Table of Contents For Ex. 1018A
`US Application 13/372,426, as filed on February 13, 2012
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`Face of application -------------------------------------------------- page 2
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`Specification --------------------------------------------------------- pages 3 - 36
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`Claims ----------------------------------------------------------------- pages 37 - 39
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`Drawings -------------------------------------------------------------- pages 40 - 43
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`(12) INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT)
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`(19) World Intellectual Property Organization .
`International Bureau
`‘7
`
`(43) International Publication Date
`14 August 2008 (14.08.2008)
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`(51) International Patent Classification:
`G01N 33/50 (2006.01)
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`
`
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`lllllllllllllllllllilllll||||||l||l||ilIllIllE|||l|||||||||||l||||E |||||l|||||||l||ll|l|||
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`(10) International Publication Number
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`WO 2008/097596 A2
`
`(21) International Application Number:
`PCT/US2008/001602
`
`(22) International Filing Date: 7 February 2008 (07.02.2008)
`
`(25) Filing Language:
`
`(26) Publication Language:
`
`English
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`English
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`(30) Priority Data:
`60/888,921
`
`8 February 2007 (08.02.2007)
`
`US
`
`(71) Applicant (for all designated States except US): BIOGEN
`IDEC MA INC. [US/US]; 14 Cambridge Center, Cam-
`bridge, MA 02142 (US).
`
`(72) Inventor; and
`LUKASHEV,
`(for US only):
`(75) Inventor/Applicant
`Matvey, E. [US/US]; 3 Louis Road, Tewksbury, MA
`01876 (US).
`
`(74) Agent; GAEIRETT, Arthur, 3.; Finnegan, Henderson,
`Farabow, Garrett & Dunner L.L.E, 901 New York Avenue,
`NW, Washington, DC 0G0l—44l3 (US).
`
`
`
`(81) Designated States (unless otherwise indicated for every
`kind of national protection available): AE, AG, AL, AM,
`AO, AT, AU, AZ, BA, BB, BG, BH, BR, BW, BY, BZ, CA,
`CN, CO, CR, CU, CZ, DE, DK, DM, DO, DZ, EC, EE,
`EG, ES, FI, GB, GD, GE, GH, GM, GT, HN, HR, HU, ID,
`IL, IN, IS, JP, KE, KG, KM, KN, KP, KR, KZ, LA, LC,
`LK, LR, LS, LT, LU, LY, MA, MD, ME, MG, MK, MN,
`MW, MX, MY, MZ, NA, NG, NI, NO, NZ, OM, PG, PH,
`PL, PT, RO, RS, RU, SC, SD, SE, SG, SK, SL, SM, SV,
`
`SY, TJ, TM, TN, TR,
`TZ, UA, UG, US, UZ, VC, VN,
`ZA, ZM, ZW.
`
`(84) Designated States (unless otherwise indicated, for every
`kind of regional protection available): ARIPO (BW, GH,
`GM, KE, LS, MW, MZ, NA, SD, SL, SZ, TZ, UG, ZM,
`ZW), Eurasian (AM, AZ, BY, KG, KZ, MD, RU, TJ, TM),
`European (AT, BE, BG, CH, CY, CZ, DE, DK, EE, ES, FI,
`FR, GB, GR, HR, HU, IE, IS,
`LT, LU, LV, MC, MT, NL,
`NO, PL, PT, RO, SE, SI, SK, TR), OAPI (BF, BJ, CF, CG,
`CI, CM, GA, GN, GQ, GW, ML, MR, NE, SN, TD, TG).
`
`Patblished;
`
`without international’ search report and to be repizlvlisited
`upon receipt oft‘r':az report
`
`
`
`(54) Title: NRF2 SCREENING ASSAYS AND RELATED METHODS AND COMPOSITIONS
`
`90
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`{P1 (57) Abstract: Provided are certain methods of screening, identifying, and evaluating neuroprotective compounds useful for {E6211-
`3 ment of neurological diseases, such as, e.g., multiple sclerosis (MS). The compounds described upregulate the cellular cytoprotective
`pathway regulated by Nrf2. Also provided are certain methods of utilizing such compounds in therapy for neurological disease, par-
`,
`ticularly, for slowing or reducing demyelination, axonal loss, or neuronal and oligodendrocyte death.
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`WO 2008/097596
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`PCT/US2008/001602
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`i‘~trf2 SCREENING ASSAYS
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`AND RELATED METHODS AND COEFEFOSITIONS
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`[0001]
`
`Provided are certain compounds for treating neurological diseases,
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`including dernyelinating neurological diseases, such as, e.g., multiple sclerosis.
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`[0002]
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`Multiple sclerosis (MS) is an autoimmune disease with the autoimmune
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`activity directed against central nervous system (CNS) antigens. The disease is
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`characterized by inflammation in parts of the CNS, leading to the loss of the myelin
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`sheathing around neuronal axons (demyelination), loss of axons, and the eventual
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`death of neurons, oligodenrocytes and glial cells.
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`[0003] An estimated 2,500,000 people in the world suffer from MS.
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`it is one of
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`the most common diseases of the CNS in young adults. MS is a chronic, progressing,
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`disabling disease, which generally strikes its victims some time after adolescence, with
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`diagnosis generally made between 20 and 40 years of age, although onset may occur
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`eariier. The disease is not directly hereditary, although genetic susceptibility plays a
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`part in its development. Relapsing-remitting MS presents in the form of recurrent
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`attacks of focal or multifocal neurologic dysfunction. Attacks may occur, remit, and
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`recur, seemingly randomly over many years. Remission is often incomplete and as
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`one attack follows another, a stepwise downward progression ensues with increasing
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`permanent neurological deficit.
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`[0004] Although various immunotherapeutic drugs can provide relief in
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`patients with MS, none is capable of reversing disease progression, and some can
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`cause serious adverse effects. Most current therapies for MS are aimed at the
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`reduction of inflammation and suppression or modulation of the immune system. As of
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`2006, the available treatments for MS reduce inflammation and the number of new
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`episodes but not all have an effect on disease progression. A number of clinical trials
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`have shown that the suppression of inflammation in chronic MS rarely significantly
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`iimi-is the sccumuietien sf disabiiity tnrnngh sustained disease pregressien, suggesting
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`that neurenai damage and infiemrnetinn are independent patnoingies. Promoting CNS
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`rernyeiinetien as a repair mechanism and etiierwise preventing axonal loss and
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`VVQ ;’.i}{}Sl{i97S§}{~S
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`§’CT.I’US20i}8:’{it}16€}2
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`neuronal death are some of the important goals for the treatment of MS. For a
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`comprehensive review of MS and its current therapies, see, e.g., McAlpine’s Multiple
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`Sclerosis, by Alastair Compston et al., 4th edition, Churchill Livingstone Elsevier,
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`2006.
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`[0005]
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`“Phase 2 enzymes" serve as a protection mechanism in mammalian
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`cells against oxygen/nitrogen species (ROS/RNS), electrophiles and xenobiotics.
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`These enzymes are not normally expressed at their maximal levels and, their
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`expression can be induced by a variety of natural and synthetic agents. Nuclear factor
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`E2-related factor 2 (Nrf2) is a transcription factor responsible for the induction of a
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`variety of important antioxidant and detoxification enzymes that coordinate a protective
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`cellular response to metabolic and toxic stress.
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`[0006] ROS/RNS are most damaging in the brain and neuronal tissue, where
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`they attack post-mitotic (i.e., non—dividing) cells such as glial cells, oligodendocytes,
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`and neurons, which are particularly sensitive to free radicals. This process leads to
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`neuronal damage. Oxidative stress has been implicated in the pathogenesis of a
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`variety of neurodegenerative diseases, including ALS, Alzheimer's disease (AD), and
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`Parkinson's disease (PD). For review, see, e.g., van Muiswinkel et al., Curr. Drug
`
`Targets CNS--Neurol. Disord., 2005, 4:267-281. An anti-oxidative enzyme under
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`control of Nrf2, NQO1 (NAD(P)H dehydrogenase, quinone (1), was recently reported
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`to be substantially upregulated in the brain tissues of AD and PD subjects (Muiswinkel
`
`et al., Neurobiol. Aging, 2004, 25: 1253). Similarly, increased expression of NQO1
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`was reported in the ALS subjects’ spinal cord (Muiswinkel et al., Curr. Drug
`
`Targets--CNS. Neurol. Disord., 2005, 4:267-281) and in active and chronic lesions in
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`the brains of patients suffering from MS (van Horssen et al., Free Radical Biol. & Med.,
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`20%, 41 31i«3‘i 1}. These ebservatiens ineicete that the Nn‘2 pathway may be
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`activated in neuredegeneretive and neiireiniiemmatery ciiseeses as an endegeneus
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`pretestive mechanism.
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`indeed, mes: -recently, it has been reported that induced
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`activation of Nrf2~dependent genes by certain eyciepenanene-beserii cempeunds
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`{i\iEPi1‘*) ccunters the texic effects ef metaboiie inhibitien and RQSIRNS preductien in
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`VVG 2€«}i)8./tl£P7596
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`PCWEJS2008/301602
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`the brain and protects neurons from death in vitro and in vivo (see Satoh et al., PNAS,
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`2006, 103(3):768-773).
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`[000?] Additionally, many publications have reported neuroprotective effects of
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`compounds in natural plant-derived compounds (“phytochemicals"), including
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`oi-tocopherol (vitamin E), lycopene (tomatoes), resveratrol (red grapes), sulforaphane
`
`(broccoli), EGCG (green tea), etc. For review, see Mattson and Cheng, Trends in
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`§\leurosci., 2006, 29(11):632-639. Originally, the action of these compounds was
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`attributed to their anti-oxidant properties. However, while most anti-oxidants are
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`effective only at high concentrations, at least some of these compounds appear to
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`exert neuroprotective effects at much lower doses. Emerging evidence suggests that
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`these compounds may exert their neuroprotective effects by activating cellular
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`stress-response pathways, including the Nrf2 pathway, resulting in the upregulation of
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`neuroprotective genes. However, the exact mechanism of action of these compounds
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`remains poorly understood.
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`[0008] To date, more than 10 different chemical classes of inducers of Nrf2
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`pathway have been identified including isothiocyanates and their thiol addition
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`products, dithiocarbamates, as well as 1,2-dithiole-3-thiones, trivalent arsenic
`
`derivatives (e.g., phenyl arsenoxide), heavy metals, certain conjugated cyclic and
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`acyclic polyenes (incluéing porphyrins, chlorophyllins, and chlorophyll), and vicinal
`
`dimercaptans. These inducers have few structural similarities. They are mostly
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`electrophiles, and all can react chemically with thiol groups by alkylation, oxidation, or
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`reduction, suggesting that the intracellular sensor for inducers is likely to contain very
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`highly reactive (cysteine) thiols. The inducers can neodify thiol groups by a variety of
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`mechanisms including: alkylation (Es/lichael addition acceptors, isothiocyanates,
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`quinones); oxidation (e.g., peroxides and hydroperoxides); and direct reaction with
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`thiol/disulfide lirikages (e.g., vicinal dithiols such as 1,2-dimercaptopropanol, lipoic
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`acid). These diverse response mechanisms provide plasticity for cellular responses to
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`a variety of electrophilic and oxidant stressors.
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`[0009] Provided are methods that comprise at least one of the following
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`methods:
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`Vt/i} 2308/'fl975§§6
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`i’(ITft}S2tBt§8/Git} 682
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`1} rnethette ef screening fer et teeet ene new eendidete cerrtpetrne fer
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`treettrtg e heuretegieet eieeese;
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`2) rnethede et evettreting neurepreteettve erepertiee et at ieeet ene drug
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`eerteieete fer treettng e heureiegteei disease;
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`3) methette of eerrieerirrg (e.g., ier hieequiveterree} et ieeet twe
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`phermeceetieei eerrreeeitiene which eernpriee terherte eete eerivetivee;
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`4} rnetheee ef treeting e netireiegicet disease by administering he the euhteet
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`in heett thereof at teeet ehe eemeeunct that is eertieiiy etructuretty eirniier te
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`DMF er Mh/i¥; end
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`5) rhethede et treating a rtetrreiegirrei eieeeee hy e centhinetien therapy thet
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`eerherteee edrninietretien et at ieeet ene tiret eerheetrnd thet uereguietee
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`the t\trt2 eethwey end et teeet ehe eecentt eempetrrict that deee rret
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`uereguiete the Nrt2 eethwey.
`
`iflfltttj
`
`in eeme errthedimertte, the neuretegieet disease is e neurectegeneretive
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`disease such as, fer exernete, ALS, Perttineene dieeeee, Aizhetrhefe disease, ene
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`t-h,rntihgten‘e eieeeee.
`
`in eerhe erhheciirnente the neureiegieei eieeeee is MS er
`
`enether eernyeiirseting hetrreiegieet disease.
`
`{G31 t}
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`in eeme ernheeirnente, the rhethede 1-ifiturther eemeriee:
`
`e} eenteeting e eeii with the test eemneene, ene
`
`h) eeterrninthg whether the Nrtz pathway ts ueregutetee in the eett.
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`tn eerne emtteciirhehte, the rhetheee they further eemririee:
`
`e) cteterrnihtng whether the test eemeetinrt stews er prevents demyetinetieh,
`
`exerrei tees, eneler heurerzet (teeth, end/er
`
`d) eeteeting the test eemeetrnd es e eenttidete fer treating netsrectegeneretien
`
`it‘? e rreureiegirset dieeese it 1) the Nrt2 eethwey ie upregeieted end 2)
`
`dernyeitrtetteh, exertet ieee, enttier neurenei eeeth ereiie prevented er
`
`stewed.
`
`{fit')'t2}
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`ire eerne ernhettintente, the rnethede 'i»3 eerhpriee eehteetihg e cett with
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`at ieeet ene test eerneeend end determining whether the Ne? eethwey ie epreguieted
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`in the eeti.
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`tr: etrch rhethede, er: npregtitetieh et the Nit? eethwey eheve e threeheid
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`VW} 2€}t}8,"t)97S§€§
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`.‘€’tC'i‘f[iS2tit)8./iit};té'{}2
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`(eg, by at ieast 30% ever a eehtrat) indicates that the at ieast aha eempetmd has at
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`ieast aha eietegieai pteperty hehetieiat in treating a hetireiegicai disease (a.g.,
`
`heureeretestive eteeetties},
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`in same emheeimehts, the eereguiaticm at the i\h'i2
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`pathway is assessed (in viva arid./er in vitre) by at ieast aha at the teiiewihg:
`
`i} exeressiee ieveis ef ehdegetteesiy prettueed ahdier exegeheueiy
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`ihtreduced hirt2;
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`ii) sehceiieiar iecatizetieh ahciiei" aesiear trahsieeatien at Nrt2;
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`iii) exeressien ieveis enema“ activity at me at mere genes under eentret at
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`Nit? {e.g., eedegeheus NQO1) at an Nrt2-tegeietee reperter gene ih an
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`artitieiat reperter sehstruct;
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`iv)
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`ieve-is et txirtfi binding te the t~t:"t2-hihtiihg Etta eiement ARE;
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`V) stahiiity et i\h"t2ft<eaet cempiexes; and
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`vi) mectittsatieh (e.g.. aikyiatieh} ieveis at Keep? aedior at ieast ens ether
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`Nrt2.’i<eap’i -asseciateizt preteihs.
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`{Sets}
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`in seme emiaeeimehts ei’ metheds ‘i-3,, the semeeuhds that are being
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`screened, evaieatett, at cerhpared eemprise at ieast ehe member at at ieast me at the
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`teiiewing eiasses at cempetihds: miid aiitytetihg agents, Miehaei adciitiea aeceptere,
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`and eemseehds that are metaheiizeci tt§:)t‘.§i”i administration te hiiishaei aeditien
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`aeceeters.
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`in same emheeimehts, the ivtiehaei additien aeeeeter has the structure at
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`Fermuia i, ii, iii, ea" iv set terth iaeiew.
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`{C3914}
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`in same emhedimehts methaci t eethprises:
`
`a) eehtactihg a seii with a tsiuraiity at test eemeetahds,
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`hi determining whether the Nit? pathway is iipreguiateci in the ceii, and
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`2:) seieetittg hem the piuraiity et semeetthds at ieast she eemeeuhd that
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`upreguiates the Nat? pathway,
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`wherein an epreguietieh at the Nrtz aathway by the seiected at ieast ehe ceeipeund
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`indicates that the seieetee at ieast aha aemeeunttt may he useftii fer treating a
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`heureiegicai disease, The eiuraiity at eempetihtts may he represented, eg., by a
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`eemhiheteriai chemieai tihrary, and the methea may he eertermee, eg, by
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`high~threughg:>tit sereettihg.
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`‘W83 2t}{l8l’{l97596
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`PC?"/'US2l)ti8it}(lltE€}2
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`[0015]
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`in some embodiments method 2 comprises:
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`a) contacting a cell with the at least one drug or drug candidate, and
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`b) determining whether the Nrf2 pathway is upregulated in the cell,
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`wherein an upregulation of the Nrf2 pathway by the at least one drug or drug
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`candidate indicates that the at least one drug or drug candidate is useful for
`
`neuroprotection in treating a human having a neurological disease.
`
`[0016]
`
`In some embodiments method 3 comprises:
`
`a) contacting a cell with a first composition comprising at least one test
`
`compound, and
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`b) comparing the level of Nrf2 pathway upregulation in the cell by the at least
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`one test compound to the corresponding level of the l\ln°2 pathway
`
`upregulation in a control cell treated with a second contpositien comprising
`
`at least one of DMF and MMF.
`
`[0017]
`
`in some embodiments of method 3, the test compound is fumaric acid,
`
`a salt thereof, or a fumaric acid derivative.
`
`In some embodiments, the first
`
`composition comprises DMF, MMF, or both.
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`In some embodiments, the dose and/or
`
`the formulation of the first composition differs from the dose and/or the formulation of
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`the second composition.
`
`[0018]
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`In some embodiments, method 3 further comprises:
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`c) comparing at least one pharmacokinetic parameter (e.g., serum-half-life) of
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`the first and the second compositions.
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`[0019]
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`In some embodiments method 4 comprises administering to the
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`mammal a therapeutically effective amount of at least one neuroprotective compound
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`having Formula I, ll, ill, or IV, e.g., a fumaric acid derivative (e.g., DMF or MMF).
`
`[0020]
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`In some embodiments method 4 provides a method of slowing or
`
`preventing neurodegeneration in a patient in need thereof, by administering the
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`compound in an amount and for a period of time sufficient to slow or prevent
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`demyelination, axonal loss, and/or neuronal death, e.g., by at least 30% relative to a
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`control.
`
`{G021}
`
`in some emhedirnents method 55 comprises:
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`\VG Ztitifi./i}9’?":“396
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`E’€?T/"flS2(3S8/'{}{i_t60?;
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`a} administering ts the manérnai a tnerapeatiaaiiy effective arncunt at at Beast
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`ans first cernpaund that apregtiiates the Na‘? pathway, and
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`b) administering a therapeutically effective amount of at least one second
`
`compound that does not upregulate the Nrf2 pathway.
`
`[0022]
`
`In some embodiments of method 5, the at least one first compound,
`
`used in step (a), is 3 compound of Formula I, II. III, or IV, e.g., a fumaric acid derivative
`
`(e.g., DMF or MMF); and the at least one second compound, which is used in step (b),
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`is an immunosuppressive or an immunomodulatory compound that does not
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`upregulate the Nrf2 pathway (e.g., by more than 30% over a control).
`
`[0023]
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`In some embodiments method 5 comprises administering to the
`
`mammal a therapeutically effective amount of a compound of Formula I, II, III, or IV.
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`[0024]
`
`in some embodiments of methods 1-5, the at least onecompound being
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`screened, identified, evaluated, or used for treating a neurological disorder is not
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`fumaric acid or its salt, or a fumaric acid derivative (e.g., DMF or MMF).
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`[0025] Other features and embodiments of the invention will be apparent from
`
`the following description and the claims.
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`BREE? BE$CRiP“i'i$i*é Q? “t“iiEgFi;G_i..._iRES
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`{@925} Figure 1 derncnstrates that DMF and Mitt? are activaters at Nrf2 at
`cnncentratiens within ciinicai expesnre range (ceiis in cutters),
`V
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`§€C3G2?}
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`Figure 2 shears resaits ei RNAE experiments,
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`E9628] Figure 3 snaws evidence at i\irt2 activation by DMF and MMF in viva.
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`{$929} Figure 4 shsws evidence at Nrf2 activaticn by DMF and MMF in viva.
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`[t’.ifi3G} Fumaric acid esters, such as EMF, have been prepesad fer treatment
`
`at MS (see, e.g., Sshimrigit at al., Eur. J. i\ieurci,, 2008, 'i3{6):8G4~tG; Drugs R&D,
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`2395, 8(4};229~3G).
`
`{M3313 Pravitied are, arneng ether things, means ter identifying ccmpaunas
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`with a new therapeutic modality useful in at least one of multiple neurological
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`indications and, optionally, complementary to other drugs for the treatment of a
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`neurological disease, including a number of currently used immunomodulators.
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`WO 2008/097596
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`PCT/US2008/001602
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`[0032] DMF is a member of a large greup of anti-oxidant molecules known for
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`their cytoprotective and anti-inflammatory properties. These molecules also share the
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`property of the Nrf2 pathway activation. Thus, the finding that DMF activates the Nrf2
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`pathway in conjunction with the neuroprotective effects of DMF further offers a
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`rationale for identification of structurally and/or mechanistically related molecules that
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`would be expected to be therapeutically effective for the treatment of neurological
`
`disorders, such as, e.g., MS.
`
`[0033] Certain terms are defined in this section; additional definitions are
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`provided throughout the description.
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`[0034] The terms “activation” and “upregu|ation,” when used in reference to
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`the Nrf2 pathway, are used interchangeably herein.
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`[0035] The terms “disease” and “disorder” are used interchangeably herein.
`
`[0036] The term “a drug for treating a neurological disease” refers to a
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`compound that has a therapeutic benefit in a specified neurological disease as shown
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`in at least one animal model of a neurological disease or in human clinical trials for the
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`treatment of a neurological disease.
`
`[0037] The term “neuroprotection" and its cognates refer to prevention or a
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`slowing in neuronal degeneration, including, for example, demyelination and/or axonal
`
`loss, and/or, neuronal and/or oligodendrocyte death. Neuroprotection may occur
`
`through several mechanisms, e.g., through reducing inflammation, providing
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`neurotrophic factors, scavenging free radicals, etc. As used herein, a compound is
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`considered neuroprotective if it (1) upregulates the Nrf2 pathway above a certain
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`threshold and (2) provides neuroprotection, regardless of possible other mechanisms
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`of action.
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`[0038] The terms “treetmerit," “therapeutic rrsetiied,” “therapeutic benefits,” end
`
`the iiite refer te ttzereeeetie es weil es ereehyiecticipreventetive measures, Thus,
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`these in need ef treatment mey include iradivieueis eireeey iievirig e seeeiiiee disease
`
`and these wite are at risk fer ecqtsiririg that diseese.
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`[0039] The terms “therapeutically effective dose" and “therapeutically effective
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`amount" refer to that amount of a compound which results in at least one of prevention
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`W0 20138/097596
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`PCT/US20lE8/001602
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`or delay of onset or amelioration of symptoms of a neurological disorder in a subject or
`
`an attainment of a desired biological outcome, such as reduced neurodegeneration
`
`(e.g., demyelination, axonal loss, and neuronal death) or reduced inflammation of the
`
`cells of the CNS.
`
`[0040]
`
`In one aspect, provided are methods of evaluating neuroprotectlve
`
`properties of test compounds, including the following methods:
`
`1) methods of screening for new candidate compounds that may be
`
`useful for treating a neurological disease;
`
`2) methods of evaluating neuroprotectlve properties of drugs and
`
`candidates that are used or proposed for treating a neurological
`
`disease;
`
`3) methods of comparing (e.g., for bioequivalence) two or more
`
`pharmaceutical compositions which contain fumaric acid
`
`derivatives;
`
`[0041]
`
`In some embodiments, methods 1-3 may coneprise:
`
`a) contacting a cell with the test compound,
`
`b) deterrraining whether the Nrf2 pathway is upregulated in the cell,
`
`and, in some embodiments, additionally performing the following step(s):
`
`c) detennining whether the test compound slows or prevents demyelination,
`
`axonal loss, and/or neuronal death, and/or
`
`d) selecting the test compound as a candidate for treating neurodegeneration
`
`in a neurological disease if 1) the Nrf2 pathway is upregulated and 2)
`
`demyelination, axonal loss, and/or neuronal death are/is prevented or
`
`stowed.
`
`Method 1
`
`[0042]
`
`In some embodiments the methods of screening for a candidate
`
`compound for treating a neurological disease comprise:
`
`a) contacting a cell with a plurality of test compounds,
`
`b) detennining whether the Nrf2 pathway is upregulated in the cell, and
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`c) selecting from the plurality of compounds at least one compound that
`
`apregulatss the Nrtz pathway,
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`wherein an upregulation of the Nrf2 pathway by the selected at least one compsund
`
`indicates that the selected at least one compound may be useful for treating a
`
`neurological disease. For example, the plurality of compounds may be represented by
`
`a combinatorial chemical library, and the screening method may be performed by a
`
`high-throughput screening as described in, e.g., High—Throughput Screening in Drug
`
`Discovery (Methods and Principles in Medicinal Chemistry), by Jtirg Htlser (ed.), John
`
`Wiley 8: Sons (2006).
`
`[0043] Combinatorial libraries of compounds are also described in, e.g.,
`
`Solid-Supported Combinatorial and Parallel Synthesis of Small-Molecular-Weight
`
`Compound Libraries (Tetrahedron Organic Chemistry) lan Salusbury (ed.), Elsevier
`
`(1998); Combinatorial Libraries: Synthesis, Screening and Application Potential
`
`(Library Binding), by Riccardo Cortese (ed.), Walter de Gruyter (1995). The libraries of
`
`compounds may be, for example, quinone libraries and other libraries as described in
`
`Mittoo, Comb. Chem. & High Throughput Screen, 2006, 9:421-423.
`
`[0044]
`
`In some embodiments, the at least one compound or plurality of
`
`compounds being screened and/or selected comprises at least one compound
`
`selected from at least one of the following groups of compounds: mild alkylating
`
`agents, Michael addition acceptors or compounds that are metabolized to Michael
`
`addition acceptors, including compounds of Formulas l, ii, iii, er EV.
`
`{$045}
`
`in seine of the embodiments, the at teest ene eemeennd is eeiected
`
`hem tenterie eeid, its seite, and temarie acid derivatives.
`
`Method 2
`
`E0046} Aise previded are metheds ei evaiueting netirepreteetive prepertiee cit
`
`at ieeet ene drug as“ drug candidate tea" treating at ieast ene neereiegieai disease.
`
`Such metheds eemerise:
`
`a} eentacting a eeit with the at Eeast ene drug er drug candidate, and
`
`h) determining whether the Nri2 pathway is eetegeiateci in the eeit,
`
`wherein the uereguietteh at the t\irt.?. pathway by the at ieast ene drug as“ drug
`
`candidate indieates that the at Eeeet ehe drag er drug candidate is neureereteetive in
`
`treating a human having at neuretegieai disease.
`
`..1g..
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`.PC'1‘/‘US2€t{}$.I'{l016ti2
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`[0047]
`
`In some embodiments, the upregulation of the Nrf2 pathway by the at
`
`least one drug or drug candidate indicates that the at least one drug or drug candidate
`
`has at least one activity selected from slowing demyelination, slowing the loss of
`
`axons, and slowing the rate of neuronal death.
`
`[0048]
`
`In some embodiments, the method of evaluating at least one drug or
`
`drug candidate comprises an additional step:
`
`c) evaluating demyelination, loss of axons, and/or neuronal death.
`
`[0049]
`
`In some embodiments, steps a) and c) are performed in vivo
`
`at least
`
`one model of a neurological disease, e.g., as described below.
`
`[0050]
`
`in ether embodiments, particularly those in which the neurological
`
`disease is multiple sclerosis or another demyelinating disease, the evaluated at least
`
`one drug or drug candidate for a neurological disease is chosen from the following:
`
`FTY720 (2-(4-octylphenethyl)-2-aminopropane-1,3-diol; Novartis); anti-lL12 antibody
`
`(e.g., ABT-874; Abbott Laboratories); GSK683699 (GSK/'l'anabe); Neurovax (Immune
`
`Response Corp.; Darlington, Curr. Opin. Mol. Ther., 2005, 7(6):598-603); anti—CCR2
`
`antibody (e.g., §V|LN 1202; Millennium); interferon [3-1a (e.g., Avonex®; Biogen ldec);
`
`anti-a4-integrin antibody (e.g., Tysabri®; Biogen ldec/Elan); anti-CD20 antibody (e.g.,
`
`Rituxan® (Biogen Idec/Genentech); TV 5010 (Teva); NBI-788 (Neurocrine); MBP8298
`
`(BioMS (see Warren et al., Eur. J. Neurol., 2006, 13(8):887-95); Mylinax (Oral
`
`Cladribine; 2-chlorodeoxyadenosine; Serono/IVAX); Teriflunomide
`
`((Z)-2-cyano-N-(4-(trifluoromethyl)pheny|)-3-hydroxybut-2-enamide; Sanofi-Aventis);
`
`Temslrolimus (Wyeth); Laquinimocl
`
`(5-chloro-N-ethyl-1 ,2-dihydro-4-hydroxy-1-methyl-2-oxo-N-phenylquinoline=3-carboxa
`
`mide; Active Biotech/'|'eva); and interferon tau (Tauferon; Pepgen).
`
`[0051]
`
`In some embodiments, the at least one drug or drug candidate being
`
`evaluated is at least one compound selected from at least one class selected from a
`
`mitd aikyiating agent, a Michael addition acceptor, and a compound that is
`
`metabolized to a Michael addition acceptor, including compounds of Formulas I, II, III,
`
`or iv.
`
`-11.-
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`WW3 2€}€}8/997596
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`PCT/USZGQS/(Bt)16{}2
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`{M3525
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`tn some at the emhediments, the Ct)f‘?t§3e3t£t‘t{§ is fttmaric acid, its satt, er
`
`a ftsmartc acéti ttertvative.
`
`Methhtt 3
`
`{M353} A339 pmvideé are methtsds at comparing (e.g., far htaequivaience) at
`
`Beast two pharmaceuttcai (.2Qt‘f‘§p()S§E§C§E't$. Such metheds mmprise:
`
`3) C(3t"t‘§3{.2‘i§§"‘tg a ceii with at teast hht-3 first compositihh camprising a test
`
`ctsmphuhd, ahd
`
`ta) mmparihg the tevet of the Nrfz pathway uhreguiatieh th the shit by the test
`
`C€}m§I3G3.§§'3<.‘§
`
`ta the ctzrrespcahdthg tevt:-:§ at the Nat? pathway upregutaticsh in a
`
`set: treated wéth at taast £3318 sectahd ctzsmphsitéhh (“cemparatcsr
`
`camhc:»stticm") chmprtsthg DMF, NEMF, GE" bath.
`
`{(36543
`
`in same emhedimehts, suhstahtiatiy fiissimitar tevets at upreguiatihh by
`
`the at teast tithe first and at teas: one secund composittmts indicate that the
`
`cemhhsiticns are hat héhequivaieht.
`
`{@1155}
`
`in same 8mb{§C§§¥‘t‘§£‘m$, the tegt mmpmahd is ttimarit: acid, its sait
`
`therein‘, 3 hsmarit: aszié tterivatéve, as" mixtures theraef. ht same embhdimahts, the first
`
`comhhsitioh cemptiseh at ieast she Qf §MF, MMF, and bath {EMF and Mh/SF. ht shma
`
`amtmdimahts, the time aE‘t{3/€35‘ the fhrmutatian at the at tgast she first GQ§‘t‘i§.'3QS§€§Q§‘E
`
`diffem htzem the dash ahélhr the tcsrmuiatthh of tha at teast (me hechnfi ctsmpcssitiah.
`
`The at iaast {N38 férst campasétéah may he a cehtrhtiect reieass campasittah such as,
`
`e.g., cmhpasitihns éestsrihect in WC 23{38iG3‘?’342.
`
`{G658}
`
`Eh same amhadisttehts, the methhd further csmprises and adcziititmai
`
`Stags):
`
`tz) mmparing at Eeast me pharmamkinetic gsarametsr at the at teast aha fimt
`
`and the at iaast aha sachhd chmposttthns.
`
`EGG5?} Pharmacckihetic parameters and methhds tar evaiuatirtg tha same are
`
`WSEE §<t‘tQWi“£ and are described in, e.g., Pharmachkthetics, Seahhd Editiah {mugs and
`
`the Pharmaceuticafi Sciences) hy it/him Gihatdi at at. (eds), tntcsrma Heaithcare {W82}.
`
`Exampies at such pharmactzhihatic parameters that can he evatuatect ihctude serum
`
`hait—ttfe-3, tztearahca, and vaiume tiistrihhtitin.
`
`“:2-
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`[0058]
`
`in some embodiments, substantially dissimilar pharmacokinetic
`
`parameter(s) of the a least one first and at least one second compositions indicate that
`
`the compositions are not bioequivalent.
`
`[0059]
`
`In some embodiments, the test compound being evaluated is a miici
`
`alkylating agent, and more specifically, a Michael addition acceptor, or a compound
`
`that is metabolized to a Michael addition acceptor.
`
`[0060]
`
`In some of the embodiments, the test compound is fumaric acid or its
`
`salt, or a fumaric acid derivative.
`
`[0561] Also provided are methods of treating a mammal who has or is at risk
`
`for developing a neurological disease, including the following methods:
`
`4) methods of treating a neurological disease by administering to the subject
`
`in need thereof at least one compound that is partially structurally similar
`
`to DMF or MMF (including compounds selected using methods 1-3
`
`described above) ; and
`
`5) methods of treating a neurological disorder by a combination therapy that
`
`includes administration of a first compound that does not upregulate the
`
`Nrf2 pathway and a second compound that ueregulates the Nrf2 pathway.
`
`Method 4
`
`[0062] Also provided are methods of treating a neurological disease by
`
`administering to the subject in need thereof at least one compound that is at least
`
`partially structurally similar to DMF and/or MMF.
`
`[0063]
`
`In some embodiments of method 4, a method of treating a mammal
`
`who has or is at risk for a neurological disease is provided. The methods comprises
`
`administering ii: the rnammei e therepeuticeiiy effective amount at at least ene
`
`neurepretective cernpeund which has Fermeie i, ii, iii, er iv, e.g., e tunieric acid
`
`derivative (e.g., iliivii-’ er MMF).
`
`[0004]
`
`in seine embedimente of rnetherzi 4, e methed ef siewing er preventing
`
`neerodegeneretien {mere speciiiceiiy, e.g., dernyeiinetien, axenei tees, endier
`
`neurenei death) in a subject in need thereet, by administering the at ieast cine
`
`rznrnpeend in en erneunt end fer a peried cit time sufficient it} its at Eeesi 0318 of stew or
`
`-13»
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`PCT/US2008/001602
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`prevent demyelination, slow or prevent axonal loss, and alow or prevent neuronal
`
`death, e.g., by at least 30%, 50%, 100% or higher over a control over a period of at
`
`ieest 5, to, 12, 20, 40, 52, 100, or 200 weeks, or more.
`
`Method 5
`
`[0065] Also provided are methods of treating a mammal having a neurological
`
`disease by combination therapy.
`
`In some embodiments such methods comprise:
`
`a) administering to the mammal a therapeutically effective amount of at least
`
`one first compound that upregulates the Nrf2 pathway, and
`
`b) administering a therapeutically effective amount of at least one second
`
`compound that does not upregulate the Nrf2 pathway.
`
`[0066]
`
`In some of embodiments of method 5, the at least one first compound,
`
`used in step (a), is a compound of Formula I. II, III, or IV, e.g., DMF or MMF; and the
`
`at least one second compound, which is used in step (b), is an immunosuppressive or
`
`en immnnomoduietory oomponnd thet does not upregiiiete the hirtiz osthwey (e.g., hy
`
`more then 33%, 55%, tQ{i% over e eontroi).
`
`iiifififi
`
`in some embodiments of method 5, the method comp