WO 2008/137318
`
`PCT/US2008/061205
`
`determinedas the aeeivy ofa positive ST control, Cys-Cys-Gilu-Leou-Cys-Cys-AamPro-Ala-
`andset te LOGSs,
`
`—
`
` ¢ Collection (Bethesda, May.
`se chioride anionseeretion can be examined usin t
`
`grownte contigeney in 24-well cultureplates with a iol mixture of Nam's {
`Dalbecer’s modified Hagle's medium (DMEM), supplemented with $24fetal calf seram and
`
`were used) at between passages 34 and 60, Chloride ian secretion is measered inthe presence of
`st articie similar the methods described in Dharmasthaphorn ot al. (1984) 7 Clin
`
` ett and Bigby (1903) AmJ Physici 264:C446-52, Grictly, an Ussing
`
`chamber is modified to allow maintenanceofthe integrity of the cell monolayers during the
`
`study, The modified chaniberis designed to minimixe turbulence created by the air HA svetem
`x
`10° P&4 cells are plated an a permeable supsart
`
`ainageto the monglayers,
`
`area} and maunained for 5-6 day before use. The supporis are suspended over
`
`OU-mm ealture dish to permit “bottors feeding’ by laying them on top ofk
`
`esenibed in Barret and Bigby supra. After cell growth, ihe entire ring
`8
`
` 4
`
`
`
`
`and hence edge damage is avoided. Mucosal and serosal reservoirs cf
`cy
`oxygenated Ringer's solution (pH 74, at 37°C!) that contained Gn millimolar}:
`
`chamber. No pressure is
`
`oxer
`
`po aa
`
`2) CL, PIGS: HOOs, 25; HoPOs, 2.4; HPO, 0.4; and glucose, 10.
`
`}} across the cell monolayer is measured by calomel! clectrads
`ot
`monitored with a potentiometer. Throughout the experiment, except for 3-10 seconds every 5S
`
`min whils the PO is being recorded, spontaneous tiasue PID is short chrouited and naliified by ex
`
`
`<2
`wutermatic volt:
`ge clasp (WPI, New Haven, CT) with Aw AgCh electrodus. Tissue conductance
`vedoe
`“
`2
`(6) is calewlated from the PD and the imposed carrent according to Ohm‘s law. The magsitade
`
`of changesin the sheet circuit current (Ise) is used as anindex of chlor
`
`
`
`Landsodiumexcretioningated
`
`49
`
`MYLAN - EXHIBIT 1022 Part 10 of 16
`3131
`
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`
`MYLAN - EXHIBIT 1022 Part 10 of 16
`
`

`

`WO 2008/137318
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`
`solated boo.
` soproximately b-S em. The loopy are injected with 8 test agent or
`
`2.18 described in Londonef al, 1907 Am J Physiol p.e3-108, The loop is
`
`ai byinjectingx
`
`vehicle. Following s recevery time of OC minutes the loope are excised. Weights are recorded
`
`ior each inop before and aller remewal af the fluid contained dhareim.
`
`‘The leneth of eachloop is
`
`alse recorded. A weight to length ratio (W/L) for cach loap is calculated to determine the efvets
`
`of test agent as commpared to vehicle. To determine the effect of a fest agent on sodihianexcretion,
`
`tQ
`
` YRad frem the isaps ie collected and profiled for cleotrebyte
`
`performed using
`
`Sxanmple 2: Animal Models af Hypertension
`
`Various animal models of hypertension can be used te screen the agents described herein fxr
`anti-hypertensive activity.
`in general, hypertension can be indaced in rats in al least four ways,
`mchading: genetically-induced, environmentally-induced, pharmacologically-imduced, and renal-
`
`induced. A variety ofradent lypertension models are described in Pinte of al. (1898
`
`i$
`
`3G
`
`ular Research 39:77-88)}, Badyal ot al. (2003 Inchan Journal of Pharmacoicg
`
`
`
`362) and the referenves cited therein. One of the most widely used rodent models of lye
`
`
`
`
`ais the Spumtaneously Hypertensive Rat (SNR). Other models miclude: (1} the iwo-kidmey
`—
`clip, (2) wansgenic rais overexpressingthe murine Ren? gene, (3) DOCA (deasycorhcasteran
`ig
`scelate}-salt made! and (4) ihe Dahl salt sensitive rat. Thus, for example, agers described
`~*~
`tx
`xerein, Can be administered to Dahli salt sensitive rats (Rapp and Dene 1985 Hypertension 7:34¢s
`
`te
`
`9} to determineeffects on blood pressure, urine volume sad urinary sodium excretion and lef
`
`for example as described in examples 4 and & heoreim).
`rentricuiar wall thickness sos
`
`
`
` isubiects were fomuale Sprague-Dawley rats which weighed between ZUK230 ¢
`
`at ihe time of experimentation. Following arrival at the animal dueiity, rats were heused imsolid
`
`dollom: cages in groups of twee, where they had unlimited access to fod and water.
`
`38
`
`3132
`
`3132
`
`

`

`WO 2008/137318
`
`PCT/US2008/061205
`
`yt
`Permmperaiure was maintained at $12°C, and lights were on a P2012 hr evele
`MOAN}.
`
`with Hehis on at
`
`Followingat least 3 days of acclimation tothe facility prior to experimentatic
` (PQ) with ofther vehicic (phosphate buffered saline) or test article, and tear
`my
`
`indi vishal metabolism cages where they had access to frod and water. The vs
`
`excreted was recorded from 0-3 hours, and 4-6 hours post dose. In addition, 0.5-].GmLurine
`angles were isken at cach of the above time points and frozen for later analysis, Urine ;
`< ay
`%
`cd for sodinmconcentration using ISEcrown-ether menibranc methodology on an
`
`Olympus AUS460 chemistry immuno analyzer (Olympus America Inch
`
`eres,ae
`
`Figures 1,2, and 3 demonstrate the effects af Lubiprosione and ST peptide ( i
`
`
`
`
`CORLC NPAQPOOYon urine sadium and urine volume m this assay
`
`aeIgpeeaes2 Pan we
`
`eal&.
`
`Salt-sensitive and salt-resistant, 4-5 week-old male Dahil rats {Brookhaven National
`
`Upton, Now York, USA} are fed with Purina rat chow with 0.4%NaCl for the frst 3-4 weeks.
`‘hereafter eail-sensitive and salt-resistant rats are randomized into two populations receiving
` jor a low-salt (0.4%NaC}} dict fora further 2 weeks. Follerwing
`this, cach poyulaticriis se
`rated Info lwo groups, onereceiving test agent in lap water and the
` given in meremental doses until systolic blood pressure fail-
`*4h
`aeomet adt%oeesa oeosgrm rs3
`sot & to g2eea yes ge
`EBoy wn
`
`At the end of the studyrats are anaesthetized with intraperitoneal ac
`
`mavke), and systalic. and diastelic blood pressures are measured direct
`S
`a
`a
`of the right fernoral artery, using ¢ Beckman R6L1 recorder, Blood (8-10 ral for determination
`
`
`
`uvity (New England Nuclear Corporation, Boston, Massachusetis, USA) and
`a
`aldosterane concentration (Diagnostic Prochacts Corporation, Los Angelos, (S.A) is obtained by
`decapitation. Mearts are removed and placed) in s Petri dish, and blood and blooc
`fhushed out with ould saline. Superficial water is removed by blotting, The whole heartis
`fa
`weighed, thereafter theatria ami the right ventricular free wall are dissected fromthe
`
`3133
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`

`WO 2008/137318
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`
`interventricular sepium. The remaining interventricular septum and the left ventricle represented
`
`lef ventricular weight, and the lef ventricular weight: body weight ratio is taken as a measure
`
`for left ventricular mass or Jef? ventricular hypertrophy.
`
`
`
`Axuimber of disorders assaciated with fluid or salt retention maybe prevented ortreated with
`x
`vat rechice sodram absorption in the intestine and/or merease anion secretion fe.g., in the
`
`us
`ful agents include: guanylate cyclase receptor C agonists, soluble guanylate
`
`cyclase modulators, proslanoids inchiding prostaglandin B and derivatives thereof, chloride
`
`channe! activators (e.g. Amitizal® Gubiprostone)), SHT4 agonists, oyelic nucleotides, laxatives,
`
`QFYR deystic fibrosis transmembrane conductance regulator} modulaters, agent
`
`cAMP levels, sodhon transport inhibitars (ee. sodium channe! inhibitors saci
`iThe
`
`neg
`3.
`resins, and combinations thereat described herein.
`
`hosphodiosteruseinhibitors, rermbibitors and aldasterone antagonials, potassium, polymer
`
`‘The agents that redacesachum absorptiin iy
`
`4 n
`
`4
`‘ %
`
`ast
`
`~ 2
`
`0
`
`
`
`the infestine and/or mercase anion scerction can be used alane or in comixmation wath one cr
`~
`gore agents useful in the treatrnent of congestive heart fathure, andor one or more ipod lowerme
`MS
`agent and/or ane or mare asti-chypertensive agents.
`
`CALLERSCREEREECE
`
`
`
`in the frestraent of congestive heart failore
`.
`mea
`yer
`scents deseribed herein oan be administered together with one ar more agents useful in the
`treatment of congestive heart failure including, for example, nesiritide, dobutamine (beta
`OY
`receptor amlagonist), milrinone (phosphodiesterase inhibtior}, Levosimendan (Sinden),
`
`
`
`N° ~[C1,2-dihydro- | -acenaphthylenyDmeihylJsdenesine,
`*
`in}, an agent which mereases the cellalar availability of adenosine,
`
`an adenosine A» receptor agoniat, an adenosine transport inhibitor, or as adenosme deaminase
`
`
`Lipid lowering agents or dilipidemia agents are those agents that act directly or Indirectlyfo
`
`reduce seram cholesters!. Such agents include, but are net Hmited to, be achi sequestrants such
`
`fad cae
`
`as cholestyranune (a styrencalvinnyibengens capolymer comaining quaternary amncnHuny
`
`3134
`
`3134
`
`

`

`WO 2008/137318
`
`PCT/US2008/061205
`
`cationic groups capable of binding bile acids, such as QUESTRAN®or GUESTRANLIORTS
`sholestyramine which aro available from Bristol-Myers Seuibb), colesevelam §ydrochloride
`une
`egy
`fsuch asWELCNGU®7
`ablets (polyaliylamine hydrochloride) ercas-linked
`a
`with epichlorahydrin
`and alkylated with J-bromodecane and (6-bromohexyl-trimethylannmontum Gramide} which are
`5
`available trom Sankyo}, colestipel (a copalymerof diethylenctriamine and i-chloro-2,3-
`epoxypropans, such asCOLESTION®tablets which are available fromPharmacia},
`Pigs
`dalkvlaminoalky! derivatives of a cross-linked dextran, LOCHOLES? DEAR-Senhadex
` 8, OLECERIDES), water soluble derivatives such as 3,3-ioene, N-
`(oysioalkyDalkylamines and polighisam, insoluble quaternized palyetyrenes,
`os
`mixtures thereof andthosebile avid sequestranis disclosed in WOOT/L1345, WO
`nataes oy¥ WS
`‘Ay
`thi wk mssLads 3
`USI OR2895, and USS7TOS188. Suitable inorganic cholesterol sequestranis include bismuth
` fate plus rnontmoriliontteclay, alumirnomnhydroxide and calcium carbonate antacids.
`
`Sin
`
`
`HMC-Codrodnetaseinhibitors are dyslipidemiic agents that can be used in therapeutic
`comlnnation with GCC receptor4
`OG
`CoA reductase inbibiters for use in therapeatic combination with a compoune
`include: atorvastatin (LIPITOR ®: disclosed in US468 1803, LISS38S8070 and
`
`
`
`
`
`cakuum (disclosed in US52739¢5)}, chhydrocompactin, disclosed {
`bervasiatin (disclosed in UISSO82859), carvastatin, cerivastatin (BAYCOL®:disclosed ia
`WISSS02 799, and USS17 7080}, crilvastatin, dalvastatin (a
`
`Puvagtall
`PSOOLSy dise
`m US4739073 and USS34772), glenvastatin, Suinsdostatia
`
`AA}, velostatin. (visinolin: disclosed in US4448784 and US44501713,
`lovastatin fmovinalin, MEVACOR®(Merck and Co.} and related compounds disclosed ix
`US422 1938), mevastatin (and related compound disclosed in L/S3983 140), compactin (and
`related camprunds disclosedin US4804770), pitavastatin ¢ also known as NK-i94, itavast:atin,
`nigvasialin, mshastatin disclosed in USS102888), pravastatin (PRAVACHOL(Bristol Myers
`
`Squibb} and related gcanpounds disclosed in US4346227), rivastatin (sodhun J-(4-fluorephenyi
`isthoxymethylpyridin-3-yl }-3 S-dibydroxyG-heptanoate), rosu
`EN
`mowsas ZD-4522 disclosed in USS260440}, atevastatin, v
`minvasistin (ZOCOR(Merck and Co.} andrelated compounds as disclosed in US4448784 and
`USSSS01 71), sirrivastatin, CLOS 1, compounds disclosed in WOU
`
`
`
`
`
`
`
`umLad
`
`3135
`
`0as
`
`Aenes
`
`wees ia
`
`hg re
`
`ged ry
`
`3135
`
`

`

`WO 2008/137318
`
`PCT/US2008/061205
`
`SSTS4, USGA ISTO, USIG86237, US4499789, LIS4346227, USSTS3I67S, UISSGI3610,
`EPOL2TQ25, and EP401226, and optical or geometric isomers thereof and nontoxic
`pharmaceuticallyacceptable salts, N-oxides, esters, quaternary arnmonium salts, and prodruge
`thereat
`In HIMO-CoAreductase inhibitors where an open-acid form can exist, salt and eater
`lorma maypreferably be formedfrom the open-acid, and all such forme are included withinthe
`meaning ofthe term “HMG-CoAreductase inhibitor" as used herein. Pharmaceutically
`soceptable salts with respect to the HMG-CoA reductase inhibitorincludes non-toxic salts of the
`compounds which sre generally prepared byreactingthe free acid with a suitable organic or
`
`inorganic base, particularty those formed from cations such as sodium, sotassitar x
`
`calctum, lithium, magnesium, zinc and tetramethvianimonium, as well aathos:
`
`pore
`
`preSeed
`
`Coa
`anlines such as anyoionia, ethylenediamine, N-methyiglacamine, lysine, ar
`emuhine,
`menzylethylenediamine, chloreprocaine, diethanolamine, procaine, N~
`e
`
` thylamine, [-p-chiarobengy!-2-pyrrclidine- i -ylamethylbanalm- idazale,
`
`Wethylaming, piperazine, and trie(hydroxymethyl) aminomethane. Further examplesofsalt
`forma of HMOMCoAreduotase inhibitors may include, but aro not Heiitedto, acetate,
`bernxoncaulfonate, bonztade, bicarbonate, bisulfate, blarirare, horate, bromide, calclum edetate,
`ramsplate, carbonate, chlorite, clavulanate, citrate, dihydrochloride, odetate, ediaviate, estolate,
`euvlate, fumarate, glucoptate, ghiconaie, glutamate, giyecilylarsanilate, hexyir
`
`hydrabaniine, hydrobramide, hydrochloride, bydroxynapthoate, iodide, isnthionate, lactate,
`lactobianate, laurnte, malate, maleate, mandelate, mesylate, meethylsulfate, mucate, napaylatc,
`
`uitratc, dleate, oxalate, pamaote, palmitate, panthothenate, phesphz
`
`polygalacturonate, salicylate, stearate, subacetate, succinate, tannat
`
`tricthiodide, and vale
`
`Other dyshipidemic agents which can be used intherapeutic combination with a GCCreceniar
`
`z
`hs tay
`
`ithibitors suchas L-659,699 ((BID-11-19'RRhyydroxy-methyt}-4-oxo-2R-
`HMG-CoA synthase
`4-undovadienois acid} and those disclosed in USS120729,
`
`
`tAfe
`
`3136
`
`3136
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`

`

`WO 2008/137318
`
`PCT/US2008/061205
`
`cholesterol absorption inhrbitora such as plant sterols, plant sianols and/or Tatty ack] estesrs a
`8
`wie, slariad esters, Gele~
`plant stanols such ag sitostano! ester used in BENHRCOL® margan
`sh eS
`‘
`ADNCHTHLOTL |inhibitors inehade
`i,
`sitesternl, and siero! glycosides such as tigqueside, Other cholesierel :
`x
`idin-2-ones; 4-blarylyl-l -phenslazetidin-2-ones; 4-chydroxyphonyljazetidineg-
`y
`anes: L4-dinhonyl-3-hydrexyaiky!-2-azetidinones, 4-biphenyl- | sphenyiacetidin-2-ones, 4-
`
` 4
`
`LADS
`
`taryiyi-]-phenviazelidin-2-canes; and 4-biphenylylaxetidinones,
`
`acyl coenzyme A -cholesteral acyl transterase (ACAT) inhibitors such as avasimibe (Current
`Opinion in Investigational Drags. 349-291-297 (2003), eflucimibe, THL-004, lecimibe, DuP-1 56
`Ae
`&
`KYSOS, SMP 7ST, CL-277.082 (lin Pharmacol Ther, 48(2):1 89-94 (19905) and the like: and
`
`poe yes,Sort
`
`those discinsed in LIS545 10379,(1V096/26948 and WOoG/TOS59:
`
`CETP inhibRers such as FIT 70S identified as in Nature
`
`~F (2000), torrefrapib
` UISZ00301 86952 and WOO0/G1 7164), CP $32,832, BAVYS3-2148, 8C
`S01, 50 78S, and the Hike including those deseribed in Current Opinion im investigabonal Unuigs.
`AHS
`201333 and those disclosedin J. Antibiot, 49(8): $15-814 (1996), and Bioorg. Med.
`
` (996) and patent publications USSS 12548, USSIATIS
`
`
`34, W994 12397, WORSETSS, WOSe1 31
`
`{PRPS 1O7, BPSTS448, DRTOTONG244, DEIOTA HIS], DELOTAL IOS,
`
`DRIP7042437, OF
`S125, DEVG627480, DETIS32188, DED
`
`squalene synthetase inhibitors sack as squalestatin-1, TAN-475, and chase discloged in
` 24, UISS712306 (coophosphono-salftmates), Biller et ai (1988) J. Med.
`i}
`3
`,
`thom., 31/1869 (e.g. isoprencid (phasphinylanethyliphosphonates}, Hiller et al (2890) Current
`Ess
`*
`PharmaceuticalDesign, 2:1, F. Ortiz de Montellanc et al (1977) J. Med. Chem. 2-242
`
`3 %
`
`tes}, Corey and Volante (1976) J. Am. Chern. Scc., 98:1
`
`diphosphate analog A and presqualenc pyrophosphate (PSQ-PP)} analogs), McCi
`
`
`nbinyinkosphonates}, Casson, TP. £.. PhD digsertatron, fur
`Dept. Med. Chem. U of Utah, Abstract, fable of Contents, pp 16, 17, 40-43, 48-31, Sun
`
`wi tA
`
`3137
`
`i3
`
`aA
`2}
`
`
`
`3137
`
`

`

`WO 2008/137318
`
`PCT/US2008/061205
`
`
`
`. Ter. Patents (1993) 861, and patent publications EPOSSTO2GA),
`
`
`
`HOGS S77AL EPQOLIT40A) RPOFOSGOTAS, EPOVOLUTISAL, and WOOD
`
`antioxidants such as probucol (and related compounds disclosed in US3674836), probucel
`derivative
`$ AG1-1087 (and other derivatives disclosed in L9S6121319 and US6i47250},
`tovopherol, asearhic acid, 6-caurotene, seleniumand vitamins such as vitamin B86 or vitamin Bi?
`
`and pharmaceutically acoeptablesalts andesters thereof
`
`PPARwagonisis such as those disclosed in USG028109 (Suoropheny] compounds), WOOQ75103
`substituted phenyipropionic compounds), WO08/43081 and fibric acid derivatives (brates)
`
`such as hecloiibrate, benxafibrate, bexafibrate (CLAS. Rewistry No. 41 S59-87-0, soe
`
`USS7S1328), biniiibrate (CLAS. Reuistry No. 69047-39-8, see BESS4722}, <iprotfibrate (CAS,
`Registry No, 32214-84-3, see US3948073), clinofibrate (CLAS. Registry No, SO298-08-2, see
`W83716583),clelibrate(suchasethyl2trlorophenoxy}-2-methyl-propionate,e.g,Airomid-
`obenzay)phenoxy}-2 mnethyl-propanoieanid, }“methylethylester, Abbott
`
`S& capsules CWoveth-A verse. atofibra
`SW capsules OW
`yeth-Ayorsd}, olalibr
`
`Trcor® snicronized fenofibrate (o-
`fibrate
`fenofibrate (such as Tricor® micrpnized fenofibrate ({2
`
`Laboratories} or Lipanthylmicronized fenofibrate (Labortoire Founier, Franec}}), semosbene,
`
`fsmn
`
`pont
`
`thy
`
`gomiitroail (such as 4-(2,S-dimethyl phenoxy}-2.2-dimethvipentanaic acid, eg. Lopid®tablets
`
`iParke Davia}, hfibrol, G4764"
`
`4, LYS1TS674 and those Abrate and Tdbrate acid
`
`O3348 1, WOR DS3907, WOOR0dSL1G,
`
`derivatives discissed in WOUSA
`Be Z Ss.aes=Nae= iepe eww oe ao3 eo yen a ba peed isaa en ” s auch as GW3965, TSOTS137, and APTCOIE?
`
` S28> axicl Thase disclosed
`
`ay US S003 0125387, WOO3AMS3 82, WO03/083352, WO0S/059874, and the ike:
`
`(human HMI44 is Genbank Accession No. AYI48384 and ratHMTAAis
`HM74 and HMI7SAA
`
`EMM.patAROGk624) receptor agonists such as nicotinic acid (siacin} and derivatives thercef
`{e.8. Compcumds comprising a pyridine-3-carhoxylate structure ar a pyragine-2-carboxylate
`structure, including act! forms, salts, esters, zwilterions umd tautorners, where aay
`ne Tmt net Hiteto taese disclosed in Wise et al (2003) 3. Biol Chem, 2 x a we on ae ones tA
`
`taybaa
`
`3138
`
`3138
`
`

`

`WO 2008/137318
`
`PCT/US2008/061205
`
`vipytascie-3-carboxyho acid and acitran (4,4-dihydro-S-methy)-4-oxo-S-phenyi-2-Soran
`
`carbexylic acid pyradine-3-acetic acidy), as well as S-meth yiricotinic acid, nicotinuric acid,
`tet
`nicertirel, nicofuranase, acipimox (S-methylpyrazine-2-carboxylic acid 4-axide), Niaspan®
`x
`(niacin extended-release rapiets, Kos} and those which can be easily ideniifk
`
`
`the art wi
`tr and agonize theHM74A or HM?4 receptor (or exam
`
`
`{3003) Nature Medicine S352 (calenummobilization assay using the HM74 receptor which
`
`1 (2003) J. Biol. Chem 278:9869 (nicotine binding and {
`Ped
`tal (2003) Biochem. Biophys. Res. Comm. 305.354 fy
`
`assay using the HM74 receptor which could be adapted to the HM744 receptor), Tenary cf al
`
`ory
`
`could be adapfed to theHM74A receptor) and US6420183 {FLIPR assays are deacribed
`generally in and may be adapted to theHM74A or HM?receptor)
`
`renin angiotensin system: inhibitors:
`
`bile acid reabsorption inhibitors (bile acid reuptake inhititors}, such as BART 1483, SC435,
`
`PHASSAGAD, S802), AZDTIOG, and the Hke:
`
`yaw
`
`CPt
`
`PRARG agonists Gneluding partial agonists} such as GW$61516, and GW SOG7345, and those
`
` WO02/46176, WOOL/076857, WOO6291, WOO8/03 2493,
`WOoe/20275 tninoline— compounds), WOS/38845 (aryl eampounds}, WCANY6316!
`
`phenols}, WO03 /024395, WOO7/28 [40 WOOLTST SF,
`
`3
`
`tap.
`
`ij é-disubstituted phenyl compounds), WOOL/G0379 (aryl compounds},WOOL/T2612 &
`WOOMLSIS? (benzoic acid compounds), and WO97/3 1907 (substituted 4-hydroxy-
`hah
`phenylalconis acid camprund}:
`
`sterol biosymbesis iniibitors such as DMP-S65-
`
`irigiveende synthesis infibiters:
`
`a
`microsumal irighyceride transport (MYTTP} inhibiters, such as Inplitapide, LABSST, ans
`
`CPS46086, ABC733, anpliapide and the He;
`
`3139
`
`3139
`
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`WO 2008/137318
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`PCT/US2008/061205
`
`NMGsd roduetase pene expression inhibitors fe.compounds that decrease HMG-CoA
`
`roductase expression by affecting (e.g. blocking} transcriptionor translation of HMG-CoA
`reguclase into protein or compounds that maybe biotransformedinte compounds that havethe
`aforementionedattributes by one or more enzymes in the cholestere) biosynthetic cascade or
`maylead to the accumulation of an isoprene metabolite that has the afnrementioned activities
`{such reguigtion is readily determined by those skilled in the art accarding to standard assays
`(Methods ofEnzymology, 1149-19 1985) anch as those disclaserl in f
`substifuted Isnosterol derivatives) and E. 1. Mereer (1993) Prog, Lip. Re:
`stercis that suppress the biosynthesis ofHMG-CoA reductase};
`
`
`aes
`
` rethonyJbenzene-methanamine hydrochlorideh
`
`pinhsbitors such as NB-598 (()-N-ethylN-(6,0-dinsethy!-2-hepten-4-y- ayl +
`
`lowdensity Hpoprotein (LDL) receptor indreers such asHOUE-402 (an imidaxzolidinyl-pyriniidine
`derivative that directly stimulates LDL reeeptor activity, see Huettinger ef al (1993) Arterioscler.
`‘ih
`Thromb. 13:)00S);
`
`eee tA
`
`Qlatelel aeprogalion inhibitors;
`
`S-L0 or FLAP inhibltors:
`
`PPAR madgistors Gncluding compounds that may have multiple funetionality for uctivating
`
`vanous combinalions of PPARaGPPAR’ and PPARS) such as these disclose
`
`
`USOlG8781, US6166049, WOOU/T240T, WOOO 18455. WwOOOI4I * ACME
`YOON:“S WOU0/23442, WO00/23445, WO00/2345 1, WOO)
`32, WUSRY236332,
`
`392, WOUG/S3563, WO00/63 153, WOU/63196, WOOK 196,
`
`
`
`S322, WO00/76313, WOOL/0d35), WOOL 1444
`WOO fT
`FP PUGd, WOOLZEISE, WOOL2TS78, WOGLIS181, WOONZS
`
`
`WOOLS238, |
`DUPOIS, WOG2OR 1428, WOH
`
`
`WOse 78 Le
`
`WCFZS SS WOOSf073 87, WOOG/T 1355, WOOT 2534, WOOT
`
`16265, WOO3 033483, WOOs/O42 194, Wood!
`
`3140
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`3140
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`WO 2008/137318
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`PCT/US2008/061205
`
`-(4-[difluvorophenyl+1 heptvtureidojethy)ipher
`
`miacin-bound chromium, as diaclosed in WOG3/0395345-
`
`substituted acid derivatives disclosed in WOO3/0401 14:
`
`apolipoprotemB imbibiters such as those disclosed in WO82/080347, WOO?,
`edt
`
`ie
`WOORO4S021, WOOHO4 TS PSs
`
`Pe:
`Sector Aa moxdniaters such as those disclosed inWOOS/OS 74517, WOO3/0475 CVGERGR I:3
`
`vE
`dval bile avic transport (IBAT") inhibitors (or apical sodium ec-dependent Mile acid transport
`
`(ASHM) inhibiiors) such as benzothiepines Gacluding 12-benzothiazepines: | 4-
`boenainhiazepines; 1 5-bersothiaxepines: 1,2, $-benzothiadiazepines);
`
`4:
`
`SPARSactivalors such as disclosed in WOOQ1/00603 (thiazole and oxavale devivat
`
`%
`
`
`
`SIR-323-4),WOOT/IR 149 (fluora, chlore and thio phenoxy phenyla
`
`
`US2083365 inon-f-oxidizablefatty acid analegues), and WOSSAMSI 5S.
`
`LPS
`
`oSnee
`
`yypertens agents
`
`tt
`
`The agents described herein can be used in therapeutic combination with one ar more anti-
`
`hypertensive agents, including bat not limited to:
`
`diurciics, such as thiagides (e.g., chiorthalidone, cyelothiazide (CAS RN 2259-96-53},
`ehlorothiagide (CAS RN 72956-09-3, which maybe prepared as disclosed in US2800194),
`dichlorophenamide, hydraflumethisxide, indapamide, solythiazide, bendroftumethaxide,
` methyelothavide, poivth
`razide, chiorthalidone, indapamide, metolazane,
`quinethazone, althiszide (AS RN SS88-16-9, which may be prepared as dis
`Patent Noa. Of }, Genathiazide (CAS RN O1-33-8, winch maybe prepare
`
`} bathlazide Gvhick may be prepared as disclosed in British Pater
`
`
`
`
`
`Lot We
`
`3141
`
`3141
`
`

`

`WO 2008/137318
`
`PCT/US2008/061205
`
`i
`and hydrachiorothiazide), loop diuretics (¢.2., bumetanide, cthacrynic acid, furosemide, and
`lorasemide}, polussiinnsparing agents (e.¢., amiloride, and triamterene (CAS Nisnber 304-01-
`
`Uy}, and aldosterone antagonista (e.g., spironolactone (CAS Number 53-O1-7 and active
`metabolites therealinchaling canrenone), epirenone, andthe like}:
`
`ut
`
`O-adrenergic blockers such as Amiodarone (Cordarone, Pacerone}, bunolol kvdrewhioris
`
`RN31969-03-8, Parke-Davis}, acebutolol (4N-[3-Acety-4-[2-h
`
`methylothyamine}propoxyphenyl|-batanamiide, or ()-3-Age
`
`Q
`on,Cat
`
`Tt
`
`Soar
`
`oN
`
`‘isepropylamine} propoxy] butyranilide}, acebutalol hydrochloride fe.g., Sectral®, Wyeth-
`Ayerst), alprenoicd hydrochloride (CAS RN 13 707-88-S see Netherlands Patent Application No,
`SAH3 O92}, alenolol (e.g., Tenarmin®, AstraZeneca), carteolol hydrochloride (e.g, C:
`
`fimiab®, Abbott}, Celiprolel hvdrachloride (CAS RN S7470-75
`
`
`¥
`ectamolol hydrochloride (CAS RN 77S90-95-5, see also US4059622), lahetalal hydrochloride
`s
`2
`iz.g.,Normodyne®, Schering), esmolol hydrochloride (ez, Breviloc®Baxter), levobataxoio!
`
`hydrochloride (c.g., Betaxon™’ Ophthalmic Suspension, Ajcon), levebonolnl hydrochloride jes.
`
`Setagan® Liquilllm®with C CAPO Compliance Cap, Allergan}, nadolol! fe.g., Nadolol,
`ue
`Corgan, Mylan}, practelal (CAS RN6673-354, see alsa US3408387), proprancie!
`hydrochlonds (CAS RN 318-98-9}, sotalol hydrochloride (e.g,Belapace AF"Serle}, timole!
`~<hmethylethy Damino}-3-[[4-4(4-morpholinyl}-12,5-thindiazol-3-yils
`Wid
`RPS
`henulydrate, (8, CAS RN 91524-16-2)}, timolol maleate (S)-1-[(1 J-dimethyletivl) amino’
`
`
`iia. (G-amerpholinyl-1,2yerthiadiaxol -3- yl] axybpepe“)-2-inttenedinate (i:1} sale, ©
`RN26021-17-3) b seproles i2-Propansl, 1-[4-{{2on
`
`
`
`batenadioate (2:1) (sald)
`
`
`
`mothanal, ao'-[{iminohis(methylene)this[6-fuoro-3 4-dihydro-, CAS RN 09200-09-6 see alse
`
`BD wast
`
`US. Pat. No. 4,654,362), cicloprolel hydrochloride, such 2-Propancl, 1-[4-{2-
`{eyclopropvimethoxyethoxyIphenoxy]-3-[ L-methylethyjamino}-, hydrochloride, ALAS. RN
`82686-79-3}, dexproprauciol hydrochloride (2-Propanol, {-[ ]-methylethy-amine]-~1-
`¥
`¥
`naphthaionyviossy)-hydrochloride (CAS RN L3071-11-9), diacetalol fydrochlornide ¢Acetamide,
`
`GO
`
`3142
`
`3142
`
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`

`WO 2008/137318
`
`PCT/US2008/061205
`
` y
`Syv“
`‘drochloride
`
`‘h
`
`s
`
`yprapyl esier, Cb) salfete (1:1) (salt), CAS RN 88844-735-)
`£
`
`~5-P]-hydresye2 6, CAS RN 75659-08-4), oxaprolal
`bydrochinride (2.Propane i-+(2-cyclohexyiphenoxy)}-3-1U1-methylethyDamine|. hydrochloride
`% dimethylethy!|vo.
`CAS RE S93
`3}, Destolol sollate (Benxoie acid, 2-furo-,3-[13-laminocarbonyiiamine|- -
`nicisiolSydrachiorideeaeesalfanamide,N-[4-[]-bydroxy-2-(methylaminopropsipt
`
`“i
`
`muntohydrochloride CAS RN 77
`
`-7}, metoprelel 2-Propanol, t-[4
`mt
`nEhoayethyphenoxy} 3-[ l-omethylethylamino}-; CAS RN 37380-58-6), metoprole! tartrate
`fsuch as 2-Propanol, |-[442-methoxyethylphenoxy]-3-[(] snethsylethylhaminoe+, «2,
`q
`Lopressora:, Novartis), pamatolal sulfate (Carbamic acid, [2-[4-[2-hydroxy-34 6 l-
`
`yii-ethyll-, methy] ester, (2) sulfate (salt} (2:73, CAS RN Propanol, 1-(2-cyclapentyiphenoxy}3-[ 1 l-dimethyle-
`
`S RN 38363-32-5}
`
`ot:
`a
`we
`ae
`~
`’
`~
`ra
`
`yaraimo]}-propoxyiphenyl]-, CAS BN 9673-35-42} gprenalal
`hydromy3
`J
`X
`hydrochloride (Propanol, 1-[(i -methyicthyamine}-3-]2-(methylihichphenoxy}, hydrochloridg,
`2
`39832434), tohunalol (Benzamide, 4-[2-[!2-hydroxy-3-(2-methyviphenes y}-
`
`
`
`olhosyl}-, CAS RN 38103-61-6), bopindolol, indenolal, pindolel fag. Visken}
`propanolsi ex, insderal, Inderal-LA), tertatolol, Coreg (carvedilol}, and tihsalal, and the Hke;
`
`calcuan channel binckers suchas besylate salt of amlodipine (such sa 3-cthyl-S-methyl-2-(2-
`
`pe
`
`ta
`
`A
`
`rteSenn
`
`aminoethosyinethyi)-4-(2-chlarophenyi}1 ,4-dihydro-d-methyl-3 S-pyridinedicarbasxy!
`
`benxenesulphonate, og, Norvasc®, Pfizer}, cloniiazem maleate {1,5-Rensothiasepind(Sh)~
`3-{acetylox y}-8-chlore-3-[2-dimethylamino)ethy!|-2,3-dihydro-2-(4-methnayoheny
`q
`3
`3
`(f<}-2-buteriedioate (1:1), see also LIS4S67195), isradipine(3,5-Pyridinedicarboxvite acid, 4-(4
`
`
`
`
`
`bono fnravany))- | 4-diydre-26-dimethyl-, methyl I-methylethy! ester, (t)-4(4
`
`benmotbrazany!i d-dibydie-2,6-dimethyl-3, S-pyridinedicarboxyinic, see alsa L/S44a6607 2}:
`
`ye (such as isiesapropyd (2- methoxyethy)) b, d- duhydro ~2,6- dimethyl -4- (3-
`3
`, Bayer), felodipine (such as ofiyy)
`
`~3,3+ pyridine ~ dicarhoxylate, e.g. Nimatap@®
`
`uo
`methy! 4-(23-dichlorapheny!}14-dihydro-2.¢6-dimethyl-3,5-pyndinedicarboxyinte-, 6...
`ae
`we
`¥
`wdend a4Ta pat mae te ieerixxtonded-Release, Astrateneca LP} nilvadipine (3,5-FPyridinedicarboxyllc acid, 2-
`
`eg fern oo
`
`Oo]
`
`3143
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`

`WO 2008/137318
`
`PCT/US2008/061205
`
`
`
`
`netermethyi-4-(3-nitrophenyl}-,3-metiry! 3-Ci metiylothy)} ester, alsa see
`peop?
`
`ipine (suchua 3\5-pyridinedicarboxylicacid, |.4-dihydra-2.8-dimethyl4-
`mirophenyl-, dimethyl ester, o.g., Procardia XL® Extended Relea
`
`
`
`hydrochloride (sack as 1,5-Benzothiazepin-4(5H}-one,3-(acetyloxy)4
`3-dihydro-2i4methoxyphenyl}-, monobydrochloride, (-+)-cis., ep, Ts
`veraparil hydrochloride (such as benzeneacetronitrile, (alpha)}-[/3-([2-43 4-dimethoxypheny!
`ethylmethylamine|propyl}-3 4-dimethoxy-{alpha}(1-methylethy!} hydrochloride, og, lseptin®
`A
`SR, Knoll Labe}, teludipine hydrochloride (3,5-Pyndinedicarboxvileacid, 2-
`oy
`! t
`iidimethylaminomethyl |4-[2-[¢1 £)-3-(1 1 -dimethylethoxy}3-oxo- i-propeny! Iphenyth-{ 4+
`
`
`
`cibydro-G-metlyyl-, Hethy! ester, monebydrochloride) CAS RN LOSTOO-03-tedotlosdil
`
`
`¥
`e
`iPhosphonie acid, [2-(2-phenaxyethyl}- | 3-propanc- diyi}bis-, telrabuty
`
`4), fosteddl (Phospharis acid, [[4-(2-benzothiazolyliphenylmethyl], diethyi ester CAS RN
`
`
`O22), aranidipine, asclnidipine, barnilipine, benidipine, bepridil, cinaldipime, clevidiping,
`efonidipme, gallopamil, lacidipine, lenuldipime, lercanidipine, monatenil maleate {1 -
`*
`
`Piperazinebutananide, N-(6,11-dihydradibensa(b,ehihiepin-1 i
`Dy-f4-fucroghenyvib,
`é-butenedioaie (1:1) Ch-N-+46,11-Dk“ydrodibenzo(.e)thiep-
`inin-d }-vi)-4-(p-fl uoropheny)-i-
`piperawinebutyramide maleate (1:1) CAS RN 132046-06-1), nicardipine, nisoklipine,
`x
`nitrondipine, manidinime, pranidipine, and the Hke:
`
` t
`
`T-shamnel calolom antagonisis such as muibefradhl,
`
`angigtensm converting enzyme (ACE) inhibiters such as benazepril, benaxepril hydrochloride
`
`{such as 3-[1 |-(ethoxvearbonyl}-3-pherny!-(18)-propyllamino|-2,3,4,5-tetralyedra-2-oxo-1H -1-
`
`(2$)-bensazepine-l-acetic acid monohydrochloride, ¢.g., Lotrel®, Novartis), captopril (suchas
`1-29}-3-mnercapio-2-methylsropionyl|-L-proline, cg... Captopril, Mylan, CAS RN G3S71-£6-2
`and others Gisclosed in US4046889), ceranapril (and others disclosed in US4452 790), ectapril
`¥
`falecepril, Dainippon disclosed in Bar. Therap. Res. 39:6
`TE LBRO): 402543
`
`(ielinan-LaRocie) dachssedin J. Carcdiovase. Pharmacol, 9:39 (7987), indalapeit (delaprii
`hydrochloride (QH-1,24-Benzothiadiazine-7-anlfonamide, 3-“bieyelo[2.2. Hbeptote2+¥l-G-
`chloro-3.4-dihydre- ,indhoxide CAS RN 2250-06-3}; disclosed in US42845081), enalapril fand
`<:
`others disciased in US4374829), enalopril, enaloprilat, fosinopril, (such ss seans-L-proline, €-
`
`a2
`
`3144
`
`eeNaat
`
`pet
`
`tet
`
`in,Nae
`
`boean
`
`3144
`
`

`

`WO 2008/137318
`
`PCT/US2008/061205
`
`
`
`reece ao
`
`LPS
`
`eyelohexy!-i-{[|2-methyl-1-(]-oxopropoxy) propoxyi(4-phenyibuty) phogp!huryllacetyl}-,
`q
`Fes
`sodiansalt, e.g¢.. Monopril, Bristol-Myers Squibb and others disclosed in
`
`x
`sosinepril sodium {L-Proline, 4-cyclohoxyl-1-[f(R}-[(1S }2-methyl-1-+}-ox- &IPTORORYSPPrOpORN},
`imidapril, indelapril (Schering, disclosed in J. Cardiovase. Pharmacol. $:643,
`lisinepel iMorck}, losinopril, maexipril, moexipell hydrochloride (8-Jsoquinolinecarhoaylic acid,
`2
`piethoxycarbony]}-3 -phenylpropyljamine} i -oxoprapsy]}-1- 2,3.4-tetrahydro-
`=
`2
`. monohydrochioride, GS} CAS RN 82586-52-3), quinapril, quinaprilat, seni
`disclosed in EP 79022 and Curr, Ther. Res, 40:74 (1986), perindopril erbumine
`(Hoeechsst)
`
`S}-t-Carboxybutyljalanylhexalydro-2-indolinerarborvilcacid, |
`as
`othy! ester, compound with terf-botvianine (lik eg. Aceon®, Solvay), perindopril (Servic
`disclosed in Bur J. chin. Pharmnacel, 31:519 1987)}}, quanipril (disclasedd in US4344ogoy,
`=
`‘\
`G
`set iraey &,ae
`C% wea ee5 fond cosSeeas%es
`er
`ssurapril (Schering, dis
`
`Pharmacel. Toxicol, SO (Supp. 3):173 (1986}}, te
`irandalapril, zofenopril fand others disclosed in 1184216906), rentlapril (fentlapril, disclosed in
`tam. Exp. Pharmacol. Physiol. 10:131 (19833), pivepril, YSO8G, teprotide (Bradykinin
`:
`‘ }, BRE1 36,378 iSmith Kline Beecham, see EPBOR22 anal
`i
`1 af
`potontiator APPOs CAS RN 3:
`ao
`iret ary
`Sad tht eed
`EPGOQGGS), Mi-838 (Chapal, see CLA. 102:T2388¥ and Jap, J, Pharmacol 40:373 (1986), CGS
`igsi4 (Ciba-GeigySy, 3-{{ f-othoxycarbonyl-3-phenyl-(18}-propyflamine)}-2.345-tetraiydra-3-ox-
`a I(Q8)-benzazepine1 acetic acid HCI, see UK. Patent No. 2108614), COS 16,617 (Cihe-
`>
`(LS }S-atine-1-carboxypentylfamino]-2,3.4~ S-tetrahydro<2-oxo-LH-I-
`x
`
`
`
`oy oe SY
`
`penizazepine- |-ethanoie acid, see LIS4473575), Ra 44570 (Hoechst, se
`
`Si 1234 (19833), R 31-220) (ofiman-LaRoche seeFEBS Lett 165-201
`
`(1984)), WY-44221 (Wyeth, see J. Med. Chom, 26:394 {i863},
`(Pharmacologist 26:243 266
`and those disclosed in LIS2003006922 (paragraph 28), US4337201, USa432073
`inhosphonanidates
`
`td Nah
`
`neutral endopeptidase inhibitors such as omapatrilat (Vanlew@), COS 30440, cadoxatril and
`
`ecadot, fagidetril Galso known as aladetril or alatriepril}, sanmpatrilet, mixangril, and
`semopatrist, AYETSSS, RR4030and those disclosed in UIS536207237
`
`US4?49688. USS552397, USSS0408G, 0
`3
`EPOSOO444, RIMMALS22 EPOS9Od44, RPOSOSG1O, EPOS343963, EPS34398, EPS34ae7,
`
`
`
`ged pon
`
`G3
`
`3145
`
`3145
`
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`

`WO 2008/137318
`
`PCT/US2008/061205
`
`endothelin antagonists such as texosentan, AS08165, and YM62869. and the Hke