate, (CI -6 alky1)2aminocarboxylate, C1-    .
. 6alkylcarboxylate, C1_6heteroaralkyl,.    aralkyi, C1.6heterocycloalkyl, and C1-
:6carbOcycloalkyl:• In:certain preferredsuch.entddiments; W is 1,3-thiaz01-4y1: ; 25    substitt:ifed Withasubstituentselected from methyl; ethyl, isopropyl, and.,
cyclopropylmethyl.
RS.is an is ON2t1zo
    &eta inveferted: such..ern bodi men ts; when-the. i sox azp,1-3-y1;:is-:    •
Substitufed,:it: is substituted.at-least at the 5,position.....-In.certain preferred embodiments„-„.•    • !
30    when the isoxazo175.-y1 is substituted,. it, is.suibstituted,at least at the 3-position.:
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WO 2009/045497    PCT/US2008/011443
Irircertain embodiments, L is .CO,= Z is-absent, and-R5 is an unsubstituted
-
:.1-n certain ernboditrients, Lis C=0; Z is absent,.and R.-3 is a substituted is'oxa2o1-3- .y1....in-certairi. such embadinients,..W:is isox&kol..--3,,y1 substituted-with wsub.stittrent-r.•
.3    ;:selected froit C16a.licy.i;-:(31:-.6alkoxy,:Ci..6alkoXyalkyl,-Ci.6hydroxyalkyl;.•earboxylic acid,    --    .:
aminocarboiyiate.Cioalkylamihocarboxylate,:(Cjalkyl)2aminocarboxylate,    •
.:...::6alkylearboxylate,•C1,_6ipteroaralkyl,:.Ci..:6arallc3/1,:Cf_6heterocycloallkyl, and ct t.6c4rbocycloalkyl::.Iii-certainpreferredstich embodiments R5,is s o xazOle-:3-yl,substituted.
.with,-,a•substiraent:kled.f.rornInethyls    isoPrOpyl,, and
. -In certain embodiments L is C=0;rZ isabsent, -and R5 is isoxazoi-..3-yl• substituted, •
•    - -with a 4= to.6:Inernberednitrogenrcnntaining'Cl.6hetetocycloalkyl.'. In certain-such
;•-• yi.rilethyl)f in!.certain.',alternative such •embodiments;    is absent,',
w
isoxazol-3--y1 sabstituted.with    , wherein W is 0, NR,. or C142, and R is H
15    or Ci....6alkyL In certain such embodiments, W is 0.
In certain embodiments, L is C=0,1; is absent, and R5 is isoxazol-3-yi 'substituted • • 5.-mernbered nitrogen-containing •C 1„6heteroaralkyl,-such as pyrazolylmethy14,:-. • . —imidazolylmethyl, triazol-5,ylmethyl, preferably. 1,244Ttriazol-,5-ylmethyl.- :• -
In certain embodiments, L is C=0, Z is absent,- and R5 is isoxazol-3-yl substituted 70.    with 01.:6alkoxy•or.Ci26alkoxyalkyI, preferably methoxy, ethoxy,• methoxyrnethy1,, or •
metlioXyethyl.
In certain embodimentS,L is C=0, Z•is absent, and R5 is isoxazol-3-y1 substituted. Ci_6hydroxyalkyl,preferably hydroxymethyl or hydroxyethyl.
. In certain embodiments, 1-...-is•C=.0a is rabsent,:and :R5.- is isoxazo1,3-y1-substituted ry    • :::with;adarbpxylic:acid;arrtinocarboxylate;
'011v1)2amino.carboxylate,,or C1_6alkylearboxylate: ..1n certain-such, embodiments, RS ms:
substituted with. ITIthyl 'carboxyl ate..or. ethyl carboxylate, preferably. -methyl carboxylate:
In'eertain•mbodirnents, L    is.absent, and R.5. is aryunsubstituted    .
isoxazol-5-yl.
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WO 2009/045497    PCT/US2008/011443
In-Certain:embodiments; L is C=0,.Z is absent, and-R5 is a. substituted isoxazol-5-
yl.-. In certainisuCh-embodiments, R.5.is isoxazo1;5-yi substituted with a substituent.    • .
..::selected    p6a11(4274.y,- C allcoxyal kyl, _6hydroxyalkyl , ;carboxylic . acid,
•    aminocarboxyIate,.Ci:6aikylarninecarbpxylate,. (ci.6alkyl)aminocarboxylate,.C:14-•
•    . 6alkylcarboxyl ate, • C 1,611eteroaralIcy1,:C1_6aralk-yl, C.i_eiheterocyclbalkyl, and C . ~6d~aT>~oc;rclaalkyl In. certain-preferred-siich 'embodiments-Rs,. is isoxazole-5,-yi substitute••: frora.methyl.,:e-thy.1,TiSopropyl, -and cycl
'";    In7certdimembodimentS.L-is,C=.:0;!Z.isabserif--end;R5.iS:.isoxazoh5-yt'substituted:
 
•    .-,with-a.4;;-,to.•64:nerribered nitrogen.,containing.C.1,6heterocyoloalkyl.-.• In certain    .
10.    -embodimentsi-R5 is isoxazol--5:yl.substituted•with azetidinylmethyl, preferably ,azetidin-1.—
.yii,rittliy1..7    ertain,alternative Sac+. embpdimenis,    Z is absolit,,a.id
;Wherein'W
•    or C1_6alk-yl. In certain such embodiments, W is 0. . •
In certain embodiments,.L is C=0,:Z is absent, and R5 is isoxazo1,5,y1 substituted 15-•    with .5-membered nitrogen-containing Ci_6heteroaralkyl, such as pyrazolylmethyl,,, .
krnidazolyirnethyl, triazol-5-ylmethyl., preferably 1,2,4•triazol-5-yirnethyl•
•    In:Certain-embodiments; L    absent, •atidR.5:isisoxazol-5-y1 substituted...
.        Cf..6alkoky.or CVGalkoxyalkyl;.preferably.metboxy;.ethoxy; methoxymethyl;-or.
methoxyethyl,
20    • • In certain embodiments, L is C=0,Z is absent, and R5 is isoxazol-5-yi.substituted •
. v,,.ith.€1•:.5hydroxyalkyl, preferably hydroxymethyl or hydrox-yethyi:
la certain embodiments, L is. C--•=0, Z•is absent; and' R5 is isoxazolt5,.-y1•.substituted • •: i c acid;. aminocarbox yi ate, C .6alkylamino carboxyl ate,. (Ci..•    tt;,7 •
- alkyl-)vartilloca. tbax yl    or CI    k yi c arboxylate: In certain such -emb °dim ents,,R.5--i S. 7' :1
.substituted:witk methyl carboxylate.or ethyl- carboxylate„..preferably.methytcarboxyl ate.
lrr-certain e•bodiments,_Z is. NR, preferably,NH. Uses of Enzyme inhibitors
ra'clerly protein degad ati On is crucial to•the•aintenatice of normal
and:the proteasome is integral , to-the Protein:degradation process. The protpasorn e:
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WO 2009/045497    PCT/US2008/011443
controls the level s-7of prOteins that are nu-Portant for cell-cycle progression and apoptosis . • no-i-rnal.anth-nalignant.cells-,- for example,-.0yclins; caspases, BCL2 and uF-kB
'(Kumatoriet•al„ Prot.: Natl•.-Acad:-SOi.•USA-.(1990)..87:7.071,7075; Alinend et    .
•    , Leukemia '(2002) -16:. 433.443): Thug,- it- is': riot surpiisin.g. tharinhibitingproteascirn . : .aCtivity.ban;transl ate.into • tberapieS to, treat arious disease; states; such . a;•.; trialignant,,  .-dtialignant 'Arid autoirt-itrune-diseases, depending,ori the,cells
" • Y    Both i i i~itro'and iii vivo models have''sliowii that inal'igriarif "cells;' .    •    . .„    _
in general, diet
-susceptible to prOteasorrieinhibitiort Iti.faCt,:proteas'orne inhibitiontas already been
••• -',yalidated, as a therapeutic-strategy for the-treatment of multiple myelorna..• This -cd:clici-be
10    'due,.-iri part, .to the.hig.hly proliferative malignant cell' s.dependency (m the proteasome •
s:Fisteingt.t: rapidly TerrioN:e    -(Rolfe (5,q;    •j:    Med.• ( 77 9.97) 75:5- 1    •
NatUre (2004) 4: 349-360). Therefore, certain embodiments of the invention relate-taa ".•
method. of treating cancers comprising administering.to a subject in need of such    .
..    •    .    .    _    .
treatment an effective amount of the proteasome inhibitor compound disclosed#1.6rein.,
15    As used herein, the term "cancer" includes, but is not limited to, blood born an solid •
tufnors. Cancer refers to disease of bloOd, bone, organs, skin tissue and the vascUlar syst6tri,    not limited to, cancers of the bladder, blciod, bone, brain4breast,
cervix, chest, colon, endrometrium,•esOphagus, eye, hoad, kidney, liver, lung, .-1;:4-_hph
-•, anodes,-mouth; neck;• ovaries-, pancreas,,prostate, -rectum, renal; skin,:stomach, testis, ,:throat3•andfuterus-.,•Specific.cancers-include;but- are not-limited-to, leukemia (acute .,
lvmphocytic leukemia (ALL), acute lyelogenous leukemia (AML), chronic lyrnphocytic leukemia (CLL), chronic myelogenousleukemia.(oa),Ilairy cell leukemia), mature B cell neoplasms (small lymphocytic lymphoma, B cell prolymphocytic leukemia-,.
lymphoplasmacytic lymphoma (such as Waldenstrom's macroglobulinemia), spienic    •
 
25    marginal zone lymphoma, plasma cell myeloma, plasmacytoma, monoclonal
immunogldbulin deposition diseases,•heavy chain diseases, extranodal Marginal zone B Cell "lymphoma (MALT lymphoma), nodal: marginal zone Y.3 cell lymphoma (1\ilq21).„ follicular lymphoma, mantle cal tymplidina, diffuse B cell tympho-ma, rfiediastinal (thymi4 I at-ge B. tell 1yr-1-Thema, intravakular large13. cell lyniphOrtiai-prirriary e;ffusion •
30        lymphoma and Burkitt lymphoma/leukemia), mature T cell and natural, killer (NK) cell., , •
neoplasms (T cell prolymphocytic leukemia, T cell large granular lymphocytic leukemia.
aggressive NK cell leukemia, adult T. cell leukemia/lymphoma, extranodal NIUT    .
•    •- • „
 
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WO 2009/045497    PCT/US2008/011443
lyrriphOrna, .enterOpathy4Ype T cell lymphOina,:hepatosplenic .T.    -- •
..NK:aelllyrnphoma,Anycosis • fungoides (Sezary•syndrorrie), primary cutaneoUs..anaplastic¬.• --10.tge.celliymphoma,.lymphomataid    angioimmunoblastic.T, cell lymphoma,
- • -.-:unspecifed.peripheratT cell:lymphoma-and -anaplaSticiarge-cell
.•    5    lymphorria.(nodula•-sclerosis,•.mied celluarity, lymphocyterich, lymphocyte:depleted...or;
t        :not depleted; nodular; lymphocytevredorninant),:-myelornatmultiple.myeloma;..indolent
rny.elorna,:srnolderingrnyeliorna),.,chronicmyeloprolifetatiye.disease, -
' • •• ' • •ii--..-t-uYelodysplasticknyelopfoliferatiye• disease, myelodysplaStic•-syndrornes;,-f •.--
•    iMmuno-defiCiencr..associated lymphoproliferative.disorders, .histiocytic and dendritic.cell..
0 ---.:iteoplasms,-Mastocytos*: chondrosarcoma-,•Ewing-sarcoma;fibrosarcoma;:maligiant • '.giant cell tutnor,:rnyeloma;bo.ne.disease;osteosarcoma, breast. cancer (hormone • .• • :••• -; • dePendent.horritorie.,independerit),,g.yneCological cancers:(cervical,•endornctrialjallopiarr.: .:•,
qtbec..gestatioitaLtrophobla.stic.diseaseoVarian,cperitoneal;.uterine,.vaginal and    •4
basal-cell carcinoma (pcc), '§quamous. cell .careinoma '(SCC),•malignant.melanorna,-• .
15    • dermatofibrosarcoma protuberans, Merkel cell carcinoma, Kaposi's sarcoma,
astrocytoma,•pilocytic astrocytoma, dysembryoplastic neuroepithelial tumor,
.oligodendrogliomas, ependymoma, glioblastoma multiforme; mixed gliomas-,
•    .. oligo.astrocytornas; medulloblastorna, retinoblastoma, neuroblastoma;.germinoma, •
teratorna malignan(thesotheliotna(peritthieal mesothelioma;. pericardial •mesothelioma;•: 20    • pleural mesothelioma), gastro-entero-pancreatic or gastroenteropancreatic neuroendocrine
tumor (GEP-NET),. carcinoid, pancreatic endocrine tumor (PET), colorectal adenocarcinoma, colorectal .carcinoma, •aggressivelneuroenclocrine tumor,
leiomyosarcomarnucinous adenocarcinoma, Signet Ring cell adenocarcinoma,.
•    thepatotcllular carcinoma, cholangiocarcinoma; hepatoblastoma, hemangioma, hepatic .:... 25    ;adenoma,• focal• nodular hypetplasia (no.ditlar: regenerative hyperplasia;harnartonna);
.    • !Amall.celLlung carcinOma(NSCLC)•(scitiairious,cellAtingCarcinoma;•radenocarcinomai.,•.,::.,    ,•.•
large :celi.lnng.carcinoma};small•cell-lung carcinomaThyroid carcinoma; prostate.canter ••• • :(bonnonesefpctoFyondrogen.indepericient;_apdrggen:dependpilt, hormone,insensitive);.--.. r. and.soft.• tissue:sarconias.(fibrosarcoma;malignant• fibrous hystiocytoma, .•
dermatofibrosarcorna, lipbsarcomay:rhab,.doinyosarcoma.leiOmyosarcoma,•"..  hemarigiosarcoma; ..synOviatsarporna; malignant peripheral nerve sheath
-tumor/neurofibrosarcama, extraskeletal osteosarcoma).    •
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WO 2009/045497    PCT/US2008/011443
-Many tuthors.of the haematopoietic.and lymphoid tissues are characterized by aru: • :increase.in cell proliferation, or.a particular type of cell. The chronic myeloproliferative...-
•    diseases".(CMPDs)areelonal-haematopoietid stern. cell disorders .characterized by -..:•. •
•    'proliferation in .the bone marrow of•one, or. more of.the. myeloid lineages, -resulting iti.:.- -..intveaseduumbers:of .'granulocytes., Ted- blood cells41nd /or platelets int:Ileperipheral..,
As,..such;,the use:of proteasomeinhibitors'for-theiteatment ofsuch-diseasesis . • aftradtive2.an.d.being:eXamined    liaeniatologica:(2007.)-92:11,2+1229.11:.
•    ;: CM12:cart include:Chronic myelogenous;leukaeiniai-.chronic.rieutrophilicieukaeriliak,'
•    .chronic eosinophilic leukaemia;-polycYthaemia 'era, chronic idiopathic myelofibrosis;-..... -10--    "essential-thrombocythaernia'and unclassifiable-chronic myelopro liferative disease An _    ,
aspect of the:invention:is the. Method of.treating.CMPD. comprising administering.ta.a--
,subjectimneed. Of suelr.treatment an effectivearnountof the proteasome
compound disclosed .herein..
.Myelodisplastic/myeloprolife,rative.diseases, such as chronic myelomonbcytic
15    leukaemia, atypical chronic myeloid leukemia, juvenile myelomonocytic leukaemia-arid
unclassifiable myelodysplastic/myeloproliferative disease, are characterized by hyperceilularity of the bone marrow due to proliferation in one or more of theMyeloid
lineages. Inhibiting the proteasome with the composition described herein, can,serve to    .
treat these,myelodisplatic/myeloproliferative diseases by. providing a subject in need of
2.0    such treatment an.effective amount or the composition_
Myelodysplastic syndroines (Nips) refer to a group of hematopoietic stem cell
•    disorders eharacterized by dysplasia and ineffective hcematopoiesis in one or more.of.the inajor Myeloid cell lines. Targeting NF.-kB with .a proteasome inhibitor in these .. hematologic malignancies indices apoptosis, thereby.killing the malignant cell .(Braun -et :25 ...    al: Cell-Death-and Differentiatiory(2006)•1.3:748-35.8.)-. A further embodiment ofthe
•    - invention-is-a -methOd AO treat-MDS comprising adrninistering to a subject in need of such - •    .
•    - treatment an effective _amount of the compound disci osed. herein. MDS includes ;•refractory .anemia;-refractory anemia with, ringed-si deroblastS„refractory cytopenia-Nvith.' .myltil ineage-dYsplasia,- refractory:anemia with excess b las ts,, un cl assi liable • . • •
•    • ..y30    myelOdysplastiosyndrome and 1-nYelodysplastic:syndrome.associated with isolated:.,
-del(5q) chromosome abnormality.
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WO 2009/045497    PCT/US2008/011443
Mastocytosis: is a proliferation ,of mast cells and their subsequent accumulation in ---one or more organ:•systems. Mastocytosis includes, but isnot limited to, cutaneous ma:stocytosis,-indolentsystemic mastocytosis•(ISM); systemic mastocytosis
•    •associateilclOnal haematological no•:Imast=cell-linea-ge diseaS“SM-Al-INMD),
,....aggressiVe SyStelniC.ipastocytOsis•:(ASM);•.mast cell.leukemia (RECL),- mast cell:sarcoma:.
: • (MQS)-and,extrcutaneousz.nastacytoma:, -..knother-embocliment bf the invention -:.:-:-4'.;::,..-:.:::-....rnethod-,:to;,treat:Inastocylosis:.dorriprising, administering atreffect amount of:the, compound:-,;
.4disclosed herein,toi-asubject diagnosed.with, mastocytosis.- •    •
:The:rif dtea§drii r e tit Os INF,:kB, Which iri turn-regUlates.genes irivOlvedin, the -immune and-inflaMMatciry response. For example,-NF-kB is required fortthe expreSsion •
.Of lheinurionoglybilin light chain n gene, the IL-2 receptor u-cliairi.gehe,    `:.lass I    .
-..-titajor:histbscompatibilitY cOmplek-gerie; land a number:of eytOkine .geneS,encoding;for.-.:n
,,.:ek:aftiPle„.IL=2;,-IU•6,,,.:grfinulocyte'dOlony-srinrulatingfactdr, and IFN.,,f3 (Palombellol-
-et/T;(1994) 7g:773=785• Thus, =in certain embodiments,•the invention relates fornethods 15    of affecting.the leVel of expression Of IL-2, MHC-I, IL-6, TNFa, IFN-p or any of the.
•    other previously-mentioned proteins, each method comprising administering to:a:subject an effectiv• amount of aprotedsome inhibitor composition disclosed herein. lncertain
•    ettibodimentS, the inventionincludes a Method of.treating an autoimtnune disea,,,',ein a. • : • marninal..cornprising:administering-e-therapeutically effective-amount' of the; compound ,: ,
•    20-    deScribed-herein:'•:An ":autoimn-aine disease- herein js.a. dis.ease or disorder ari sing fronr.-1--
and directed •against an individual's own tissues. Examples of autoimrnune diseases or -disorders include,-.but :are not limited to, inflarnmatoryresponses such as•inflamrriatory . skin diseases including psoriasis and:dermatitis- (e.g. atopic dermatitis); systemiu:    - -
•    ,scleroderrna and sclerosis; responses associated with inflammatory bowel disease (such as
•    25    .CrolVs disease 'anO-Lacerativaccilitis); respiratory distress syndrome (including
resPiratory; distress-syndrome; .ARDS); dermatitis; meningitis; encephalitis; uveitis; -•-. !-,colitis;.•glomerulonephritis; -allergic.conditions such- as:eczema and asthma and-other • :•tondifionsinvolving -infiltration ,of T cells and chronic inflammatory responses;., ,a therOsclerosis; .1 eukocyte adhesion, deficiency;. rheumatoid -arthritis; systemic lupuS,.-
.    • -3.0-    erythematostis-(S,4-4. diabetes mellitus (e.g.:Type l.:di'abetes mellitus or irisulin,deperident,--•
..;:diabetes4iellitiS); Multiple-sclerosis; Reynatad-Ts- syndrome; autoimrnune :thyroiditis;
.allergic.encepbalornyelit•is; Sjorgen'S•syndrorne;. juvenile onset diabetes;and immune::
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WO 2009/045497    PCMS2008/011443
responses associated -with-acute:and. delayed. hypersensitivity mediated by cytokines .,::,-..TAymphocYtes:- typically:found tuberculosis, .sarcoidosis, polyrayositis,
and yasculitis; pernicious' anemia:.(Addison'S..disease); diseases involving -leukocyte,
•    s.ystern•(.(.3N.S)- nftarn raatory disorder;    tipl organ    -
"::yridrortip;-hemolytic.anemia.(inOudirig; but not limited.to cryoglobinemia or COom.bs: F.,•,---..-.::.:,..7.--positive5anemia)i?myasthenia..gravis,,,-antigen-antibodrcomplextnediated,diseases;:.antii:
glomenilar:b.asement m:embrane-.disease antiphospbal pid. syndrome; allergic neuritis : ~L::~ ° ; a;:.. ; >;::
.,Grayer'.-disease; • Larriberp•ton. myasthenic.:syndromes„ peinphigoid
,...,...a.utoinOiurre poi yendocrinopathies Reiter's di sease:-.stiff-marr-syr;drorne; -13,6heet-,diseaSe;. pr7-giant-ciell-rarteriti8;Tirrrnrone:complex,liephritis.,:tgA-nephropathy,...:IgM-polyneuropathies;--    77!
'.•immune-thrombocytopenic p.urpura..,(ITP)-or.antoirnmune thrombocytopeni a.
:.:The immune: sy.Sterri screen&fottautd.logous:'cells. that are:virall    -    •
,.,y-2,undergone,oncogeniOransfOrrnatiomor•present -utifaritiliar peptides, on.their".surface:%.-.-,,,.i,i,.:.-
Thitrac,ellular proteolysis-generate small- peptides for-presentation:to    h.o.cytesto ..•
. -15 - induce MH.0 class.1-Mediated immune responses: Thus, in certain embodiments, the .
invention :relates to a method.of using the compound as an irrimunomodulatory. agent for -
•    . • inhibiting or altering antigen,presentation in a• cell, :comprising exposing the cel“or    .
. ...administering to. a subject) to the-compound described :herein, . Specific.embodinients , •    „.;iriclude.a_rnethod.of treating graft or transplant-related:diseases„ such as -gra ft7y.erssus-host:
2.0 .    disease. oT.h9st ,yersus,graft . di seas e. in...a „mammal,- comprising administering a    •
.therapeutically effective amount. of the compound. described herein— The term "graft". as .
used herein refers to biological material derived from a donor for transplantation into, :
: • recipient..7Graftsincl ude..such-di v.erse.materi al. as, for example,.isolated    • :-
cells; tissue such as. the amniotic membrane of a newborn, bone man-ow, hematopoietic - •
•    , ..•precursor cells,...and ;ocular. issue.„--suche as :Carneallissue., and prganS• Such: as. skiry heart:, diver„Ispleen .pancreasi.thyn3idJobe. lung, kidney,:tubular organs .(e,g,,,intestine; blood    .
•    ,• •,. •    ,,svesselsocesophagirs):„The :tubular organs-can .be used.tb.replace:damaged portions:of
;,e.sophagf-tit. blood- vesSelsor.bile-dtict:The -skin..grafts.-;carr.be: used not -only for bums,:birt.. also.as...a :dressing 10 .datnaged.intestine_ or :to.: Closeertain. defects -such.. as :di aphragrnatic. -;•,hernia. The-.gr.-aft-is-derived: froin 'any mammalian source; 4ieltiding.human,whether from    :
orAiying.donors,‘:In:some cases, -the donor and recipient is. the -same    - •
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WO 2009/045497    PCT/US2008/011443
•    -Preferablythe graft is bone marro*...or an orgartsbchas heart and the donor of the.graft: and the hOst are matched for HL.A class II antigens.
Histiocytic 'and dendritic tell neoplasms are derived from phagocytes and - • •
•    • accessory'eellS, which have MajOrroles .imthe.procesSiiigand 'presentation of antigens:to 5. ''.1y.iiiphocyte:' Depietinithe ProteaSomeebntentiin-dendritic cells haS been Shown to alter iieif'antigenindneed;reskinses-:(dliapaite 'et    OanCer-Res:.:(2006)    - .
- .    Thus, atiothr.'erribodftrienif-the nvention comPtiSeS'adrninistering an 'effectiVe;:anibUnti.::::,-:,
'    • •    •    •    ,    •    •    .    ,    •    -    .    •,    .    •    ...    .
,:Of the aegrfpo'sitiOti..disCloSed herein .to. 6-.SubjeCt'WitlihiStibt-Ytie Or-dendritic
•    rieOpia.sin:; 4-listibeytic.and. dindritite,cell neopla-sixis riciude.r istioc,rtic
•    1 0 • 'Langerhans cell histioc,ytosis,,Langei-hans,cell sarcorria., interdigitating dendritic cell.    _
•    Saitornali:urifor• foilk;ula•-dendriti,..,    saltOrrlatU➢rsOr afzd    spc-.6 fi ed dallf.17-1    ,
sarcoma.
qjf•Alie.proteasbrhe    'Veen §howrifo. be -b-ri6ficia.l. to trea:t.dispases..
whereby a cell type is proliferating and immune disOrders; thus, an embodiment ofthe.
15    invention includes the treatment of lymphoproliferative diseases (LPD) associatedwith
primary immune disorders (PID) comprising administering an effective amount of the disclosed compound to a subjectin need thereof. .The-most common clinical settings of.:. immunodeficiency asso ci ated v;iith an increased incidence of lymphoproliferativ:e. .
kid,f174,Cell neoplasms and lyniphonias,‘ are priinary.' inirnutiodefiCieneYSOdrorneS'and'other:primaryininnirie diSorders,- infection with She.* •
human immunodeficiency virus. (HIV), iatrogenic immunosuppression in patients who -have-received solicl'orgari or•one matrow allograftS, and iatrogenis innmanosuppressi on
associated with methotrexate. treatment:: Other.PIDs commonly associated with L.PDs.,- but not limited 16, are ataxia telangiectasia (AT); Wiskott-Aldrich syndrome (WAS), .
-25    coinniOn'VatiableiMinunodeficieney(CVID), SeVere combined immunodeficiency.
lympbciProliferative disorder -(XLP); Nijmegen.breakage syndrome-,
(NBS); 'hyper-1'04 sYndrome,':and-•utoirnmund lYmphoprOliferative syndrome (ALPS).
Additional: embodiments. of,the.inverition relate to-methods for.affecting the proteasome,dependent-sregulation of onOoproteins and.methodsPf treating or .• 30, - ••- ,cancer.growthi• each method comprising exposing a cell (in vivo; e.g:, in •a subject, or
Vitro) to the .protea.sorne. inhibitor cafriposition:diklosed herein. HPV-1 0 and HPy--.18:-.
derived - Ed proteins stimulate ,ATR4and ubiquitin;dependent•conjpgation.and:degradation
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WO 2009/045497    PCT/US2008/011443
•    of ':053 •in-crude,reticulacytelysates.: •Th'e-recessive oncogene p53..has. been. shoWn10 • • • accumulate at the-nonperinissiye temperature in .aceltline with•-a mutated. tbermo)abile    :•.
- -EL.-Elevated levels, of p53 may lead to apoptosis.. :Examples of. proto-oncoproteins •
•    d egadd:by 'the Athiquitin. system include .c,•Mos, c-Fos, and c-Jun. •ln certain .• • einbodiments.,:the inyentionxelates•b -a..methodlor...treating p53.7related g.poptosis,: - .::comprising.'administering aisubjectan•effectiyeam punt of aTroteasorn.e composition disclosed .herein._.
- r    Another.aspect.ofthe'inVentiot Telate.8 to the.use.of-proteaSoine..inhibitOr,
• :;... •    „-.    -...compoSition&discloSed.herein'for,theAreatment of oeurodegencrative
•    I a • canclitions,including,..but,not limited to stroke; schemic damage to the nervons,syF.;tem.,:, „.
-.214:aral-tranina,(e:g.,.tperaissive brizin..c.ktinago;.:Tinal    injvcy,, and traania6r.-;.d;ama&:.::•to-::
,..,,.:-..the:-nervous-system); rnultiple!scierosi&•and.:otheri.mmune,mediated -0.ni:Ilain•-.BarreisynclrOMeanchitS"-Yal*4';',acutemotor..allon4;netiropathy;acute ;inflammatorrklemyelinating. polyneurop4thy,:and..Eisher Syndrome), MY/AIDS..
15 .    . dementia .complex;. axonomy, diabetic neuropathy, Parkinson's disease, Huntington's •
disease,. multiple. sclerosis, bacterial, parasitic,. fungal; and viral meningitis, encephalitis, vascular. dementia; Multi-infarct dementia, Lew y. body. dementia, frontal lobe dementia
•    such. a& P icic' s :diseas.e;:subcorti cal dementias (such as Huntington or progeSsiVe.. •    ..-:. •
supranuelearpalsy.);.focat cortical, atrophy, syndromes. (such. as primary:aphasia),
20 ...,..‘metabolicttoxicdementias (such as, chronic,hypothyroi di Sra or B.1 2„deficiency.),,anci• dementias •caused by infections (such as syphilis or chronic meningitis).
•    :Alzheimer's disease is •haracterized by•extracellular deposits of13.-amyloid protein . .• -fp-AP) in,senile plaques. raid •cerf.,-bral    1.3•AP.is a. peptide fragment of 39 to
,arnino acidS:derivedirom amyloid protein. precUrsor (APP). At least .three isotbrms of 25 -...:APP-arekntAtiV(695;175tarid.770.arriiiio acid's).•='Alternativesplicing'ofmRNA
•    :generates' the, iSofortris ithrinal,iiisoeessing,affeots . pOrti tin.o f the ft-AP -sequencer thereby • ......preventinglhegeneratiOn of    It is believed •that abnormal protein processing-by be,
proteasbare.ecinteibutos..to the 'abundahceof LAP.itt:the Alzheircier brain: The ARP.,
•    ---processing•enzyme in.,rats--.contairs'about ten d ifferenti subnnits, (22-•kDa-32.-k Da), The.-25.?    „
:30. •
..:::■,vhichis:-:.identioalloAhe.13,..sub.-imitbf human acropai (Koj Una; S . • et al.:,
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WO 2009/045497    PCT/US2008/011443

Sae, (l'992).-304:.:57:60). TheAPP.-process•ing enzyme cleaves at the.G.
:Lys16,borid; idthepreSence -ofcalciumion;. the enzyme also cleaves-rat-the Met-h--Asp'' ..-„ • :bond, and-the-AspAla' bonds •toireleaSe the extracellular domain of !3.-AP.:
-    : One aspect of the invention; therefore, •relates Ito a•method of treating Alzheimer's,•.    -• •• •
- disease; i,-::orriprising administering to a -sUbject ari:effective amount of the proteasom6 . ,    •
,. inhibitor, corriPositiondiSClosed'herein: :.Such treatment includes. reducing thexate•of_13.,,-..
AP -prOcesSing;iredticing:the rate of p-NP:plaque forination,. reducing the rate of•P;A:P± • getieration,.and redUeing the clinidal signs of AlZheinieris- disease.

.ei.cessiVe..and persistent fon:nation .of fibrous connective-tissue,. . 1.0 - • • resultingfroakthehyperproliferative growth:of fibroblasts and is associated
activation. ofthe.T.GF7:13:signalingpathway.:. FibroSis;inVolves extensive
=extraeellular matrix;
and camoccur.within virtually any-tisSue or across several different::
fiSsues.: •Normally,-: the. level of intracellular signaling proteih (Smad) that activate,    . ,
transcription of target:genes upon TGF-13.stimulation is regulated. by.proteasome activity -
15- ..    2000),• However, .accelerated degr, adation.of the TGF-f3 signaling cornponents
has been observed in fibrotic conditions; such as -cystic fibrosis, injection- fibrosis;::
•    -endomYocardial; fibrosis,. idiopathic pulmonary fibrosis, •nyellofibrosis, retroperitoneal • • 'fibrosis, progressive Massive fibrosiS;-nephrogenic-systemic fibrosis.. Other conditions:. .    •    .    •    .    _
that are often:associated. with fibrosis include 'cirrhosis, diffnse parenchymal lung disease;: 20    post-vasectomy pain.syndrome,,tuberculsis;.sickle-,celtanenha and rheumatoid -arthritis.,
•    .An embodiment of the invention :is themethod of treating a fibrotic or fibrotic-associated •
•    condiiionConipriSing• administering-an effective amount of the corriposition described heeeiri tb subject• in 'need of such treatment.
•    - • The freatment of burn victims i's.ofien. hampered by fibrosis,_thus, in certain 25    embodiments,. theinventi on relates to the-topical dr•systemic administratidn- of the,    • • •••
...inhibitorsq6.freat, bums: :Wound- ClOsure-following surgery. is often .associated with :dis    gars,!which•iri aY be prevented by inhibitiOrrof fibrosis. Thus. in certain ,•-•
.einbodiritentS;thE inVentiOn.relates: to. a method.-fOrthe prevention or reduction of . scarring.
OverprOduCtiOn of:lipopol-ysaccharide. (LPS)-induced cytoki nes such as
.-considered to Ilexen tra    •he'protesses associated with septic shock. „Furthermore;
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WO 2009/045497    PCT/US2008/011443
,'-gerrerallyaccepted.that thefitat,-step.,iri the 'activation' of cells by..LPS is the.binding.of :
to specific :membrane receptors: The .a- -and-P,subunits. of the 20S proteasome
....ccmplex.havebeen.identified-as'LPS-bindiTig proteins,,.suggestinithat the LPS7induced'T
.::signal.transduction.rnay be- an -irriportant therapeutic target yin the treatment or-prevention:... o-f-sepsis.(Qureshi,N,'et,ak,../..fmmurz..(2003).1'7I•:: I-515-1525). Therefore, in certain: mbodiments;:the,proteasome inhibitor Coniposition may be used for the inhibition of
•    TNEn. to prevent and/o•Areat septic shock.    •
 
,,,,Ischernia and reperfusion injur•:results,hrhypoxia, a condition in,whioh there,is
•    5    .deficieripi,of:oxy.genzeactinag the.tissues ofl-he body._ This, condition, causesincreased ,
14'    degradation of IK-Ba, thereby resulting in -the.activaltiOn of ,NF-KB: (Koorig et: al.,1994),    •
l't 'has-been der' riodstrated that fhe sc\.,erity ofinjury resulting hypoxiF1 can be tedii.cted. with-theadniinistratiOn. of a:profeasorneinhibitor4Gao et al.; 2000;,BaO-et
.!!..',7.1*.    'al .,:2003) 'Therefore;. certain enibodiments 6f:the-invention relate :to -a
treating an isehemic condition..or reperfusion injury comprising administering to; a- subject . •
15 , in need of suchlreatment an _effective amount of the proteasome inhibitor compound disclosed herein. Examples of Such conditions or injuries include, but are not limited to, acute'coronary syndroine (vulnerable plaques), arterial occlusive disease (cardiac,    .
cerebral, peripheral arterial and vascular- occlusions), atherosclerosis (coronaryielerosis, .
•    . _coronary artery:disease), ,infarctions, heart failure,,pancreatitis,,myocandial hypertrophy,„ _ 20    stenosis, and restenosis.
NFr-ic13 alSo binds specifically to the. HIV-enhancer/promoter.. When compared td
•    the-Nef ofmac239, the HIV regulatory protein Nef of pbj14 differs by two amino acids in,. -
7-the region which controls:protein kinase binding. -It is believed that the protein-kinase
signals the phosphorylation of IkB, triggering I►.13 degradation through the ubiquitin¬25, ..,: prOteasorne pathway. : -After: degradation, NF-03 is released intb the-nueLdus; thus, .1, enhancing the transcription' of HIV: (Cohen, 31'; Science, (1995) 267:960):4In certain emhodinientai the invention Eelatesr to a rnethOd for inhibiting or reducing: HIV infection-
in- subject;. or.a-method for decreasing the level of; iralgene expression, each inetho,i •
.
`comprisingadministeringto the'subject. aneffeCiVe=a-mount of theproteasome inhibitor ,
.30 .: . composition disclosed herein.
'Viral infections: contribute to -the 'pathol ogy *Of many diseases. :Heart 'conditions ',such •as ongoing myocarditis:and dilated eardiomyopathy have been linked-to the... .
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WO 2009/045497    PCT/US2008/011443
cogsAckievirus    cornparative whole,genome:microarray anal yses. of infected
t mouse bearts;:specificiroteasorne subunits mere uniformly u.pregulated..irr
•    • whichdeveloped• chronic myotard•itis (Szalay et al,. Am J•Pathol 168:1542,,52,' 2004- • Some virusesattilize-the ubiquitin-proteasome systerain theviratentry step wherethe • - -.:viruslis.released fron-i.theendosomeinto•the:cytosOl.:::The.rriouse-
 hepatitis.virus.. Ml V...
helongs to..the!-Coron'avirfriae:-farnily,:'whiclralso..includes the.severeactite respiratory:.:;
-•r:skrIcirotr (SARS-)::coronvirus„-T.YU:arid;Lar-(1'Virot.,7,9:6-.44,;_64.8,-.2005)•demonstratedAhat;:,
• :; treatrnent,o'feells,infected.,with:MEIV: with apr.oteasorne:inhibitor resulted in a decrease:,7.-.7
in. iral'replication, oorrelating.withLreduced viralliter.as compar,ed,b3..that.•f unti:eat,2d
•    --: 'cells; zfbeihurnairhepatitis,B- virus-SHBV)altteinberof-the .1-lepadnaviridae
requires    encoded .ierrki.elop-proteinsto.pIopa.gat. Inhibiting the, •
.••••.:';:proteasome.degr:adationpathw.ay:causes.a,sig,rtifiCant reduction in-the:amountotsegreted.,:
::envelope proteins: (Smsek:.et al J. VuoL 79 294442920 2005)    addition
•    other hepatitis viruses    andE):yri ay also utilize the. ubiquitinTroteasorne.•'
.15 degradation pathway for secretion, morphogenesis and pathogenesis.. Accordingly;. in • . . -certain embodiments; the invention relates-to a• method for.treating viral infection,. such as SARS- or hepatitis A,B; -C, .p and E; comprising contacting a cell • with (or administering. to a subject) .aneffective. amount :of the compound disclosed herein.
In-:certain embodiments,,the disclosed.compositions may be useful for. the.....
20:        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: (Paugam .et    Trendsparasitol. 2003, 19(2): 55-59)..•:.
Furthermore, entarnoebaspecies:have..been.shown.tolose encystation capacity when-
exposed.toproteasome inhibitors, (Gonzales, et al.,, Arch. Med.. Res. 1997,28, .Spec No.: . • 25: hi.certain• sticlyernbodiments,• the' administrative protocols for.th eprotea softie ::• ..• •.. inhibitor .compositions are useful :for the treatment of parasitic infections in humans, ,:. •
.catis-ec17by.:.a protozoarEparasite,sblected from .Plasmodium
inalariae;',.and_P::a\fale,:,.which.caiise,rnalaria)y:Trypanosorna
which causes: gh'agas7..diSeaSe;'.and T,brudeiwhiCh= causes' A frican: sieepirig.:1:•;
30 • • sickness), Leishmania sps.'(in•cluding-L.    dOno.Vani, .L.infanturn,
mexiCana,,etc.:),Tneurnocystis•carinii (a:protozoan:known to cause pneumonia inAIDS,.• •
and other- inimunnsuppressed patients),;Toxoplasrria• gond Ent amOeba• histol yti    .•
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WO 2009/045497    PCT/US2008/011443
Entamoeba invaderis, and Giardia lamblia:- lo certain embodiments, the disclosed:
proteasome-inhibitor compositions -areuseful for'the treatment of parasitic infectionsin,:, -animals_and livestock-caused by 'a protOzoan parasit• selected fi-orn-Pla,smodiunt-hennani,. -CryptosporidiurnsPs.c.Echinococcus.ganulosus•;.--Eirraeria    Sarcc cystis
arid NeuroSpbra crassa:: Other compciunds.that.act-aslproteasorneinhibitors 'the: treatment of paraSitiodiseasesi,are,described in.WO '98/10779; which is incorporated-.,:.-.--
.herein:in its entirety; -    .    • 7
lirceitain embodiinents411eproteasofne inhibitor compositions, inhibitproteaSorne,..--, aetivity .in a. ijraSite .Withotit    Ted:,13    d, cells .and white ,bio odcel I s ,    .certa    .

'such. embodiments;:the4ong.half-life of blood. cells: may provide prolonged rprotection: ,itgat    lg'-'cl„Kroctsmes    parasites.      cx;rti-ii.31
inhibitor COmpei gitiiins ay.provi de prolonged protection.':    regard to:
chernOprOphylaxiS:against futtrefinfection.    •
'Prokaryotes have what is equivalent to the eukaryote 20S proteasome particle.
15        •Albeit, the subunit composition of the prokaryote 20S particle is simpler than that of
eukaryotes, it:has the ability to hydrolyze peptide bonds in a similar manner. For
example, the nucleophilic attack' on thespeptide bond occurs through the threonine residue .
on the N-terminus of.the413.-subunits. Thus, an embodiment of this invention relates.to a'
treating'prokargoti&infectiOns; comprising 'administering to a ,subj ect
-‘ .7•..effectiVe.knount of the    dbinpOsitibildisclosed hereirl...PrOkaryotic
. 'infections may:include diseases caused by tither mycobacteria (such as tuberculosis,...,
. leprosy or. Buruli Ulcer) or archaebacteria.
- „- It lias'alSo: been dertionstrated that inhibitors that bind to the 20S. proteaSome -
•    Stimulate bone fonnationl-in-bone organ cultures..:Furthermore,.when such in"; ibitors have.: 25 . • • .beenidrninistered:systernicalfy tanlice, certain'proteasome inhibiters-in.creaseci bone i-olurne..and-•bbne.forination :rates even 70%-{Garretct R; et 61,, J.    (2093),..;
.1.771-74782), 'therefore. suggesting -that .the. uhiquitin-proteasome, machinery regulates:. .,oSteohlastdiffcrentiatinit'antkbon:e'forniation,-,Therefore;.-the.discloSed proteasome c:'.. inhibitor -corn poSi tion-. may be useful-in, th eltreatm ent and/or prevention„of-diseases,-,:, • , 30. -associated With bone loss, such as osteoporosis:
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WO 2009/045497    PCT/11S2008/011443
•    .Thus;: in certain embodiments, the invention relates" to a method for treating a •
disease oncordition selected -fromeancer; autoimmune disease,: waft ox transplant-related
neurodegenerative disease,• fibrotic-associated condition,- ischemic.-related, cOnditionsinfettiOn (viral;.parasitie-orprokaryotic,)••and diseases associated with bone •    - ,••-••
5    Toss,' cornpii sing administering a crystall in:.:compound o Formula ,(11).
    -':binpoundsfprepared- as- desCribed herein cati.be administered in various:forms;    •
depending•rohthe disotderlto be treated andltheage;•condition, and bOdy weight Of,the:-.‘ - :••!-    -•'.'-patieritaS,is'welt-knowit-in :the att:'F-op..exatriple; where the compounds are to
•    :•.•adtriinistered birally;,V,.y.frig.y,baorrnulate.;d:s.tablets,..capsuies -granules, powd4tflsi: of:    • F-
•    ,--•    • . syrups-- or-for•parenteral administrati on, :they.. may. -be .-forrnuiated as injections    •..•
inteailki:Isfail.ar,    -subc4inanotxts), drop .i..n&c:ic,ta.1-,-,pepacations,,o17-.... . .
.•-•,...-.7...:".,:"-",..1.-2:suppositories:-.-JF-or    therophthalmiomneons:mernbraneTrouteitheyniarbo,—,:;,i)p,,
as:eye drops.Or. eye...ointments.-•JhesefOrrnulations: canbeprepared.by
•    .coaverttiohal means,: and if desired,-the active ingredient may be mixed with any 15    • conventional. additive or.excipient, 'such as -a binder, adisintegrating agent, a lubricant.;••a- .
corrigent, a solubilizing agent, a:suspension aid, 4n..emulsifying agent, .a coating agent,
. cyclodeXtrin, and/or:a buffer. Although the dosage. will vary depending on .the syrriptos-,
•    - ace, and • body weight of the patient,-the nature and-severity of the clisordeT is be,treatea'•Pr. prevented,- the .routepf.administrationand:theform. of thedrug,, in general,. a. daily...dos age;, :
•    20• . ,offrom 0:01:to:2OOG-mg of the compound-,is.recornm.ended for an.adult.human patient,: . and this.-may be administered in a-single dose or in divided doses. The amount of active.. .. ingredient vinicheanbe c(?mbined "with. a carrier material to prociuce. a single dosagekiln!
•    'will generally be- that. amount -of thecompound which produces-a therapeutic effeet.
•    T.he.preeise time of administration and/or amount' of the composition ;'lat will •
•    25 - • yieldthemos(effectiNt results.iri terms of efficacy of :treatment in    •    .••,
•    -depend -upon-the'activity, pharinacokinetics; and bioa mail ati lity of a particular! compound;, cOrdition of the patient (iac•iding age,,sex. -disease type and stage, gene-ral-
•    :physical conditieri;•respo.nsiveness.-.to-azi-,/en dosage:. and type Ofinedi cation),..ronte•
•    :..administration; -etc.':-I-lowev.er,:the.abovegui•delineScan• be used aSth-eb asisfor.fi n ,tuning -tne,treatment:eg,v determining the. Optimum, ;Lim eand/or. amount of    ni strati on, :;•
require.no more than •routine experirnentatio-n-consisting of monitoring-thec.,--.:Irr:r.:-,-;-:.-;:..-.•
•    isubjectand-adjusting the dosage and/or timing.    . c., •
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WO 2009/045497    PCT/US2008/011443
The phrase '`pharniaceutiCally.acceptable.is .employed herein to. refer to those -compositions; andlOr doSage.forrns:-which.arewithin. the scope of: . • •
•    •;- •ourid•medical judgmen.4'snitable- for: use in- contactwith.thetissu.es. ofluunatrbeings.,and.2,: : Nexcossive.toxicity;.- irritation; allergic response, or other. probl ern op    :;.
- -.5    2:•eb-Mplication, COITIMertsuiate..with .a reasonable:benefit/risk
--Thephrsepharrna.ceukicallyracceptable:carrier". as:used -herein-means' a,    •
.--...pharrnaceuticaltr:acceptablermaterial;';compoSitionAivehicle,:::•shcir. as a liquid or:solid
•    filleri,cliluenf,,ekeipient;-Scilvent:Orencapsulatin.g-materi    -Each earriet---.must
acceptab iiilhe..sense. of being -coMpatiblevith .the o cher ..ingredients. of the, y ,,fonnula•tion and..not injurious, to.rthe•patient:- . Some.examples of:Materials •which.can SerVe

•    il pilaltrilaottitiv:4113r:ac:14,13ti,blo    x    6. d    f,..1.).1sugarz„Klcia, as •    ...•    .•    •
.i...,:sticrose;-_-(2).-starchespstichaS:cern...statch;qpotatolstarch;a.nd::substitutedor
•    • ethyl .•cellulose.,.-ancLcellulose- acetate;- (4). p owdered tragae.anth; (5) .rrialt; (6) gelatin;• (7).
•    15:    talc; (8) excipients, such as •cocoa butter and supposi.toty. waxes; (9) oils, such as peanut
cottonseed oil,.safflower.oil, sesame oil, olive:oil, corn. oil,. and soybean oil; (10) •
•    glyeols., such as propylene:glycol; .(1:1).polyols, such as glycerin, sorbitol, mannitol, and . 'polyethylene glycol; (12): esters, such. .ethyl-oleateand • ethyl laurate; (13) .agar;-:(1 bufferingagents:,; such.as magnesium hydroxi and aluminum, hydroxide;. (15): al gini
. ,. 20 ...    acid; (16).:pyrogen-free•water;•17) isotonic saline; •.1.8) Ringer' s :solution; (1,9)..ethyl
•    alcohol; (20) phosphate..buffer solutions; and (21) 'other non-toxic compatible substances . -.en ployedin.pharmaCeuti cal- formulations:- In, certain embodiments, pharmaceutical
•    ...compositions-of the present    are..non-pyrogenic,.    do not. induce. significant:    --
temperature 'elevations when administered to a patient.
.    -Th-etemi -'..pharrnaceutiCally:acCeptabl e    refers to .the relatiVel ymon4oXic;    .
•    • :inorganic. arkt urganic, acid additiott salts-,of the inhibitor(s)_    :salts:con. be Prep ?.red-.:.
•    :in sit during Abe final: isolation and:purification Of the inhibitor(s),, or by—separately  .:..:.redetinga-pUrified-.inhibitor(s)*•initS!freeyb.ase 4.01m:with- a:suitable- organic:oz - ,••• • • • '•••`: ,    ...aoici;:arkl':iSolating•the: salt' thus. foimed;:aepresentatiYe :salts_ include the hydro b rernid-e;'•
•    • 3.0-• .....--hydrOchlaride,..:Sulfateithistilfate;•phosphate,cnitrate;t acetate,- valerate;
•    stearate,-...1.auratei:benzoatejactate;:.phosphatetosylate
sn.ceinate;.tartraterfaphthylate,. rn esylate;glucoheptonate-,,.lactobion    sulp henate.- •    •
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WO 2009/045497    PCT/US2008/011443

. salts, and amino acid,Salts,and.the like., (See, for example, Berge et al. (1977) "Pharmaceutical Salts"; ,J. Pharm. Sci. 66: 1-19.) -
In other c'ases,,theq.ahibitors useful in the methods of the present :mention. may -': contain one or more acidic functional •gfaups and, thus, are capable of forming. .phar macttifically acceritablesalts with pharmaceutically acceptabie. bases: Thellern:.
•    stihatirraceuticall yaceeptable'salts' in theseinstanCesrefers to the relatively non-tozc-iC.:,.;-, .inorganiCand-organi&base addition. :salts ofan;    :These salts can    • •
..13repared in • Situ dui=ingthe.- final isblation-,andlydrification•oftheinhibitor(s),-O)
separately.seacting•the purified -inhibitor(s) nits. free.;.aci dlovrt:w ith    „,
1,0 . • . • as the hydroxide, carbonate or- bicarbonAe:of, a pharmaceutically. ac,ceptable r".1,t-ta1.cation, ,
TA-with' -a .37,1,•Nai-rnaccutically  .:::tertiary•ami-ne'_',:R:epreientatiie;_alkali;or;aikaline",earth,saltsinclude.the.lithiurn,':sodiutn, '-:.pOtassiutn,;:,caltiumnagnesiurtis'andaluminurtr.s-alts;sand,thelike:    .    171';
organic amines .useful--for the:fonoation ofbaSeaddition-salts-include ethylarnine,
.15    .diethylamine; ethylenediamine, ethanolamine, diethanolamine, piperazine; and the like:
(see, for example, Berge et al., supra).
Wetting agents, emulsifiers,.and lubricants, such as sodium lauryl sulfate and .-inagnesium stearate,as well .as coloring agents,,release agents, coating agents, sweetening;,flavoringyarroLperfuming a:gent's,,p-resetvative's and antioxidantS can-also. he: 220    present in the compositions.
, Examples. of pharrnac,.enti cal ly acceptable antioxid.ants include: (1) water soluble-antioxidants, such as ascorbic acid, cySteirie hydrochloride, sodium bisulfate; sodium . ' inetabisulfite,.;,_;odiurnsulfite and.thelike.,..(2) oil se,luble antioxidants,:such.as ascorbyl.•,,
•    pahnitate, butylated•lydroxyanisole.(BHA),•butylated•hydroxytoluene (BHT), lecithin;.'..--,proPyl.gallate,alpha;:tocopherol; and •the:like and,(3)metal:chelating agetits;•such4s,
Acid; 'ethyleti&li:arnirie tettaaeetfa acid:(EDTA),,:sorbito"i„ tartaric acid, ,,rid the like.
_    -    Formulations suitableforOral.administratidninarbe in theform of capsules,    .:,-t
•    ,. :--caclietS,    s; tablets;.lozenges7•(using allavered basis, usually sucrose and. aca cia
tragacanth);. powders, granules; or as a solution or a:suspension in• an,aqueousor nom,: :    •
•    aqueous liquid,,oras an:.oil: in-water or water-in-oil,liquid emulsion, or as an elixir.op,'
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WO 2009/045497    PCT/US2008/011443
-sYrup; or. as pastilles (using an inert matrix,.-such as- gelatin and glycerin; or sucrose-and- -
•    -•:acacia)-and/or-as mouthwashes; :and. the-like, each-containing- a predetermined .amount of    •
..an-inhibitor(s).as- an .activeingredient_ A,.comp.osi•tion.may alto: ;e• administered as: a bolus, electuary, or paste,
•    '-andfor any.olthefolloiArig:-(1.),fillers'or extendett-Such a.s •starcheS,:cyclodextrinS,,...,,-,:,...-..--:i. ,    •    actose,• sucrose,..glucoSe;;;Mannitol-,.and/Or„silitic-,acidf•(2) binders, s-siCh.as,-
•• carboxymethylcelluloSe: alginates; aelatin polyvinyl-pyrrolidone; sucrose,-
•    43)-1,1)h,    agtti    CIO    ;7.
:...carbonateipotato
-......2...,,;,.....,...,,,,:„.,...T...-,:...4:45):solutiOrkretardingr-agentS-;:?such•as;paraffirtv(6);absotptiorracceleraters;...stich:ass',.t..;•:e.e:-,...,'.:.;:.'N:::1-„:- quatemaryarmnonium compounds;.(7). wetting agents, such -as, for example,
    15    alcohol and glycerol menostearate; (8) absorbents; such as kaolin and b-entonite.clay; (9)    •
lubricants, such a- talc; calcium stearate, magnesium stearate, solid polyethylene glycols.; sodium lauryl. sulfate; and mixtures thereof; and (10) coloring agents. In the caseof
•    capsules, tablets-and pills,. the pharmaceutical colin.:,osition.sma7,/ also comprise ,agents.,.Solid.cornpositions ofasimilar.type,may also be,employedas.fillers in soft .and-
..    :hard-filled.gelatin.•capsules..usingsuch excipientsas :lactose or milk .sugars, as well!as    . •    •
'high molecular weight polyethylene glycols, and the like.
A tablet may be.macle by.: corripresSion or molding, optionally with one or more ,,e...T•essor:✓ ingredients,. • Compressed tablets hay be prepared usiwz bind (t'
•    gelatin • or hyth-Oxypropylin.ethyl cellulose); lubricant, inert diluent, preser    •
... • disintegrant-. (for -exarrip le; -sod ium starch .glyco I ate. or.cro ssA inked. sodium- carbokyrnethyl: _i:w..11ulose);:„surfaCe.,active..or:.dispersing agent: -Molded tables may be made by. a suit4bl• machine-•lnixture. of the powdered -inhibitor(s)) moist6ned. with a. inert liquid:
diluent.
and•other soliddosageforms; such .-as drageeS•capsules,
:granulesupinay optionally be spored or prepared with;;coatings andshel I s; :such as:enter): c
Coatings- and ‘Other cOati ngs    I -known.in- the ph drin aceuti ca I -fon-n ulat ng. art., ,They-rnar
also be formulated so:a's to provide.slowor. controlled -release of the aativeingredient.,;- ;.:
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WO 2009/045497    PCT/US2008/011443
•    ..thereinusing; for •exarnple;••hydroxypropylmethyl •cellulose in varying proportions-to c..provid•thed:',.sired release prdfile,•:other polymer matrices; liposomes; and/or , .microspheres. Theymay.hesterilized,by; for'example; filtration through a bacteria,
•    „retaining filter, -er-by in corporating steriliZing- agents- in the form. of sterile solid
I.411*-.1- can-be dissolved in sterile. water,-•or some.other sterile injectable •
. ,medium inimediatelybefore use These: compositions may also optionally. contain: :•.o.Pacifying.agentS-and'.may,be,ofia compositionthatthey.release the acti ye ingtedient(S); :;:..7....
•    • onlyi;or preferentially in acertairi portion•ofthe gaSttointestinal tract; optionally;:in.a.:
pi y.d.; manner .Examples of embedding compositions IvIiieb can be ust3ti xrclzri
107-polytrieric-substances-and: waxes.- The-active ingredient can also -beirrmiero-ericapsulated-- .
-    approy...riate,--kyith one, or more of the above,described excipients.
tiral:adminiStration:includepharrriaceutically ,;;.ethnISions;,tnicrotrnulsions;:solutitins, -suspensiOnS,•••syrupsand-elixirs'..:iln ad clition
•    -active ingredient, the liquid dosage forms-may contain inert 'diluents .cornrrionl y
    15    the art; such as, for example, water or other,solvents,•solubilizing agents; and ernulsifie,rs•-
•    such as ethyl -alcohol, isopropyl alcohol, ethyl Carbonate, ethyl acetate, benzyl alcohol,: benzyl benzoate, propylene glycol, 1,3-butylene glycol; oils. (in particular, cottonseed;:..-. goundnut,.corn, -germ, olive; castor, :and sesame oilS); glyeeroti tetrah/d.rofitryl-•;;AcolF).:01;
    ,    .polyethylene,glycols• and,fatty„acid esters of sorbitan, .andmixtures thereof_ •    :t    .
    '20• • .    Besides. inert -diluents; •the:oral- coiripos iti ons' can,al Soiriclude. adjuVants -such as ..
•    • wetting•agents, emulsifying •and suspending agents, sweetening, flavoring, coloring,:
•    perfuming, and preservative agents.
•    -•    : .Su.spensiOn. inaddition to the active. inhibitoi:(s) ttay-contairt suspending. as.touts
as; for example; ethoxylated .isosteapyl- alcohols; polYoxyetivlene sorbitol and sorbitan
•    25, ,:.,.esterS,::rnicrOcrystallinecelluloSe,..aluttintirrinietahydroxide.;.bentonite,
••• tragacanth,.and mixtures thereof.
Form nlati CMS for •reCtal- or- vaginal administration n=ay •bepresented -as a •• suPPOsitory;•Whi ch: m ay be prepared by. rnixing .one.ot more    b tor(s). with on_e:orniore•
-suitable nonirritating exci pietits-ot:earriers comprising;, for •example;'cocoa butter, :',:.polyethrYleneglyCol,. a 'suppository Vtirai or-a_salacylate, which is solid at i'dorn. • -•
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WO 2009/045497    PCTIUS2008/011443
   
•    • temperature, but liquidatbody temperature and, therefore, will melt ,;n- the rectum. or vaginal .cavity and release the active agent.
•    FormulatioriS• which, are, suitabl e for vaginal adniinistration -also    pessaries;!
.    .
tampons, .crearns,, gels, pastes,..foarns, or. spray formulations containing such carriers-as..:
are known in the art to be appropriate.'•
...DosagelOrMS,fOr.thetopical. or transdertnaladministration.of an- inhibitor(s), , ..•
include powders sprays ointments pastes creams lotions gels solutions patches and:
•    ...inhalants ;. The.aetiVe component rnaYbern.fxed under sterile Conditions with a . •.. . y acceptable carrier; and    any-07es ervatives buffersi. or propella.nts.
which Tnay. be required,
•    •    ointm•ents;:pastes, -creamS; and ,gels .thay. contain, in• addition to .inhibitor(s); 
excipients;',such as- aninial.and vegetablefats,',oils;iwaxes,,:paraffins,..starch,:tragacanth„,.:•,:..;....,:. bellulose.derivatives; polyethylene -glycols;.•silicones bentonites,- silicic :acid; talc, and. • .    .
Zinc oxide, or mixtures thereof.
15    :Powders and sprays can contain, in addition to an inhibitor(s), excipients such.as
lactose; talc,•silicic acid, aluminum hydroxide,. calcium silicates, and riolyamide pow,der, or mixtures of these. substances. Sprays .can additionally contain customary. uropeilant,r,:- • such: as chlotofluorohydrocarbons and volatile unsubstituted hydrocarbons, such as. - •- butane and propane.
2,0    • . The inhibitor(s) can-be alternatively. administered by aerosol. ThiS: is. •
accomplished. by. preparing an aqueous. aerosol,. liposental preparation, or solid. particles; • s
•    :.containing the composition: • .knonaqueous    fluorocarbon propellant) suspension...
'could be usect Sonic nebulizers are preferred because, they minimize exor:+sing, the agent • .    shear, which can result:in degradation of the compound. •
25    y,- an- aqueous.aprosol is.thade.by..fonnulating.ari aqueous...solution or,
..suspension of the aitent together'  th: conventional pharrn' aceutic.al ly accepta-Ole..carriers. • . . • ',and• stabilizers...:ThetarFier•and stabilizett Vary with•the• requirements 6f4bc., parricUlar.
' •
composition, but typically include nonionic    (Tweens,,Pluronics,•sorbitan
•    zstersjecithiri,•Qrectrop.hors); pharmaceutically acceptable co-solvents. such ..as  30    polyethylene glycol; .innocuous,proteins like serum-albumin, oleicacid,-Tarnino acids•such,
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WO 2009/045497    PCT/US2008/011443 •
as glycine,:buffers, salts, sugars, or; sugar alcohols. Aerosols generally are prepared.from isotonic solutions.
.--Transdermal.patehes ha-Ve the added advantage of providing controlled- deliVery of :an inhibitor(s).to the body: Such :dosage fonns can be. made by dissolving or. dispersing
5:    = die agent in the-proper medium: Absorption enhancers can also be used to increase the- 
t=flux, of the iniiibitor(s).across-theskin.. The rate of such -flux can be' controlled by,,eithei-:-. providing- a•rate-controlling membrane or•dispersing the.inhibitor(s) in x.polymet.matrix,...
or gel.
-.'-...-Pharrnaeeiatical.compositioris-Of.this.inVention suitable .forparenteral:-
-10    adrifinistration comprise.one-or. more inhibitors(S)•in combination with one or.1110.1-e'
pharthateutically acceptable sterile aqueous ,o.r.noilaqUeou:s.olutiotis, dispersionsy.. suspensions.or-emulsions;-oLsterile.powders .which may befreconstitutedintoSterileA:.7.-1,.. injectable-solutions Ordfspersions just.prionto
hufferS;babteriostats; solutes which render the*formulation isotonic with the blbod'of the • . 15    intended recipient or suspending or thickening agents.
Examples of suitable aqueous and nonaqueous carriers which may be employed in. . the pharmaceuticmi compositions of the invention include water, ethanol, polyols (s!..v.:;11.- as glycerol, propylenetglycol, polyethylene glycol, and the•ike), and suitablemixtures.,...:, , ;.... thereof, vegetable oils, such as olive oil, and injectable organic esters, Such as ethyl, • •
20-    !oleate. Proper fluidity:can be maintained, for example, by the use of coating materials,
. such as lecithin; by    maintenance of the required particle size in the case    .    ..•
dispersionS, and by the'USe Of surfactants.

     These compositions may also contain adjuvants such-as preservatives, wetting agents, emulsifying agents, and dispersing.agents. Prevention.of the. action of    .•
microorgariisms.may be enstiredby the inclusio . of various antibacterial-and-antifimgal agents, for example:, paraben,..chlorobutanol, 'phenol sorbic .acid;.and-thelike. •it mayalso
.,... be desirableto.inchide:tonicity-adjusting,agents,- such as sugars; sodium. eliloricle.;:and.the,..-,:l like into thecompositions. :In ,additionyprolonged absdrption of.theinjectable....:    „.:.
..--:pharmaceutical' form may-be brought about .b.y-the inclusion ofagents which delay
-. 3    -.absorption such. as.aluminum monostearate and gelatin:

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WO 2009/045497    PCT/US2008/011443
•    •    .    •    •• . ,Inisoine za:ses,•-in order to prolotig the effect of a .drug,. it is desirable to slow the .1 . absorption•of the dru•g-frorn subcutaneous or intramuscular injection:. For example,:'7.7:. .7.7. ...• .7% - : :delayed absorption. of a-pareriterally :adminis.tered drug form is accomynished by;
•    ;dissolving cir:stispending the drug •in- ari- oil    • .
•    7.- • - - • Injectable depot forths ar. made by 'formirignlicrocapstlernatric.:eS- Of, illhib itOr(S) ,    in.Wiodegradablepblytters.Such.as:polylattide-polyglY.colide, Depending on the.ratio-of..1;
•    volprier,dtie:tature; ofthe- paTti cularpblyaner
•    7. ,...-,release-- ean'be torttrolled:- Exatnples -of other•bi o degr-adabl e• poi ym ers °include :
,......p,oly(orthoesterS).and-poly(anhydrides)., Depot_injectable formulations. are.alS,Q.pi!Qpated
. • .    . • • • • • by.entrapping'the drug inliposomes or.mictoemplsions..whichare compatible- with body •    .
tissue,

:liepreparat.ions.ofagetitsiday be given orally..P
they are administered in tablets or capsule form, by injection, inhalation, eye lotion,: • . ointment, suppository, infusion; topically by lotion or ointment; and rectally by suppositories: Oral administration is preferred.
. • • The phrases ."parenteral administration" and administered ed parenterally". as:: used • -  ...:--..%-....••herein--Means.mOdesofadministration,other.than,enteral and,topical.administrationi..•7‘.7.,:

usually by. injection, and includes, without limitation, intravenous, intramuscular,
•    • •20-.    • sintraitterial;,intrathecal;•intracapsulacintraorbital, intracardiac, .intraderrnal; - • -    • .7.-• • -. •
iitrapelitorical-,:transtracheal.•,.subcutaneous; ,subc-ntitnilat intrnartic.t.t•lar,    ,.•,    •
•    'subarachnoid, intraspinal and intrastemal injection, and infusion,    •••
•    The phrases .".systemic administration,' 'adrnimstered systemi cally„" "peripheral
•    .    ., a.drnipistratioe añ&administred 'peripheral I y7?-as used. herein mean -the admini stra.ti on ., • 
-25. • s Ofa:ligand,,drug, or other material other than.directly into the central nervous systent, _• .s-uchthat.itenters the patient's syStern andithifs;:is7subject'to metabolism and_other, like,-    .:7    •
processes,.for;exartiple; subcutaneous administration:    ,    :.7
- ..• :t .    These,i nhib i tors(s).. m ay be, ethyl inistered. to. • hum anS and other• animals for I erap
y any suitable:.route-of adminiStration, including orally nasally, as by for example', a
: .30    spray; rectally,,intravaginally, parenterally,,intfacisternally, and topically,. as by.po.wders3,7,:-.
ointments.or drops,-including buccally and sublingually.    •    •.-.    ,
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WO 2009/045497    PCT/US2008/011443
Regardless of .the -route of administration- selected, the inhibitor(s), which may be •    •
-useclimazsuitablehydrated:fornyandlor the pharmaceutical compositions of thepresent invention, .are.fermulated into pharmaneutically acceptable. dosage,forms by. conventional :.:methods known to those of skill in the art.
- .• Actual dosa.ge levels of the •active ingredierits.4i. the pharmaceutical compositions
    this ,ihV-entionmayh'e.':variedso,as.to-obtain.arrgmountoftheactive...ingredientwhichiS-::,::;    ;'"ts •
effeetiVe.AO;aehieVellie•desired,theimpeutic,respanseloraparticularTatient;:icomfpOSition;-;:::::..-:.;•. and:mode- of administration,. withoutbeingtoxic%ta.the-patient:- :    •
.- •    •-::PPlie:cOneentratiori of acilis.elesedCOmPOUnlin:a:pha-rritaceutically.a.CeePtable:-..•....F!;:-,--
    ;.: .1 0..    !.-rriixture .v.vilIVary-deperiding: on 'several. 'faCtorS,- including the-dosage• of the .cornpOund
adminiSteted,.the,phannacokinetic 'characteristics of the, compound(s)
die tOute,of.administratiom.ifrigerieral,:the.compoSitions, f,thisr.irivention..may-be^4--il>vi:.,.,14...:•
solution containing:aboht4.„1''40%-rw/vtof.a.,ceiripotind-disclos.ed:.::::,-?.::',..,
. herein, among other 'substances, for parenteral administration. Typical. dose .ranges.:are, ••
    1 5 :    •from about 0.01. to about .50 mg/kg of body:.weightsper day,-given in 1-4 'divided doses... ..
... Each divided dose-may contain the same or different *compOunds • of the invention: - The . • . • dosage will...be,an effective amount depending on several factors including' the. overall
health ,pf. a patient; and the formulation and route: of administration of the selected compound(s).
    20    The, term."Cv,alkyl"- refers' to substituted-or unsubstituted.satitrated hydrobarbort
•    .groups,..inclUding straight,,chain alkyl and branched-chain .alkyl groups: that contain from...,.    .-•
•    x toy carbon's:in the chain; including haloalkyl groups such as trifluoromethyl ..and.2;2;211
„Coalkyl: indic•ates..a hydrogen vThere.the.group.is    a•terrninal:Positio.n,...,
a bond if internal:, :The.term s .'c,-,_;,al-kenyl."-..and..`;C2.jyalkynyr. refer to substituted or-• .; • .25.-:',:),7,.:.unsubstituted:linsatpratect:aliphatic groups 'analogou.s.;.inlength and possible suhstitution.to........-.. J..,..the.alkyls.described ab.ove, but    _containat least. one double or triple bond respecti:\%ely:....::::
The.Hterm-:',alko.Xy??,referst-to.an .alkyl,group:hay. ing an,...oxygemattached'fbereto,,:,:::,,:-.,:— •    „,
,.Representative.alkoxy...groupOnclude-itiethoxy.„-etho'xy,...propoXy,-
is. Wig,. hydrOcarb oils covalent] y linked hy.-an.oxygen....Accordin.g1 •the '.......,Substituent of an- al icyl.that renders.:that•allc)).1- an 'ether is -.or.resernb l es: an ..alkox
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WO 2009/045497    PCT/US2008/011443
Thelterm `CIL6alkoxyalky.1" refers to-a C1:6alkyl group substituted with an alkoxy • -group, thereby forming an ether.
•    The term•--"Cf..6aralkyr,-as Used. herein, refers-to .a Cl..6ailkyl group Substituted with an aryl group.
- The terms "amine" and "amino" are art-recognized and refer to both unsubstituted • andsubstituted aminestand Salts. thereof e:g4a-rhoiety that CM be.represented by:the.. -'• ;: general formulae:
—N    or —N.....R10
Aso    Riff
-::whereintR9.-,-:R`?.:and-Ri-g:.each independently represent &hydrogen, an
10    ;1•:;(61--i2),,;,.-R8,..'ot--0.:and..R191taken,tagether  with theNatom to which they 'are attached    :'•

complete a heterocycle having from 4 to 8 atoms inlhe ring structure; R8 represents an
•    aryl, a cycloalkyl, a cycloalkenyl, a heterocycly1 or a polycyclyl; and m is zero or an integer from I to 8. In preferred embodiments, only one of R9 or R") can be a carbonyl, e.g., R9, Rig, and-the nitrogen togetherdo not form an irnide. In even more preened
15 • • embodirrierits, R9 ande (and optionally R1° )-each independently represent a kydrogen,
-• an alkyl; an alkeny1;-of?(CH-7),„=R8-..--In• certain-embodiments, the amino group basic, meaning the protonated form has a pKa > 7.00.
•    The. terms "amide" and "amino" are art-recognized as an amino-substituted carbonyl andincludes• a'moiety.that can be represented by the general formula:'
0    Rio
20    R9
wherein le.,,R'':are,aS defined above: • Preferred embodiments of the amide will not • :
•    'include imides which may-be unstable: •
    --Used: herein includes    and 7-Membered- substituted    ,
,unsubstinited single-4ing-.arOmati -groups'in!which.eath atom.of the ring is carbon:, Th alSo.-includes .polyCycke ring sYstems:having two or more .cyclic.-rings m• :which-two,or:more •earbons,are:cornmon to: two -.adjoining-rings .wherein at. least one...Of the •-•.-
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WO 2009/045497    PCT/US2008/011443
.; rings- is-aromatic,..e:g.i4heother cyclic-rings can be cycloalkyls,. cyclo alkenyls, ; • .cycloalkynyls-,-,aryls,:-.hetero    and/or heterocyclyls::. Aryl- groups• include'benzerie,•
Oen anthrene, phenol-, aniline,-and    ..    •
-The tenns•`.`carbocyCle" .and "..carbocycly17, 'as- used:herein, refer to ••23. •non-,.arorriatie•-..
•    • - substituted or unsubstituted ring    of the ring is..carbOn.. The teats.
..-...::-"carbo-Cyclenj'and:iccoarbocycly)1.!.als-o/includepolycyclio ring ysterrishaving.two-or.rric>re,:-.:„,,-.*.....,-,,,
rings in which. two- ormore,carbon s: are coiMnon•-to ..tWo adjoiningrings wherein:at'
,leaSt:orie::of 'the rings fiS,carbo.cyclic,'6:g.; dae.other.cyclic iingS can be cycloalkyls;. • . .cyclealkeny.ls,•cycloalkynyls,:aryls;:beterbaryls,-.an.d/or heterocyclAs.
- -10 •    ....Theterrn."aarbonyl".isi..art-recognized and..includf.-;s-such:rnoieties as can- be••, •
represented by the general formula:,
0
'Rig'
wherein 'X is .a bond or represents an oxygen or a sulfur, and R11 represents a hydrogen, an alkyl., an alkenyl, -(CH2),,-R8 or a pharmaceutically acceptable salt, R represents a 15    hydrogen, an alkyl', an alkenyl or -(C1-12),,--R8, where m and R8 are as defined above.
Where X is an oxygen and R" 'or-RI i s4-iothydrogen, the,formula represents an,'"ester".-
,    .
. .Where-.X an. oxygen; 'anti R".is. a -hydro gen,: .th e formul a represents a "carboxylic acidl?. - . !Thezterms-tlieteroaryP. includes-substituted ofunsubstituted aromatic
•    inemb.eredTing structures,.more preferably 5-.-to.6-inembered rings, whose ringstructures
20    include one to fOur hetereatoms.-. The term•`.`heteroaryl""also includes polycyclic ring„
-• 'systems having-two or-more cyclic: ringsi whi ch two or more carboils.are e.-.:criantdri::to., • !.
two adjoiningrings.Whereiri atleast. on e the:rings is. eteroaromatic;.    .
 cytaic ri figs cab be cyclOalkyls„ cycloalkenyls;'.c'yclo•alkynyls,•:aryls, .heteroaryls,• and/or
lleterocyelylsT, kleteroaryl ;groups-include,: for :example, pyrrole,. furan, thiophene,-.• 95 . .oxazole„ thiazole, tri azo le, -„pyrazoIe, .:pyridine.: pyrazine, pyridatine and;.. pyrimidine, and the like.
!7"    z. Jhelerm.-!lieteroatorr: as:used herein means.an. atOrn of p.n..y•elenien .other than:. •.:.
-,:carbon_orliydrogerr.:,PreferredheterpatomS:are nitrogen; oxygen; phosphorus!and.sul fur:
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WO 2009/045497    PCT/US2008/011443
"-The-terms "heteroCyclyr or "heterocyclic group" refer -to Substituted or    •• • .;:--
unsubstituted rion-:atomatic.3--to:10-mertibered-ring Structures,-rnote preferably
•    -..rnembered.ririgs:whoSering .structures include one.-tO. four heteroatorns: The term. terms-
•    .,• • ."heterecycly1'.'or.'heterbcyclic.-group"-alSo include:polycyclic.-ririg.systerris.havingtwo. or    - '
 
•    -• • • more .cy.cliurings inythich;two,of-mor,z:carbons..are.comrnon to two adjoining:rings    •
whcrecn at leastorie.of,thexings L%:hetero-Cyclick.e.g.;:tp other cyclic nngs can be
 
dyeoally;    .ad/or
•    includ,- for examplei'piperidine, piperazine, pyrrolidine, • morpholine, iactones,.lactams, and the    .    • ;
.1 • .-;•, The term'    refers-to - a -C. alkyl. group substituted- with a.. • • •    . • -    • - • .
hydioxy group.
.Theternispolycyclyr-or."polyc.yclic7:.refer-lo.two.or More rings....(e;g4.1
•    •'; •    •cydlOalkenyls,,aycloalkynyls.;; aryls; ;:heterbarYls;:andtor
- which two or more. carbons are 'common to two adjoining rings, e.g..; the rings arefused .• . • -
•    15    -rings". Each of the rings of the' polycycle can be substituted or unsubstituted; -.;
.: •        • The terrn,"proteasonie" asused herein.is Meant to include immuno- 
constitutive proteasornes.,
,:.• The •terin "substantially pure" as Used hereila„.refers to a.crystalline polymorph:that :    • .1
 is-greater than.90%.pure, meaning that contains less than. .10% of any other compound,
20    including the-corresponding:ainorphous compound. Preferably, .the crystalline polyrnorph •
. • is greater than95% pure; or even greater than 98% pure.
.    The terra "sUbstituted" refers to moieties having stibstituents replacing liydrogerc.
' on one or more carbOns of the backbone. It will-be understood that "substitution".-or • !`substinitedwith7includes.the implicit proviso* that such substitution is in accordance',:1,-,1.-    E
•    2 5,    • • with: perm itted valence of the sUbstituted atom, and- the substi tuent, and that the.    ,
•    substitution results' in a stable compound, e.gl, whi'ch does -not spontaneously undergo    ;    • - ,
; .• -- • --    ‘,..transfon-nation.such,as:by rearrangernent,.cyclization„elimination;• etc. As used
- : •-the‘ term "•:'-substittitecr isAcontemPlated    include;a14iermisSiblesubstituents of organic,:-
.    .
•    ,;.;. . .    • compounds: An ,a.broad: aspect the. perm issib e substiMents ncl ud tycl i c and -eye] c,
30 :    branched and unbranched, •carbocYclic and 'heterocyclic; aromatic and non;- aromatic
' •    ;.    --substituents.of-organic compounds. •The permissible.substituents can be:one-or more- and
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WO 2009/045497    PCT/US2008/011443
the -Same or,differentforappropriate-organib.-„ebmpounds.. •_Forpurposes.:of
    r.:.•the;heteroatoms.such as-nitrogemmayhave. hydrogen .substituents.end/or.-any permissible •    .
subs tituents: of organi c.,c.otripounds described hereiniwhich satisfy the. valences of the''
•    ,beteit atoins•.:-Substituents.ean:..inolude;.forrexample,.:a-halogen, a hydroxyl,. a. carbonyl—    • •
•    • -(sueb.,as:-a .carboxyl-,,an.Alkox'yearb.ony.t, a.forrny1,- or:an .ac.y1); a-thiocarbonyl.-(suCh.as -:..tbioesterya,thioacetate, orLthiofor,mate);:an 4k.oxyl,raphos.phoryL.
azido; • a;sulfhydrykAn!alkylthio,...k.SUlfate, sulforiate;•a
-sulfony1;.a.heteroeycly1.,..an.aralkyl;•-• an 'arom atic Qr. beteroarom atie 4t-Wiety. . It, win be •    :
understood-by those-skilled, in-the-art:that the-moieties-substituted -on the hydrocarbon,-•- -    -. •
: • !chairi din themselves be Substituted, if appropriate.
...-.A•f.!theraP.eutically:effeetivelamount:?:-ofa compound.with respeCt .to methotottr.eatmenti7efers•toan:arriountof;the:cOn    yound(S),in_apreparation    '47n",
when administered: as-part of a desired. dosage regimen (to a mammal, -preferably..a....
1.5    human) alleviates a symptom,- ameliorates a condition, or slows the onset. of disease • 1
-. conditions according to clinically acceptable standards for the disorderor condition:to be
treated or the cosmetic purpose; .e.g„ at a•reasonable benefit/risk ratio ap.plicabl e: to. any
•    medical treatment. •
Theyterinnhioethei-P.-tefers.-to an:alkyk group, .as -defined. above.,.having a. Sulfur.
, 90 .        prefenrecLernbOdimentS; the "thioethee! is repfesented.•by,-S-:: _
. alkyl. .Representative.thioethergroups include methylthio, ethylthio, and the like.
     . As used herein, the term "treating". or "treatment includesTeversing, reducing, or
• :-:arresting the s    car.sigtis, and .Underlying pathology. of a condition.in.nranno. .
ci imprOve or stabilize a sUbject's condition. •

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WO 2009/045497    PCT/US2008/011443
Exemplification
Example 1
Synthesis of Compound 1
SynthesisSynthesis of (B)
HyOrcix:ybenztriazole(1-10B1)..(10:81-gi 80_t . mrna)- and, D1EA (200.0.-mmol:
25:85g; 3S rrtif)' -Was added lo. a'SolutiOn of N130c lencine (50.0.thmo4.1:1.56'.g)    .
phenyl alan ine rneth.Nd ..eSter (50.0..mmbl, 1 0.78.g)    Of,DMF. The mixture...*ag- •
•••-: 10    octal ed;to :0-?C . in am ice.-watet: bath -and berizotria-401,1:-y1 oxytri S(di methyl amino),    -
pho.sphonium;likafluorb.pliosphate(BOP),(80:&rornoL. 35,38 g) was.:added,in•soieral
• : portions•over fi Minutes:: • Thereactio mwas:plaeed.Ainder an atmosphere of argomand•,-
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WO 2009/045497    PCT/US2008/011443
•    • stirred :overnight: • The reaction was diluted with brine-(1000. mL) and extracted with •-
•    x 200 mL): •. The organiclayers Were' .combined and Washed with water (10-x.••...
•    - 100 InL).arid.brine X.150..rilL) and dried. over MgSO4, The •Mg804.w as removed .by -filtration and th.e.volatiles-remo-ved-under .reduced-pressure.    (A.) -(18•117--g). -To, a.50
•    .,:m11; :0-9Gcbaled solution of 80% TFAID524-was added SOcNtiLeiiPheQ.Me (45.86'.
•    -i-rnol,,; ;1.8 g)., ..-ThesOlution: was. stirred- andsllowed. to• wann•te -.morn '.temperature -aver 2. • '.t-,•-....:,...-.:-..hr...:7:11.1e-.NrolatileS,WereTemoved-lintierredliced...pre'ssure to -giV.e-    :
.,:(4-5186    -g).5.:DMF:(500-'mL),t1OBT f1.3:3"7:rntrial; 9.:91`'x) and    - -
.32.0.itiL): were-then ,added .ti) Abe    Th 6mixture was.- cabled to is AC
1-0    •ice,waterhath ati&BOTT.7-3-.3••rnmot- 32-.45•g) 'was -.added in several-partions-o-verfrve-
    ,Thi-T:.teaction:was.plaeed.iiridey avE.;con and•-31,1owed to warm to.roam-    .
overnight... The rea ai o rrikas' diluted -with-    (1500-mL) .and eXtracted. with    ,
3.00 ni,L).,.77T11.8 organic Myers Were.coinbinedand:Washed With:H20 (6..x.:30 O. rrrL) d • . brine (1 x'300 triL). an•driedover MgSO4.. The Mg8.04:was removed by-filtration and.- •
. 15    the volatiles-removed:under reduced pressure to .give. a yellow solid. Et0H (200 mL,
95%) was then added to the yellow solid and the mixture was heated to.65 °C to dissolve. all. of the solids. • The solution was then added to 1000 mL of chilled H2O and the •
.. resulting .preCipitate collected to give (8) (21.59 g).
Synthesis of (C) .
.20.-    was.m-bee-dWith•TF:A/DCM. (80%) and_wasi"stirred at roof''•
- temperature for. I hr., at which.-time thernigturemas _concentrated and placed .under.high.
•    vacuum for 2 hr giving the TFA salt of theri--peptide :amine. • To a 0 °C solution of the'. • •-
•    .T•l$A.. salt (L80. rosno1).    ( I    DJE A (3.6 mrriol, 0.7    ,fallowed.by
chlOroacetyl.chloride•(2.7 mrnol, 0.215 mL). The reaction was allowed to warm to RT -
•    .-;25, • .• while stirring .overnight-under-an ;.atmosphereof nitrogen,: The•mixture Was thendihited
(1i5-rrif.):arid. extracted: withEt0A.c (3.x 4.5' mL): The organic layers:
-- -Combined, :washed. with:f1,0:(2.xl.,5-m.L).and brine(2-x 1-5 .mL) and dried over    •
The. N a2SOW, Was...rernaved..loy :filtration and.the-voiatiieg removed _under reduced _Press nr.e.:.
•    ::.    .:The.,crudetriaterial Wasrsus.pended in E".t0AC and: filtered to:give. (C) (0.640 g):
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WO 2009/045497    PCT/US2008/011443
Synthesis of (D)
. .K1'(0,019 rnmol, .0.0032.g) and mbrpholine (0.1 l mmol; .0.0096 g) were added. to- •    •
a-sdhitiori of (C) (0:094 riimol; 0.050 g) THE (10 naL) and-the mixture was stirred • : -.•
•    overnight-under an •atmosphere of nitrogen, • The volatiles were removed under reduced:: - -. -pressure and. the.crude -material taken up•in Et0Ac (15 m14,• washed with WO :(2x1.0 nth), •
and -brine_ (2X.10:inL) .and 'dri ed.o•er :Mg 8 04:.    gS 04- w as. reirioved by'filtration and
-the.Volatiles removed finder reduced press-hi-el° -giN2e7:(D)2:;1-•
Synthesis (1(E)
ds-ad ded :to: a gurry..of-(1)).(0.094-mitiol), ita:44fiLL.O.f¬.    -    : Me01-18-1,0 cooled to-0    .Alfter •2 :at    reiction    :with: 20
 
and..diluted further witly1„0:.mL..1410..,. The.pH of The reacti on mixture-was..,  adjusted,ta;3.•,tivi,th:]:N    •and.dri ed,    , 7 -.;
Th-e-MgSO4-Was removed by -filtration .and; the volatileS were rernoved-tinder re diked.
pressure to give (E):
15    Synthesis of Compound .1
(E) (0.082 mmol, 0.046 g), DIEA ,(0.328 mmol, 0.057 mL) and HOBT;(0,133
nimol; 0.018 g) were added th: a stirred solution of:(F) (0.082 mmol) in DMF (ZiAL)-. The.
•    :, mixturevas.cobled to. 0.:?C in ,aa.ice..bath    g)    .. •
several.-portions: •he•mixture:was:stirred at 5..c10.finder an atmosphere-of argon
•    20    overnight. The reaction was then diluted with H2O- (15 mL) and extracted with Et0Ac..
- The organic layer, was. Washed :with. Water, .sat: NaHCO3,.and brine and dried over • - anhydrous. MgSO4: The. IVIgS0.•‘• was remoyedby filtration .and. the volati 1:0,1 reirthvi...;d:*. under reduced pressure to give cOmpound 1' (0.034g) (IC50 20S CT-1_,,,.100nM; IC50 Cell- • Based CT-L<100nM).
25    Example 2
     7    Compound, 1. (1 .-o-g)_ was: dissol vedin ethanol- (16•mL) heated.tO 80.°C... Water
m14.was :then, saowlyadded an d. thecle•ar ,soludon; was allowed. to cool to ambient-•; temperattim and:- the §otut ion; was brought:1o. stipersaturati on,by -evaporating of 10 TriL:of: , 'selvent with.:conipressed air ,,The:resul•ting crYstals-.were -tilteted,. washed .with:gl.r.nt
•   

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WO 2009/045497    PCT/US2008/011443
,deionized water,methanol, and:dried under vacuum for 12 hours to provide crystalline:    •.•
. • cortmonnd -1.'(,0.9,g) with a melting point of 212 °C.: • :.
•    • -    •• • The characteristic DSC: curve- of the samples is- shownin .Figure '1 as recorded- on
• t• •    • T-11/V1nstriinients•Differential Scanning Caldrii-neter:2920at•a.heating rate óf.10    •
5    Eatainpk.3_
,    •    7 •-•    7
Compound 4 -(11.0 •-g). was dissolved in acetonittil (1,7- triL), heated - to 80 °C Water•    • .
..:-,c,-„,-.7,,•-•-...-..•,;,•(8~..trih)-,.,Was,then,slowly-added-and the clear.solutiow:was.allowed...to..cool-to-arnbierit.,-,..;••-,•::::;.;.•',..,':!•,,:••,:.••••:.•
•    • .temperature and•the solution:w as brought. to.,:supersatutation by.evaporating -off .1•0.ro:L,of.
•    '..•§Olvent4vitlucompressed    T.he•resillting- oryStals:Vere•filtered; _washed -w ith•
,12- hours tp.pra)zi.cie.•mystatiihe::-:.t.-.!.
cpki.Und,1•-..(0.‘85 g):rwith..a.rrielting• point of 212  Example 4    „ •    • .    •••'

s. Compound 1(1.0 g) *as' dissolved in ethanol (17 mL) heated to 80 °C. Water (5 • mL).was then slowly added and the clear solution was allowed to cool to ambient
15    temperature' and the solution was brought to supersaturation by evaporating off I 5rnL of
solvent With compressed air. The resulting crystals were filtered, washed with R.-.7AL, 1:1 ••• _ : deionized water-ethanol, anddried under vacuum.for-12 hours to provide crYstailine - • . ,
- 7. •cOmpound 1(082 g),with-a melting•point of 212 °C.    .-    ..    • • -•- •
Example 5
    „'-Cornpoun1il1(1_0 g) -was iiissolved in ethyl acetat6. (30 niL) •Lvfat.ed.to 80 '.C..- •-••,    •
Water (5 mL) was then slowly added and the clear solution was allowed to cool to    • : •
•    -arnbic-i:nt temperature.:and-the. solution was brou•ght•to:supersaturation by evaporating off    - • •-•
••    •    • • • • 90:Tilli, of solvent.with-compressed    The 'resulting crystals were filtered, washed, With'. ?:-:• ••.    .; • :•,.:
- %.:..;5-Tn-1, • ethyl:;acetate,' ariddried. Under -vacuum' for:12:hours-to provide
•    . 25 • • . -110".'60. gywith a melting point of 212C. . •

Example 6    ..•
•    '    gYwas.-...dis•solVed ih Cthadi(i'5 friL)-...heated to-80. °C..•.•Water-.(5
L).wasithen, slowl.yaded -,and•t heti ear'solution•Was• allowed • tOlcooh-to    •
•    ••-•• ,tertip.erature•andthe,-solut ion was brought to .superSfattiratiOri-by evaporating off 1.0.rilLof..7,-.- 0    Solyent4.itlycompretsed_ air The resulting crystals were filtered washed with •10-.•mt;.:4,7_4:•;.:    '
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WO 2009/045497    PCT/US2008/011443
.deionized. water-ethanol,. and. dried under-vacuutn,for 12 hours to provide crystalline... compound -1 (0.54 g) with a melting point of 212
Example 7
r'oTPA    - • •
•    CbztIN    TFA-H2N
 H2, Pd/C , •    .• 0
.    (G)    • -(F)
•••••••;.--    0
0    0 ;
    Ph    (E)
HOBT, BI    H 7.1
D1EA,
7     

 
Ph
5    Synthesis of (F)

1 0.
O. V    Compotind (G);(0.43 g) was prepared •according to U.S. Pat. Ap.plicaticitaia. . :2005-0256324• and was added to a flask along ith Pdte. (10% wt, 0.10 g) fo4tmed
.slow:addition of;TFA.(3.5-mL)...-The flaSk.Was evacuated and-back-flushed witii.hydrogen•'z%
gas three times and then the reaction mixture was stirred under one atmosphere of — •
•    - hydrogen -at,roont temperature for two•tours:. The reaction mixture was .then filtered. • throughCelite .eind the-ill:v:1k. was .concentrated under reduced•pressure.
. Dichlotorriethane-(25'mt) was.added and the volatiles removed under reduced pressUrc.- • syrup was dried.•under high vacuum. to. a colistan•t-    t..
.• syrup wan then transferred.to•50 mL volumetriellask.and rinsed-with 8.5    diethyl•.ether

15    tO yield Crystalline compound (F) (0.33g).
Avelthesis ofCompound I
-A .10.-Int volumetric flask was charged. WithJ,-,hydrOx:yti-enzotriazole "(HOBT,, 0;54
and N;N.,NTW‘tetrarnethyl,s0(1H-benzotriazol-'71--ypuronium .hexatIvorophosphate
g):and diluted to .50' ml..; withDME...This stock 'solintidn oftoupling -20    • I'd-agents Was 0...40 M for both HOBT and HBTU.
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WO 2009/045497    PCT/US2008/011443
-(E) -(0:41 '6,- (E) (0.33g),.• and the. coupling reagent stock solution (2.7 mL), were added to :a 1.0 mL volumetric-flask and the mixture-was-.cooled to 0 °.C.. DMA .(0.56 mL) .  was then added .drapwise to.the cooledsolution.. The mixture was allowed to stinat for 60 minutes. and-was th.en.quen•chedby the:additi OTT Of saturated sodium.bicarhort
..(15 11.11)..Tht=orrixture-    diluted Withoethyl..acetate (35 mL) and .thelayelis separated:`
The: organi layer    vvashed .saturated_sOdium*.bieat:b:onate (3 .x-15 mL); 'brine: (2 :
int) -afid-dri ett.overrsodiumsullate:'.11.6' soditim:sulfate wag. remo.vedhyliltration,and. the::
rernov;ed:under_reduce&presSurelo.give -a-thick:syrup :whith:Was further-dried under high v,i0.1t1171). 40" giVe:f.i: cride compound 1 as..a..foam
10    Example 8
.1:    .compoUndl (0.590 g) was completely dissolved in methanol:CIA.
•    •    •,;:?1,stirrineandheatingin an oil batht(80.3q)..and deionized :water; (17 mL) ,Was. added:.
dropwiSe. -The. rnikture-Was‘:seeded...with:.Crystalline.compound 1;. stirred:and alloWedto. • -: slowly evaporate for 12. hours to approximately 20 mL to precipitate compound:II.. The.
15        • suspension was filtered, washed with 1:1 deionized water-methanol (4 mL), and:dried
under. vacuum for 12 hours at room temperature to yield compound 1_ .as
(0.25:g):* The-crystallization was.repeated. two additional times to yield. crystalli ne. compound 1 (0.13 g).
Crystalline compound 1 (0.3 g) was disSolved in isopropanol (15 mL) by stirring and heating in an oil bath (807°C): The solution was concentrated under redueed pressure- • 7.%F.:<15:1ICE:    5-ri4L,• Deioni.zed water (20 rAL) :was •ly.iick-ii. added
- • suspension-was rigorously stirred for 1 hour. The glassy precipitate was filtered, rinsed With:deloniZed.Water.(25,mL).;f:tnd .dried •to yield .-amoiphOu-s'..c...aartpo.and.S -(:0 3, g)....
•    •    The characteristi c.D S curve; of the- amorphons sample is-shown•in Figure-7 • •
.125    whiCh.: wa. s-tecordectOri a TA Instruments Differential Scanning Calorimeter 2920. at.a.::
...heatiitg rate of I:QC/minute for    aniOrphous. form. of CompoUnd 1.
.:•::.-:The•Characteristic. X-ray diffraction:pattern of the iannorphous poWder ifi.shown • fFigUre '8,, and was teddrded.: on,th S hi ni ad2u. :X RD-.16000-un d er Cif Ka- radiation., frol.tage: 'and'enrient Set-at.40..kV. and .40:mA,-. divergence -and scattering. slits set at 'Iand.receiv ing
slit set-ate:15. min:-Nal scintillation detecto•uSed.for diffracted radiation;. .a  continuous..scan'at39/min.(0:4'see/0:02?.step):froin-2.5. to:40°,20 was-used; samples were
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WO 2009/045497    PCMJS2008/011443
placed in an.aluminurn holder with silicon insert; and data collected and analyzed .with XRD-6100/7000 v.5.0].
Example 9 Synthesis of (F)
flask-•was-cliarged iNitli.0 3); andethyl a oetate.(400--raL) and the. solution. was_."
-.;cooled    15--minutes-With,;Sfirring:.:..Tritluoroacetic acid (200 mI.,),:waSf-, •
dropWise-,.:maintaining an. intern al:teinp.etatufapf less than d 0°C. 1?d/c.(3.:64)
' 'added:in one;Torti    nd the flask -cknas. purged ,under.high-vacillirn and refilled with:.
:hydrogen three•times-." -After- 2- hours,-.the :reaction. Was,filtered throughCelite ancLther----;,-
10    filtrate evaporate() under reduced pres'sure.to a thick 9range oll which we -swirled,v,-tntIy
di eth ykether.. :As:•the, flask was sWirlekl.fine:crystals formed.. ThellaSk-WaS ...allowed to 'sit atroOni:terriPerature;- and . rapid_ crYstallizatidn    •-.-
-:arribient;teMperatiire; the flask was capped-tightlyand placed in the freezer Overnight-(<•
5 °C). The resulting crystalline solid was filtered and washed with ice cold ethyl. ether
15    (50 mL) and dried under high -vacuum. Fine .white crystals (14.1 g; melting point: 137 •
°C) of (F) were obtained.
Synthesis of Compound 1
    ,A flask 'Was: chargedWithfF).(10g),*(E) (153. g), FIBTLI (15.3-g),--1-10Bt-(5.-5 g),    •
•    :and DMF (300m4)..•:The mixture,was stirred vigorously -until dissolvedand. was -placed in    •
20    art NaCl/ice., bath.(-5 °C). After 15 minutes, DIEA (7.1 m1_,) was added dropwise over
•    : <10 Minutes,.maintaining.anintemal temperature of less than -3 °C. After addition w-as, .
•    complete, the.reaction.rnixture was stirred in th9 bath for ryne.hour avid    quenched by
addition of .saturated,N aHCO3 (aq.) (200 triL). • The slurry was extracted with ethyl • -
.acetate:0-.5.1.,) and th6 organic layer was Washed 'with sat. NaHCO3(aq) (2 .x 300 inL); - and sat. Ng-1-(aq.) (200:mL), and then dried over MgSO4.
The organic layer was concentrated. tn.-5:0 mL under reduced. pressure and
-rnethylethy1ketone:.(20.0 mL) was-:added, .and.-the.soluticn. was again conCentrated,tri-,50.: .
MethylethY1 ketone1(.125 iiiL),WaS:acided again.; and• the sblution waS.stirred'
bath- (80 °C)'Until.ellear.-• 1The:Solutioii.was then alloiuec to cool and Was -seeded with. pure ; ;30    crystalline.CompoUnd 1: The-mixture-NvaS•-.Stirred •fon.2. hours at 25 °C and then OveriiiAht:..
•    - at ,O.Pa:.?The, whitesbli &precipitate. was.ftitered -and:Washed With ice cold methyl ethyl - !.: - •:, •
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WO 2009/045497    PCT/US2008/011443
ketone (300 mL) to give white Solids. The solid was dried.underbigh vacuum at ambient .•
tent perature to-a constant weight to yield-13.5 g of pure compound 1:
Example 10
 

 
HOBT, HBTU
BIER, ONIF
 
5    Synthesis of (F)
•    -A. flask was charged with (H) (100 g) [see:.•Bioorg. Med. Chem. Letter 1999; 9, •
•    2283-88]; and dichlorornethane (300 mL) under.nitrogen and the solution ww.,i coOlii'd it :•,anice.bath    :.•Trifluoroacetic -acid- (136,9'mL)- was-added dropwise with stirring    •
at 0-10-°C; after which the reaction mixture was removed from the ice bath and stirred at -•
 
10 • room :temperature forl-boiifs.. Methyl tertzbutyl. ether (300 -mL) was then; added -and.
•    1.11J, Of SOlventwas-emaporated AiticlOtr., re d.w.:.:(3..pt--(-.;:ssui-e. • MIRE (7.00 tr.11,) was •themackied. --
via addition funnel and the soliition stirred for.20minutes at 20 °C, then heptanes (1000 ••
.111Q were -added ..wi thiri..10._rainuteS, and the reaction mixture cooled to 0-5 'C. • Ted .•
.:.reaction-mixture-was stirred. for 30 minutes, and:then:the-solids.were filtered,, ii.11:sedwith:..,, • ..1-5.'    . cold heptaneS-(0..5!?C,.!3.•x.•100;m1;) and. dried 'under-high:vacuum to the. constant: weight.::-.-:•
t~r•ielai 90.69 g of (F) as a white solid.
•    Synthesis of-componnd
A solution of.,(9•(137..-5,3'g) in DMF (900 mL). waS.cooledin a.NaCIRce bath to;-2
1- BTL-1-(13.g:06..g);H.OBT,(5-5..90.0-,..(F)'(90:00.g)-aild ice cold•DMF (180.ML):wet0  20,    then added.tObe:sOlutibrifdllewedby addltioh.Ofteat D1EA•(67A 9 g,. 509.66barrip1)via,,, ;:•;
.,,-:a.droppingfunnel- at •a.rate.suchthat the internaF.temperature remained at
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WO 2009/045497    PCT/US2008/011443
 
two hours neat iseproiSy.lethylamine.(24,0 g). was added via.-a dropping -funnel.- The Inixture.was stirredat-0-9C-until conversion9.9%,....The reaction mixture was:then  transferred.portionwise.into a dropping funnel.and slowly•added•to an ice cold halff. : saturatedNal-ICO3-solution (3,6 L).(internal.terhperature maintained at 2&°C): -The-
. 5 .    resulting slurry was .stirred    a mechanical . stirrer for 30 minutes and the solids- vv-,:ere    :„•
thenfilferedandthe filtertakewashed ..with ice-cold rwater -(2:x '-1350;m14,- The
.+    were then diSsolVedirrldichlordmethane:(2:RtLyandthe-orgarii-ephase was:extractedtWith-.,
r •    water: (portions= of27:00,ML)Until'irelatiVeipercerit&-ea
H•PLC (200 UL:Solutioti for HPLC• sample).; The Organic. phase..Wasfiltered: through a-.
 

 
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WO 2009/045497    PCT/US2008/011443
. • The resulting compound 1:(17.3. g) was completely dissolved in methanol (275
nth). by stirring-and h.eating in oil bath (batbset at,8-5,-,C; mixture temperature less than-76-5  9C)..Deioni.Zedwater (75' mL) was-added •dropwise•over15, minutes, and the clear-mixture, ..
•    wa.s. allowed :to.    torOonitemperature, Seed •crystals of compound 1-were added tc;.the
.    alined.75.•,-Olution;•and the mixture-was allowed-to slowly.concentrate.under a stiear►    • • • •
•    :.:::.:compressed ai•to approximately•250 mL.ovev9 hOurs.,The-crystals- were. thenAltered. - f.-and•washed.witliA-,1,--deionized:Wateriniethanot
•    • Under vacuum. for l2 hours'.at:221-9Cqcc yi eld,crystalline-compound    410
Example 12'
10 • •    Crude' cdriappund.1 (.12.1:g) was corripletelY 'dissolved in methanol.,(50 n1L) by .
,..;    Stirring-and heating in oil bath '(bath set at.85 •°C; mixture. temperatureJess Allan .65    • - •••
The:clear solution Was.allowed•td COCA :to rain temperature. and.Seed-crYStals .,:.compourid•l-were•-added.,to,theirsolution.:,.The inikturemag,allowed to. -Crystallize,over
. three hours at room temperature. -The resulting solid was washed with 1:1 deionfzed - . 15 . water-methanol (500 mL), filtered5.and- dried under vacuum for 12 hours. to yield .crystalline- compound 1 (9.4.g).
• lExarreple 13
Synthesis of (F)
•    flask.was, charged-with (H) (1 gyand ethyl-.acetate (20-mL) and the-solution was 20    • cooled in an ice bath for 15 minutes with stirring. • Trill uoroacetic acid (10 mL) was-then .
.'added dropwise, while -maintaining an internal ternperature -of less than. 3 'C..- stirring at 0-DC-.fort. hours,-the reaction was -.allowed to-warm to, ambient temperature and--was stirred for-two-additional hours.. The solution was then evaporated under-reduced • y pressure-to 'a.thick'colodess oil:- :This .crude mixtur& WaS..swirledgently
- t- -diethyl ether and•s:the-Solution.was swirled, fine crystals formed., •After230' min.utes at •
ambient ..ternperature, the flask Was•capped tightly and-placed-in Ithe.freezer overnight:
The• Fesulting crystalline solid was filtered and washed- with ice cold diethyl ether,••and
•    •• then dried or Itigh.v4cuum to-a constant:Weight to give fine white crystals of (F). (670 mg)..
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WO 2009/045497    PCT/US2008/011443
Example 14
Synthesis of Compound 1
Compound (E.). (14.2.g)• fIBTU (14.3 g), HOBT (5.1 g) and DMF (300 mL), were .
tidded.to.(F) and the mixture was stirred at room temperature to complete dissolution....
The reaction was cooled in ice- bath•or 15 minutes; and. DIEA (32 mL) was added bver.15.._. .;-minutes While-maintaining an.:interrial temperature.•ofless mikturewas,then.stirred-at.0.-for One'hounbefore it was quenched with
s-odiurn bicarbonate.(200.mL) .The mixture. was extracted with ethyl acetate (1.5IL); and
 organic layer:was ,Washed:w.ith.saturated,sodiuMbicarbonate. (2-x300.mIL),and,:-. 10'• deionized crater .(1 X.200 mL).• The.eombined'aqueous wash was extrf.,:ctcdwith :eth.y1. •:
acetate (200.mL)- and the organic layers were:ocimbiried (I L;).:. •
',:The.coribinedbrganiclayerS.:(1,.74.;)_Weretobnceritrated under TedUced:pressure, 100.-MIL:f011oW.ed-by addition of rnethanal:(200-fripy and theIrriikture.Wa-s -again' • concentrated to 1.00•inL. Additional methanol    mL) was added, deionied.water (75
•    15 •    mL). was slowly added with .stirring, and the mixture concentrated to 300 mL. The clear .
solution was seeded with. crystalline compound' l,. stirred and allowed to slowly concentrate under -a strearn of compressed air to about 200. mL. The off•whiteAolid•was . washed' unti•solidterned white and.filtrate•tumedrclear with:a 4:1:.deionized methanol (2 L)    deionized water-methanol (500 mL). The resulting solid was
•    20 • :dried-unde• vacuum•for- 12-hours at 22 to provide- compound 1 (16.8 g):,    •
•    . CompoUnd ,l was coMPletely.dissolved in ethanol (200 rriL) by stirring and • :• - .heating in oil bath•(bath set.at•85_".C; mixture.temperature less. than.65 °C). The.clear  solution was alloWed to. cool to room temperature and seed crystals of compound 1 were added:to-the stirred solution; and the. mixture,Was-flushed with air. and allowed to 25 • •    :crystallie,.,' The Inixture was then-filtered; Washed.Withl.:-1 deionizedwater-ethanol. (-200
and dried unde•vacuum..•for.12 hours atroom temperature to yield-10.2 g
crystalline eornpoUnd,-1.
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WO 2009/045497    PCT/US2008/011443
Example 15
Synthesis of (F)
A 500mL•flask was equipped.with a:mechanical stirrer, thermocouple, cooling
(G):(12.5 g)-waidissolvedin ethyl.acetate-(125mL) and the clear solution Was
cooled. to 045:G folliowed -by slowa.dditiori of .triflueroacetic acid (375- mL) such-that the . 4-.-0internaltemp'erattire.was:rriaintained.below's1.0.M.•Aftet warmin gto, room tempe.rature-;.•,--,....:f.•4:•..',
•    54Y614/0(1,25..g)Lwas•addect aridghe f.reaCtion. mixtureAnder .an• atm osphere ofhydrtagerkr.-; for 2 hotds.:::The-reactiOnrnixture.was,filtered through. a glass -fiber and rinsedwith ethyl. .ia:cetate.(50.inL)?..'::The:filtrate-masgherrc-oncentrated..Under roduced• pressure go-yield:a)
1.0 -.•MTBE:.(50:triL)-was . added tughe-oil. and co-evaporated to .yell ow oil: at 12-5..,9C.: •
•    MTBE (60miL)•.,wasagainadded'and the•mixture waS:'.coOled,-La-.,-.1 0 7..'C.;-.and. stirred Tot,60: •
-minutes-1-1-eptans020::rnL) Were-.then-iStirred.rnixturand stirring::
 ,   
:; was' contiutidd-at --t4:01?C'-for. an:additional:15    leeted .by
`filtration and the.crystals were rinsed with heptanes (2.xA0 mL) and dried under high . -.15    vacuum at-room temperature (22 ?C) to a constant weight (10:1 g).
Synthesis of compound 1
.A -flask equipped with a=mechanical stirrer; thermocouple, cooling bath,-•-rkitrogen• inietind drYing.fdbe was cliaried"with:DNIF;.(F) •13:9 g), (E) (244.8 g), HBTLT (242,8 g),.and HOBT (86.5 g) and the,mixture was stirred,.and cooled to.0,5 °C. DIEA (156 mL)
•    20 .. was then added slowly over at least 30 minutes, While maintaining temperature between . 0;5-PC: :The reaction .mixtureNoaststireed.at 0-5 .?(.7.! for one hour and was :hen poured: into , a-vigorously stirred .saturated solution-of sodium bicarbonate (3630 mL) and. ethyl. acetate_  (900 nit): Additional ethyl acetate.(2000-ML) waSadded to extract die product andlthe
••. Organic layer.was separated:. The:aqueous layer wasghen- extracted. with..etbyl -acetate,.
.25 :•-_-:--(1930.mt)-..-The organic phases-were combinedand•wkshed with saturated ,solution Of ,-
sodiurn'bicarbeinafe.;(2420.1nLy andhrine..(2420.mL)-;:dried over magnesi    . •
r. g);•filtered•throiigh glass-fiber filte,tand    with ethyl .acetate (2.x -360:rnI.-).,.
.:;:methanol .(7251n L) :was. added and•coevaporated. under, reduced preSsure to yield,serni, . 30. • solid compound 1:-    KvaS. dissolVed,in.metbanel (5320 fuL) anchthe'
solution. Wasstirred:while water (21.3.0 mL.). w as. added .oven•twentyrninutes.
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WO 2009/045497    PCT/US2008/011443
addition of water was complete,' approximately. 0.3 g ofpure crystalline seeds --w.ere..added
• and . the .inethan ol/w ater soluti on was stirred for. three hours. The resulting .crystalline. ..white solid was isolated,.byfiltration and-tho fine. white-crystalline product was rinsed
'Lwith$a methanol/water solution .(1.21; 1200 -mL).: The resulting solid was rinsed. with'
5 •    methanollwatey' solution :(.1,:1:,---1200 mL) and the -crystalline product was: poured onto •
,.:drYinglray and•dried-iota constant weight underthigh.!vacuum.at.2:71°C.under..nitrogen-'
-    .
-7:bleed to=Yielttcrystalline compound -1 (230 g).
Example 16
Synthesis of (F)
.A.:100trai; three,neck:roiind bOliorri flask_ Was charged with (61(5- g) and 1 .• ..• :.:•dichlotOmitharie-(15..rriL): :Themixture-was:8tirred.iintil the solids had disSolved, ;and, ,thentil-aCe&ii-itafpicebath.. After20 tininntes-,Itheinternal-teinperainii had -..,a-rieFtrifltioikiacelic:aid'Wasadded drOpWfse oVer 5i rn =After the addition W as: eonipldte,..:•'-"L'-.  the flask was allowed to warm to room temperature.. After 2 hours, MTBE was added to
15        :the flask (35 •iL) and-the mixture was cooled .in an ice bath, wherein (F) began to.
.crystalliZe. during cooling.. HeptaneS (65 _mL,) were then added to the flask dropwisoover;
15 min and-the-flask was placed in. the freezer (-5 °CY. After 1 hour, thesolid .productlwas: collected- and w:ashedmith•heptanes:(10 niL) te-provide 4.57..,g of Example 17
20    Synthesis of Compound 1 Citrate Salt
 
Compound 1 (1.0 g) .and •citric acid (2.7 g) were diSsolved in THE (75    and
lacetonitrile    solution was :then stirred for 2 hours at room ternperature;..    • -:
-.,Which:time-a.whitoprecipitate formed-. The flask was then cooled to -10 .°C.and stirred, .
overnight:•-The:Solids:weee filtered and ..washed with 100 pl. ijicetobitrile.to.give 11...,52:.g.„, ? 5. • • •.:"-of the citrate salt of compound 1.
 
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WO 2009/045497    PCT/US2008/011443
Example 18
Synthesis Of (H) and (Q)

•    A suspenidn of dimethyl hydioxylaminehydrochlo6de (10.53 g. 108 namol) in DCM (270 mL) under an atmosphere of argon was stirred vigorously for 0.5 hours-
. • . 'followed by:additiori of TEA: ('0.92.-g; 14.754nL, 108 minol) Via addition funnel:. A    :
Sbhition of Bo.c4•,encine-01-1.(25:0 g,108.rinTiO1) itiDCM (270.mL)•waSeobled to 0•?..0 :.• •
llowedhy dropwise addition of isdbutylehloroforrnate (14.71 g,: 1.3:984n 108;:inmol).-:•.- •
•    : via..additioniunnel.:,Thesnixture,was thrth et .wole0; to::-;20 °C and NMIYI.(10.92 gy 11:87 ,•. . -rri4.i08,inrnol). was added-Nia addition funnel-at su,eb al-ate.to maintain _tine. it    •    •
.ternperature.below.-:1.04?C: -Aftenstirring-foi- minutes at.-20 ?C; the previousirpre.pared:::.
,    dimethylhYdroiylarnine solution:was added via:a-Wide-bore Teflon cannula. , The reaction
: 1.5 •    w.as:removed;•frofir the•cooling.bath 'and Alto w ed -to warm' to. rooirr temperature,- :     

.•    The.mixture wastheirdi lute& with water_ (1 00 :rnL) and-stirred for.15. minutest.' ..•
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WO 2009/045497    PCT/US2008/011443
The layers wereseparatedand the. aqueous. layer extracted with DCM (2 x '50 -rnL):-.. The. • organiclayers.Werecombined;-wasbed.With 1 N 1-1C1•(4.-x, -.150 mL),. water (I.    : • .
•    • sat•NaHCO7i .(2..k.1001nL),:brinc• (1- x.250 mL) and dried-aver Na2SO4... The Na2SO4fwaS •••
•    .reinoved.by•filtration.and--theeia    retnov,ed de•-educed prQssure..to give (1) (28.05 1:
5 .. g, 102 rnmol).
Synthesis of (1)    •

•    '.    :(I)    g, 56 A- inrricil,),.in .104 ML•of dry. THF,, under
•    %atmosphere of argon was_added- iSopropenyl. rilaiesiurn bromide (164 miL,-; (1s.6q;:0:.5.1Vi :solutionkin;THR)1.drOpWise    Therate
§. adjusted :sidob lhat the internal.reactiOn.teniperatUre was maintained' belrAw    : • •
After six .hours- the reaction-mixture waS'pehred:intd-250 nil., of •sat,,N1-14C1: and 500,a-riL:-.    •
--Wet    stirringfor,30:minutes the aniXtnrebecame,clear ,and. theyolatiles:were!:,'....',.
—.removed under redneed•preSsure-and,th:e.crude::material diluted withEt0Ac.:(200:mEi)..,-
The.layers were separated and the aqueous layer extracted with EtOAc (3 x. 150 ml...); the
    15 .    organic layers•were Combined; washed with water (2 -x 1-50- mL), brine (2 x 150 mL). and .
•    - dried •over MgSO4. : The MgS0;4 ‘vas.rernov.ed:by-filtration• and the volatiles removed: . under. reduced preSsure. Purification .by flaslychrornatogr aphy (15:1 hexanes/. Et0Pic) gave.(1) as a solid (7.5.g, 29.37 mmol).
Synthesis of (K) and (L)
    20 .    •    To a 0 °C• solution of (1)(5.0.g, 19.58 mmol) in 200 tril, of Me01-1. was added •
-.CeC1:3■A1-17,0 .(8.75-g„.23,50.mrnol):- -The Solution w    rid& an atmosphere otf • -
__argon until the CeC13,71-120 was completely dissolved. To.-this solution was added ...
NaB1-14•(0.88 g,    10 portiOns ove,..2-n-iinuteg.. *The reaction was then 'stirred- .
. under-:an atmosphere. argo•-for.6 hours, :The• reaction was quenched: at -0,9C
    25' 7:    1,1:/i.th:approkitnah-Ay    nil; of •glacial*1-10A:b-:and ,afteit• 3.0: 1Tli tes of additional •stifting-at,...
•    O'"C:the:raixturt;!..h.ocofite clear.l'.The'volatiles`.Were.renr6V,TA *Linder reduced presvilte:arkt% th6-rerrialhingt■illakeb    .Was washedWith -waw    •
th.E.;),;.brine,(-2: x 100 in.L)) ,and.dried. ay.& :Mig,SCV The-MgSO4-Was removed-by
-filtration    .vo.latiles:remdved under.rednced•pressure giving (K) and..(L),.cas a
Of..ditereonierS. 4:5:1• as deterainied by
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WO 2009/045497    PCT/US2008/011443
•    Synthesis of (M), (N), (0) and (P)    ,
'    • , • • To a Solinion-of(K).and(L).(0.025 g, 0.097.'inmol) irtDCM (1 mL) Was added    •
••     rtzCPBA.(0:018'g,- 0.107 -mm01): The mixture was stirred at room temperature -for. one, .
•    . hour-af:Whieh time the mixture was diluted with sat..NaliCO3 (5 rriL): The layers were
•    •• •
:5 • separated and'the aCitiecitts layerextractedwith DCM (2 x.2 mL). -The.orga.nic
-    -    Were--combined and Washed'with water (2x- 5;m13);..brine (2 5rnL),and•dried.over.,.....,;. ;    ,,z,
•    - . JI::•1\11-gSO4-. -:The MgSO4 was removed by filtration andtheT.volatiles removed under'rediteed:F
-    • '    •    ./    pressure to give an oil. • Synthesis of(H). and (Q).

10.    .• To a-solirtion:of-DeSs:MartinPetiOdinaffe(0.023 g; 0.055•mmol)    mL    ;    ::•:- ,
.• at5?.CWas-:added.a,:mixture of(M), (N),:(0);:and:.(P):(0.010 g 0 037 mmol) as a solution
in MeCN:.(1-..inE).:The'inixtuie.-waStPlaced-under!ank:affhoSphete.-,Of argon Undallowed:tti::,2::?:''r
•    -    •    Warn:ii: to :thoin.-terriperattite. while. stirring '0:Veil-light.: When Cbinplete, White,precipitate
had formed and the reaction was cooled in an ice-bath and diluted With 2 mL Sat. .    e    • - •
15    NaHCO3. The.mixture was diluted with 10:mL of Et0A.c and the solids removed.by
filtering through. a:plug of Celite. The mixture was transferred to a separatory funnel and.    •
the layers separated. The aqueous layer-w.as extracted with EtOAc (2 x 5 mL) and the-%  organic•layers-,combined; washed withwaterf (a x-,5;:inL)- and thine (1:x 1 0-mL) and then  dried over NaySO4. The Na,SO4 was removed by filtration and the volatiles removed
20 • • under reduced pressure.to give arnixture of,(1-1) and (Q) as a light, yellow'oil.    : • • • •

Example 19    .    •    '• •    •    •    •
Alternate Synthesis of (H) and (0) -    - - .
 

 
Alternate Synthesis of (H) and (0)    •
25.    , 2. ,•:••    :-.5_-solutiorrOf (R).. (Q.2.00-g; 'a..78..mr091) in.pyridirie (3    L) was 'added •    4
- -    .•.. 10% aqueous:N..a0CI ;(1..5mL)dropwise,:a.t:. a rate such that the internal; reaction    ;
teiliperature rerned bel Ow    .-After,the addition of Na0C1 was complete, th    .;    .t
- reaction flask was placed in a.0.°C bath and stirred for tw.o hours.. The mixtur.e was.then    : .
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WO 2009/045497    PCT/US2008/011443
diluted with Et0Ac (10 ML), washed with water (2 x 10 mE), brine •(2 x:10 mL) and dried : .over-Na2aa4. The.NatSO4 was removed by filtration-and the volatiles removed under • • reduced.pressure to-give the crude mixture of (H) and (Q). .Purification by flash
:chfoniatography (20:1;hexanesPEt0Ac) gave (1-1)-as.an- oil (0.059 g; 0.216 inmolyand :(Q). -:. • as a solid (0.023 g, 0.085 nunol).
Example 20
SYnthesis of CoMpotind 1
 

 
•    • •    HBTU, HOBT
DIEA, MeCN

 
Ph
Synthesis of (F)
•    10, - •    -    To a •10 it-IL-round bottbined flask was added (H)'(0.050 g, 0.18 rornol) aridDCM'
•    . :(0.80n1L). •Themixturewas Co. oled•to 0 '"C and neat TFA (0.20 mL) was added,. dropwise. After the addition of TFA was-complete the flask was allowed to warm to rC40:1"a temperature. whiletstirring for one hour. .•Threvollatiles were then remoVed under.; • .: • - •
•    reduced pressure.andthe restiting•oil was.ichasedvith.ECM    x 2) ancl :he volatiles
15 * removed under reduced pressure.
Synthesis of Compound- 1
inli_roundhottomed-•flask containing (F) was added-(E}    GA 5-
HOBT..(0.031,.g;0:23Minol);_and HBTU (0.087. g,    ,
.,and the tnixtUre was 'Cooled tO:.0.    To-this mixture WasslowlY- added DIEA (0:077.g,• 
20 • - . 0..1 04.1TIL, -0:6 rnmol).andtherniXture-was allowed: to 'stir at 0°C for one-hour before-
•    -.- quenching 'with. Saturated:NakICQI:(5..M.L);..,Theinixture was diluted 'with_Et0Ac:(1..-5-•triL)..--.,.- .
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WO 2009/045497    PCT/US2008/011443
and the layers• were separated:',Theorganic layer was washed. with saturated NAHC0i:(3.. • . x..5 nik;),trine -(2 .x. 5- mL)-and dried`overNa6SO4: The Na2304 was removed by--filtration..., andthe ••olatiles removed..under.reduced..pres sdreto .givr, a thick oil: -.To the flaSk. •
..:••17-.eontaining the, oil was :added D.C191•'(    .and the jilaced Under high-vacuum • while,.    ”:
giving•Conwound 1 (0.100 g,    as a foam:    .. .
Example 21

     Alternate Synthesis of (S)
To a 10 1rkL round bottomed flask was added (G) (0.055g, OA 8 rnmol)iformic
(2-m1")•,. and, Pd;C: (3% wt, 0.05 g)._ Once-the tleprotectitin was cleemed•Lernpfeke:by TLC and1CMS, the.vdiatil.es were-removed under reduced pressure. The .oil was chased . With.Dervi.(2 mi., x 2)and the Volatiles' removed under reduced pressure.
Synthesis of Compound. I

•    .TOja:10 inL.y.-ound bottomed: flask 'c:oritaining (•) was added: (E) (0:085.g; 0.15.
•    .r4-0-nol); MeCN (2,0 m.L), HOBT (0.031 g, .0.23 .mmoI), 1-1BTU •(0,087 g, 0:23 .rnrnol), and
•    7- th'.wi.s•ebble&f6    . To :thiSinixture':NiTas-.SIowly -added DI E.    gi• 104 V:_k
int;.0.6,mtnO1)::The'rnixtire WasAlidn alloWed-to-stitat,0:?.0 for, 60minutes andm quen-chetliby/the:addition.of =satUrated.NaHCO (5 'rals):-., The mixrure*Was
.1Z
20    andithe-layerS separated-:-:;The :organic:layer was,washed With, saturatedt;;:-A,
.Nal-1e0:'(3;:x"-5,m7L).,,-brine.:(2':i 5 ifiL). and dried oi,ferNa.2$04. -The
- 88 -
 
WO 2009/045497    PCT/US2008/011443
 
. by:filtration, -and the:volatiles.rerrioyed under Teduced;presSure- to give: a thick    TO:the-
-.•:t1 ask containing the .6i1 Was,added.:DCK(1.-triL)- arid thernixture placedunder:high vacuum.While-swirling.giving Compound..1. as :a foam-.
Exathple 22 Synthesis qf
 

 
. Minter.:(2)4,:alL)-was.added.to a:three neck:flask eutrippe,d-with:ZI17=`nani cat. stirrer, 'atradditionfunnel,.and_athermocouple with.display
temperature of --5:16.0‘9c'..'..SOlid..caleium hyPeCh10:ri•te.:(107,8;-748-Mmbi),,wa'sAherradded,-':. 10    • •.ovevaPproximately.5 minutes, While:the: tertiperature of the mixture-is triaintainecbat-    • •..
•    approximately -5°C to 0°C. The mixture.w'as then. further cooled to -10°C to -5°C and .stirred for 10 minutes followed by addition of NMP•(1000 mL) via addition funnel:at a
•    rate to maintain internal temperature between -10-•.°C.-to -5 °C. The reaction shirry. was then stirred at -10 °C for 15 Minutes. (1:4 (47.8 g, 187 mmol) was dissolved iti-NIVIP (40 ... 15    .....:0mL)-and.added dropwisetath.e -reaction. mixture. while maintaining.the internal:_
•    temperature between    °C and•-t0    The'reaction-mixture'was then stirred at 'Pe to •
•    0 PO until the reaction- Was‘cornplete• by•TLar. Upon ntaction. completi    the mixtui-e was •
. quenched by slow addition,of 1.0 J.y1.:-.-iodiurn thinFulfate:solution .(500.mI.,),: maintaining
•    an internal temperature of-10 CC to *-5 C. Ethyl Acetate (1000 mL)-was then addedythe • 0 • layers'.were :ileparated -and the -aqueouslay.&; was :extracted ..twirx.rno,re. The (....ornbined..
•    . organic.layers were. washethwith.watei: (500. mL) ands-brine (500 -mt), dried. over
•    magnesiuin sulfate•filtered and-.concentrate• underTedOced pressure
whicirwas    hex anes-.(6:004.n.fl)'-a.ndifiltered .thro•gh. aplug: of silica to: prOVide:
4.H) as a pale yello-wpil.(20,3 g).
25
Equivalents
r,Those.skilled' i.n the' artzwilhrecogni ze;, or.-.be able' to: ascertain,using.no; more- than,.
-.1:,...-,,routine.ekperimentation,-numerons equiptalentsi to-The• compounds 'and-methods    -7
- 89 -
 

WO .20091045497    PCMS2008/011443
     .. • thevead    bed;l er i s Such. rqUiVitientsere-conaidered.to• be within theseope. of
invention and are covered by-the following_

• • .Ali titheabo-tie,cited references: and•publications are hereby incoiperated by •
 
We Claim:
1.    A method for preparing a crystalline compound of Formula (II)
0
07—\N
(II)
5    comprising (i) preparing a solution of a compound of Formula (II) in an organic
solvent; (ii) bringing the solution to supersaturation to cause formation of crystals; and (iii) isolating the crystals.
2.    A method of claim 1, wherein the organic solvent is selected from acetonitrile, methanol, ethanol, ethyl acetate, isopropyl acetate, methylethyl 10    ketone, and acetone, or any combination thereof.
3.    A method of claim 2, wherein the organic solvent is selected from acetonitrile, methanol, ethanol, ethyl acetate, and methylethyl ketone.
4.    A method of any one of claims 1 to 3, wherein bringing the solution to supersaturation comprises addition of an anti-solvent, allowing the solution to 15    cool, reducing the volume of the solution, or any combination thereof.
5.    A method of claim 4, wherein bringing the solution to supersaturation comprises adding an anti-solvent, cooling the solution to ambient temperature, and reducing the volume of the solution.
6.    A method of claim 4 or 5, wherein the anti-solvent is water.
20    7.    A method of any one of claims 1 to 6, further comprising washing
the crystals.
 
-91-
 
8.    A method of claim 7, wherein the washing comprises washing with a liquid selected from anti-solvent, acetonitrile, methanol, ethanol, ethyl acetate, methylethyl ketone, acetone, or a combination thereof.
9.    A method of claim 8, wherein washing comprises washing with a
5    combination of an anti-solvent and the organic solvent.
10.    A method of claim 9, wherein the anti-solvent is water.
1 1 .    A method of any one of claims 1 to 10, wherein isolating the crystals
comprises filtering the crystals.
12.    A method of any one of claims 1 to 11, further comprising drying
10    the crystals under reduced pressure.
13.    A crystalline compound having a structure of Formula (II)
 
14.    A crystalline compound of claim 13, having a DSC thermogram
15    substantially as shown in Figure 1.
15.    -A crystalline compound of claim 13, having a melting point of about 205 to about 215 °C.
16.    A crystalline compound of claim 15, having a melting point of about 211 to about 213 °C.
20    17.    A crystalline compound of any one of claims 13 to 16, having an
XRPD pattern substantially as shown in Figure 2.
 
    18.    A crystalline compound of any one of claims 13 to 16 having 20
values 6.10; 8.10; 9.32; 10.10; 11.00; 12.14; 122.50; 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; 39.46.
5    19.    A method for preparing a crystalline salt of a compound of Formula
(II)
 
(II)
wherein the salt is selected from a citrate, tartrate, trifluoroacetate,
10    methanesulfonate, toluenesulfonate, chloride, and bromide salt; and
the method comprises (i) preparing a solution of a compound of Formula (II) in an organic solvent; (ii) adding an acid selected from citric, tartaric, trifluoroacetic, methanesulfonic, toluenesulfonic, hydrochloric, and hydrobromic; (iii) bringing the solution to supersaturation to cause
15    formation of crystals; and (iv) isolating the crystals.
20.    A method of claim 19, wherein the organic solvent is selected from diethyl ether, THF, acetonitrile, and MTBE, or any combination thereof.
21.    A method of claim 20, wherein the organic solvent is a mixture of THF and acetonitrile.
20
22.    A method of any one of claims 19 to 21, wherein bringing the solution to supersaturation comprises slow addition of an anti-solvent, allowing the solution to cool, reducing the volume of the solution, or any combination thereof.
- 93 -
 
23.    A method of claim 22, wherein bringing the solution to supersaturation comprises cooling the solution to ambient temperature or lower.
24.    A method of any one of claims 19 to 23, further comprising washirig the crystals.
5    25.    A method of claim 24, wherein the washing comprises washing with
a liquid selected from diethyl ether, THF, acetonitrile, and MTBE, or any combination thereof.
26.    A method of claim 25, wherein washing comprises washing with
acetonitrile.
10    27.    A method of any one of claims 19 to 26, wherein isolating the
crystals comprises filtering the crystals.
28.    A method of any one of claims 19 to 27, further comprising drying the crystals under reduced pressure.
29.    A crystalline salt of a compound having a structure of Formula (II)
15    o N     

wherein the salt is a citrate salt.
30.    A crystalline salt of claim 29, having a DSC thermogram substantially as shown in Figure 11.
20    31.    A crystalline salt of claim 29, having a melting point of about 180 to
about 190 °C.
-94-
 
32.    A crystalline salt of claim 31, having a melting point of about 184 to about 188°C.
33.    A crystalline salt of any one of claims 29 'to 32, having an XRPD pattern substantially as shown in Figure 12.
5    34.    A crystalline salt of any one of claims 29 to 32, having 2e values
4.40; 7.22; 9.12; 12.36; 13.35; 14.34; 15.54; 16.14; 16.54; 17.00; 18.24; 18.58; 19.70;
19.90;    20.30;    20.42;    21.84;    22.02;    23.34;    23.84;    24.04;    24.08;    24.48;    24.76;    25.48;    26.18;
28.14;    28.20;    28.64;    29.64;    31.04;    31.84;    33.00;    33.20;    34.06;    34.30;    34.50;    35.18;    37.48;
37.90;    39.48.                                           

10    35.    Use of a crystalline compound of claim 13, in the manufacture of a
medicament for treating a disease or condition selected from cancer, autoimmune disease, graft or transplant-related condition, neurodegenerative disease, fibrotic-associated condition, ischemic-related conditions, infection (viral, parasitic or prokaryotic) and diseases associated with bone loss.
15
 
1/12
(6/M) M0131V3H
 
SUBSTITUTE SHEET (RULE 26)
 
2/12
 
11
0    0
0    0
0    0
,:t    r■I
(Sd)) AlISNRNI
0'
SUBSTITUTE SHEET (RULE 26)
4/12
In    0
TIVDS A AdV81198V
 
SUBSTITUTE SHEET (RULE 26)
 
WO 2009/045497    PCT/US2008/011443
5/12
31VDS A AdVH1182JV
SUBSTITUTE SHEET (RULE 26)

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