Us 2019 / 0262353 A1

US 20190262353A1 ( 19) United States (12 ) Patent Application Publication (10 ) Pub. No. : US 2019 /0262353 A1 YONG et al. (43 ) Pub . Date : Aug . 29 , 2019

( 54 ) TREATMENT FOR PROGRESSIVE Publication Classification MULTIPLE SCLEROSIS (51 ) Int. Cl. (71 ) Applicant : UTI LIMITED PARTNERSHIP , A61K 31/ 55 ( 2006 . 01 ) Calgary (CA ) A61K 31 /404 ( 2006 .01 ) (72 ) Inventors : Voon Wee YONG , Calgary (CA ); A61P 25 / 28 (2006 .01 ) Simon FAISSNER , Bochum (DE ) ; ( 52 ) U .S . CI. Marcus KOCH , Calgary (CA ); Nathan CPC ...... A61K 31/ 55 (2013 . 01 ) ; A61P 25 / 28 James MICHAELS, Kamloops (CA ) ( 2018 .01 ) ; A61K 31 /404 ( 2013 .01 ) (21 ) Appl. No. : 16 / 343, 818 (22 ) PCT Filed : Oct. 24 , 2017 (57 ) ABSTRACT ( 86 ) PCT No. : PCT/ CA2017 /051269 In one aspect , there is provided a method of treating , $ 371 ( C ) ( 1 ) , prophylaxis , or amelioration of a neurological disease by (2 ) Date : Apr. 22 , 2019 administering to a subject in need thereof one or more Related U . S . Application Data compounds described herein . In a specific example , the ( 60 ) Provisional application No . 62 /412 ,534 , filed on Oct . neurological disease is multiple sclerosis (also referred to as 25 , 2016 . “ MS ” ) . Patent Application Publication Aug. 29, 2019 Sheet 1 of 23 US 2019 / 0262353 A1

3000 Iron mediated neurotoxicity * * * *

* * * *

* * * Neurons 1000 fitini TTTTTTTToitain | 1 | ControlFeSO4DMSO 50 WMAtenolol PimozideFosfomycinKetoprofen MefloquineQuinapril Liothyronine'5-Chloroindole Ranitidine Bumetanide TolfenamicTranexamic acid acid ketoprofenDequaliniumPyridostigmineCyclosporine Paromomycin Labetalol Tenoxicam Mitoxanthrone Trazodone FeSO450 PM Control Fes0 , 50 PM FeSO , 50 UM + Indapam

Control Feso , 50 UM FeSO , 50 UM + Indapam

FIGURE 1 Patent Application Publication Aug. 29, 2019 Sheet 2 of 23 US 2019 / 0262353 A1

??? - iron Antiaggregation Antihypertensive

Antipsychotic Tricyclic . Antibiotic antidepressant

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. A . v DipyridamoleClopidogrelClarithromycinCefaclor vErythromycin Rifampin*Loperamide Ketoconazole Labeta?ol Methyldopa Metoprolol Atenolol Carvedilol IndapamideMefloquine Primaquine Mitoxanthrone LevodopaTrimeprazine Chlorpromazine Clozapine Periciazine FlunarizineDimenhydrinateDiphenhydramine PromethazinePhenazopyridine Yohimbine MemantineLiothyronine Clomipramine Desipramine Doxepin ImipramineTrimipramine X Live cell imaging neurotoxicity

Legend : FeSO4

%Plpositiveneurons en Indapamide + Feso : * terowaniaintegrirana erDet Desipramine + FeSOA -

...... wwerwe . www . Boswwwwwwwwwwwww 0 1 2 3 4 5 6 7 8 9 10 11 12 Time th ] FIGURE 2 Patent Application Publication Aug. 29 , 2019 Sheet 3 of 23 US 2019 / 0262353 A1

Rotenone mediated neurotoxicity C Drug * Rotenone Antipsychotic Antihypertensive Tricyclic antidepressant * * * *

???? www %controlneurons cmcm.

. FinW.WWW .

ways . . DipyridamoleClopidogrelCefaclor Labetalot Methyldopa Metoprolol Indapamide Mefloquine PrimaquineMitoxantrone TrimeprazineChlorpromazine Clozapine Periciazine FlunarizinePromethazine MemantineLiothyronine Clomipramine Desipramine Imipramine Trimipramine Doxepin Control Rotenone 10 UM + Indapamide Rotenone 10 uM We

3 . S

FIGURE 3 Patent Application Publication Aug. 29 , 2019 Sheet 4 of 23 US 2019 / 0262353 A1

HORAC HORAC 15001 - Blank 1500 , + Blank www > Die schem Indapamide * Clomipramine 1000 RFU RFU 500 5007

Time (min ) Time (min )

. Tricyclic Antihypertensive Antipsychotic antidepressant VE * * *

HORACGAE - cost

Galicacid Clopidogrel CefaclorMethylodopa Metoprolol "Atenolot Indapamide Primaquine 4 Clozapine DoxepinImipramine Dipyridamole MitoxantroneMitoxantroneChlorpromazineTrimeprazineop PericiazineFlunarizineDimenhydrinate Promethazine MemantineLiothyronine Clomipramine Desipramine Trimipramine FIGURE 4 Patent Application Publication Aug. 29 , 2019 Sheet 5 of 23 US 2019 / 0262353 A1

T - cell proliferation

150 -

Antipsychotic Tricyclic antidepressant Ž %activatedT-cells(cpm) .# . # . # # SRO ttt sk

# tu

* * * * * * * * X N DMSO Primaquine Clozapine Doxepin Non-activated ActivatedDipyridamole Clopidogrel Cefaclor XLabetalolMethyldopa MetoprololIndapamide Mefloquine Mitoxanthrone TrimeprazineChlorpromazine Periciazine FlunarizinePromethazine MemantineLiothyronine Clomipramine Desipramine ImipramineTrimipramine aCD3 /aCD28 activated

FIGURE 5 Patent Application Publication Aug. 29, 2019 Sheet 6 of 23 US 2019 / 0262353 A1

Neuroprotection by clomipramine b Clomipramine interacts with iron Live cell imaging neurotoxicity

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LOW

?? 100M ControlOSNO Fesod Control(wash )DMSO FeSO4 SO HIMClomi +(washiFier *Fe oui ) 4 ( awendung Clomi(wash ,readded )+Fe . e $ o wos 2hPro -incubation Clomi *fa muco mini Clom 2 UM T - cell proliferation G1 phase S - aseud

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100IN 500 nM Non-activated Activated DMSO Non-activated Activated Non-activated Activated" Clomipramine dwoo auwe dituoo ULJE

B - cell proliferation B - cells TNF - 0 150007

0000- Lud Pgimi 2004 ControlPane Activated auguedwoo Activated BUDEJOWO Activated FIGURE 6 Patent Application Publication Aug. 29, 2019 Sheet 7 of 23 US 2019 / 0262353 A1

EAE - Delayed treatment

* * * * * * * * - Vehicle * * * * + Clomipramine Clinicalscore * * *

????????????????????????? Annas on ooo N N P G A B on Days post MOG

Treatment period FIGURE 7A Patent Application Publication Aug. 29, 2019 Sheet 8 of 23 US 2019 / 0262353 A1

Treatment period 100

Sumofscores VehicleClomipramine FIGURE 7B Patent Application Publication Aug. 29, 2019 Sheet 9 of 23 US 2019 / 0262353 A1

# - Corrigasmine Clinicalscore A

Days post immunization Treatment period

Sum of scores Weight day o Weight day 16

huwe .

S AN weight[9] Sumofscores wwwiiwiwiwilini [?]??????. .

CCL2 6 R ALLS LAHAHALAAMCHLULA RelativeRNAexpression RelativeRNAexpression RelativeRNAexpression WWW WA

Serum Spinal cord f Clomipramine DMCL P = 0 . 7026 SpinalcordfuM

500 000 15000 50 100 150 20 Serum Inha Serum nM )

Clomipramine wwwwwwwwwwwwwwwwwwwww

FIGURE 8 Patent Application Publication Aug. 29, 2019 Sheet 10 of 23 US 2019 /0262353 A1

Clomipramine LETTT

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ibak1

ANSMOLOS|018

Microglial actlvation Axonal damage H3E 3 ax ?????????? Microglial activation / axonal damage

SPT F . 30 Somman * Rankordar Raskorder Axoraldomage RES randum VE ST BURA Microglial adlikako

FIGURE 9 Patent Application Publication Aug. 29, 2019 Sheet 11 of 23 US 2019 /0262353 A1

Chronic EAE b treatment initiated at remission Chronic EAE - treatment from clinical onset + Vehicle * Clomipramine

Score Clinicalscore olub

+ Clomipramine minimaal p * 0 .0175 T TTTTT Days post Immunization Days after immunization Treatment Treatment Biozzi ABH EAE * * 2 - 0 .0062 1 - Vehicle II . Clomipramine ) 144Well Clinicalscore Disability AssessiveShowVIN MOG immunization O ......

A First relapse Remission Chronic phase Days post immunization . .. phase . Treatment Time FIGURE 10 Patent Application Publication Aug. 29, 2019 Sheet 12 of 23 US 2019 /0262353 A1

5 -Chloroindole - Atenolol - iron 150 , drug

H X %controlneurons - H# HEH HCWmo TAS HOTI RHHRI@ - IMMT It OCHT HEIM HGTH- SET oc AcetaminophenAcetylcysteine AcyclovirAllopurinol Almotriptan Altretamine Amikacin sulfate Amlodipine besylate AmoxicillinAmphotericin Antipyrine B Aspirin Atenolol 5-Chloroindole Acebutolol-2-Carboxylic hydrochloride acid Acetazolamide AmantadinehydrochlorideAmiloride AmiodaroneAmitriptylinehydrochloride hydrochloride hydrochloride

FIGURE 11A

Atorvastatin - Carbachol irondrug

%controlneurons

C# HEW nm - C DC HEM BRINA HEH! IMIMI HOCH - 0 - 0 x 1 del II COM FW 1 HMC-70 Bupropion V Captopril AtorvastatincalciumAtovaquone Azathioprine AzithromycinBenazepril BenserazideBacitracin hydrochlorideBaclofen hydrochloride Bethanechol Benztropine Brompheniramine chlorideAh Bezafibrate Bisacodyl maleate Budesonide Bumetanide Candesartan Busulfan cilextil Carbachol FIGURE 11B Patent Application Publication Aug. 29, 2019 Sheet 13 of 23 US 2019 /0262353 A1

Carbamazepine - Clozapine drug XHD

X %controlneurons XHULIN X i EI 11HOC# BEKER H7? HEI IA FT?? CefaclorCefadroxilCephalexin Hc CimetidineCiprofloxacin Clemastine Clopirogrelsulfate Clozapine CarbamazepineCarvedilolChlorpheniraminetartrate (SChlorpromazine )maleate ChlorpropamideChlorthalidone ClindamycinClarithromycin ClomipraminehydrochlorideClonidine hydrochloride hydrochloride CloxacillinClotrimazole sodium

FIGURE 11C iron Colchicine - Dipyridamole drug %controlneurons HEWA ECO Cresol HOCIO DanazolDapsone Digoxin CyclobenzaprineColchicineCromolynCyclophosphamidehydrochloride sodium hydrateCyclosporine DextromethorphanDesipramineDequalinium hydrochloridechloride hydrobromide DiclofenacDiazoxide DiltiazemsodiumDiphenhydramine Diflunisal Diphenylpyraline hydrochloride Dimenhydrinate hydrochloride hydrochloride Dipyridamole

FIGURE 11D Patent Application Publication Aug. 29, 2019 Sheet 14 of 23 US 2019 /0262353 A1

Disopyramide - Fosfomycin drug X - iron

H %controlneurons

HCM HCM HEN ? XHWHINE 1 HC HCHHI HOCI HH-0 Hmmun ERRIA HLEHR DisopyramideDoxepinDoxycyclinephosphate hydrochloride Doxylaminehydrochloride Edrophonium succinate EnalaprilErgonovine chloride ErythromycinEthambutol maleate maleate hydrochloride estolate Ethosuximide EtodolacDEWOEzetimibe FamciclovirFlunarizine Famotidine FluphenazineFenofibratehydrochloride Fluoxetinehydrochloride HORREO Flurbiprofen Fosfomycin

FIGURE 11E

Furosemide - Isosorbide Dinitrate drug iron

%controlneurons HERRAM COMMUNE HEM XHOVIH HERRAM ICH FurosemideGemfibrozilGliclazide Glyburide GuaifenesinHydralazine HaloperidolHexylresorcinol HydroxychloroquineHydrochlorothiazide hydrochloride Hydroxyzine Hydroxyureasulfate Imipramine pamoate Ibuprofenhydrochloride IndapamideIpratropium Indomethacin bromide Irbesartan IsosorbideIsoniazid dinitrate

FIGURE 11F Patent Application Publication Aug. 29, 2019 Sheet 15 of 23 US 2019 /0262353 A1

Ketoconazole - Mefenamic acid drug XL = iron X %controlneurons K HC XH?? HCI LIM LEAN ECH CHIO Lisinopril HADIO, KetorolacKetoconazoleKetoprofentromethamineKetotifenLabetalol hydrochloride fumarate LactuloseLeucovorin Lansoprazole calcium LiothyronineLevodopa LevofloxacinLoperamide sodium hydrochloride LoratadineMaprotiline LoxapineLosartanLovastatin succinatehydrochloride MebendazoleMefenamic acid

FIGURE 11G

Mefloquine - Mycophenolic acid drug = iron

%controlneurons XW XHT HEM XHBOAT HE =H HEW www . * XKOCIo wu. HMC- II OHEM IEEECHI Mefloquine HOWO MinoxidilHCho HOCHO MemantineMeloxicamhydrochloride MercaptopurineMethazolamide MethenamineMethocarbamol Metoclopramide Methotrexate Methoxsalen Methyldopahydrochloride MetoprololMetolazoneMidodrine MetronidazoleMitoxanthronetartrate hydrochloride Moxifloxacinhydrochloride ModafinilhydrochlorideMycophenolic acid

FIGURE 11H Patent Application Publication Aug. 29, 2019 Sheet 16 of 23 US 2019 /0262353 A1

Nabumetone - Periciazine drug pengiron

%controlneurons K

UNT RIT ETHNIE COM HPRIME Hm201- INIKILAN42 HEWI 17: 12 - 13 Nadolol Naproxen(+ )Nifedipine Orlistat CHO NaloxoneNabumetoneNaltrexonehydrochloride hydrochloride Neostigmine bromide Nilutamide NimodipineNitrofurantoin NylidrinNorfloxacin OlmesartanNortriptylinehydrochloride medoxomil Orphenadrine ParomomycinOxcarbazepine citrate Pentoxifyllinesulfate Periciazine

FIGURE 111

Perindopril - Propafenone drug results iron

H XHRA %controlneurons - KICH HINDUHI Monu HETERNIH - KHIR-TO ARMO KEHH. HITE PerphenazinePhenelzinePhenytoin sulfate sodium Pimozide Pindolol Piroxicam PregabalinPrimidone Probenecid KEtimo PerindoprilPhenazopyridineerbumine hydrochloride Pioglitazone Potassiumhydrochloride p- Aminobenzoate Pravastatin Primaquinesodium diphosphateProcainamide ProchlorperazineProcyclidinehydrochloride Promethazine Propafenoneedisylatehydrochloride hydrochloride hydrochloride

FIGURE 11J Patent Application Publication Aug. 29, 2019 Sheet 17 of 23 US 2019 /0262353 A1

Propanolole - Sulfasalazine drug - iron

%controlneurons LE HC HOCH HCHINI HTIN HA LIH HIHNO AMANTHARAANHC? PyrantelpamoatePyrazinamide Quinine sulfate RanitidineRifampin Rifaximin RosiglitazoneRiluzole SildenafilSpiramycin HA PropranololPseudoephedrinehydrochloride Propylthiouracil(+hydrochloride/ - ) Pyridostigmine Quinapril bromidehydrochloride RY SertralineRosuvastatinhydrochloride Spironolactone SulfamethoxazoleSulfasalazine

FIGURE 11K

Sulfinpyrazone - Trimethoprim drug 150 iron

H %controlneurons XHB KW

HET HC# CH - - - HER HEMA EU HLR-O OMENI ECH HOE HEIN Sulindac Thioguanine TopiramateTrandolapril SulfinpyrazoneSulfisoxazoleTetracycline Tenoxicamhydrochloride Theophylline ThiothixeneTimolol maleateTolbutamide Tolfenamic acid TranylcypromineTranexamicTrazodone TrifluoperazineTrihexyphenidylacidhydrochloride sulfate hydrochloride Trimeprazinehydrochloride ' Trimethoprimtartrate

FIGURE 11L Patent Application Publication Aug. 29, 2019 Sheet 18 of 23 US 2019 /0262353 A1

Trimipramine - Zolmitriptan X drug A iron %controlneurons H HE HEI-? 1909 JUS CI HTTC. TrimipraminemaleateVancomycin ValsratanUrsodiola hydrochloridesodium Verapamil Yohimbine Venlafaxine hydrochloride hydrochloride Zidovudine Zolmitriptan(AZT )

FIGURE 11M Patent Application Publication Aug. 29, 2019 Sheet 19 of 23 US 2019 /0262353 A1

a Demyelination detected by b Volume ofmyelin loss after lysolecithin eriochrome cyanine (blue ) * 2400 R Young * Aging Volume(*10%um") # 1000 R

+ 400 R Leson Epicenter Young 4hours 24hours 72hours C Depiction of the average myelin loss per group over time, at various sections from the lesion epicenter

4 hours 24 hours 72 hours 1359 425 1757 100 100

2 1 . . . wwwwwwwwww 50 5069 Caudat Dicontor Rostre Epicenter Caudal Epicenter Restral TAAFIGURE 12 Patent Application Publication Aug. 29, 2019 Sheet 20 of 23 US 2019 /0262353 A1

a SM1312 labeling of axons within the lesion of lysolecithin -demyelinated mice b Lower % spared axons in aging mice at 72h Young Aging p = 0 .0164

NAWM .

. %SMI312+axonsparing

2 Injured Young Aging

FIGURE 13 Patent Application Publication Aug. 29, 2019 Sheet 21 of 23 US 2019 /0262353 A1

Young Naive Aging Naive Young LPC Aging LPC

DOSSIEREN BABIESSERE

#Dutugis Lesiginating Bh50m Het 7* $ 6signaling TREMisigoating Dendtekscet essaturaliose RAGsignaling MIFtagation Loukocytestate extravasatoo inmunity signating ' MlFediatasfglucocorticoidunese soeineregras P?????Our ???thisk Busku w941 catyce G2 ONA damage thskpionaton butoanesa jert l ProductionofOO eitricAZ and reactive Oxyge Species interconnages ' wwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwww www . www . www . me Be bagai desatero???A- ?SA325 bosted?NA?10? CPR Foy Receptor- mediated Phagocytosis in Alacsophagas and Monocytes for CP L Signaling EP : Leukocie Extravasatoa Signating gp91phox

CF MFRegualion of imata mully CPMF- mejated Glancanicod Regulator .

. . CA TOHke Receptor Signaling FPKM Extracelular CP : Rac Signalng . . . .. Cytoplas CRE L- a Sanning CPE TREMT Signaling CF Oandrile CosMasturlar YoungNaive Aging Naive YoungLPC CP Call Cycle: Q2MDNA Damage Checa ant Regulation CPCD Signaling in T Helper Calls CYBB CPK note d Patern Recognition scecices in Kecogaitian of Bacteris and Viruses

FIGURE 14 Patent Application Publication Aug. 29, 2019 Sheet 22 of 23 US 2019 /0262353 A1

Young Aging 15001

1000 phoxgp91 CD45 6p91phox#vearea

Young Aging

Malondialdehyde P - 0 . 0002 Malondialdehyde#vearea

Aging

Malondialdehyde GO 22

FIGURE 15 Patent Application Publication Aug. 29, 2019 Sheet 23 of 23 US 2019 /0262353 A1

lesionvolumex100un 59 wwwww

Coro Indapamide Control Indapamide

my e Lipid peroxidation p = 0 .04

15 ! P = 0 .06 a Malondialdehyde*area(uma) Numberofaxonsperfieldview : $ baby Control Indapamide Control Indapamide Demyelinated vehicle Demyelinated + indapamide

FIGURE 16 US 2019 /0262353 A1 Aug . 29 , 2019

TREATMENT FOR PROGRESSIVE amount of clomipramine, or a functional derivative thereof, MULTIPLE SCLEROSIS and a therapeutically effective amount of indapamide , or a functional derivative thereof. CROSS REFERENCE TO RELATED [0010 ] In one aspect there is described a method of APPLICATION treating progressive multiple sclerosis comprising adminis tering to a subject in need thereof, a therapeutically effective [0001 ] This application claims priority to U . S . 62 /412 , amount of indapamide , or a functional derivative thereof. 534 , filed Oct . 25 , 2016 , the entire contents of which is [0011 ] In one aspect there is described a method of treat incorporated by reference in its entirety . ing progressive multiple sclerosis comprising administering to a subject in need thereof, a therapeutically effective FIELD amount of indapamide, or a functional derivative thereof, [ 0002] The present disclosure relates generally to com and one or more of hydroxychloroquine , minocycline , or pound ( s ) , composition ( s ) , and method ( s ) for treatment for clomipramine . progressive multiple sclerosis in a subject . [ 0012 ] In one example said multiple sclerosis is primary progressive multiple sclerosis . BACKGROUND [0013 ] In one example said multiple sclerosis is secondary [0003 ] Multiple sclerosis is a multifactorial inflammatory progressive multiple sclerosis . condition of the CNS leading to damage of the myelin sheath [0014 ] In one example said multiple sclerosis is progres and axons/ neurons followed by neurological symptoms sive relapsing multiple sclerosis . (Ransohoff et al. , 2015 ). Approximately 85 % of multiple 10015 ] In one example said treatment further comprises sclerosis patients present with a relapsing -remitting pheno administering a therapeutically effective amount of Laqui type and the majority of these evolve to a secondary nimod , Fingolimod , Masitinib , Ocrelizumab , Ibudilast , progressive disease course after 15 -20 years . Ten - 15 % of the Anti - LINGO - 1 , MD1003 (high concentration Biotin ) , patients experience a primary progressive disease course Natalizumab , Siponimod , Tcelna ( imilecleucel - T ) , Simvas with slow and continuous deterioration without definable tatin , Dimethyl fumarate , Autologous haematopoietic stem relapses . cell transplantation , Amiloride , Riluzole , Fluoxetine, Glati [ 0004 ] While there have been tremendous successes in the ramer Acetate , Interferon Beta , or a functional derivative development ofmedications for relapsing -remitting multiple thereof. sclerosis during the last decade, nearly all studies conducted [ 0016 ] In one example said subject is a human . in progressive multiple sclerosis have failed such as the [0017 ] In one aspect there is described herein use of one recently published INFORMS study on the sphingosine - 1 or more of dipyridamole , clopidogrel, cefaclor, clarithromy phosphate inhibitor fingolimod (Lublin et al ., 2016 ). The cin , erythromycin , rifampin , loperamide , ketoconazole , reasons for the lack of medications in progressive multiple labetalol, methyldopa , metoprolol, atenolol, carvedilol, indapamide , mefloquine, primaquine, mitoxanthrone , sclerosis are manifold . levodopa, trimeprazine , chlorpromazine, clozapine , perici azine , flunarizine, dimenhydrinate , diphenhydramine , pro SUMMARY methazine , phenazopyridine, yohimbine, memantine , liothy [0005 ] In one aspect there is described herein a method of ronine , clomipramine , desipramine , doxepin , imipramine , treating progressive multiple sclerosis comprising adminis trimipramine , or functional derivative thereof, for the treat tering to a subject in need thereof, a therapeutically effective ment of progressive multiple sclerosis in a subject . amount of one or more of dipyridamole , clopidogrel , cefa [0018 ] In one aspect there is described herein use of one clor, clarithromycin , erythromycin , rifampin , loperamide , or more of dipyridamole, clopidogrel, cefaclor , clarithromy ketoconazole, labetalol, methyldopa , metoprolol, atenolol, cin , erythromycin , rifampin , loperamide , ketoconazole , carvedilol , indapamide, mefloquine , primaquine, mitoxan labetalol, methyldopa, metoprolol, atenolol, carvedilol, throne , levodopa , trimeprazine, chlorpromazine , clozapine , indapamide, mefloquine , primaquine , mitoxanthrone , periciazine, flunarizine, dimenhydrinate , diphenhydramine, levodopa , trimeprazine , chlorpromazine , clozapine , perici promethazine , phenazopyridine , yohimbine , memantine , azine , flunarizine , dimenhydrinate , diphenhydramine, pro liothyronine , clomipramine , desipramine , doxepin , imip methazine, phenazopyridine , yohimbine , memantine , liothy ramine , trimipramine, or functional derivative thereof. ronine , clomipramine , desipramine , doxepin , imipramine , [0006 ] In one aspect there is described herein a method of trimipramine, or functional derivative thereof, in the manu treating progressive multiple sclerosis comprising adminis facture of a medicament for the treatment of progressive tering to a subject in need thereof, a therapeutically effective multiple sclerosis in a subject. amount of clomipramine , or a functional derivative thereof. [0019 ] In one aspect there is described herein use of [ 0007 ] In one aspect there is described herein a method of clomipramine , or a functional derivative thereof, for treating treating progressive multiple sclerosis comprising adminis progressive multiple sclerosis in a subject in need thereof. tering to a subject in need thereof, a therapeutically effective [0020 ] In one aspect there is described herein use of amount of imipramine , or a functional derivative thereof. clomipramine , or a functional derivative thereof, in the [0008 ] In one aspect there is described herein a method of manufacture of a medicament for treating progressive mul treating progressive multiple sclerosis comprising adminis tiple sclerosis in a subject in need thereof . tering to a subject in need thereof, a therapeutically effective [0021 ] In one aspect there is described herein use of amount of trimipramine , or a functional derivative thereof. imipramine , or a functional derivative thereof, for treating [0009 ] In one aspect there is described a method of progressive multiple sclerosis in a subject in need thereof. treating progressive multiple sclerosis comprising adminis 0022 ] In one aspect there is described herein use of tering to a subject in need thereof , a therapeutically effective imipramine, or a functional derivative thereof, in themanu US 2019 /0262353 A1 Aug . 29 , 2019 facture of a medicament for treating progressive multiple [ 0036 ] In one example the subject is a human . sclerosis in a subject in need thereof. [0037 ] In one aspect there is described herein a method of [0023 ] In one aspect there is described herein use of identifying a compound for the treatment of progressive trimipramine , or a functional derivative thereof, for treating multiple sclerosis , comprising : selecting one or more com progressive multiple sclerosis in a subject in need thereof. pounds from a library of compounds that prevent or reduce [0024 ] In one aspect there is described herein use of a iron -mediated neurotoxicity in vitro , therapeutically effective amount of trimipramine, or a func [0038 ] selecting one or more compounds from step ( a ) that tional derivative thereof, in the manufacture of a medica prevent or reduce mitochondrial damage in vitro ; selecting ment for treating progressive multiple sclerosis in a subject one or more compounds from step ( a ) for anti - oxidative in need thereof. properties , [0025 ] In one aspect, there is described a use of clomip [0039 ] selecting one or more compound from step ( a ) for ramine , or a functional derivative thereof, and a use of ability to reduce T -cell proliferation in vitro , optionally , after indapamide , or a functional derivative thereof, for treating step ( a ) , selecting a compound from step ( a ) which is progressive multiple sclerosis in subject in need thereof. predicted or known to be able to cross the blood brain [0026 ] In one aspect there is described a use of clomip barrier , or having a suitable side effect profile , or having a ramine , or a functional derivative thereof, and a use of suitable tolerability . indapamide , or a functional derivative thereof, in the manu [0040 ] In one aspect there is described herein a kit for the facture of a medicament for treating progressive multiple treatment of progressive multiple sclerosis , comprising : one sclerosis in subject in need thereof. or more of dipyridamole , clopidogrel, cefaclor , clarithromy [0027 ] In one aspect, there is described a use of indap cin , erythromycin , rifampin , loperamide, ketoconazole, amide , or a functional derivative thereof, for treating pro labetalol, methyldopa , metoprolol , atenolol , carvedilol, gressive multiple sclerosis in subject in need thereof. indapamide , mefloquine , primaquine , mitoxanthrone , [ 0028 ] In one aspect, there is described a use of indap levodopa , trimeprazine , chlorpromazine , clozapine , perici amide, or a functional derivative thereof, in the manufacture azine, flunarizine, dimenhydrinate , diphenhydramine, pro of a medicament for treating progressive multiple sclerosis methazine, phenazopyridine, yohimbine ,memantine , liothy in subject in need thereof . ronine , clomipramine , desipramine , doxepin , imipramine , [0029 ] In one aspect, there is described a use of indap trimipramine , or functional derivative thereof and Instruc amide , or a functional derivative thereof, and one or more of tions for the use thereof . hydroxychloroquine , minocycline , or clomipramine, or a [ 0041 ] In one aspect there is described herein a kit for the functional derivative thereof, for treating progressive mul treatment of progressive multiple sclerosis comprising : a tiple sclerosis in subject in need thereof. therapeutically effective amount of clomipramine , or a func (0030 ) In one aspect, there is described a use of indap tional derivative thereof, and instructions for use . amide , or a functional derivative thereof, and one or more of [0042 ] In one aspect there is described herein a kit for the hydroxychloroquine, minocycline , or clomipramine, or a treatment of progressive multiple sclerosis comprising : a functional derivative thereof, in the manufacture of a medi therapeutically effective amount of imipramine , or a func cament for treating progressive multiple sclerosis in subject tional derivative thereof, and instructions for use . in need thereof. [ 0043 ] In one aspect there is described herein a kit for the [0031 ] In one example said multiple sclerosis is primary treatment of progressive multiple sclerosis comprising : a progressive multiple sclerosis . therapeutically effective amount of trimipramine, or a func [ 0032 ] In one example said multiple sclerosis is secondary tional derivative thereof, and instructions for use . progressive multiple sclerosis . [0044 ] In one aspect there is described a kit for the [ 0033 ] In one example said multiple sclerosis is progres treatment of progressive multiple sclerosis comprising : a sive relapsing multiple sclerosis . therapeutically effective amount of clomipramine, or a func [0034 ] In one example further comprising a use of a tional derivative thereof, a therapeutically effective amount therapeutically effective amount of Laquinimod , Fingoli indapamide , or a functional derivative thereof, and instruc mod , Masitinib , Ocrelizumab , Ibudilast , Anti - LINGO - 1 , tions for use . MD1003 (high concentration Biotin ), Natalizumab , Siponi [0045 ] In one aspect there is described a kit for the mod , Tcelna ( imilecleucel- T ) , Simvastatin , Dimethyl fumar treatment of progressive multiple sclerosis comprising : a ate , Autologous haematopoietic stem cell transplantation , therapeutically effective amount of indapamide , or a func Amiloride , Riluzole , Fluoxetine , Glatiramer Acetate , Inter feron Beta , or a functional derivative thereof, for the treat tional derivative thereof , or a functional derivative thereof , ment of progressive multiple sclerosis , primary progressive and instructions for use . multiple sclerosis , or secondary multiple sclerosis . 0046 ] In one aspect there is described a kit for the [0035 ] In one example further comprising a use of a treatment of progressive multiple sclerosis comprising : a therapeutically effective amount of Laquinimod , Fingoli therapeutically effective amount of indapamide , or a func mod , Masitinib , Ocrelizumab , Ibudilast, Anti - LINGO - 1 , tional derivative thereof, and one or more of hydroxychlo MD1003 (high concentration Biotin ) , Natalizumab , Siponi- roquine, minocycline , or clomipramine , or a functional mod , Tcelna (imilecleucel - T ) , Simvastatin , Dimethyl fumar derivative thereof; and instructions for use . ate , Autologous haematopoietic stem cell transplantation , [0047 ] In one example said multiple sclerosis is primary Amiloride , Riluzole , Fluoxetine , Glatiramer Acetate , Inter progressive multiple sclerosis . feron Beta, or a functional derivative thereof, in the manu [0048 ] In one example said multiple sclerosis is secondary facture of a medicament for the treatment of progressive progressive multiple sclerosis . multiple sclerosis , primary progressive multiple sclerosis , or [0049 ] In one example said multiple sclerosis is progres secondary multiple sclerosis . sive relapsing multiple sclerosis . US 2019 /0262353 A1 Aug . 29 , 2019

[0050 ] In one example further comprising one or more of age as shown after normalization to the control neurons ( A ) . Laquinimod , Fingolimod , Masitinib , Ocrelizumab , Ibudi- The rescue effect was however small . Treatment with rote last , Anti - LINGO -1 , MD1003 (high concentration Biotin ), none induced marked morphological changes with retraction Natalizumab , Siponimod , Tcelna (imilecleucel - T ) , Simvas of cell processes ( B ) . The scale bar shows 100 uM . Shown tatin , Dimethyl fumarate , Autologous haematopoietic stem are normalized data of mean : SEM of 1 - 3 experiments each cell transplantation , Amiloride , Riluzole , Fluoxetine , Glati performed in quadruplicates . Two -way analysis of variance ramer Acetate , Interferon Beta , or a functional derivative ( ANOVA ) with Bonferroni multiple comparisons test as thereof. posthoc analysis vs. rotenone: * p < 0 . 05 ; * * p < 0 .01 ; * * * p < 0 . [ 0051 ] Other aspects and features of the present disclosure 001; * * * * p < 0 .0001 . will become apparent to those ordinarily skilled in the art [ 0056 ] FIG . 4 _ Scavenging of hydroxyl radicals in a upon review of the following description of specific embodi biochemical assay. The anti -oxidative capacities of selected ments in conjunction with the accompanying figures. compounds that reduced iron mediated neurotoxicity were analyzed using the hydroxyl radical antioxidant capacity BRIEF DESCRIPTION OF THE DRAWINGS (HORAC ) assay . Panel A shows a representative experiment 00521 Embodiments of the present disclosure will now be depicting the decay of relative fluorescence units (RFU ) described , by way of example only , with reference to the over 60 min for indapamide , gallic acid (GA ) and the control attached Figures . (blank ). ( B ) The upward shift of the curve for clomipramine [ 0053 ] FIG . 1 — Screening of generic compounds to pre in the HORAC assay indicates an anti -oxidative effect that vent iron mediated neurotoxicity . Shown is an example of a is even stronger than gallic acid . HORAC gallic acid equiva screening of drugs to identify those that prevent iron medi lents (GAEs ) were calculated by the integration of the area ated neurotoxicity to human neurons. Neurons were pre under the curve of the decay of fluorescence of the test treated with drugs at a concentration of 10 uM , followed by compound over 60 min in comparison to 12 . 5 uM gallic acid a challenge with 25 or 50 uM FeSO4 after 1 h . In this and blank . Shown are data of n = 3 - 4 independent experi experiment, several compounds (yellow bars) prevented ments ESEM , with each experiment performed in triplicates against iron mediated neurotoxicity ( A ) . Values in A are ( C ) . The antipsychotics showed strong anti - oxidative effects , mean : SEM of n = 4 wells per condition . One -way analysis of as demonstrated with HORAC GAEs of > 3 . Data points > 1 variance ( ANOVA ) with Bonferroni posthoc analysis vs . represent anti - oxidative capacity ( the gallic acid effect is 1 ) , iron : * p < 0 .05 ; * * p < 0 .01 ; * * * p < 0 .001 ; * * * * p < 0 .0001 . Rep O represents no antioxidative properties , and data < 0 show resentative images show the control and iron treated neu pro - oxidative effect . RFU : Relative fluorescence units . Two rons, as well as the prevention of neurotoxicity by treatment way analysis of variance (ANOVA ) with Dunnett ' s multiple with indapamide ( B bright field , C fluorescence micros comparisons test as posthoc - analysis ( a , b ); the first signifi copy ) . Neurons were detected by anti -microtubule -associ cant time point vs . gallic acid is depicted as * . One -way ated protein - 2 (MAP - 2 ) antibody. The scale bars depict 100 analysis of variance (ANOVA ) with Dunnett ' s multi com um . parisons test as post -hoc analysis vs . gallic acid . * p < 0 .05 ; [0054 ] FIG . 2 — Summary of compounds that attenuate * * p < 0 .01 ; * * * p < 0 . 001; * * * * p < 0 . 0001. iron mediated neurotoxicity. Shown are all 35 generic drugs [0057 ] FIG . 5 - Effects on proliferation of T - lymphocytes . that prevent iron mediated neurotoxicity ( A ). The number of The tricyclic antidepressants (clomipramine , desipramine , neurons in each well of a given experiment was normalized imipramine, trimipramine and doxepin ) reduced prolifera to the number of neurons of the respective untreated control tion of T -cells markedly (p < 0 . 0001) . Data were normalized condition ( 100 % ) . The corresponding FeSO2 treated condi to counts per minute ( cpm ) of activated control T - cells . tion ( red ) was also normalized to the respective control. Shown are data pooled from 2 independent experiments Some of the major drug classes are depicted in the figure . each performed in quadruplicates. Data are depicted as Shown are the mean SEM of 2 -4 independent experiments , mean + SEM . One - way analysis of variance (ANOVA ) with performed in quadruplicates ( thus , 8 - 16 wells per treatment Dunnett ' s multiple comparisons test as post -hoc analysis across experiments are depicted in the figure ). Panel B compared to activated splenocytes . * p < 0 .05 ; * * p < 0 . 01 ; shows the results from live cell imaging of neurons chal * * * p < 0 . 001 ; * * * * p < 0 . 0001 . lenged with FeSO4 in a concentration of 50 UM . Upon [ 0058 ] FIG . 6 Clomipramine reduces iron neurotoxicity pre -treatment with indapamide or desipramine 1 h before the and proliferation of T - and B - lymphocytes . Clomipramine addition of iron , the number of propidium - iodide positive attenuated iron mediated neurotoxicity in a concentration cells was significantly reduced after 7 . 5 h and even below dependent manner from 100 nM (p < 0 .005 ) ( A ) . Washing the level of the control condition after 12 h , suggesting a away clomipramine led to cell death by iron , but this effect strong neuroprotective effect. Live cell imaging was per could be prevented after pre - incubation of clomipramine formed over 12 h , where images were taken every 30 min . with iron , suggesting a physical reaction between clomip The time- point from which significant changes were ramine and iron ( B ) . Live cell imaging studies show that the observed is marked with a symbol ( # control; + DMSO ; * increasing accumulation of PI - positive neurons exposed to indapamide ; ~ desipramine ) . Shown are means SEM of n = 3 iron over time was prevented by clomipramine ( C ). Clo wells per condition . Results were analyzed with a two -way mipramine furthermore reduced the proliferation of T - lym ANOVA with Dunnett ' s multiple comparison as post - hoc phocytes (D ) , reflected by a reduction of cells in S - phase and analysis . an increase in the G1- phase of the cell cycle ( E , F ). Prolif [0055 ] FIG . 3 — Prevention of mitochondrial damage eration of activated B - Cells was reduced by clomipramine induced by rotenone . Some of the generic drugs that pre from 2 uM (G ) , correspondent with reduced TNF - a release vented against iron mediated neurotoxicity were tested ( H ) . Data are shown as quadruplicate replicate wells of an against mitochondrial damage to neurons . Some com - individual experiment that was conducted twice ( A , D , E , F ) , pounds , such as indapamide, prevented mitochondrial dam once ( B ) of three times ( G , H ) ; panel C represent triplicate US 2019 /0262353 A1 Aug . 29 , 2019 wells of one experiment. Results are mean + SEM . One -way glial activation ( lbal stain , c ) , which was accompanied by analysis of variance ( ANOVA ) with Dunnett ’ s multiple axonal damage with formation of axonal bulbs ( indicated by comparisons test as posthoc analysis compared to the an arrow , Bielschowsky stain , e ) Clomipramine treatment FeSO4 or activated condition ( a , b , d -h ) and two- way analy reduced microglial activation concomitant with preserved sis of variance ( ANOVA ) with Dunnett ' s multiple compari axonal integrity ( d , f ) . This was reflected in a blinded rank sons test ( c ) : * p < 0 .05 ; * * p < 0 . 01; * * * p < 0 . 001; * * * * p < 0 . order analysis ( h , i ) . Infiltration and microglial activation 0001. positively correlated with axonal damage ( , k ) . cle and d / f [0059 ] FIG . 7 – Clomipramine initiated from day 5 delays are adjacent sections . Images are shown in 20 - and 40 - times the onset of EAE clinical disease . Female C57BL /6 mice original magnification . The scale bars show 100 um . Non ( age 8 - 10 weeks ) were treated with clomipramine IP (25 parametric two- tailed Mann -Whitney test (g -i ) and non mg/ kg ) or PBS ( vehicle ) from day 5 after induction of parametric two - tailed Spearman correlation with 95 % con MOG - EAE ( a ) . The disease onset was delayed and from day fidence interval (j , k ). Significance is shown as * * p < 0 .01 ; 11 the clinical course differed significantly ( p < 0 . 001 ) . Even * * * p < 0 . 001. tually, clomipramine treated mice also developed the same [ 0062 ] FIG . 10 — Clomipramine improves the chronic disease burden as vehicle - treated mice . The overall disease phase of EAE . a ) Female C57BL / 6 (age 8 -10 weeks )MOG burden is shown in panel b ) . N = 8 vehicle and n = 8 clomip immunized mice were treated with clomipramine IP ( 25 ramine EAE mice . Data are depicted as mean : SEM . Two mg/ kg ) or PBS ( vehicle ) from remission after the first way ANOVA with Sidak ' s multiple - comparisons test as relapse , and this did not affect disease score between the post -hoc analysis ( a ) and two - tailed unpaired non -paramet groups ( n = 10 vehicle , n = 10 clomipramine ) . B ) In a second ric Mann -Whitney test ( b ) . Significance is shown as * p < 0 . experiment, MOG - immunized C57BL /6 mice were treated 05 ; * * p < 0 .01 ; * * * p < 0. 001 ; * * * * p < 0 .0001 . from onset of clinical signs . Here, clomipramine reduced the [0060 ] FIG . 8 - Early clomipramine treatment suppressed clinical severity of the first relapse (day 14 - 20 , p = 0 .0175 , EAE disease activity . Female C57BL / 6 mice ( age 8 - 10 two -tailed Mann -Whitney t - test ) and of the second relapse at weeks ) were treated with clomipramine IP (25 mg/ kg ) or the late chronic phase ( day 42 - 50 , p = 0 .0007 , two - tailed PBS ( vehicle ) from the day of induction of MOG -EAE ( day Mann -Whitney t - test ) ( n = 5 vehicle , n = 6 clomipramine ) . 0 ). From day 11 the clinical course differed significantly Note that an initial two -way ANOVA with Sidak ' s multiple ( p < 0 .05 ) ; while vehicle - treated mice accumulated progres comparisons test of the experiment from day 13 to 50 was sive disability , clomipramine treated mice remained unaf not statistically significant, since vehicle - treated mice spon fected even up to the termination of experiment when taneously remitted to a very low disease score between days vehicle - treated mice were at peak clinical severity (paralysis 25 and 42 , so that differences with the treatment group could or paresis of tail and hind limb functions , and paresis of not be detected . Hence , we analyzed differences of the acute forelimbs) (A ) . The overall burden of disease per mouse was and chronic relapse phases outside of the period of remis plotted in panel B , while the relative weight of mice , sion , using Mann - Whitney t - test. c ) Using Biozzi ABH reflecting general health , is shown in panel C . In the lumbar mice , treatment from onset of clinical disability showed a cord , at animal sacrifice (day 15 ) , there was a significant positive effect on the chronic phase ( p = 0 .0062 , two - tailed upregulation in vehicle - EAE mice of transcripts encoding Mann -Whitney test) ( n = 5 vehicle , n = 5 clomipramine ) . Ifng , Tnfa , 11 - 17 and Col2 compared to naïve mice , whereas When a two - way ANOVA with Sidak ' s multiple - compari clomipramine treated mice did not show these elevations sons test was used , the results were not significant since the ( D ). Levels of clomipramine and the active metabolite individual variability of mice in either group in any given desmethylclomipramine in serum and spinal cord at sacrifice day was very high for this model in our hands. d ) A summary ( e ) are consistent to concentrations reached in humans. of the effect of clomipramine when treatment is initiated at There was a strong correlation of serum levels of clomip the onset of clinical signs. ramine and desmethylclomipramine with spinal cord levels ( f) . Data in panel D are RT -PCR results , with values nor [0063 ] FIG . 11 Shown are all 249 generic compounds of malized to Gapdh as housekeeping gene and expressed in the iron mediated neurotoxicity screening ( A - M ) . The num relation to levels in naïve mice . N = 8 ( vehicle ) and n = 7 ber of neurons left following exposure to each compound (clomipramine ) EAE mice . Data are depicted as was normalized to the number of neurons of the respective mean : SEM . Two - way ANOVA with Sidak ' s multiple - com control condition . The corresponding iron situation was also parisons test as posthoc analysis ( A ) , two -tailed unpaired normalized to the respective control (red ). Compounds non - parametric Mann - Whitney test ( b ) , two - tailed unpaired which exhibit significant protection are highlighted in yel t -test ( C , E , F ) and one -way ANOVA with Tukey ' s multiple low and marked ( X ) . Shown are the means = SEM of 1 - 4 comparisons test as post - hoc analysis ( D ) . Correlations were experiments , performed in quadruplicates each . calculated using a linear regression model, dotted lines show [0064 ] FIG . 12 shows Lysolecithin deposited in the ven the 95 % -confidence interval ( f) . Significance is shown as trolateral white matter of the mouse spinal cord produces a * p < 0 .05 ; * * p < 0 .01 ; * * * p < 0 .001 ; * * * * p < 0 .0001 . larger volume of demyelination in aging 8 - 10 month versus [0061 ] FIG . 9Reduced inflammation and axonal dam 6 weeks old young mice . Panel a shows the greater spread age upon clomipramine treatment. Vehicle - treated animals of demyelination (loss of blue in the ventrolateral white had marked parenchymal inflammation , indicated by an matter ) across multiple sections rostral ( R , numbers are um arrow ( a ) , whereas clomipramine - treated animals only had distance ) from the lesion epicenter (which is the bottom low meningeal inflammation ( b ). This was reflected in better most section here of a representative young and aging histological scores ( g ) evaluated by a previously described mouse ), which manifests as a larger volume of myelin loss method (Goncalves DaSilva and Yong, Am J Pathol 174 : in aging mice ( b ). * p < 0 .01 ; * * p < 0 .001 . Panel c represents 898 - 909 , 2009 ) ( a , b : Hematoxylin / eosin and luxol fast blue , the average myelin loss rostral and caudal to the epicenter in HE & LFB ). Vehicle - treated animals had pronounced micro both age groups. US 2019 /0262353 A1 Aug . 29 , 2019

[0065 ] FIG . 13 shows Greater axonal loss following lyso disseminated encephalomyelitis , acute demyelinating optic lecithin demyelination in aging mice . a ) Axons are visual neuritis , demyelinative transverse myelitis , Miller - Fisher ized by an antibody to neurofilaments (SM1312 ) in normal syndrome, encephalomyelradiculoneuropathy, acute demy appearing white matter (NAWM ) and in the lesion , with elinative polyneuropathy, tumefactivemultiple sclerosis and fewer axons spared in lesions of aging samples at 72 h (b ). Balo ' s concentric sclerosis. Note that the data in panel b represent remaining axonal [0073 ] In a specific example , the neurological disease is number in the injured ventral column expressed as a % to the progressive multiple sclerosis . counts in the uninjured ventral column . Two - tailed t - test. [0074 ] In a specific example , as described herein there is [0066 ] FIG . 14 shows RNAseq data of 3 day laser- micro provided a treatment for progressive multiple sclerosis in a dissected lesions that homed onto NADPH oxidase . a ) Heat subject . map ( 3 samples/ group , where each sample is a pool of 5 [ 0075 ] As used herein , " progressive” multiple sclerosis mice ) after lysolecithin (LPC ) lesion in young and aging refers to forms of the disease which progress towards an mice . b ) Upregulation of canonical immune -associated path ever -worsening disease state over a period of time. Progres ways in aging vs young mice that converge , through Inge sive multiple sclerosis includes, but is not limited to , for nuity Pathway Analysis , into NADPH oxidase 2 subunits. d ) example , primary progressive multiple sclerosis , secondary The RNAseq levels of the catalytic subunit of NADPH progressive multiple sclerosis , and progressive relapsing oxidase 2 , gp91phox ( also called CYBB ) are selected for multiple sclerosis . display . * p < 0 .05 . 10076 ] These subtypes may or may not feature episodic [0067 ] FIG . 15 shows higher expression of gp91phox and flare -ups of the disease , but are each associated with malondialdehyde in aging lesions . a, b ) The catalytic subunit increased symptoms, such as increased demyelination or of NOX2 , gp91 phox , is readily found within CD45 + cells in pain and reduced capacity for movement, over time. aging but not young demyelinated lesions (d3 ) . ( c, d ) Simi [0077 ] The term “ subject ” , as used herein , refers to an larly , malondialdehyde as a marker of oxidative damage is animal , and can include, for example , domesticated animals , in aging lesion associated with MBP + myelin breakdown . such as cats , dogs , etc ., livestock ( e . g ., cattle , horses , pigs , [ 0068 ] FIG . 16 shows indapamide treatmentof aging mice sheep , goats , etc . ) , laboratory animals ( e . g . , mouse , rabbit , after lysolecithin injury results at 72 h in a smaller demy rat, guinea pig , etc . ) , mammals , non - human mammals , pri elinated volume, less axonal loss , and lower lipid peroxida mates, non -human primates , rodents, birds , reptiles , amphib tion . Indapamide ( 20 mg/ kg ) was given ip immediately after ians, fish , and any other animal. In a specific example , the demyelination , and once / day 24 h apart for the next 2 days , subject is a human . and mice were then killed on day 3 . Impressively , indap 10078 ] The term " treatment” or “ treat” as used herein , amide reduced the volume of demyelination ( a , b ) and pre refers to obtaining beneficial or desired results , including served axons ( c , d ) , likely through the reduction of free clinical results . Beneficial or desired clinical results can radical toxicity as manifested by the lower accumulation of include , but are not limited to , alleviation or amelioration of malondialdehyde in demyelinated mice . one or more symptoms or conditions, diminishment of extent of disease , stabilized ( i . e . not worsening ) state of DETAILED DESCRIPTION disease , preventing spread of disease , delay or slowing of [0069 ] In one aspect, there is provided a method of treat disease progression , amelioration or palliation of the disease ing , prophylaxis , or amelioration of a neurological disease state , diminishment of the reoccurrence of disease , and by administering to a subject in need thereof one or more remission (whether partial or total ) , whether detectable or compounds described herein . In a specific example , the undetectable . “ Treating ” and “ Treatment” can also mean neurological disease is multiple sclerosis (also referred to as prolonging survival as compared to expected survival if not “ MS” ) . receiving treatment. “ Treating ” and “ treatment” as used [0070 ] The term “ multiple sclerosis ” refers to an inflam herein also include prophylactic treatment. For example , a matory disease of the central nervous system (CNS ) in subject in the early stage of disease can be treated to prevent which the insulating covers of nerve cells in the brain and progression or alternatively a subject in remission can be spinal cord are damaged . This damage disrupts the ability of treated with a compound or composition described herein to parts of the nervous system to communicate , resulting in a prevent progression . wide range of signs and symptoms, including physical, [0079 ] In some examples, treatment results in prevention mental, and psychiatric or delay of onset or amelioration of symptoms of a disease [0071 ] In one example , as described herein there is pro in a subject or an attainment of a desired biological outcome, vided a treatment for multiple sclerosis in a subject. such as reduced neurodegeneration ( e. g ., demyelination , [0072 ] As used herein , “ multiple sclerosis ” includes mul axonal loss, and neuronal death ), reduced inflammation of tiple sclerosis or a related disease , and optionally refers to all the cells of the CNS , or reduced tissue injury caused by types and stages of multiple sclerosis, including , but not oxidative stress and / or inflammation in a variety of cells . limited to : benign multiple sclerosis , relapsing remitting [ 0080 ] In some examples, treatment methods comprise multiple sclerosis , secondary progressive multiple sclerosis , administering to a subject a therapeutically effective amount primary progressive multiple sclerosis , progressive relaps of a compound or composition described herein and option ing multiple sclerosis , chronic progressive multiple sclero ally consists of a single administration or application , or sis , transitional/ progressive multiple sclerosis , rapidly wors alternatively comprises a series of administrations or appli ening multiple sclerosis , clinically -definite multiple cations. sclerosis , malignantmultiple sclerosis , also known as Mar [0081 ] The term “ pharmaceutically effective amount” as burg ' s Variant, and acute multiple sclerosis . Optionally , used herein refers to the amount of a compound , composi " conditions relating to multiple sclerosis ” include , e . g . , tion , drug or pharmaceutical agent that will elicit the bio Devic ' s disease, also known as Neuromyelitis Optica ; acute logical or medical response of a tissue, system , animal or US 2019 /0262353 A1 Aug . 29 , 2019 human that is being sought by a researcher or clinician , for [ 0094 ] In some example , the multiple sclerosis is second example , the treatment of progressive multiple sclerosis . ary progressive multiple sclerosis . This amount can be a therapeutically effective amount. [0095 ] In some example , the multiple sclerosis is progres [ 0082 ] The compounds and compositions may be pro sive relapsing multiple sclerosis . vided in a pharmaceutically acceptable form . [ 0096 ] The compounds and / or compositions described [ 0083 ] The term “ pharmaceutically acceptable ” as used herein may be administered either simultaneously ( or sub herein includes compounds, materials , compositions, and / or stantially simultaneously ) or sequentially , dependent upon dosage forms (such as unit dosages ) which are suitable for the condition to be treated , and may be administered in use in contact with the tissues of a subject without excessive combination with other treatment( s ). The other treatment ( s ), toxicity , irritation , allergic response, or other problem or may be administered either simultaneously ( or substantially complication , commensurate with a reasonable benefit / risk simultaneously ) or sequentially . ratio . Each carrier, excipient, etc . is also “ acceptable ” in the [0097 ] In some example , the other or additional treatment sense of being compatible with the other ingredients of the further comprises administering a therapeutically effective formulation . amount of Laquinimod , Fingolimod , Masitinib , Ocreli [0084 ] In one example , there is provided a method of zumab , Ibudilast , Anti -LINGO - 1 , MD1003 (high concentra treating progressive multiple sclerosis comprising adminis tion Biotin ) , Natalizumab , Siponimod , Tcelna ( imilecleucel tering to a subject in need thereof, a therapeutically effective T ) , Simvastatin , Dimethyl fumarate , Autologous amount of one or more of dipyridamole , clopidogrel , cefa haematopoietic stem cell transplantation , Amiloride, Rilu clor, clarithromycin , erythromycin , rifampin , loperamide, zole , Fluoxetine, Glatiramer Acetate , Interferon Beta , or a ketoconazole , labetalol, methyldopa , metoprolol, atenolol, functional derivative thereof. carvedilol , indapamide, mefloquine , primaquine , mitoxan [0098 ) The actual amount( s ) administered , and rate and throne , levodopa , trimeprazine , chlorpromazine, clozapine , time- course of administration , will depend on the nature and periciazine, flunarizine , dimenhydrinate , diphenhydramine , severity of progressive multiple sclerosis being treated . promethazine , phenazopyridine , yohimbine , memantine , Prescription of treatment, e . g . decisions on dosage etc . , is liothyronine , clomipramine , desipramine, doxepin , imip within the responsibility of general practitioners and other ramine , trimipramine, or functional derivative thereof. medical doctors , and typically takes account of the disorder [ 0085 ] In a specific example , there is provided a method of to be treated , the condition of the individual patient, the site treating progressive multiple sclerosis comprising adminis of delivery, the method of administration and other factors tering to a subject in need thereof, a therapeutically effective known to practitioners . amount of clomipramine, or a functional derivative thereof. 10099 ] The formulation ( s ) may conveniently be presented [ 0086 ] In a specific example , there is provided a method of in unit dosage form and may be prepared by any methods treating multiple sclerosis comprising administering to a well known in the art of pharmacy. Such methods include subject in need thereof, a therapeutically effective amount of the step of bringing the active compound into association imipramine , or a functional derivative thereof. with a carrier, which may constitute one or more accessory [ 0087] In a specific example , there is provided a method of ingredients . In general, the formulations are prepared by treating multiple sclerosis comprising administering to a uniformly and intimately bringing into association the active subject in need thereof, a therapeutically effective amount of compound with liquid carriers or finely divided solid carriers trimipramine, or a functional derivative thereof. or both , and then if necessary shaping the product . [0088 ] In a specific example, there is provided a method of [0100 ] The compounds and compositions may be admin treating multiple sclerosis comprising administering to a istered to a subject by any convenient route of administra subject in need thereof, a therapeutically effective amount of tion , whether systemically /peripherally or at the site of indapamine , or a functional derivative thereof. desired action , including but not limited to , oral ( e. g . by [ 0089 ] In a specific example , there is provided a method of ingestion ) ; topical ( including e . g . transdermal, intranasal, treating multiple sclerosis comprising administering to a ocular , buccal, and sublingual) ; pulmonary ( e . g . by inhala subject in need thereof, a therapeutically effective amount of tion or insufflation therapy using , e . g . an aerosol, e . g . indapamine , or a functional derivative thereof, and one or through mouth or nose ) ; rectal; vaginal; parenteral, for more of hydroxychloroquine , minocycline , or clomip example , by injection , including subcutaneous , intradermal, ramine , or a functional derivative thereof. intramuscular , intravenous , intraarterial, intracardiac , [0090 ] The term “ functional derivative ” and “ physiologi intrathecal, intraspinal, intracapsular, subcapsular, intraor cally functional derivative” as used herein means an active bital, intraperitoneal, intratracheal, subcuticular , intraarticu compound with equivalent or near equivalent physiological lar, subarachnoid , and intrasternal; by implant of a depot/ for functionality to the named active compound when used example , subcutaneously or intramuscularly . and / or administered as described herein . As used herein , the [0101 ] Formulations suitable for oral administration ( e. g ., term “ physiologically functional derivative ” includes any by ingestion ) may be presented as discrete units such as pharmaceutically acceptable salts , solvates, esters , prodrugs capsules , cachets or tablets , each containing a predetermined derivatives , enantiomers , or polymorphs . amount of the active compound ; as a powder or granules; as [0091 ] In some examples the compounds are prodrugs . a solution or suspension in an aqueous or non - aqueous [0092 ] The term “ prodrug ” used herein refers to com liquid ; or as an oil - in - water liquid emulsion or a water -in - oil pounds which are not pharmaceutically active themselves liquid emulsion ; as a bolus; as an electuary ; or as a paste . but which are transformed into their pharmaceutical active [0102 ] Formulations suitable for parenteral administration form in vivo , for example in the subject to which the ( e . g ., by injection , including cutaneous , subcutaneous, intra compound is administered . muscular , intravenous and intradermal) , include aqueous [0093 ] In some examples , the multiple sclerosis is primary and non - aqueous isotonic , pyrogen - free , sterile injection progressive multiple sclerosis . solutions which may contain anti- oxidants, buffers , preser US 2019 /0262353 A1 Aug . 29 , 2019 vatives , stabilisers , bacteriostats , and solutes which render sis , primary progressive multiple sclerosis , secondary pro the formulation isotonic with the blood of the intended gressive multiple sclerosis , or progressive relapsing multiple recipient; and aqueous and non -aqueous sterile suspensions sclerosis . which may include suspending agents and thickening [0109 ] In one aspect there is described a kit for the agents , and liposomes or other microparticulate systems treatment of progressive multiple sclerosis comprising : a which are designed to target the compound to blood com therapeutically effective amount of indapamide, or a func ponents or one or more organs. Examples of suitable iso tional derivative thereof, and instructions for use . tonic vehicles for use in such formulations include Sodium [0110 ] In one aspect there is described a kit for the Chloride Injection , Ringer ' s Solution , or Lactated Ringer ' s treatment of progressive multiple sclerosis comprising : a Injection . therapeutically effective amount of indapamide, or a func [ 0103 ] The formulationsmay be presented in unit- dose or tional derivative thereof , and one or more of hydroxychlo multi- dose sealed containers, for example , ampoules and roquine , minocycline , or clomipramine ; and instructions for vials , and may be stored in a freeze -dried ( lyophilized ) use . condition requiring only the addition of the sterile liquid 10111 ] A kit may also include one or more of a container , carrier, for example water for injections, immediately prior a buffer, a diluent, a filter, a needle , or a syringe . to use . Extemporaneous injection solutions and suspensions [0112 ] To gain a better understanding of the invention may be prepared from sterile powders, granules, and tablets . described herein , the following examples are set forth . It Formulations may be in the form of liposomes or other should be understood that these example are for illustrative microparticulate systems which are designed to target the purposes only . Therefore, they should not limit the scope of active compound to blood components or one or more this invention in any way . organs . [0104 ] In another aspect, there is described a method of EXAMPLES identifying a compound for the treatment of progressive [0113 ] In the following examples , standard methodologies multiple sclerosis , comprising : selecting one or more com were employed , as would be appreciated by the skilled pounds from a library of compounds that prevent or reduce worker . iron -mediated neurotoxicity in vitro , selecting one or more compounds from step ( b ) that prevent or reduce mitochon [0114 ) Materials and Methods drial damage in vitro ; selecting one or more compounds [0115 ] Cell Culture and Treatment of Human Neurons from step ( a ) for anti - oxidative properties, selecting one or 10116 ] Human neurons were isolated from brain tissues of more compound from step ( a ) for ability to reduce T - cell therapeutically aborted 15 - 20 week old fetuses, in accor proliferation in vitro , optionally , after step ( a ) , selecting a dance with ethics approval of the University of Calgary compound from step ( a ) which is predicted or known to be ethics committee , after written informed consent of the able to cross the blood brain barrier, or having a suitable side pregnant donors . Neurons were isolated as previously effect profile , or having a suitable tolerability . described ( Vecil et al ., 2000 ) brain specimens were washed in phosphate buffered saline (PBS ) to remove blood , fol [0105 ] Methods of the invention are conveniently prac lowed by removal of meninges . Tissue was mechanically ticed by providing the compounds and / or compositions used dissected , followed by digestion in DNase ( 6 - 8 ml of 1 in such method in the form of a kit. Such a kit preferably mg/ml ; Roche ) , 4 ml 2 . 5 % trypsin and 40 ml PBS ( 37° C . , contains the composition . Such a kit preferably contains 25 min ) . Thereafter , the digestion was stopped by addition of instructions for the use thereof. 4 ml fetal calf serum (FCS ) . The solution was filtered [0106 ] In one example , there is described a kit for the through a 132 um filter and centrifuged three times , 1 , 200 treatment of progressive multiple sclerosis , comprising: one rpm , 10 min ) . Cells were cultured in feeding medium of or more of dipyridamole , clopidogrel, cefaclor, clarithromy minimal essential medium (MEM ) supplemented with 10 % cin , erythromycin , rifampin , loperamide, ketoconazole , fetal bovine serum (FBS ) , 1 uM sodium pyruvate , 10 UM labetalol, methyldopa , metoprolol, atenolol, carvedilol, glutamine, 1x non -essential amino acids, 0 . 1 % dextrose and indapamide, mefloquine, primaquine , mitoxanthrone , 1 % penicillin /streptomycin ( all culture supplements from levodopa , trimeprazine , chlorpromazine, clozapine, perici Invitrogen , Burlington , Canada ). The initial isolates of azine , flunarizine , dimenhydrinate , diphenhydramine , pro mixed CNS cell types were plated in poly - L - ornithine methazine, phenazopyridine , yohimbine , memantine , liothy coated ( 10 ug /ml ) 175 flasks and cultured for at least two ronine , clomipramine , desipramine, doxepin , imipramine , cycles (Vecil et al. , 2000 ) in medium containing 25 uM trimipramine, or functional derivative thereof; and instruc cytosine arabinoside ( Sigma- Aldrich , Oakville , Canada ) to tions for use . inhibit astrocyte proliferation and to deplete this major contaminating cell type . For experiments , the neuron - en [0107 ] In another example , the kit further comprises one riched cultures were retrypsinized and cells were plated in or more of Laquinimod , Fingolimod , Masitinib , Ocreli poly - L -ornithine pre - coated 96 -well plates at a density of zumab , Ibudilast, Anti - LINGO - 1 , MD1003 (high concentra 100 , 000 cells /well in 100 ul of the complete medium supple tion Biotin ), Natalizumab , Siponimod , Tcelna ( imilecleucel mented with cytosine arabinoside . Medium was changed to T ) , Simvastatin , Dimethyl fumarate , Autologous AIM V® Serum Free Medium ( Invitrogen ) after 24 h . After haematopoietic stem cell transplantation , Amiloride, Rilu a period of 1 h , respective drugs were added in a concen zole , Fluoxetine , or a functional derivative thereof; and tration of 10 uM , followed by application of FeSO4 after 1 instructions for use. h or 24 h , or the other toxins after 1 h . All conditions were [0108 ] In one example there is described a pharmaceutical performed in quadruplicates . A day later cells were fixed composition comprising clomipramine, or a functional using 4 % paraformaldehyde (PFA ) and stored in PBS in 4° derivative thereof, for treating progressive multiple sclero C. US 2019 /0262353 A1 Aug . 29 , 2019

[0117 ] We note that in tissue culture , the toxicity of iron to tration of 1 uCi per well, and cells were harvested after 24 neurons begins immediately . Thus, it has been our experi - hon filter mats . Mats were then evaluated for radioactivity ence that pretreatment with test protective agents is neces ( counts per minute ) using a liquid scintillation counter. sary . With the continuous insult that occurs in multiple sclerosis , a pretreatment paradigm with test compounds [0124 ] Activity on B -Lymphocytes against iron neurotoxicity in our experiments can be justified [0125 ] Venous blood from healthy volunteers was as that simulates the protection against the next injury in the obtained and peripheral blood mononuclear cells (PBMCs ) disease . were isolated by Ficoll gradient centrifugation (1800 RPM , 30 min ). From PBMCs, B -cells were isolated by positive [0118 ] Drugs tested were contained within the 1040 - com selection with CD19 directed microbeads (Stemcell Tech pound NINDS Custom Collection II, which was purchased nologies ) . Purity was assessed by FACS after staining for from Microsource Discovery (Gaylordsville , Conn . , USA ) CD19 (Stemcell Technologies ) . Cells were plated in a con and used as previously described (Samanani et al. , CNS & centration of 2 .5x105 cells/ well in X - VIVOTM medium neurological disorders drug targets 12 : 741- 749, 2013 ) . (Lonza ) supplemented with 1 % penicillin / streptomycin and Briefly , there were 80 compounds located in specific wells 1 % Glutamax and treated with drugs for 1 h . Cells were then on each plate ( e . g . B07 ) . 3607 would thus refer to position activated with 10 ug/ ml IgM BCR cross - linking antibody B07 of plate 3 . Each compound was supplied at a concen (XAL ) ( Jackson ImmunoResearch ) , 1 ug/ ml anti- CD40L tration of 10 mM dissolved in DMSO . and IL - 4 20 ng /ml for 24 has previously described (Li et al. , Science Translational Med 7 :310ra166 , 2015 ) . Conditioned [0119 ] The iron stock solution was prepared using 27 . 8 mg media were harvested after 24 h for ELISA . Medium as well iron ( II ) sulfate heptahydrate (FeSO4 ) (Sigma - Aldrich , Oak as respective drugs were re - added followed by application of ville , Canada ), 10 ul of 17 . 8M sulfuric acid and 10 ml 3H - thymidine in a concentration of 1 uCi per well to deionized distilled water . After filtering with a 0 . 2 um filter, investigate proliferation . After 24 h , cells were harvested on FeSO4 was added to cells in a final concentration of 25 -50 filter mats and after drying counts per minutes were mea uM in a volume of 50 ul medium to the cells . Rotenone was sured using a liquid scintillation counter . dissolved in dimethyl sulfoxide (DMSO ) and used in a final [0126 ] Flow Cytometry concentration of 10 uM . 10127 ]. Two days after activation and drug treatment sple [0120 ] Hydroxyl Radical Antioxidant Capacity (HORAC ) nocytes were harvested , washed with PBS followed by Assay resuspension in PBS with 2 % FBS . Cell cycle analysis was [0121 ] Selected compounds that prevented iron mediated performed taking advantage of propidium iodide staining neurotoxicity were analyzed for their antioxidative proper (50 ug/ ml ) using an established protocol (Besson and Yong , ties using the hydroxyl radical antioxidant capacity 2000 ) . Cells were washed in cold PBS and resuspended in PI/ Triton X - 100 staining solution ( 10 ml 0 . 1 % ( v / v ) Triton (HORAC ) assay, in accordance with the procedure outlined X - 100 in PBS with 2 mg DNAse - free RNAse A and 0 . 4 ml in Cíž et al. 2010 (Food Control 21: 518 -523 , 2010 ) . In this of 500 ug /mIPI ) , followed by incubation at 4° C . for 30 min . assay , hydroxyl radicals generated by a Co ( II ) -mediated Stained cells were analyzed on a FACSCaliburTM with the Fenton - like reaction oxidize fluorescein causing loss of software CellQuestTM (BD Biosciences ) . Cell cycle analysis fluorescence (Ou et al. , J Argricultural Food Chemistry was conducted using the software ModFit LT, version 3 . 3 50 :2772 - 2777 , 2002 ). The presence of an anti - oxidant (Verity Software House Inc. ) . reduces the loss of fluorescence and this can be monitored [ 0128 ] FACS Gating Strategy. every 5 min over a period of 60 min with a Spectra Max [0129 ] Cells were identified by gating into the lymphocyte Gemini XS plate reader (Molecular Devices , Sunnyvale , population , followed by single cell gating to exclude dou Calif ., USA ) and the software SoftMax Pro version 5 . For blets and aggregates. This was followed by identification of monitoring fluorescence , we used an excitation wavelength the GO /G1 population and processing with the software of a = 485 nm and an emission wavelength of À = 520 nm . ModFit LT, version 3 . 3 ( Verity Software House Inc . ) to [0122 ] Proliferation of T- Lymphocytes calculate the percentage of cells in different cell cycles . [0123 ] A previously published protocol was used for iso [0130 ] Intracellular staining was performed following lating and activating T - cells (Keough et al . , Nature Comm fixation and permeabilization of splenocytes using the Fixa 7 : 11312, 2016 ) . Spleens from female C571316 mice were tion /Permeabilization Solution Kit (BD Biosciences , Mis sissauga , Canada ) , followed by staining with anti - human / harvested and after mechanical dissociation the cell suspen mouse phospho - AKT (S473 ) APC antibody , anti -human / sion was passed through a 70 um cell strainer and separated mouse phospho -mTOR (S2448 ) PE - Cyanine7 antibody and by Ficoll gradient ( 1800 RPM , 30 min ) . Splenocytes were anti- human /mouse phospho -ERK1 / 2 ( T202 /Y204 ) PE anti plated (2 .5x105 cells in 100 ul/ well ) in anti -CD3 antibody body ( all eBioscience , San Diego , Calif . ) . Stained cells were coated 96 -well plates ( 1 , 000 ng ml- plate - bound anti- CD3 analyzed on a FACSCaliburTM with the software Cell and 1, 000 ng ml- anti -CD28 suspended in media ) to acti QuestTM (BD Biosciences ). vate T - cells . Directly before plating , wells were treated with [0131 ] Immunocytochemistry and Microscopy respective drugs in a final concentration of 10 uM . Cells [0132 ] Staining was performed at room temperature . A were cultured in RPMI 1640 medium , supplemented with blocking buffer was first introduced for 1 h followed by 10 % FBS , 1 uM sodium pyruvate , 2 mM L -alanyl - L incubation with primary antibody overnight in 4° C . Neu glutamine , 1 % penicillin / streptomycin , 1 % HEPES and 0 . 05 rons were stained using mouse anti -microtubule -associated mM 2 -mercaptoethanol (all supplements were from Invit protein - 2 (MAP - 2 ) antibody , clone HM - 2 (dilution 1 : 1 ,000 ; rogen ). After 48 h , ' H - thymidine was added in a concen Sigma- Aldrich , Oakville , Canada ). ( Table 3 ) US 2019 /0262353 A1 Aug . 29 , 2019

TABLE 3 (SCH ) in PBS before injection . SCH was used in a concen tration of 6 . 6 mg/ ml emulsion each for 2 injections (days 0 Antibody Company Catalog Species Dilution and 7 ). Ibal Wako 010219019 - 18741 Rabbit 1 :250 [0140 ] The number of animals was chosen according to MAP - 2 . clone HM - 2 Sigma- Aldrich M4403 Mouse 1 : 1. 000 experience with previous experiments (FIG . 7 : 8 / 8 (vehicle / clomipramine ) ; FIG . 8 : 8 / 7 ; FIG . 10 a ) 10 / 10 ; b ) 5 /6 ; c ) 5 / 5 ) , and animals were randomized after induction of EAE . [0133 ] Primary antibody was visualized with Alexa Fluor Animals were handled according to the Canadian Council 488 or 546 - conjugated secondary antibody ( dilution 1 :250 , for Animal Care and the guidelines of the animal facility of Invitrogen , Burlington , Canada ) . Cell nuclei were stained the University of Calgary . All animal experiments received with Hoechst S769121 (nuclear yellow ) . Cells were stored ethics approval ( AC12 -0181 ) from the University of Cal in 4° C . in the dark before imaging . gary ' s Animal Ethics Committee . Mice were scored daily [0134 ] Images were taken using the automated ImageX using a 15 -point scoring system , the investigator was not press® imaging system (Molecular Devices , Sunnyvale , blinded (Giuliani F , Fu SA , Metz LM , Yong V W . Effective Calif. ) through a 10x objective microscope lens, displaying combination of minocycline and interferon - beta in a model 4 or 9 sites per well . Images were analyzed with the software of multiple sclerosis . Journal of neuroimmunology 165 , MetaXpress® (Molecular Devices, Sunnyvale, Calif. ) using 83 - 91 (2005 )) . the algorithm “multiwavelength cell scoring ” (Lau et al ., [0141 ] Histological Analyses Ann Neurol 72 :419 -432 , 2012 ) . Cells were defined accord [0142 ] One h after the last administration of clomipramine ing to fluorescence intensity and size at different wave animals were anesthetized with ketamine /xylazine , blood lengths. Data from all sites per well were averaged to one was taken by an intracardiac puncture for serum , and ani data point. mals were then subjected to PBS - perfusion . Spinal cords [0135 ] Live Cell Imaging and cerebella were removed . The thoracic cords were fixed [0136 ] Neurons were prepared as described above . in 10 % buffered formalin , followed by embedding in par Directly after the addition of FeSO4 to healthy neurons, the affin . Cervical and lumbar cords were snap frozen . Tissue live cell -permeant Hoechst 33342 ( 1 : 2 diluted in AIM - V was further processed as previously described 52 . Briefly , medium , nuclear blue; ThermoFisher Scientific , Grand the thoracic spinal cord was cut longitudinally from the Island , N . Y. , USA ) and the live cell- impermeable propinium ventral to the dorsal side with sections of 6 um thickness . iodide ( PI, 1 : 20 diluted in AIM - V medium ) were added in a Sections were stained with hematoxylin / eosin , lbal to visu volume of 20 ul ( Sigma - Aldrich ) . In compromised cells , PI alize microglia and Bielschowsky ' s silver stain to visualize could now diffuse across the plasma membrane . Live cell axons . Sections for lbal and Bielschowsky ' s silver stain imaging was performed using the automated ImageXpress® were blinded , before images depicting area of maximal imaging system under controlled environmental conditions microglial activation or axonal damage were chosen for ( 37° C . and 5 % CO2) . Images were taken from 9 sites per blinded rank order analysis by a second investigator. well at baseline and then every 30 min for 12 h . After export [0143 ] PCR with MetaXpress® , videos were edited with ImageJ (NIH ) 10144 ] Lumbar spinal cords were harvested , snap frozen in in a uniform manner. Nuclei were pseudo colored in cyan , liquid nitrogen and stored in - 80° C . Samples were homog Pl- positive cells in red . enized in 1 ml Trizol followed by the addition of 200 ul [ 0137 ] Experimental Autoimmune Encephalomyelitis chloroform . The suspension was shaken , centrifuged ( 11 , (EAE ) 500 RPM for 15 min at 4° C . ) and the RNA - containing 10138 ] EAE was induced in 8 - 10 week -old female upper phase was transferred into a new tube and precipitated C57BL / 6 mice (Charles River , Montreal, Canada ) . Mice with equal amounts of 70 % ethanol. RNA was extracted were injected with 50 ug of MOG35 - 55 ( synthesized by the using the RNeasy Mini Kit according to the manufacturer ' s Peptide Facility of the University of Calgary ) in Complete instruction ( Qiagen ) . RNA concentrations were measured Freund ' s Adjuvant ( Thermo Fisher Scientific ) supplemented using a Nanodrop ( Thermo Fisher Scientific ) . cDNA prepa with 10 mg/ ml Mycobacterium tuberculosis subcutaneously ration was performed using the RT2 First Strand kit ( Qia on both hind flanks on day 0 . In addition , pertussis toxin ( 0 . 1 gen ) with 1 ug of RNA according to the manufacturer' s ug / 200 ul; List biological Laboratories , Hornby, Canada ) instructions . Real time PCR was performed using the Quant was injected intraperitoneal ( IP ) on days 0 and 2 . Animals Studio 6 Flex (Applied Biosystems by Life Technologies) were treated with clomipramine (25 mg/ kg ; 100 ul of 5 with FAST SYBR Green and primers for Gapdh ( Qiagen ) as mg/ ml solution ) by IP injection by IP injection from day o housekeeping gene , Ifn - y (Qiagen , QT01038821 ) , Tnfa or day 5 (FIG . 7, 8 ), from day 30 at remission ( FIG . 10a ), or (Qiagen , QT00104006 ), 11 - 17 (SABiosciences , from 13 at onset of clinical signs (FIG . 106 ) . The solution of PPM03023A - 200 ) and Cc12 (Qiagen , QT00167832 ) . Rela clomipramine was prepared daily in fresh PBS . tive expression was calculated using the AACT method with [0139 ] The Biozzi ABH mouse model (Al - Izki et al. , Gapdh as housekeeping gene . Data were normalized to gene Multiple Sclerosis 17 : 939 - 948 , 2011 ) was used as a model expression in naïve mice . of progression . EAE was induced in Biozzi ABH mice aged [0145 ] Liquid Chromatography -Mass Spectrometry 8 - 10 weeks by the subcutaneous application of 150 ul [014 ] The assay is a modification of the liquid chroma emulsion in both sides of the hind flanks. The emulsion was tography -mass spectrometry (LC -MS ) assay of Shinokuzack prepared as follows : Stock A consisted of 4 mlof incomplete et al. (Forensic Science International 62 : 108 -112 , 2006 ) . For Freund ' s adjuvant mixed with 16 mg M . tuberculosis and 2 preparation of samples, 100 ul of ice cold methanol were mg M . butyricum . One mlof stock A was mixed with 11 .5 added to 100 ul of serum in each sample after addition of the ml incomplete Freund ' s adjuvant to become stock B . Stock internal standard maprotiline . The tubes were vortexed and B was mixed in equal volume with spinal cord homogenate left on ice for 10 min followed by centrifugation at 10 ,000xg US 2019 / 0262353 A1 Aug . 29 , 2019 for 4 min . An equal amount of distilled water was added to f0149 ] Calibration curves consisting of varying amounts each supernatant. Spinal cord samples were each homog of authentic clomipramine and N - desmethylclomipramine enized in 10 volumes of ice -cold 80 % methanol . Twenty ul and the same fixed amount of maprotiline as added to the of o - phosphoric acid were added to all samples after addi samples being analyzed were run in parallel through the tion of internal standard (maprotiline ) . The tubes were procedure described above and the ratios of clomipramine vortexed and left on ice for 10 min , followed by centrifu and N - desmethylclomipramine to maprotiline were used to gation at 10 , 000xg for 4 min and an equal volume of determine the amount of drug and metabolite in the serum distilled water was added to each supernatant. and spinal cord samples . [ 0147 ] An HLB Prime uelution plate was employed for [0150 ] Statistical Analysis sample cleanup for both serum and spinal cord samples . [0151 ] Statistical analysis was performed using the Graph After running the supernatants described above through the pad Prism software version 7 (La Jolla , Calif. , USA ). For wells, all wells were washed with 5 % methanol in water and cell culture experiments , one -way ANOVA with different allowed to dry completely before elution with 100 ul 0 .05 % posthoc analyses was applied , as stated in the respective formic acid in methanol: acetonitrile ( 1 : 1 ) . The eluents were figure legends. EAE scores were analyzed using two -way transferred to low volume ul glass inserts (Waters , Milford , ANOVA with Sidak 's multiple comparison as posthoc Mass. , USA ) and 10 ul from each eluent were injected into analysis . Statistical significance was considered as p < 0 . 05 the LC -MS system . ( * ) , p < 0 .01 ( * * ), p < 0 . 001 (* * * ) and p < 0 . 0001 ( * * * * ). All [0148 ] Analysis was performed using a Waters ZQ Mass experiments were performed in quadruplicates, if not oth detector fitted with an ESCI Multi -Mode ionization source and coupled to a Waters 2695 Separations module (Waters ) . erwise specified . Mass Lynx 4 . 0 software was used for instrument control, [0152 ] Results data acquisition and processing . HPLC separation was per 10153 ] Protection Against Iron and Rotenone Neurotoxic formed on an Atlantis dC18 ( 3 um , 3 .0x100 mm ) column ity ( Waters ) with a guard column of similar material . Mobile [0154 ] Of the 1040 compounds available in the NINDS phase A consisted of 0 .05 % formic acid in water and mobile Custom Collection II , we first conducted a search of avail phase B was composed of 0 . 05 % formic acid in acetonitrile . able information to exclude those that were either experi Initial conditions were 80 % A and 20 % B at a flow rate of mental, agricultural, not available as oral drug , not listed at 0 . 3 mL /min . A gradient was run , increasing to 80 % B in 15 Health Canada, steroid hormones or veterinary medications . min ; this was followed by a return to initial conditions . The Moreover , we omitted those that were not known to cross the column heater and sample cooler were held at 30° C . and 4° blood - brain barrier. We note that while we selected drugs C . respectively. Optimized positive electrospray parameters that are orally available , for ease of use , this does not imply were as follows: Capillary voltage 3 .77 kV ; Rf lens voltage that injectable medications would not be effective medica 1 . 2 V ; source 110° C .; desolvation temperature 300° C . ; cone tions in progressive multiple sclerosis , as illustrated by gas flow (nitrogen ) 80 L / h ; desolvation gas flow (nitrogen ) ocrelizumab recently (Montalban X , et al. Ocrelizumab 300 L /h . Cone voltage was varied for each compound : versus Placebo in Primary Progressive Multiple Sclerosis . N clomipramine 25 V ; N -desmethylclomipramine 22 V ; and Engl J Med 376 , 209 - 220 (2017 ). Out of the original list , 791 maprotiline 25 V . The m / z ratios for clomipramine, N - des compounds were thus excluded and 249 were selected for methylclomipramine and maprotiline ( internal standard ) further testing . The detailed information of each of the 249 were 315 , 301 and 278 respectively . compounds are provided in Table 1 US 2019 / 0262353 A1 Aug . 29 , 2019

BiochimBiophys Acta944:383 (1988) REFERENCES USAN,INNAdvDrugResBAN,JAN16:309(1987) USAN,INN USAN,INN USAN,INN USP,INN USP,INNBAN,JAN USP,INN BAN,JAN USP,INN BAN,JAN USP,INN BAN,JAN USP,INN USP,INN USP,INN USP,INN BAN,JAN BAN,JAN USP,INN BAN,JAN STATUS experimental BAN BAN BAN BAN BAN USPJAN, BAN SOURCE synthetic synthetic synthetic synthetic synthetic synthetic synthetic synthetic synthetic synthetic semisynthetic synthetic synthetic synthetic synthetic semisynthetic

gly)antagonist( antianginal, inhibitor,diuretic antiparkinsonian;treatmentofdrug inhibitor,diuretic vasodilator,Ca antigout, receptoragonist 681.78adrenergicagonist, channelblocker antiarrhythmic antipyretic anhydrase antiglaucoma 136.11antihyperuricemia, 335.475HT1B/2D extrapyrimidal coronary 195.61NMDAreceptor 372.90antihypertensive, antiurolithic 187.71antiviral, induced reactions 266.09Na+channel blocker 567.06Cachannel MolWtBIOACTIVITY 151.17a 222.25carbonic 163.20mucolytic 225.21antiviral 210.28antineoplastic 781.77antibacterial 313.87antidepressant 365.41antibacterial

TABLE1 FORMULA C9H6CINO2 C18H29CIN204 C8H9NO2 C4HÔN40382 C5H9NO3S C8H11N503 C5H4N40 C17H25N302S C9H18N6 C10H18CIN C22H47N502152 C6H9C12N70 C25H30C1I2NO3 C20H24CIN C26H31CIN208S C16H19N305S

34381-685, 154323-576 39831-555, 17440-834. 111470-996 61336-707, 30937517- 59277-893 665-667,768 37517-285 [anhydrous], 2609-463 549-188,50 26787-780 103-902 616-911 315-300 645-056 2016-888 1951-253 [amitriptyline] [acebutolol] 59-665 amantadine][ [amikacin] [amiloride] [anhydrous] cas# 94-5 48-6 position plate 402006D09 402011A11 402001E04 402001B02 402001DO8 402008C03 402001F11 402009A06 402007C07 402001HO2 A03402001 402001B03 402013B04 G03402001 402009G05 402001D02

MOLENAME 5-CHLOROINDOLE 2-CARBOXYLIC ACID ACEBUTOLOL HYDROCHLORIDE ACETAMINOPHEN ACETAZOLAMIDE ACETYLCYSTEINE ACYCLOVIR ALLOPURINOL ALMOTRIPTAN ALTRETAMINE AMANTADINE HYDROCHLORIDE AMIKACIN SULFATE AMILORIDE HYDROCHLORIDE AMIODARONE HYDROCHLORIDE AMITRIPTYLINE HYDROCHLORIDE AMLODIPINE BESYLATE AMOXICILLIN

ID 01502057 01500665 01500101 01500102 01500105 01503603 01500108 01505204 01503065 01500110 01500111 01500112 02300165 01500117 01505202 01500120 US 2019 / 0262353 A1 Aug . 29 , 2019

REFERENCES JMedNewEngl 296:784(1977) USAN,INN USANINN, USANINN, USAN,INN USANINN, USP,INN BAN,JAN USP,INN BAN,JAN USP,BAN USP,INN BAN,JAN USP,INN USP,INN BAN,JAN USPINN, USP,INN BAN,JAN USP,INN BAN,JAN BAN,JAN BAN,JAN USP,INN BAN,JAN USP,BAN BAN,JAN USP,INN BAN,JAN USP,INN BAN,JAN. STATUS JAN BAN BAN BAN JAN BAN componentof SOURCE Streptomycetes nodosus synthetic synthetic synthetic synthetic synthetic synthetic semisynthetic Bacillus licheniformissubtilis andB synthetic synthetic Madopa (Hoffmann LaRoche) synthetic synthetic synthetic synthetic synthetic semisynthetic

antipyretic, antineoplastic, reductaseinhibitor (skeletal) antiinflammatory 582.71antihyperlipidemic, 366.85antipneumocystic, 277.27immunosuppressant, 460.96ACEinhibitor, antihypertensive 180.16analgesic, 266.34betaadrenergic blocker HMGCOA antimalarial antirheumatic 213.67musclerelaxant inhibitor MolWtBIOACTIVITY 924.10antifungal 188.23analgesic 749.00antibacterial 1422.73antibacterial 293.71decarboxylase 405.52anticholine 196.68cholinergic 361.83antihyperlipidemic 361.40cathartic 435.32H1antihistamine 430.55antiinflammatory

TABLE1-continued FORMULA C47H73N017 C11H12N20 C9H804 C14H22N203 C33H33CaFNO5 C22H19C103 C9H7N702S C38H72N2012 C66H103N17016S C10H12CINO2 C24H29CIN205 C10H16CIN305 C21H27NOSS C7H17CIN202 C19H2OCINO4 C22H19N04 C20H23BrN204 C25H3406

134523-038, 29122-687 134523-005 95233-184 83905-015, 117772-700 86541-744 41859-670 [brompheniramine]51333 -223 51372-293 1397-893 1405-874 1134-470 132-172,86 590-636,674 980-712,86 446-866 322-350 [benztropine] 603-509 [11(grb), 16(gra)] [11(grb), 60-800 50-782 [atorvastatin] [dihydrate] [bethanechol] a)[/R] 13-5 38-4 22-6 16(gr platepositioncas# 402001B04 402001F04 402013D06 402006CO2 402008H05 402008F11 402001A05 402008B05 402001B05 402001C05 402009E05 402001D05 402001H05 402001A11 402006A09 D06402001 402012D11 402011F03

AMPHOTERICINB MOLENAME ANTIPYRINE ASPIRIN ATENOLOL ATORVASTATIN CALCIUM ATOVAQUONE AZATHIOPRINE AZITHROMYCIN BACITRACIN BACLOFEN BENAZEPRIL HYDROCHLORIDE BENSERAZIDE HYDROCHLORIDE BENZTROPINE BETHANECHOL CHLORIDE BEZAFIBRATE BISACODYL BROMPHENIRAMINE MALEATE BUDESONIDE

IDID 01500122 01500128 01500130 01501127 01503722 01504210 01500133 01503679 01500134 01500135 01505200 01500137 01500142 01500146 01502046 01500147 01503985 01500813 US 2019 / 0262353 A1 Aug . 29 , 2019

REFERENCES USAN,INN USAN,INN INN,USP BAN,JAN USP,INN USP,INN BAN,JAN USAN,INN USP,INN BAN,JAN USP,INN BAN,JAN USP,INN BAN,JAN BAN,JANUSP,INN BAN,JAN USPINN, BAN,JAN USP,INNBAN,JAN USP,INN USP,INN BAN,JAN USP,INN USP,INN BAN,JANUSP ,INN BAN,JAN USP,INN INN,USP BAN,JAN USAN,BAN STATUS BAN BAN BAN,JAN BAN BAN semisynthetic; SOURCE synthetic synthetic synthetic synthetic synthetic synthetic synthetic synthetic semisynthetic semisynthetic semisynthetic synthetic synthetic synthetic synthetic synthetic synthetic Streptomyces erythreus synthetic U-21251

alkylatingagent receptorantagonist inhibitsprotein 182.65cholinergic,miotic antipsychotic antihypertensive fungicide synthesis 246.30antineoplastic, 610.68angiotensin1 217.29antihypertensive 236.28analgesic, anticonvulsant blocker 318.87antiemetic, 338.77diuretic, 331.35antibacterial, 461.45antibacterial, FORMULAMolWtBIOACTIVITY 364.42diuretic 276.21antidepressant 556.57betaadrenergic 367.81antibacterial 363.39antibacterial 347.40antibacterial 390.87antihistaminic 276.74antidiabetic 252.34antiulcerative 747.97antibacterial 459.97antihistaminic TABLE1-continued C17H2ON2055 C13H19C12NO C6H140682 C33H34N606 C9H15NO3S C6H15CIN202 C15H12N2O C28H32N2010 C15H14CIN304S C16H17N305S C16H17N3045 C20H23CIN204 C17H19CIN2S C10H13CIN2035 C14H11CIN2045 C10H16N6S C17H18FN303 C38H69N013 C25H30CINO5 C18H34C12N2O5S ?chlorpheniramine] 16(gra)[/S] 31677-937, 59-7139481 70356-035, 66592-878, 119922-899 23325-782, 21462-395, [monohydrate], 51372-282 28395-031 34911-552 62571-862 72956-093 53994-733 50370-122 [anhydrous], 15686-712 113-928,132 51481-619 85721-331 81103-119 15686-518 58207-195 [11(grb), 298-464 [hemihydrate] [bupropion]55-981 51-832 (carvedilol) [anhydrous] [anhydrous] 50-533 94-202 77-361 22-9 platepositioncas# 402006F06 402008G10 402001F06 402009B04 402005F03 B07402001 C07402001 402009F03 402005D06 402001G07 H11402012 402001D09 402001E09 402001F09 402001A07 402005G03 402008E03 402009HO2 402001D10 402001F10

MOLENAME BUMETANIDE BUPROPION BUSULFAN CANDESARTAN CILEXTIL CAPTOPRIL CARBACHOL CARBAMAZEPINE CARVEDILOL TARTRATE CEFACLOR CEFADROXIL CEPHALEXIN CHLORPHENIRAMINE (S)MALEATE CHLORPROMAZINE CHLORPROPAMIDE CHLORTHALIDONE CIMETIDINE CIPROFLOXACIN CLARITHROMYCIN CLEMASTINE CLINDAMYCIN HYDROCHLORIDE

ID 01502004 01504174 01500152 01504261 01500682 01500158 01500159 01504257 01500771 01500163 01502028 01500183 01500184 01500185 01500187 01500684 01503614 01504231 01500191 01500193 US 2019 / 0262353 A1 Aug . 29 , 2019 14 Aug. 29 , 2018

REFERENCES JAmChemSoc 74:487(1952) HelvChimActa 60:1568(1977)

USP,INN BAN,JAN INN,USP USP,INN USP,INN BAN,JANUSP,INN BAN,JAN USP,INN USP,JAN USP,INN BAN,JAN USP,INN USP,INN BAN,JAN USP,INN BAN,JAN USP,INN BAN,JAN USP,INN INN,BAN USP,INN BAN,JAN USP,INN STATUS BAN BAN BAN NF,JAN BAN JAN BAN

synthetic; BAQD-10 SOURCE synthetic synthetic synthetic synthetic semisynthetic synthetic Colchicum autumnale coaltar synthetic synthetic synthetic Tolypocladium inflatum synthetic synthetic synthetic synthetic

alkylatingagent leprostatic, antigoutagent (skeletal) aggregation 512.34antiasthmatic, antiallergy 279.10antineoplastic, suppressant herpetiformis suppressant 351.32antidepressant 266.56antihypertensive inhibitor 344.85antifungal 399.45antimitotic, 311.86musclerelaxant 1202.64immunosuppressant 337.47anteriorpituitary 248.31antibacterial, dermatitis 302.85antidepressant FORMULAMolWtBIOACTIVITY 419.91platelet 457.87antibacterial 326.83antipsychotic 108.14antiinfectant 527.59antiinfectant 352.32antitussive TABLE1-continued C19H24C12N2 C9H10C13N3 C16H18CINO6S2 C22H17CIN2 C19H17CIN3NaO5S C18H19C1N4 C22H25NO6 C7H8O C23H14Na2011 C20H22CIN C7H17C12N2O3P C62H111N11012 C22H27NO2 C12H12N2025 C30H40C12N4 C18H23CIN2 C18H26BrNO

113665-842 15826-376. 6055-192,50 [dextromethorphan] 18323-449 4205-918, 23593-751 7081-449, 16110-513 6202-239, [cyclobenzaprine] 59865-133 17230-885 6700-341, [anhydrous], [clindamycin] 303-491 [clomipramine] 4205-907 642-784 5786-210 1319-773 303-537 522-510. 6707-580 [dequalinium]58-286,50 [desipramine] 125-699 125-713 [clonidine] [anhydrous] 64-868 [cromolyn] [anhydrous] 80-080 17321 0-18 47-5 platepositioncas# 402012GO2 402001C06 402008E05 402013H06 402001B11 402005HO3 402001D11 402001H11 402002A02 402011HOS 402002D02 402007B03 402002G02 402002H02 402007A09 402002D03 402002G03

MOLENAME CLOMIPRAMINE HYDROCHLORIDE CLONIDINE HYDROCHLORIDE CLOPIDOGREL SULFATE CLOTRIMAZOLE CLOXACILLIN SODIUM CLOZAPINE COLCHICINE CRESOL CROMOLYN SODIUM CYCLOBENZAPRINE HYDROCHLORIDE CYCLOPHOSPHAMIDE HYDRATE CYCLOSPORINE DANAZOL DAPSONE DEQUALINIUM CHLORIDE DESIPRAMINE HYDROCHLORIDE DEXTROMETHORPHAN HYDROBROMIDE

ID 02300061 01500198 01503710 01500200 01500201 01500685 01500205 01500209 01500210 01503207 01500213 01502202 01500220 01500222 01503127 01500227 01500233 US 2019 / 0262353 A1 Aug . 29 , 2019

J.ChemSoc1930: REFERENCES 508;1954:2012 USP,INN USP,JAN USP,INN BAN,JAN USP.INN, BAN,JAN USP,INN BAN,JAN USP,INN BAN,JAN USP,INN BAN,JAN USP-XXI, INN,BAN USP,INN BAN,JAN USP,INN BAN,JANUSP ,INN USPINN, USP,INN USP,INN BAN,JAN USP,INN BAN,JAN JAN BAN STATUS BAN BAN BAN USP,INNergotand ConvolvulvaceaeBAN,JAN semisynthetic; synthetic; SCH-6783; NSC-64198 lanataor SOURCE synthetic synthetic Digitalis D.orientalis Lam, Scrophulariaceaesynthetic synthetic synthetic synthetic synthetic synthetic synthetic GS-3065 synthetic synthetic synthetic spp

diuretic,activates Kchannelsand AMPAreceptors blocker,coronary antiinflammatory hypnotic 492.53ACEinhibitor, antihypertensive 441.49oxytocic,5HT antagonist 230.67antihypertensive, 250.20analgesic, vasodilator vasodilator 388.47antihistaminic, inhibitor MolWtBIOACTIVITY 318.14antiinflammatory 780.96cardiacstimulant 450.99Cachannel 469.98antiemetic 291.82antihistaminic 317.86antihistaminic 504.64coronary 437.48antiarrhythmic 315.85antidepressant 480.91antibacterial 201.70acetylcholinesterase TABLE1-continued FORMULA C8H7CIN202S C14H10C12NNO2 C13H8F203 C41H64014 C22H27CIN2045 C24H28CIN503 C17H22CINO C19H24CINO C24H40N804 C21H32N305P C19H22CINO C22H25CIN208 C21H28N205 C10H16CINO C24H32N209 C23H27N306

15307-796 22494-424 20830-755 33286-225, 42399-417 [diphenylpyraline] 1229-294, 25127-315 17086-281, 76095-164, 75847-733 3737-095 1668-195 4698-399 562-107,469 116-381,312 129-511,60 523-875 147-240 132-183147 [doxepin], [(1/Z)-isomer] 564-250 [doxylamine] [edrophonium] ergonovine][ [diltiazem] 58-322 isomer], [anhydrous] [enalapril] [(/E) 402013A02364-987 20-6 21-6 48-1 79-7 platepositioncas# 402002B04 402002G04 402002H04 402012611 402002B05 402002D05 402002E05 F05402002 402002H05 402013F09 402011F09 G08402013 402010H07 402011B05 402002H06

MOLENAME DIAZOXIDE DICLOFENAC SODIUM DIFLUNISAL DIGOXIN DILTIAZEM HYDROCHLORIDE DIMENHYDRINATE DIPHENHYDRAMINE HYDROCHLORIDE DIPHENYLPYRALINE HYDROCHLORIDE DIPYRIDAMOLE DISOPYRAMIDE PHOSPHATE DOXEPIN HYDROCHLORIDE DOXYCYCLINE HYDROCHLORIDE DOXYLAMINE SUCCINATE EDROPHONIUM CHLORIDE ENALAPRIL MALEATE ERGONOVINE MALEATE

ID 02300206 01500237 01500245 01500247 02300214 01500251 01500256 01500258 01500259 01500261 01500264 01500266 01500267 02300219 01501214 01500277 US 2019 / 0262353 A1 Aug . 29 , 2019 16

688(2001) REFERENCES INN,BANMetabolism50: USAN,INN USAN,INN USAN,INN USAN,INN USAN,INN USP,INN BAN,JAN USP,INN BAN,JAN USP,INN BAN,JAN USP,INN USP,INN BAN,JAN BANINN, BAN,JAN USP,BAN USP,INN BAN,JAN USP,INN BAN,JAN USP,INN USP,INN BAN,JAN USP,INN BAN,JAN USP,INN BAN,JAN USP,BAN USPINN, USP,INNBAN,JAN USP-XXII, STATUS BAN BAN BAN BAN JAN BAN BAN JAN BAN INN synthetic; SOURCE Streptomyces erythreus synthetic synthetic synthetic synthetic synthetic synthetic syntheticsynthetic synthetic synthetic synthetic Streptomyces spp synthetic SE-1702 synthetic synthetic synthetic synthetic semisynthetic semisynthetic synthetic

(tuberculostatic) topicalantiseptic 244.27antiinflammatory, analgesic antihypertensive 250.34antihyperlipoproteinemicsynthetic antipsychotic 433.96antimalarial,lupus suppressant 409.44sterolabsorption inhibitor 330.75diuretic, 375.87antidyskinetic, 194.28anthelmintic, MolWtBIOACTIVITY 1056.41antibacterial antibacterial277.24 141.17anticonvulsant 287.36antiinflammatory 321.34antiviral 337.45H2antihistamine 360.84antihyperlipidemic 477.43vasodilator 345.80antidepressant 510.45H1antihistamine 176.12antibacterial 323.42antidiabetic 494.01antihyperglycemic 198.22expectorant 196.64antihypertensive 297.74diuretic TABLE1-continued FORMULA C52H97NO18S C10H26C12N202 C7H11NO2 C17H21NO3 C24H21F2NO3 C14H19N504 C8H15N70253 C20H21C104 C26H28C12F2N2 C17H19CIF3NO C22H28C12F3N3OS C15H13F02 C3H5C104P C12H11CIN2055 C15H2203 C15H21N3035 C23H28CIN305S C10H1404 C21H23CIFNO2 C12H1802 C8H9CIN4 C7H8CIN30452 C18H28CIN305S

163222-331 104227-874 26472-479, [hydroxychloroquine] 1070-117,74 30484-776, 134-361,114 41340-254 76824-356 49562-289 52468-607 54910-893 5104-494 25812-300 21187-984 10238-218 304-201,86 747-364,118 [erythromycin] 146-565 136-776 [hydralazine] [ethambutol] [flunarazine] 23112-90 93-141 77-678 5(acid) 54-319 52-868 58-935 positioncas# 07-8 55-5 54-4 42-3 plate 402012G05 402002F07 402012E11 402005B09 H05402009 402009F05 402005HOS F09402005 402011B02 402012H03 402005G08 402002F08 402006DOS 402002HOS 402002C09 402008A10 A09402010 402002G09 402002C10 402002F10 402002B11 HYDROCHLOROTHIAZIDE402002C11 HYDROXYCHLOROQUINE402012C11 MOLENAME ERYTHROMYCIN ESTOLATE ETHAMBUTOL HYDROCHLORIDE ETHOSUXIMIDE ETODOLAC EZETIMIBE FAMCICLOVIR FAMOTIDINE FENOFIBRATE FLUNARIZINE HYDROCHLORIDE FLUOXETINE FLUPHENAZINE HYDROCHLORIDE FLURBIPROFEN FOSFOMYCIN FUROSEMIDE GEMFIBROZIL GLICLAZIDE GLYBURIDE GUAIFENESIN HALOPERIDOL HEXYLRESORCINOL HYDRALAZINE HYDROCHLORIDE SULFATE

ID 01501176 01500288 01502196 01501005 01505203 01505201 01501003 01501010 01500993 01504173 01500994 01500308 01502039 01500310 01500313 01504145 02300229 01500321 01500325 01500330 01500334 01500335 01503978 US 2019 / 0262353 A1 Aug . 29 , 2019

REFERENCES USAN,INN USAN,INN USP,INN USP,INN BAN,JAN USP,INN BAN,JAN USP,INN BAN,JAN INN,USP BAN,JAN BAN,JAN USP,INN USP,INN BAN,JAN USP,INN BAN,JAN USP,INN BAN,JAN USP,INN BAN,JAN USP,INN BAN,JAN USP,INN BAN,JAN USP,INN BAN,JAN USP,INN STATUS BAN USP,JAN BAN BAN BAN SOURCE synthetic synthetic synthetic synthetic synthetic synthetic synthetic synthetic synthetic synthetic synthetic synthetic synthetic synthetic synthetic synthetic synthetic

antipyretic, receptorantagonist diphosphate antihypertensive357.80antiinflammatory , analgesic 76.06antineoplastic, antiarrhythmic inhibits ribonucleoside reductase763.29anxiolytic, antihistaminic 365.84diuretic, 412.37bronchodilator, 428.54angiotensin2 137.14antibacterial, tuberculostatic 364.88adrenergicblocker MolWtBIOACTIVITY 206.29antiinflammatory 316.88antidepressant 236.14antianginal 531.44antifungal 254.29antiinflammatory 376.41antiinflammatory 425.51antiasthmatic 342.30laxative 369.37antiulcerative

TABLE1-continued FORMULA CH4N202 C44H43CIN208 C13H1802 C19H25CIN2 C16H16CIN3035 C19H16CINO4 C20H30BrNO3 C25H28N60 C6H7N30 C6H8N208 C26H28C12N404 C16H1403 C19H24N206 C23H23NOSS C19H25CIN203 C12H22011 C16H14F3N302S

58560-751 66985-179, 22254-246 138402-116 6527742-1 22071-154 74103-074, 74103-063 34580-148, 34580-137 32780-646, 36894-696 103577-453 127-071 [hydroxyzine] [(+/-)mixture] [imipramine] [anhydrous] 4618-182 68-882 53-861 54-853 87-332 [ketorolac] [ketotifen] [labetalol] 402002H1110246-750, 402003C0215687-271, 113-520,50402003D02 49-7 26807-658402003E02 platepositioncas# 402002G11 F02402003 402013F04 402009H03 402003A03 402003D03 G03402003 402006C06 402012D10 402005A02 402007C10 402013F10 402008F06

MOLENAME HYDROXYUREA HYDROXYZINE PAMOATE IBUPROFEN IMIPRAMINE HYDROCHLORIDE INDAPAMIDE INDOMETHACIN IPRATROPIUM BROMIDE IRBESARTAN ISONIAZID ISOSORBIDE DINITRATE KETOCONAZOLE KETOPROFEN KETOROLAC TROMETHAMINE KETOTIFEN FUMARATE LABETALOL HYDROCHLORIDE LACTULOSE LANSOPRAZOLE

ID 01500344 01500345 01500347 01500348 01500349 01500350 01500354 01504259 01500355 01500358 01500362 01501215 01503925 01500668 01503243 01500363 01503926 US 2019 / 0262353 A1 Aug . 29 , 2019

PNAS77:3957 (1980);IntJ717 Oncol12: 39:1653(2000)BAN REFERENCES (1998) USAN,INN USAN,INN USAN,INN USAN,INN USAN,INNNeuropharmacol USP,INN BAN,JAN USP,INN BAN,JAN BAN,JANBAN,USP USP,INN BAN,JAN USP,INN BAN,JAN USP,INN USP,INN USP,INN BAN,JAN USP,INN BAN,JAN USP,INN BAN,JAN STATUS JAN BAN BAN BAN USP BAN BAN USAN seedlings, synthetic;L- SOURCE synthetic Viciafaba Sarothamnus otherspp&,palnts synthetic isomer synthetic synthetic synthetic synthetic synthetic synthetic synthetic synthetic synthetic synthetic synthetic synthetic synthetic

antidotetofolic ATIangiotensinII acidantagonists reductaseinhibitor (skeletal) 511.51antianemic, 672.96thyroidhormone blocker 422.92antihypertensive, antagonist404.55antihyperlipidemic, HMGCOA 241.29antiinflammatory, analgesic 152.18antineoplastic, purine MolWtBIOACTIVITY 197.19antiparkinsonian 361.38antibacterial 405.50ACEinhibitor 513.51Cachannel 382.89H1antihistamine 445.91antipsychotic 313.87antidepressant 295.30anthelmintic 378.32antimalarial 351.41antiinflammatory 215.77musclerelaxant antimetabolite

TABLE1-continued FORMULA C20H21CaN707 C9H11NO4 C18H20FN304 C15H1113NNa04 C21H31N305 C29H34C12N202 C22H23CIN202 C22H23CIN60 C24H3605 C22H24CIN305 C20H24CIN C16H13N303 C15H15NO2 C17H16F6N20 C14H13N30452 C12H22CIN C5H4N4S

124750-998, 138199-710 83915-837, 76547-983 835,34552-53179-116 79794-755 114798-264 75330-755 27833-643, 10347-816, 10262-698 31431-397 53230-107 38-771125 19982-082 6112-761,50 1492-188 55-061,6893 [liothyronine] [anhydrous] [loperamide] 1977-102 maprotiline)( [anhydrous] 59-927 [losartan] [loxapine] 61-687 02-3 44-2 platepositioncas# 402003HO3 402010HO8 402009A04 402006B09 402006D06 402013A06 402008F05 402009D04 402008D07 402012H10 402003D04 402005H10 402013B02 402007E07 402008C10 402005H11 402003E05

MEFENAMICACID MOLENAME LEUCOVORIN CALCIUM LEVODOPA LEVOFLOXACIN LIOTHYRONINE SODIUM LISINOPRIL LOPERAMIDE HYDROCHLORIDE LORATADINE LOSARTAN LOVASTATIN LOXAPINE SUCCINATE MAPROTILINE HYDROCHLORIDE MEBENDAZOLE MEFLOQUINE MELOXICAM MEMANTINE HYDROCHLORIDE MERCAPTOPURINE

ID 01500364 02300205 01504260 01502047 01501217 02300241 01503712 01504268 01503977 02300242 01500373 01501110 01501103 01503070 01504150 01501121 01500387 US 2019 / 0262353 A1 Aug . 29 , 2019

REFERENCES USAN,INN USAN,INN USP,INN BAN,JAN USP,INN USP,INN BAN,JAN USP,INN BAN,JAN USP,BAN USP,INN BAN,JAN USP,INN BAN,JAN USP,INN BAN,JAN USP,JAN USP,INN BAN,JAN BAN,JAN USP,INN USP,INN BAN,JAN USAN. STATUS JAN JAN BAN BAN USAN BAN

synthetic; CRL-40476. CEP-1538 SOURCE synthetic synthetic synthetic synthetic synthetic synthetic synthetic synthetic synthetic synthetic synthetic synthetic semisynthetic synthetic Penicillium brevicompactum andother Penicillium spp antirheumatic, pigmentationagent antialopeciaagent anhydrase (urinary) (skeletal) folicacidantagonist antihypertensive417.46antihypertensive, antianginal 290.75antihypertensive, 209.25antihypertensive, inhibitor 241.25musclerelaxant 454.45antineoplastic, 216.20antipsoriatic, 365.84diuretic, vasoconstrictor 236.27carbonic 140.19antibacterial 211.22antihypertensive 336.26antiemetic 171.16antiprotozoal 517.41antineoplastic 320.35antineoplastic MolWtBIOACTIVITY 273.36 465.96antibacterial

TABLE1-continued FORMULA C5H8N403S2 C6H12N4 C11H15N05 C20H22N805 C12H804 C10H13NO4 C14H23C12N302 C16H16CIN303S C19H31NO9 C6H9N303 C12H19CIN204 C9H15N50 C22H30C12N406 C15H15NO2S C23H29CIFN304 C17H2006

41372-081, 54143-576, 56392-177. hydrochloride] 70476-823, 186826-868 7232-215 [anhydrous], [metoclopramide] 17560-519 37350-586 69198-103 3092-179, 42794-763 38304-915 65271-809 68693-118 24280-931 554-574 97-0100 532-036 298-817 555-306 [anhydrous] 364-625 [metroprolol] 443-481, [mitoxantrone] 59-052 [metronidazole [midodrine] platepositioncas# 402011G10 G05402003 402003A06 402003B06 402003C06 402003E06 402003F06 402012F11 G06402003 402003H06 402012A08 402003BO7 402007F11 402010F05 402009A05 402005A03

MOLENAME METHAZOLAMIDE METHENAMINE METHOCARBAMOL METHOTREXATE METHOXSALEN METHYLDOPA METOCLOPRAMIDE HYDROCHLORIDE METOLAZONE METOPROLOL TARTRATE METRONIDAZOLE MIDODRINE HYDROCHLORIDE MINOXIDIL MITOXANTHRONE HYDROCHLORIDE MODAFINIL MOXIFLOXACIN HYDROCHLORIDE MYCOPHENOLICMIL ACID

ID 01503252 01500394 01500397 01500398 01500400 01500403 01500410 02300325 01500411 01500412 01503257 01500415 01503278 01505361 01504303 01500674 US 2019 / 0262353 A1 Aug . 29 , 2019

839:209(1999); 127:605(1999) REFERENCES BrainRes BritJPharmacol 31:65(1997) USAN,INNPharmacotherapy USAN,INN USAN,INN USP,INN BAN,JAN USP,INN BAN,JAN USP,INN BAN,JAN USP,INN BAN,JAN USP,INNJAN BAN, USP,INN BAN,JAN USP,INN USP,INN JANBAN, USP,INN BAN,JAN USP,INN BAN,JAN USP-XII, USP,INN STATUS USP BAN BAN INN,BAN BAN BAN BAN SOURCE synthetic synthetic synthetic synthetic synthetic synthetic synthetic synthetic synthetic synthetic synthetic synthetic synthetic synthetic synthetic synthetic

inhibitorprodrug, analgesic, antagonist antipyretic antihypertensive (peripheral) antihypertensive inhibitor, antiobesity (skeletal), blocker 230.27antiinflammatory, 558.60AngiotensinII 495.75reversiblelipase 228.29antiinflammatory 309.41betaadrenergic 363.84narcoticantagonist 341.41morphine 303.20cholinergic 317.23antiandrogen 238.16antibacterial 263.39antidepressant 461.52musclerelaxant antihistaminic MolWtBIOACTIVITY 346.34 418.45vasodilator 319.34antibacterial 335.88vasodilator

TABLE1-continued FORMULA C15H1602 C17H27NO4 C19H22CINO4 C20H23NO4 C14H1403 C12H19BrN202 C17H18N206 C12H10F3N304 C21H26N207 C8H6N405 C16H18FN303 C19H21N C19H26CINO2 C29H30N606 C29H53N05 C24H31NOS

42200-339 51481-608 16676-292, 16590-413 22204-531 21829-254 63612-500 66085-594 17140-817 70458-967 144689-634 96829-582 4682-364,83 357-084, [dihydrate], 114-807,59 894-713,72 1400-619 orphenadrine][ 465-656 [naloxone] [naltrexone] [neostigmine] 67-209,54 sodium], [monohydrate] [nortriptyline] 402012A0942924-538 99-4 87-5 nitrofurantoin[ 69-5 98-7 platepositioncas# 402012B07 402003E07 402012C07 402003G07 402003A08 402003C08 402012D02 402008A03 402003D08 402003B09 402003D09 402003G09 402009B06 402009G04 402003A10

NAPROXEN(+) MOLENAME NABUMETONE NADOLOL NALOXONE HYDROCHLORIDE NALTREXONE HYDROCHLORIDE NEOSTIGMINE BROMIDE NIFEDIPINE NILUTAMIDE NIMODIPINE NITROFURANTOIN NORFLOXACIN NORTRIPTYLINE NYLIDRIN HYDROCHLORIDE OLMESARTAN MEDOXOMIL ORLISTAT ORPHENADRINE CITRATE

ID 01503650 01503260 01500422 01503262 01500425 01500428 01500431 01504152 01503600 01500433 01500440 01500442 01500445 01505205 01504300 01500447 US 2019 / 0262353 A1 Aug . 29 , 2019 21

REFERENCES USAN,INN USAN,INN USAN,INN USP,INN USP,INN BAN,JAN BAN,JAN USP,INN BAN,JAN USP,INN USP,INN USP,INN BAN,JAN USP,INN BAN,JAN USP,INN BAN,JAN BAN,JAN USAN,INN STATUS BAN BAN USAN BAN BAN USP,JAN BAN USP

synthetic;CI- synthetic; S9490-3, McN-A2833 CS-514;SQ SOURCE synthetic Streptomyces rimosis paramomycinus synthetic synthetic 109 synthetic synthetic synthetic synthetic synthetic synthetic synthetic synthetic synthetic 31000 1008 antiarrhythmic, ACEinhibitor antianginal, reductaseinhibitor 713.72antibacterial, antiamebic 441.62antihypertensive, 274.26anticonvulsant, antieleptic 248.33antihypertensive, antiglaucoma agent 175.23ultravioletscreen 446.52antihyperlipidemic, HMGCOA MolWtBIOACTIVITY 252.28antipsychotic 278.31vasodilator 365.50antipsychotic 403.98antipsychotic 249.70analgesic 234.28antidepressant 461.56antipsychotic 392.91antidiabetic 331.35antiinflammatory 159.23anticonvulsant TABLE1-continued FORMULA C15H12N202 C23H47N5018S C13H18N403 C21H23N3OS C23H43N305 C21H26C1N3OS C11H12C1N5 C8H14N2045 C15H11N2NaO2 C28H29F2N30 C14H20N202 C19H21CIN203S C15H13N3045 C7H6KNO2 C23H35Na07 C8H17NO2

[paromomycin], 107133-368; 148553-508 1263-894, 82834-160 [phenazopyridine] 13523-869 111025-468 36322-904 81131-706 2622-266 136-403,94 156-514, 630-933,57 2062-784 7542-372 [paromomycin, replaced] 6493-056 (perindopril) (pioglitazone) 150-130 59-041 58-399 [phenelzine] [phenytoin] 402009DO328721-075 78-0 71-8 41-0 (acid) platepositioncas# 402007B11 402012E08 402008B07 402009H06 402011HO3 402003C11 402003D11 402003G11 402006HO2 402013C08 402009B05 402013D09 402001C03 402010A06 402010D06

POTASSIUMP MOLENAME OXCARBAZEPINE PAROMOMYCIN SULFATE PENTOXIFYLLINE PERICIAZINE PERINDOPRIL ERBUMINE PERPHENAZINE PHENAZOPYRIDINE HYDROCHLORIDE PHENELZINE SULFATE PHENYTOIN SODIUM PIMOZIDE PINDOLOL PIOGLITAZONE HYDROCHLORIDE PIROXICAM AMINOBENZOATE PRAVASTATIN SODIUM PREGABALIN

IDID 01504243 01503228 01503611 01503936 01505212 01503934 01500473 01500476 01500485 01501134 01500488 01504401 01500491 01500113 01505803 01505816 US 2019 / 0262353 A1 Aug . 29 , 2019

REFERENCES USP,INN USP,INN BAN,JAN USP,INN BAN,JAN USP,INN BAN,JAN USP,INN USP,INN BAN,JAN USP,INN BAN,JAN USP,INN BAN,JAN INN,USP BAN,JAN USP,INN USP,INN BAN,JAN USP,INN BAN,JAN USP,INN BAN,JAN USP,INN STATUS BAN JANUSP, BAN BAN BAN USP,JAN SOURCE synthetic synthetic synthetic synthetic synthetic synthetic synthetic synthetic synthetic synthetic synthetic synthetic synthetic synthetic synthetic Cinchonaspp

antipsychotic,treatmentof antianginal, antiarrhythmic ACEinhibitor skeletalmuscle 564.15antiemetic, vertigo 295.81antihypertensive, 123.12antibacterial, tuberculostatic 474.99antihypertensive, 422.50antimalarial, relaxant MolWtBIOACTIVITY 455.34antimalarial 218.26anticonvulsant 285.36uricosuric 271.79antiarrhythmic 323.91anticholinergic 320.89antihistaminic 377.92antiarrhythmic 170.23antihyperthyroid 201.70decongestant 594.69anthelmintic 261.12cholinergic

TABLE1-continued FORMULA C15H27N309P2 C12H14N202 C13H19N045 C13H22CIN30 C22H30CIN306S3 C19H30CINO C17H21CINAS C21H28CINO3 C16H22CINO2 C7H1ON2OS C10H16CINO C34H30N2065 C5H5N30 C9H13BrN202 C25H31CIN205 C20H26N206S

1257-789,84 1508-765,77 22,-734183 22204-246, 82586-558, 614-391,51 [prochlorperazine] 54063-535 318-989,525 345-788,90 [pseudoephedrine] 15686-836 [pyridostigmine] 85441-618 6119-706, [anhydrous], [primaquine] [procainamide] maleate], [procyclidine] 58-333,60 promethazine][ propafenone][ [propranolol] 804-637 130-950 57-669 58-388 51-525 [pyrantel] 98-964 101-2 [quinapril] [quinine] positioncas# 402004D0263-456,90 34-6 402013C04125-337 06-9 02-6 [prochlorperazine 37-2 87-7 66-6 82-4 97-5

plate 402013C09 402013D05 402004E02 402013D10 402004G02 402008A07 402013B07 402011B07 402004B03 402004C03 402011C05 402007A10 402007H07 402004G03

QUININESULFATE MOLENAME PRIMAQUINE DIPHOSPHATE PRIMIDONE PROBENECID PROCAINAMIDE HYDROCHLORIDE PROCHLORPERAZINE EDISYLATE PROCYCLIDINE HYDROCHLORIDE PROMETHAZINE HYDROCHLORIDE PROPAFENONE HYDROCHLORIDE PROPRANOLOL HYDROCHLORIDE (+/-) PROPYLTHIOURACIL PSEUDOEPHEDRINE HYDROCHLORIDE PYRANTEL PAMOATE PYRAZINAMIDE PYRIDOSTIGMINE BROMIDE QUINAPRIL HYDROCHLORIDE

ID 01500500 01500501 01500502 01500503 01500505 01500507 01500510 01503935 01505270 01500515 01500516 01500517 01500518 01503240 01503076 01500524 US 2019 / 0262353 A1 Aug . 29 , 2019 23

Pharmacol6:177 Lett140:225 76:844(1992); FundamClin 91:3401(1969) (1995) (1992); Anesthesiology (1992) REFERENCES Drugs46749:USANINN, USAN,INNJAmChemSoc USAN,INN USAN,INNNeurosci USAN,INN USAN,INN USAN,INN USAN,INN USP,INN BAN,JAN USAN, USP,INN BAN,JAN USP,INN BAN,JAN USP,INN USP,INN BAN,JAN USP,INN BAN,JAN USP,INN BAN,JAN BAN,JAN USPINN, BAN,JAN USP,BAN STATUS BAN BAN BAN BAN BAN BAN BAN BAN JAN

semisynthetic; Camelia,thea L-5103,Ba 41166/E, NSC-113926 SOURCE synthetic semisynthetic synthetic synthetic synthetic synthetic synthetic Streptomycesambofaciens synthetic synthetic synthetic synthetic synthetic synthetic synthetic Streptomyces spp Paullinia ??????

(tuberculostatic) synthesisinhibitor glutamaterelease Crohn'sdisease antiamebic, 785.90antibacterial,RNA 5HTuptake antipneumocystis 234.20anticonvulsant, inhibitor 342.70antidepressant, inhibitor474.59impotency therapy 253.28antibacterial, 398.40anticolitisand 480.91antibacterial, antirickettsial MolWtBIOACTIVITY 314.41H2antihistamine 822.96antibacterial 357.43antidiabetic 481.55antihyperlipidemic 843.07antibacterial 416.58diuretic 404.49uricosuric 267.31antibacterial 356.42antiinflammatory 337.38antiinflammatory 180.17bronchodilator TABLE1-continued FORMULA C13H22N403S C43H58N4012 C43H51N3011 C8H5F3N2OS C18H19N3035 C22H28FN3065 C17H18C13N C22H30N604S C43H74N2014 C24H32045 C10H11N3035 C18H14N4055 C23H20N203S C11H13N3035 C20H17FO3S C13H11N30452 C22H25CIN208 C7H8N402

287714-144, 79559-970; 139755-832 59804-374. 5967-840,58 66357-355 13292-461 80621-814 1744-225 38194-502 122320734- 147098-20 8025-818 723-466 599-791 127-695 64-755,60 [tetracycline] 2(Casalt) 79617-96 52-017 57-965 [anhydrous] 2(base) 54-8 55-9 platepositioncas# 402006F03 402004A04 402010B03 402010D05 402009C04 402009A07 402009D09 402008D09 GO2402008 402004G04 402004F05 402004H05 402011A10 402011B08 402004B06 402007D09 402004C06 402004D06

MOLENAME RANITIDINE RIFAMPIN RIFAXIMIN RILUZOLE ROSIGLITAZONE ROSUVASTATIN SERTRALINE HYDROCHLORIDE SILDENAFIL SPIRAMYCIN SPIRONOLACTONE SULFAMETHOXAZOLE SULFASALAZINE SULFINPYRAZONE SULFISOXAZOLE SULINDAC TENOXICAM TETRACYCLINE HYDROCHLORIDE THEOPHYLLINE 01501151 01500529 01505321 01505348 01504263 01505213 01505262 01504099 01503423 01500539 01500550 01500552 01500554 01500555 01500556 01503142 01500566 01500568 US 2019 / 0262353 A1 Aug . 29 , 2019 24

REFERENCES USAN,INN USAN,INN USP,INN USP,INN BAN,JAN USP,JAN USP,INN BAN,JAN INN,BAN INN,BAN BAN,JAN USP-XXI, INN,BAN USP,INN BAN,JAN USP,INN BAN,JAN USP,INN BAN,JAN USP,INN BAN,JAN INN,USPBAN,JAN USAN,JAN STATUS BAN JAN BAN SOURCE synthetic synthetic synthetic synthetic synthetic synthetic; RWJ-17021 synthetic synthetic synthetic synthetic synthetic synthetic synthetic synthetic synthetic

antimigraine, GABA-Aagonist, glutamatereceptor AMP/kinate antagonist, ACEinhibitor 167.19antineoplastic, purine blocker 261.71antiinflammatory, analgesia339.37anticonvulsant, carbonic anhydrase inhibitor 430.55antihypertensive, 337.94anticholinergic, antiparkinsonian MolWtBIOACTIVITY antimetabolite443.63antipsychotic 432.50betaadrenergic 270.35antidiabetic 157.21hemostatic 231.27antidepressant 408.33antidepressant 480.43antipsychotic 448.54antipruritic 290.32antibacterial 410.52antidepressant

TABLE1-continued FORMULA C5H5N5S C23H29N30252 C17H28N4075 C12H18N203S C14H12CINO2 C12H21NOSS C24H34N205 C8H15NO2 C9H13NO4S C19H23C12N50 C21H26C12F3N3S C20H32CINO C22H28N206S C14H18N403 C24H30N204

13492-018, 25332-392, 91524-162 13710-195 97240-794 87679-376 [tranylcypromine] 19794-935 440-175,117 [trifluoperazine] 4330-998, 41375-660 [replaced],84 521-788,739 5580-030 [hemihydrate] 3313-266 1197-188 7081-369 [replaced], 155-099 [trimeprazine] 738-705 [trimipramine] [trazodone] 52-493 [/Z] 26921-175,402004H06 [timolol] 402004G06154-427, 402011C045591-457, 89-5 96-8 71-9 402004A0764-777 platepositioncas# 402006F05 402010G05 402009F09 402006G07 402004C07 402007HO8 402004A08 402004B08 402004C08 402004E08 402012E04

MOLENAME THIOGUANINE THIOTHIXENE TIMOLOL MALEATE TOLBUTAMIDE TOLFENAMIC ACIDTOPIRAMATE TRANDOLAPRIL TRANEXAMIC ACID TRANYLCYPROMINE SULFATE TRAZODONE HYDROCHLORIDE TRIFLUOPERAZINE HYDROCHLORIDE TRIHEXYPHENIDYL HYDROCHLORIDE TRIMEPRAZINE TARTRATE TRIMETHOPRIM TRIMIPRAMINE MALEATE

ID 01500573 01500576 01500578 01500581 01501198 01505801 01505264 01502026 01500584 01503121 01500591 01500592 01500593 01500595 01503117 US 2019 / 0262353 A1 Aug . 29 , 2019

244:181(1936); Alkaloids2:406 Drugs21:90 91:10071986() 1950:1534; PharmacolRev35:143(1983) REFERENCES USP,INNHoppeSeyler'sZ (1981); Gastroenterology JChemSoc (1952); BAN,JANPhysiolChem USAN,INN USAN,INN USAN,INN USP,INN BAN,JAN USP,INN BAN,JAN USP,INN BAN,JAN STATUS BAN BAN USP BAN

synthetic; CGP-48933 SOURCE bearbile Streptomyces orientalis synthetic synthetic Corynanthe spp synthetic synthetic

blocker,mydriatic LD50(rat)890 inhibitor, blocker,coronary vasodilator, inhibitor,antiviral 5HT[1B/1D] 392.58anticholelithogenic; mg/kgip antihypertensive 491.08adrenegicblocker, Cachannel agonist 457Angiotensin.51II antiarrhythmic antidepressant 287.36antimigraine, MolWtBIOACTIVITY 1484.76antibacterial 277.41antidepressant 390.91alphaadrenergic 267.25RTtransferase TABLE1-continued FORMULA C24H4004 C24H28N5NaO3 C67H77C13N8024 C17H27NO2 C27H39CIN204 C21H27CIN203 C10H13N504 C16H21N302

137862-534 139264-178 1404-939. 99300-784, 93413-695 52114,152- 30516-871 128-132 1404-906 [vancomycin] (valsartan) venlafaxine][ [verapamil] 65-190 53-9 platepositioncas# 402004D09 402009E06 402004E09 402008F10 402013B03 402005B02 402012B03 402009C10

ZIDOVUDINE[AZT] MOLENAME URSODIOL VALSARTAN SODIUM VANCOMYCIN HYDROCHLORIDE VENLAFAXINE VERAPAMIL HYDROCHLORIDE YOHIMBINE HYDROCHLORIDE ZOLMITRIPTAN

ID 01500605 01505209 01500607 01504171 02300307 01500663 01502109 01505281 US 2019 / 0262353 A1 Aug . 29 , 2019 26

[0155 ] The 249 compounds were first tested against iron TABLE 2 - continued toxicity to human neurons in culture . Neurons were pre Drug % Iron % incubated with each compound for 1 h followed by appli control control cation of FeSO4. Ferrous iron ( 25 and 50 uM ) is very toxic Name (mean ) SEM (mean ) SEM to neurons, with > 80 % loss of microtubule - associated pro CIMETIDINE 34 . 38 11 . 74 13 .93 4 .80 tein - 2 MAP2( ) - labeled neurons by 24 h in most experiments CIPROFLOXACIN 40 . 99 8 .29 37 . 73 10 .54 compared to the control condition ( Table 2 ) . CLARITHROMYCIN 55 .09 13. 17 20 . 83 6 . 28 CLEMASTINE 6 . 19 0 . 36 5 .03 0 . 78 CLINDAMYCIN 63 . 15 12 . 24 20 .06 4 . 31 TABLE 2 HYDROCHLORIDE Drug % Iron % CLOMIPRAMINE 107 . 30 11 . 31 18 . 45 4 . 73 control control HYDROCHLORIDE Name (mean ) SEM (mean ) SEM CLONIDINE 7 .47 3 . 10 5 .03 0 . 78 HYDROCHLORIDE 5 - CHLOROINDOLE - 2 37. 30 5 . 87 17 .33 1 . 12 CLOPIDOGREL SULFATE 53. 53 9 .02 19 . 15 5 . 36 CARBOXYLIC ACID CLOTRIMAZOLE 12 . 36 4 . 00 40 . 33 10 . 95 ACEBUTOLOL 49. 02 13 .89 26 . 23 8 . 69 CLOXACILLIN SODIUM 28 .43 10 . 84 12 . 54 3 .49 HYDROCHLORIDE CLOZAPINE 101 . 15 8 . 52 9 . 41 3 .67 ACETAMINOPHEN 35 . 10 22 .07 34 . 50 15 . 86 COLCHICINE 3 . 12 0 . 41 5 . 03 0 . 78 ACETAZOLAMIDE 23 .56 19 . 82 34 . 50 15 . 86 CRESOL 6 . 04 1 . 15 5 .03 0 .78 ACETYLCYSTEINE 21 .67 18 . 23 34 . 50 15 . 86 CROMOLYN SODIUM 5 . 44 1 . 11 5 .03 0 .78 ACYCLOVIR 73 . 72 4 . 53 37 .73 10 . 54 CYCLOBENZAPRINE 98 . 36 12 . 76 35 .62 8 . 16 ALLOPURINOL 25 . 48 19 .62 34 . 50 15 . 86 HYDROCHLORIDE ALMOTRIPTAN 94 . 44 13 .68 42 . 76 12 .68 CYCLOPHOSPHAMIDE 6 . 39 1 . 16 5 . 03 0 . 78 ALTRETAMINE 4 . 20 0 . 12 3 . 19 0 . 14 HYDRATE AMANTADINE 52 . 56 21. 57 34 .50 15 .86 CYCLOSPORINE 11 . 48 0 . 85 17 . 33 1 . 12 HYDROCHLORIDE DANAZOL 4 . 37 0 . 23 5 .03 0 .78 AMIKACIN SULFATE 35 . 92 20 . 79 34 . 50 15 . 86 DAPSONE 18 . 59 5 .43 7 .08 2 . 23 AMILORIDE 37 . 14 21. 42 34 . 50 15 . 86 DEQUALINIUM 10 . 47 0 . 33 17 . 33 1 . 12 HYDROCHLORIDE CHLORIDE AMIODARONE 71. 35 16 .08 28 .43 6 . 81 DESIPRAMINE 84 . 38 4 .66 4 .02 0 . 70 HYDROCHLORIDE HYDROCHLORIDE AMITRIPTYLINE 34 .81 17 . 72 34 . 50 15 . 86 DEXTROMETHORPHAN 3 .49 0 .76 5 .03 0 . 78 HYDROCHLORIDE HYDROBROMIDE AMLODIPINE BESYLATE 93 .08 16 . 11 42 .76 12 . 68 DIAZOXIDE 80 . 86 7 . 81 40 . 33 10 . 95 AMOXICILLIN 6 .41 3 .65 34 .50 15 . 86 DICLOFENAC SODIUM 5 . 92 1 . 18 5 . 03 0 . 78 AMPHOTERICIN B 3 .41 1 . 33 34 . 50 15 . 86 DIFLUNISAL 4 . 12 0 . 53 5 .03 0 . 78 ANTIPYRINE 2 . 11 0 . 66 34 . 50 15 . 86 DIGOXIN 8 .91 1 . 80 20 .06 4 . 31 ASPIRIN 68 . 20 21. 69 40 . 33 10 . 95 DILTIAZEM 86 .04 11. 77 35 .62 8 . 16 ATENOLOL 43 . 42 12 . 08 14 . 41 3 . 42 HYDROCHLORIDE ATORVASTATIN 68 .87 4 . 37 37 .73 10 . 54 DIMENHYDRINATE 36 .53 5 . 32 4 . 02 0 . 70 CALCIUM DIPHENHYDRAMINE 74 . 72 6 . 44 4 . 02 0 .70 ATOVAQUONE 67 . 74 8 . 78 27 . 13 6 . 35 HYDROCHLORIDE AZATHIOPRINE 4 .65 3 .62 34 . 50 15 . 86 DIPHENYLPYRALINE 4 .61 0 . . 9696 5 .03 0 .78 AZITHROMYCIN 56 . 76 20 . 02 37 . 73 10 . 54 HYDROCHLORIDE BACITRACIN 5 .04 0 .51 5 . 03 0 . 78 DIPYRIDAMOLE 165 .07 14 . 85 13 . 26 2 .59 BACLOFEN 35 . 79 22 .09 34 . 50 15 . 86 DISOPYRAMIDE 4 .63 11 .. 12 5 . 31 0 . 25 BENAZEPRIL 72 . 19 14 . 31 42 . 76 12 . 68 PHOSPHATE HYDROCHLORIDE DOXEPIN 76. 91 17 . 10 20 .02 5 . 71 BENSERAZIDE 15 . 89 4 . 35 20 .06 4 . 31 HYDROCHLORIDE HYDROCHLORIDE DOXYCYCLINE 12 .70 4 .50 26 . 23 8 . 69 BENZTROPINE 11 .43 6 . 78 34 . 50 15 . 86 HYDROCHLORIDE BETHANECHOL 15 . 99 9 .09 34 .50 15 . 86 DOXYLAMINE 82. 41 12 . 13 28 .43 6 . 81 CHLORIDE SUCCINATE BEZAFIBRATE 35 . 54 14 . 52 14 . 27 4 . 70 EDROPHONIUM 44 .00 12 . 26 26 . 23 8 .69 BISACODYL 93 . 29 8 . 87 20 .06 4 . 31 CHLORIDE BROMPHENIRAMINE 79 .88 7 . 42 35 .62 8 . 16 ENALAPRIL MALEATE 40 . 97 12 .64 26 .23 8 .69 MALEATE ERGONOVINE MALEATE 42 . 73 12 . 37 8 . 53 2 .85 BUDESONIDE 70 .02 7 .41 48 . 89 3 .07 ERYTHROMYCIN 56 . 71 14 . 49 18 . 45 4 .73 BUMETANIDE 29 . 38 8 . 82 11 . 56 2 . 85 ESTOLATE BUPROPION 55 . 54 4 .03 37 .73 10 . 54 ETHAMBUTOL 3 .72 0 . 94 5 . 31 0 . 25 BUSULFAN 13 . 35 7 . 31 34 . 50 15 . 86 HYDROCHLORIDE CANDESARTAN 35 .48 4 . 57 12 .68 ETHOSUXIMIDE 74 . 29 18 . 77 35 .62 8 . 16 CILEXTIL ETODOLAC 34 . 42 10 . 33 13 . 93 4 . 80 CAPTOPRIL 35 . 34 7 .07 25 . 52 4 . 20 EZETIMIBE 50 .46 10 . 96 42 . 76 12 .68 CARBACHOL 8 . 87 3 . 78 34 . 50 15 . 86 FAMCICLOVIR 91. 00 12. 00 42. 76 12 .68 CARBAMAZEPINE 13 .31 4 .07 34 . 50 15 . 86 FAMOTIDINE 25 .23 9 . 73 13 .93 4 . 80 CARVEDILOL 159 . 69 10 . 42 20 .83 6 . 28 FENOFIBRATE 24 .43 7 . 32 13 . 93 4 . 80 TARTRATE FLUNARIZINE 126 . 36 9 . 16 9 . 86 2 .61 CEFACLOR 89 . 86 3 .78 9 .41 3 . 67 HYDROCHLORIDE CEFADROXIL 9 . 46 3 .53 34 . 50 15 . 86 FLUOXETINE 81 . 41 11 . 56 35 . 62 8 . 16 CEPHALEXIN 38 . 87 4 . 33 40 . 33 10 . 95 FLUPHENAZINE 12 . 13 4 . 32 25 .52 4 . 20 CHLORPHENIRAMINE ( S ) 52 . 32 9 . 27 20 . 06 4 . 31 HYDROCHLORIDE MALEATE FLURBIPROFEN 4 .63 0 . 44 5 . 31 0 . 25 CHLORPROMAZINE 98 .76 4 .92 17 . 35 9 .79 FOSFOMYCIN 31. 24 9 . 29 11 . 56 2 . 85 CHLORPROPAMIDE 5 . 32 1 . 13 5 .03 0 .78 FUROSEMIDE 3 . 96 0 . 74 5 . 31 0 . 25 CHLORTHALIDONE 7 . 66 2 . 15 5 . 03 0. 78 GEMFIBROZIL 5 . 05 0 . 73 5 . 31 0 . 25 US 2019 / 0262353 A1 Aug . 29 , 2019

TABLE 2 - continued TABLE 2 - continued Drug % Iron % Drug % Iron % control control control control Name (mean ) SEM (mean ) SEM Name (mean ) SEM (mean ) SEM GLICLAZIDE 47 . 31 5 . 08 37 . 73 10 . 54 NADOLOL 52 . 39 11 .65 35 .62 8 . 16 GLYBURIDE 45 . 24 1 . 39 48 . 89 3 . 07 NALOXONE 69 . 47 3 . 48 48 . 59 19 . 48 GUAIFENESIN 3 . 28 0 . 30 5 .31 0 . 25 HYDROCHLORIDE HALOPERIDOL 6 . 12 1 . 05 5 . 31 0 . 25 NALTREXONE 39 .55 4 . 02 35 .62 8 . 16 HEXYLRESORCINOL 71. 52 8 . 88 20 .06 4 . 31 HYDROCHLORIDE HYDRALAZINE 10 . 15 3 . 05 2 . 76 0 . 97 NAPROXEN ( + ) 25 .64 4 . 24 48 . 59 19 . 48 HYDROCHLORIDE NEOSTIGMINE BROMIDE 44 . 83 5 . 13 48 . 59 19 . 48 HYDROCHLOROTHIAZIDE 2 .55 0 . 37 5 . 31 0 . 25 NIFEDIPINE 14 .68 1 . 31 48 . 59 19 . 48 HYDROXYCHLOROQUINE 75 . 87 15 . 95 35 .62 8 . 16 NILUTAMIDE 48 .72 14 .62 35 . 62 8 . 16 SULFATE NIMODIPINE 62 .84 14 . 99 37 .73 10 . 54 HYDROXYUREA 3 . 31 0 . 45 5 . 31 0 . 25 NITROFURANTOIN 17 . 84 1 . 31 48 . 59 19 . 48 HYDROXYZINE 4 .01 1 . 05 5 . 31 0 .25 NORFLOXACIN 13. 59 2 . 17 48 . 59 19 . 48 PAMOATE NORTRIPTYLINE 18 . 16 2 . 89 48 . 59 19 . 48 IBUPROFEN 2 . 48 0 . 52 2 . 96 0 .78 NYLIDRIN 50 .07 12 .07 22 . 92 8 . 49 IMIPRAMINE 106 . 49 7 . 76 13 . 26 2 . 59 HYDROCHLORIDE HYDROCHLORIDE OLMESARTAN 55 .85 6 . 20 42. 76 12 . 68 INDAPAMIDE 126 . 12 2 . 79 1 .58 0 .63 MEDOXOMIL INDOMETHACIN 4 .52 1 . 34 2 .96 0 . 78 ORLISTAT 1 . 00 0 . 10 42 . 76 12 . 68 IPRATROPIUM BROMIDE 63. 39 20 .68 40 .33 10 . 95 ORPHENADRINE 54 .50 11. 76 22 .92 8 .49 IRBESARTAN 60 . 93 3 . 13 42 . 76 12 .68 CITRATE ISONIAZID 2 .41 0 . 55 2 . 96 0 .78 OXCARBAZEPINE 38 . 18 5 . 58 42 . 76 12 . 68 ISOSORBIDE DINITRATE 1 . 92 0 . 38 2 . 96 0 .78 PAROMOMYCIN 35 .29 3 . 13 17 .33 1 . 12 KETOCONAZOLE 108 . 35 2 . 80 1 .58 0 .63 SULFATE KETOPROFEN 44 . 26 11 . 34 14 .41 3 .42 PENTOXIFYLLINE 66 .47 12. 78 35 .62 8 . 16 KETOROLAC 52 . 39 14 . 15 35 .62 8 . 16 PERICIAZINE 81 .97 11. 21 19 . 15 5 . 36 TROMETHAMINE PERINDOPRIL 49 . 36 15 . 80 26 . 23 8 . 69 KETOTIFEN FUMARATE 1 . 77 0 . 83 25 . 52 4 . 20 ERBUMINE LABETALOL 54 . 37 11. 87 23 . 26 5 . 81 PERPHENAZINE 78 .78 17 . 35 18 . 45 4 . 73 HYDROCHLORIDE PHENAZOPYRIDINE 101. 46 8 . 17 24 . 35 12 . 85 LACTULOSE 80 .82 19 . 43 40 .33 10 . 95 HYDROCHLORIDE LANSOPRAZOLE 63 .87 1 . 81 37 . 73 10 . 54 PHENELZINE SULFATE 46 .68 8 . 55 48 . 59 19 .48 LEUCOVORIN CALCIUM 24 . 84 21 . 16 2 . 96 0 . 78 PHENYTOIN SODIUM 30 . 13 8 . 56 48 .59 19 . 48 LEVODOPA 81 . 18 3 . 71 26 .23 8 . 69 PIMOZIDE 31. 41 0 . 74 17 . 33 1 . 12 LEVOFLOXACIN 56 . 44 8 . 21 37 . 73 10 . 54 PINDOLOL 62. 64 22 .61 40 . 33 10 . 95 LIOTHYRONINE SODIUM 141 .46 10 . 60 12 . 35 2 .03 PIOGLITAZONE 84 . 58 14 . 90 42 . 76 12 .68 LISINOPRIL 48 .67 16 .98 26 . 23 8 .69 HYDROCHLORIDE LOPERAMIDE 55 . 50 12 . 86 20 . 02 5 .71 PIROXICAM 36 . 16 6 .65 40 . 33 10 . 95 HYDROCHLORIDE POTASSIUM 44 . 46 7 . 50 20 . 06 4 . 31 LORATADINE 44 . 26 3 . 86 37 . 73 10 . 54 P - AMINOBENZOATE LOSARTAN 35 . 45 4 . 03 42 . 76 12 . 68 PRAVASTATIN SODIUM 40 .51 11 . 81 26 . 23 8 .69 LOVASTATIN 32 . 18 10 . 01 37 . 73 10 . 54 PREGABALIN 47 . 81 15 . 36 26 .23 8 . 69 LOXAPINE SUCCINATE 65 .91 8 . 00 40 .33 10 . 95 PRIMAQUINE 89 .07 4 . 70 24 . 35 12 . 85 MAPROTILINE 0 .61 0 .29 2 . 96 0 . 78 DIPHOSPHATE HYDROCHLORIDE PRIMIDONE 45 . 23 5 . 07 40 . 33 10 . 95 MEBENDAZOLE 2 . 48 0 . 44 25 . 52 4 . 20 PROBENECID 71. 46 10 . 59 40 . 33 10 .95 MEFENAMIC ACID 57 . 21 4 . 90 40 . 33 10 . 95 PROCAINAMIDE 64 . 28 12 . 63 40 . 33 10 . 95 MEFLOQUINE 47 .01 9 . 07 12 . 35 2 .03 HYDROCHLORIDE MELOXICAM 59 . 46 11 . 56 37 .73 10 . 54 PROCHLORPERAZINE 4 . 88 0 . 44 20 .06 4 .31 MEMANTINE 53 . 24 12 . 40 9 .41 3 .67 EDISYLATE HYDROCHLORIDE PROCYCLIDINE 95 . 64 22 .09 40 .33 10 . 95 MERCAPTOPURINE 1 . 73 0 . 37 2 . 96 0 . 78 HYDROCHLORIDE METHAZOLAMIDE 54 .29 17 . 70 35 .62 8 . 16 PROMETHAZINE 105 . 40 7 .03 7 . 52 3 . 06 METHENAMINE 1 . 94 0 . 04 2 . 96 0 . 78 HYDROCHLORIDE METHOCARBAMOL 0 . 84 0 . 22 2 . 96 0 .78 PROPAFENONE 51 . 34 6 . 56 37 . 73 10 . 54 METHOTREXATE 43 .34 19 . 47 48 . 59 19 . 48 HYDROCHLORIDE METHOXSALEN 59. 05 18 . 46 48 . 59 19 . 48 PROPRANOLOL 66 .49 4 . 12 40 . 33 10 .95 METHYLDOPA 101. 58 5 .66 24 . 35 12 . 85 HYDROCHLORIDE ( + / - ) METOCLOPRAMIDE 37 . 87 2 . 00 48 . 59 19 . 48 PROPYLTHIOURACIL 35 . 91 2 . 49 16 .53 1 . 48 HYDROCHLORIDE PSEUDOEPHEDRINE 26 . 74 3 . 16 14 . 94 2 .65 METOLAZONE 68 .98 14 . 60 26 . 08 5 . 27 HYDROCHLORIDE METOPROLOL 71 . 55 16 . 46 24 . 35 12 . 85 PYRANTEL PAMOATE 34 . 17 3 . 87 12 .67 2 . 66 TARTRATE PYRAZINAMIDE 67 . 20 5 . 41 48 . 89 3 . 07 METRONIDAZOLE 27 .08 2 . 88 48 . 59 19 . 48 PYRIDOSTIGMINE 35 .78 3 . 60 17 . 33 1 . 12 MIDODRINE 53 .69 6 .41 35 .62 8 . 16 BROMIDE HYDROCHLORIDE QUINAPRIL 41. 55 4 . 83 17 .33 1 . 12 MINOXIDIL 33 . 66 3 . 31 48 . 59 19 . 48 HYDROCHLORIDE MITOXANTHRONE 52 .54 4 . 13 10 . 26 2 .72 QUININE SULFATE 21 . 34 4 . 35 14 . 94 2 .65 HYDROCHLORIDE RANITIDINE 40 . 18 8 . 86 17 .33 1 . 12 MODAFINIL 43 . 94 14 . 98 26 .23 8 .69 RIFAMPIN 95 . 53 5 . 13 7 .52 3 .06 MOXIFLOXACIN 51 .59 4 .29 37 . 73 10 .54 RIFAXIMIN 53 . 80 18 . 27 26 . 23 8 .69 HYDROCHLORIDE RILUZOLE 56 .54 15 . 74 26 .23 8 .69 MYCOPHENOLIC ACID 45 . 69 11. 70 12 .07 3 . 12 ROSIGLITAZONE 77 .63 8 .97 42 . 76 12 .68 NABUMETONE 48 .91 8 .07 35 .62 8 . 16 ROSUVASTATIN 35 .43 3 . 92 42 . 76 12 . 68 US 2019 / 0262353 A1 Aug . 29 , 2019 28

TABLE 2 - continued additive effects of minocycline and hydroxychloroquine in models of multiple sclerosis : Prospective combination treat Drug % Iron % ment for progressive disease ? Multiple sclerosis (Hound control control mills , Basingstoke , England ), 1352458517728811 (2017 ) . Name (mean ) SEM (mean ) SEM [0157 ] Live cell imaging over 12 h supported the neuro SERTRALINE 24. 23 4 .43 42. 76 12. 68 protective effects of drugs . We selected indapamide and HYDROCHLORIDE SILDENAFIL 49 . 31 2 . 47 37 . 73 10 . 54 desipramine for live imaging studies. FIG . 2b shows that SPIRAMYCIN 63. 97 11. 40 37 .73 10 . 54 while the number of neurons with intracellular propidium SPIRONOLACTONE 37 . 11 9 . 86 8 . 83 2 . 55 iodide (PI ) , a dye that leaks across a compromised plasma SULFAMETHOXAZOLE 16 . 23 2 .22 14 . 94 2 . 65 membrane , in response to FeSO4 exposure increases pro SULFASALAZINE 23 .36 2 .42 14 . 94 2 .65 SULFINPYRAZONE 46 .51 1 . 33 48 . 89 3 . 07 gressively over 12 h , this was significantly attenuated by SULFISOXAZOLE 38 .28 12 . 78 26 .23 8 .69 indapamide and desipramine. SULINDAC 34 .51 7 .87 11 . 70 2 . 97 [0158 ] The 35 hits were further narrowed concerning their TENOXICAM 25 .92 3 . 53 17 . 33 1 . 12 ability to cross the blood - brain - barrier according to drug TETRACYCLINE 25 . 04 6 . 42 11 . 70 2 . 97 HYDROCHLORIDE bank .ca , their side effect profile and tolerability . Although THEOPHYLLINE 23 .29 5 . 80 14 . 94 2 .65 antipsychotics are not well tolerated they were further THIOGUANINE 21 . 73 3 . 99 14 . 94 2 .65 included in the screening due to their good blood - brain THIOTHIXENE 6 . 80 1 . 01 26 . 23 8 .69 barrier penetrance . Out of these , a group of 23 compounds TIMOLOL MALEATE 11 . 07 1 . 14 14 . 94 2 . 65 TOLBUTAMIDE 9 . 09 2 .06 14 . 94 2 .65 was chosen for their ability to prevent mitochondrial damage TOLFENAMIC ACID 40 . 26 2 . 90 17 . 33 1 . 12 using rotenone , which inhibits the electron transfer from TOPIRAMATE 46 .07 15 . 57 26 .23 8 .69 complex I of the respiratory chain to ubiquinone . Rotenone TRANDOLAPRIL 72 . 30 6 . 44 42 . 76 12 .68 induced strong neurotoxicity to neurons (FIG . 3 ) . The tri TRANEXAMIC ACID 36 . 26 2 . 56 17 . 33 1 . 12 TRANYLCYPROMINE 21 . 59 3 . 15 14 . 94 2 .65 cyclic antidepressant trimipramine , the antipsychotics clo SULFATE zapine and periciazine , promethazine and the anti -hyperten TRAZODONE 25. 93 8 . 38 10 . 26 2. 72 sives labetalol, methyldopa and indapamide reduced HYDROCHLORIDE neurotoxicity while clomipramine trended towards a protec TRIFLUOPERAZINE 4 . 42 2 . 01 14 . 94 2 .65 HYDROCHLORIDE tive activity (FIG . 3A ) . The effect size of rescue by medi TRIHEXYPHENIDYL 30 .57 5 .61 14 .94 2 .65 cations was , however , small. Of note , rotenone induced HYDROCHLORIDE marked morphological neuronal changes with retraction of TRIMEPRAZINE 73 .31 7 . 34 7 . 52 3 .06 TARTRATE neurites (FIG . 3B ) . TRIMETHOPRIM 13 . 96 3 .09 14 .94 2 .65 [0159 ] Hydroxyl Radical Scavenging Capacity of Medi TRIMIPRAMINE 88 .62 11 .61 18 . 45 4 . 73 cations MALEATE [0160 ] The biochemical cell free hydroxyl radical antioxi URSODIOL 24 .62 2 . 10 14 . 94 2 . 65 dant capacity (HORAC ) assay investigates the prevention of VALSARTAN SODIUM 64 .68 10 .94 42. 76 12 .68 VANCOMYCIN 11. 70 8 . 21 12 . 95 5 . 13 hydroxyl radical mediated oxidation of to fluorescein in HYDROCHLORIDE comparison to the strong anti -oxidant gallic acid . The gen VENLAFAXINE 72 .52 10 . 20 37 .73 10 . 54 eration of hydroxyl radicals by a cobalt - driven Fenton - like VERAPAMIL 71 .08 13 . 71 40 . 33 10 . 95 HYDROCHLORIDE reaction oxidizes fluorescein with progressive loss of fluo YOHIMBINE 100 .09 4 . 40 9 .41 3 .67 rescence . The presence of an anti - oxidant reduces the loss of HYDROCHLORIDE fluorescence over time. As noted in FIG . 4A , gallic acid ZIDOVUDINE [AZT ] 66 .49 7 . 87 35 .62 8 . 16 reduced the loss of fluorescence (upward shift ) compared to ZOLMITRIPTAN 54. 88 8 . 92 42. 76 12. 68 a blank Fenton - driven reaction that is without antioxidant , while indapamide has an even higher activity . [ 0156 ] An example of iron toxicity and a drug screen is 10161 ]. We compared the area under the curve of test shown in FIG . 1 . Of all drugs tested , 35 compounds showed compounds to that elicited by gallic acid to obtain the gallic statistically significant protection from FeSO4-mediated acid equivalent (GAE ). A GAE of 1 represents hydroxyl neurotoxicity (FIG . 2a ). Of these , antipsychotics such as radical scavenging capacity similar to that of gallic acid , clozapine or periciazine , and tricyclic antidepressants such while a compound without anti - oxidant activity would pro as clomipramine or desipramine , exhibited strong protec duce a GAE close to 0 . Some of the compounds tested tion , as shown after normalization across at least 2 - 4 experi exhibited stronger antioxidative properties than gallic acid ments ( n of 4 wells of cells per experiment per test condi with HORAC -GAEs > 1 (FIG . 4C ). These included indap tion ) to the number of neurons of the respective control amide (mean HORAC -GAE 4 . 1 ; p < 0 .05 ; one -way analysis conditions (FIG . 2A ) . For example , while the average loss of of variance ( ANOVA ) with Dunnett ' s multiple comparisons neurons over 24 h in response to FeSO4 was 85 . 5 % (i .e . test as posthoc analysis vs . gallic acid ), mitoxantrone (5 . 6 ; 14 . 5 % of surviving neurons compared to 100 % of controls ) , p < 0 . 001) , chlorpromazine ( 5 . 9 ; p < 0 . 001) , clozapine ( 4 .6 ; clomipramine at 10 um completely prevented neuronal loss p < 0 .05 ) and trimipramine (4 . 2; p < 0 .05 ). Although not sta ( 107. 3 % of controls ) . Other categories of medications with tistically significant compared to gallic acid , clomipramine neuroprotective actions against iron included anti -hyperten had a HORAC -GAE of 2 . 1. Regarding the comparison to the sives and some antibiotics . We note that minocycline , an blank situation ( i. e . no anti -oxidant present) , there was a antibiotic that reduces the conversion of a first demyelinat significant upward shift by clomipramine of the slope over ing event to clinically definite multiple sclerosis in a Phase 60 min (p < 0 .0001 ) (FIG . 46 ). Thus, although clomipramine 3 clinical trial was not included in the 1040 compounds ; in lacked significance against the strong anti - oxidative gallic a separate study , we find minocycline to completely prevent acid , the compound exhibited strong antioxidative effects iron neurotoxicity as well ( Faissner S , et al. Unexpected against the blank situation in the absence of any anti US 2019 /0262353 A1 Aug . 29 , 2019 29 oxidant ) . Interestingly , the tricyclic antidepressant desipra sought to evaluate the effect of clomipramine on B - cell mine had strong oxidative effects (HORAC -GAE - 5 . 00 ; activation . BCR / anti -CD40L / IL - 4 activation of B - cells p < 0 . 0001) . increased their proliferation and production of TNF - a ( FIG . [0162 ] Proliferation of T -Lymphocytes is Reduced by 6G , H ) and these were reduced in a concentration - dependent Antidepressants manner by clomipramine from 2 uM . [0163 ] We tested the capacity of compounds to affect [0168 ] We then investigated clomipramine in acute EAE . T - cell proliferation ( FIG . 5 ) . Splenocytes activated by anti Therapy with clomipramine from day 5 after induction of CD3 /anti - CD28 to trigger the proliferation of T - cells had MOG - EAE delayed onset of clinical signs by 2 days with a reduced incorporation of " [ H ] - thymidine upon treatment significantly better early disease course between days 11 and with dipyridamole (mean reduction 89 . 3 % ; p < 0 .0001 ; one 18 (FIG . 7A ) , which was reflected in an overall lower burden way ANOVA with Dunnett ' s multiple comparisons test as of disability (FIG . 7B ) . However, eventually , clomipramine post -hoc analysis compared to activated splenocytes ) , cefa treated animals succumbed to EAE and increased disability clor (23 % ; p < 0 .01 ), labetalol ( 26 . 8 % , p < 0 .0001 for this and (FIG . 7A ) subsequent compounds listed here ) , mefloquine (62 . 3 % ) , [0169 ] We then sought to investigate whether initiation of mitoxantrone ( 99 . 7 % ) , trimeprazine ( 43 . 3 % ) , chlorpromaz treatment from the the day of MOG - induction could ine ( 99 . 4 % ), periciazine ( 28 % ), promethazine ( 74 . 6 % ) , clo improve the outcome of EAE . Remarkably , early treatment mipramine (68 . 2 % ) , desipramine (92 . 2 % ) , imipramine ( 66 . initiation completely suppressed the manifestations of clini 4 % ) , trimipramine (54 % ) and doxepin ( 85 . 3 % , all p < 0 . cal signs (FIG . 8A ). While most animals in the vehicle group 0001 ) . Of note , methyldopa and memantine increased had a high disease burden , as shown by the sum of scores for proliferation (methyldopa 41. 4 % , p < 0 . 0001; memantine each individual animal (FIG . 8B ) and weight loss (FIG . 8C ) , 17 . 5 % , p < 0 .05 ) . Mitoxantrone and chlorpromazine , how this was profoundly ameliorated in treated mice over the ever , had toxic effects (data not shown ). course of study. PCR analyses of the spinal cord revealed [ 0164 ] Focus on Clomipramine In Vitro and in Acute and that the significant elevation in vehicle -EAE mice of tran Chronic EAE scripts encoding Ifng , Tnfa , Il - 17 and Cc12 were abrogated 101651. We selected clomipramine for further study as it is in clomipramine -EAE mice ( FIG . 8D ) . a well -tolerated antidepressant and crosses the blood -brain [0170 ] FIG . 11 (Panels A - L ) shows all 249 generic com barrier very well (drugbank .ca ). Moreover, in our assays, pounds of the iron mediated neurotoxicity screening . The clomipramine showed strong effects against iron mediated number of neurons left following exposure to each com neurotoxicity (mean anti -microtubule -associated protein - 2 pound was normalized to the number of neurons of the (MAP - 2 ) positive cells normalized to control of 107 . 3 % , respective control condition . The corresponding iron situa representing complete protection against iron toxicity ) ( FIG . tion was also normalized to the respective control ( red ) . 2 ) , had anti -oxidative properties (HORAC -GAE 2 . 1 where Compounds which exhibit significant protection are high the effect of the anti -oxidant gallic acid is normalized at lighted in yellow and marked ( X ) . Shown are the 1 )( FIG . 4) , and reduced T- lymphocyte proliferation (by means + SEM of 1 - 4 experiments, performed in quadrupli 68 . 2 % ) ( FIG . 5 ) . We began with a concentration response cates each . with the intent of investigating lower concentrations since [0171 ] Investigation of serum levels of clomipramine and plasma concentration in human of clomipramine as an its active metabolite , desmethylclomipramine (DMCL ), in anti - depressant average 122 ng /ml ( 387 nM ) (Rodriguez de mice sacrificed 1 h after the last of 16 daily clomipramine la Torre et al. , 2001 ) , but can peak to more than 600 nM in injections showed mean concentrations of 751 nM and 101 some individuals ( Thoren et al ., 1980 ) . FIG . 6A shows that nM , respectively (FIG . 8E ) . The corresponding mean spinal clomipramine had a progressive significant increase in neu cord levels were 28 uM and 1 . 5 uM ; a similar high brain to roprotection against iron toxicity from 100 nM . The effect plasma ratio of clomipramine was reported by Marty et al . was mediated in part by chelation with iron , as washing (Marty H , et al . Compared plasma and brain pharmacoki away clomipramine from neurons led to cell death , while netics of clomipramine and its metabolite demethylclomip pre - incubation with iron before application to neurons ramine in two strains of mice (NMRI and CD1) . Fundamen totally preserved neuronal viability ( FIG . 6B ) . We were able tal & clinical pharmacology 6 , 49 - 57 ( 1992 ) . ) in mice to observe the protection by clomipramine in a live - cell injected with a single 8 mg/ kg clomipramine IP . There was imaging study , in which the increasing number of PI a strong correlation of serum and spinal cord levels for both positive neurons over time in response to iron was attenu clomipramine and desmethylclomipramine across mice ated by clomipramine (FIG . 6C ) . (FIG . 88) . [0166 ] T -lymphocyte proliferation was reduced in a con [0172 ] Histological analysis of the spinal cord showed centration - dependent manner by clomipramine but signifi profound parenchymal inflammation in vehicle treated ani cant reduction occurred only from 5 uM ( p < 0 .01 ; one -way mals with a histological score of 4 . 3 , whereas clomipramine ANOVA with Dunnett' s multiple comparisons test as post treated animals only had few inflammatory cells in the hoc analysis compared to activated T -lymphocytes )) (FIG . meninges ( score 1 . 7 ; p < 0 . 001; non - parametric two - tailed 6D ) . This was reflected by a cell cycle arrest with more cells Mann -Whitney test ) (FIG . 9a , b , g ) that were inadequate to in G1 (p < 0 .05 ) and less in the S -phase (p < 0 .05 ) from 2 uM produce clinical manifestations as noted in FIG . 8a . Infil (FIG . 6E , F ) . tration in vehicle treated animals was accompanied by [ 0167 ] Due to the growing knowledge about the impor massive microglial activation , whereas clomipramine treat tance of B - cell follicular structures for progressive multiple ment prevented microglial activation , as assessed by lbal sclerosis (Romme Christensen et al. , 2013 ; Magliozzi R , et staining ( p < 0 .01 ) (FIG . 9c, d , h ) . Furthermore , clomip al. Meningeal B -cell follicles in secondary progressive mul ramine treated animals had significantly less axonal damage tiple sclerosis associate with early onset of disease and ( p < 0 .01 ) ( FIG . 9e , f, i ) . Infiltration and microglial activation severe cortical pathology . Brain 130 , 1089 - 1104 (2007 )) , we correlated with axonal injury (Spearman r = 0 .7599 , p < 0 .01 ; US 2019 /0262353 A1 Aug . 29 , 2019 30

Spearman r = 0 .774 , p < 0 .01 , respectively ; non -parametric [0178 ] One pathogenic hallmark important for the pro two - tailed Spearman correlation with 95 % confidence inter gression ofmultiple sclerosis is iron mediated neurotoxicity . val) (FIG . 9j, k ) . Iron accumulates in the CNS age - dependently (Stephenson 10173 ] We next set out to investigate the effect of clomip et al ., 2014 ) and iron deposition concomitant with T cell ramine in chronic EAE . We first evaluated clomipramine infiltration and the expression of inducible nitric oxide initiated only after the first relapse when mice were in synthase in microglia in the deep gray matter correlates with remission (day 31) . In our hands, using the more sensitive progression and is associated with neurodegeneration 15 - point EAE scoring system ( rather than the conventional (Haider et al. , 2014 ) . The deposition of iron amplifies 5 -point scale ), MOG - EAE mice can be documented to inflammation and exacerbates mitochondrial dysfunction undergo a second relapse after a remission period . Clomip through oxidative stress , eventually leading to neurodegen ramine did not affect the severity of the second relapse when eration (Friese et al. , 2014 ) . Targeting iron is thus considered initiated in mice at remission ( FIG . 10a ), likely because a promising therapeutic approach in progressive multiple substantial neural injury had already occurred from a pro sclerosis . We investigated the potential of promising generic longed EAE course . compounds to prevent iron mediated neurotoxicity . Out of [ 0174 ] In another experiment, we treated MOG - immu 249 compounds screened , 35 medications which prevented nized C57BL /6 mice from the first onset of clinical signs against iron mediated neurotoxicity were in the drug classes ( day 13 , FIG . 106 ) . Treatment with clomipramine attenuated of antidepressants ( n = 5 ) , antibiotics ( n = 4 ) , antipsychotics the marked rise in clinical disability and had a significant ( n = 3 ) , antimalarials ( n = 2 ) and others. Some of the drugs had positive effect during days 14 - 20 ( p = 0 .0175 ; non -parametric consistent outstanding neuroprotective effects , and these included antipsychotics and tricyclic antidepressants . The two - tailed Mann - Whitney test ). During remission , likely high number of antipsychotics and antidepressants as posi because the severity of disability was low , the vehicle and tive hits in the screening was striking . In addition to the clomipramine treated groups did not differ . Disease was then rescue effect against iron mediated neurotoxicity , several followed by a second increase in clinical scores in vehicle drugs showed promising results in other modes of toxicity ; treated mice , which was prevented by clomipramine (days these were desipramine , clozapine, indapamide and labetalol 42 -50 ; p = 0 .0007 ) . which were active against damage to the mitochondrial [0175 ] Another model of chronic EAE , thought to model respiratory chain . Data were corroborated by the investiga secondary progressive multiple sclerosis ( Al- Izki S , Pryce tion of antioxidative potential and the influence on spleno G , Jackson S J, Giovannoni G , Baker D . Immunosuppres cyte proliferation . Clomipramine showed outstanding sion with FTY720 is insufficient to prevent secondary pro effects in several in vitro settings such as against iron gressive neurodegeneration in experimental autoimmune mediated neurotoxicity , hydroxyl scavenging capacity , and encephalomyelitis . Multiple sclerosis (Houndmills , Basing inhibition of T- and B -cell proliferation ; in mice, clomip stoke, England ) 17 , 939 - 948 ( 2011 ) ; Hampton DW , et al. An ramine suppressed occurrence of disease in EAE com experimental model of secondary progressive multiple scle pletely , concomitant with reduced transcripts of chemotactic rosis that shows regional variation in gliosis , remyelination , and inflammatory cytokines in the spinal cord , reduced axonal and neuronal loss . Journal of neuroimmunology inflammation , microglial activation and preservation of 201 - 202 , 200 - 211 (2008 ) ), is immunization with spinal cord axons. Moreover, clomipramine ameliorated clinical signs in homogenate in the Biozzi ABH mouse . Clomipramine treat chronic EAE in two different EAE models, C57BL / 6 and ment was started at the onset of clinical signs where it Biozzi ABH mice . reduced clinical severity throughout the period of treatment [0179 ] The work presented here constitutes a systematic ( p = 0 .0062 ) ( FIG . 10c ) . approach to identify generic compounds that could be useful 0176 ]. In summary , clomipramine reduced clinical sever for the treatment of progressive multiple sclerosis . First, we ity in acute and chronic EAE in two differentmouse models . focused on ameliorating major hallmarks of progressive FIG . 10d schematizes that the initiation of clomipramine multiple sclerosis such as iron -mediated neurotoxicity , oxi treatment from onset of clinical signs of EAE attenuates the dative stress and immune cell proliferation . Second , we clinical disability observed during relapses or in chronic chose generic drugs which are available as oral formula disease . tions. The drugs have a well -known safety -profile , as there exists long - lasting experience in research and clinical use . DISCUSSION [0180 ] Some of the compounds that prevented iron -medi [ 0177 ] Unlike relapsing - remitting multiple sclerosis , trials ated neurotoxicity in our screen have been described previ in progressive multiple sclerosis have largely failed so far . ously to have neuroprotective properties and will be high One important explanation is the lack of directed actions of lighted here , as they may be of interest not only to medications against features that drive the pathophysiology progressive multiple sclerosis but also other CNS disorders of progressive multiple sclerosis , and the lack of consider with neurodegenerative features . Strong neuroprotective ation of penetration of agents into the CNS . The latter is effects were induced by tricyclic antidepressants . The anti important as the blood -brain barrier appears relatively intact depressant desipramine has been used in a Huntington ' s in progressive compared to the relapsing -remitting form disease model where it inhibited glutamate - induced mito ( Lassmann et al. , 2012 ) 5 , and pathogenic processes ongoing chondrial permeability at the concentration of 2 uM and led within the CNS may not be amendable to periphery - acting to reduced apoptosis of primary murine neurons (Lauterbach medications . To circumvent these challenges , we have EC . Neuroprotective effects of psychotropic drugs in Hun employed bioassay screens that model aspects of progres tington ' s disease . International journal of molecular sciences sive multiple sclerosis . Moreover, we have opted to test 14 , 22558 - 22603 ( 2013 ) ; Tang T S , et al. Disturbed Ca2 + generic medications that have data of good access into the signaling and apoptosis of medium spiny neurons in Hun CNS . tington ' s disease. Proceedings of the National Academy of US 2019 /0262353 A1 Aug . 29 , 2019 31

Sciences of the United States of America 102 , 2602 - 2607 for all three , drugbank .ca ) as well as clomipramine (97 .9 % ( 2005 ) ) . Furthermore, desipramine induces the anti - oxida chance for entering the CNS according to drugbank .ca ) . tive enzyme heme- oxygenase 1 in Mes23 . 5 dopaminergic Thus , we did not explore their utility in EAE . cells and increases Nrf2 accumulation in the nucleus, thus [0185 ] Mitoxantrone is used in some countries as a treat preventing neuronal cell death mediated by rotenone and ment for progressive multiple sclerosis , but has so far not yet 6 -hydroxydopamine ( Lin H Y , et al. Desipramine protects been described as being neuroprotective . Although the neuronal cell death and induces heme oxygenase - 1 expres blood -brain -barrier permeability probability is poor sion in Mes23 . 5 dopaminergic neurons. PloS one 7 , e50138 ( 0 .7979 ) , it may be postulated that the effect in progressive ( 2012 ) . multiple sclerosis , in addition to its toxic effects on T - lym [0181 ] Besides desipramine , other tricyclic antidepres phocytes , is induced by its capacity to limit iron -mediated sants had strong effects against splenocyte proliferation . neurotoxicity . Indapamide exhibited strong neuroprotective Imipramine, which showed good neuroprotective properties , effects against iron toxicity in culture , which has not yet enhances PEP - 1 -catalase in astrocytes , leading to neuropro been described previously . More interestingly , indapamide tection in the hippocampal CA1 region in an ischemia model also overcomes mitochondrial damage . As indapamide has (Kim D W , et al. Imipramine enhances neuroprotective no effect on T- lymphocyte proliferation , the drug may not effect of PEP - 1 - Catalase against ischemic neuronal damage . overcome acute -EAE , but may be interesting in longer term BMB reports 44 , 647 -652 ( 2011 ). ) Additionally , it prevents multiple sclerosis models such as the Biozzi ABH mouse apoptosis of neural stem cells by lipopolysaccharide , medi model , which shows immune cell - independent neurodegen ated by the brain derived neurotrophic factor (BDNF ) and eration 35 and a chronic disease course 22 . mitogen - activated protein kinase (MAPK ) pathway (Peng C [0186 ] As noted in FIG . 17 , indapamide alleviates oxida H , et al. Neuroprotection by Imipramine against lipopoly tive stress observed in the spinal cord following demyeli saccharide- induced apoptosis in hippocampus - derived neu nation induced by lysolecithin in this area . Specifically , the ral stem cells mediated by activation of BDNF and the lysolecithin injury to the spinal cord particularly in aging MAPK pathway. European neuropsychopharmacology : the 8 - 10 month old mice ( thought to reflect middle age in journal of the European College of Neuropsychopharmacol humans, an age commonly associated with progression of ogy 18 , 128 - 140 (2008 )) . Another novel compound recently disability in primary progressive and secondary progressive developed , quinpramine , which is a fusion of imipramine MS ) led to the activation of NADPH oxidase , whose acti and the anti -malarial quinacrine , decreased the number of vation has also been noted in MS particularly in progressive inflammatory CNS lesions , antigen - specific T - cell prolifera MS (Haider L , Fischer M T , Frischer J M , Bauer J , Hoft tion and pro - inflammatory cytokines in EAE ( Singh MP, et berger R , Botond G , Esterbauer H , Binder CJ, Witztum J L , al. Quinpramine is a novel compound effective in amelio Lassmann H , Oxidative damage in multiple sclerosis rating brain autoimmune disease . Exp Neurol 215 , 397 - 400 lesions ., Brain 134 : 1914 - 1924 , 2011 ) . Treatment with inda (2009 ) . ) . pamide reduces oxidative stress -mediated lipid oxidation as [0182 ] Due to structural similarities between clomip indicated by measurement of malondialdehyde expression ramine , imipramine and trimipramine it may be speculated within the demyelinated lesion , and resulted in reduced that these compounds may be relevant for trials in progres myelin and axonal loss caused by the lysolecithin (FIG . 17 ) . sive multiple sclerosis . Furthermore , we showed previously [0187 ] We opted to test clomipramine in the acute -EAE that doxepin reduces microglial activation to 46 % without model due to its strong effects on immune cells , its antioxi inducing toxicity ; clomipramine , however, did not have dative properties and its prevention against iron mediated microglia inhibitory activity 14 . In the synopsis of effects neurotoxicity . Clomipramine is a tricyclic antidepressant contributing to progressive multiple sclerosis , tricyclic anti which is used to treat depression , obsessive compulsive depressants are interesting for further development and disorder and panic disorders , usually in a dosage of 100 - 150 might even be suitable as combination therapy with other mg/ d , sometimes up to 300 mg/ d . It inhibits serotonin and compounds targeting features of progressive multiple scle norepinephrine uptake . Clomipramine reduces the seizure rosis. threshold and overdose can lead to cardiac dysrhythmias , [0183 ] Some antipsychotics also displayed strong protec hypotension and coma ( drugbank .ca ) . Usually, clomip tion against iron and oxidative stress . Clozapine has been ramine is well tolerated , but side effects include amongst described to reduce microglial activation through inhibition others increase in weight, sexual dysfunctions, sedation , of phagocytic oxidase (PHOX ) - generated reactive oxygen hypotension and anticholinergic effects such as dry mouth , species production , mediating neuroprotection (Hu X , et al. sweating , obstipation , blurred vision and micturition disor Clozapine protects dopaminergic neurons from inflamma der (according to the manufacturer leaflet) . Clomipramine tion - induced damage by inhibiting microglial overactiva crosses readily into the CNS with a probability to cross the tion . Journal of neuroimmune pharmacology : the official blood brain barrier of 0 .979 according to predicted ADMET journal of the Society on NeuroImmune Pharmacology 7 , ( absorption , distribution , metabolism , excretion , toxicity ) 187 - 201 ( 2012 ) ) . The strong anti -oxidative properties of features ( drugbank . ca ) . Clomipramine reduces the produc clozapine in the HORAC assay support these results . Due to tion of nitric oxide and TNF - a in microglia and astrocytes the side effect profile with enhanced risk of agranulocytosis , (Hwang et al. , 2008 ) ; the authors reported neuroprotective we refrained from usage in EAE ; nevertheless, in multiple properties in a co - culture model of neuroblastoma cells and sclerosis patients with psychiatric comorbidities and eligible microglia . Clomipramine increases the uptake of cortisol in for antipsychotic treatment, it may be reasonable to use primary rat neurons (Pariante et al. , 2003 ) and promotes the clozapine . release of glial cell line -derived neurotrophic factor in [0184 ] With regards to liothyronine , atenolol or carvedilol glioblastoma cells , suggesting a protective effect on neurons that prevented iron -mediated neurotoxicity beyond levels of (Hisaoka et al. , 2001) . The drug has been also studied in controls , these do not penetrate the CNS (probability of 68 % experimental autoimmune neuritis , where it decreases the US 2019 /0262353 A1 Aug . 29 , 2019 32 number of IFN -y secreting Th1 cells and ameliorated the 52679 ) , we found that an identical lysolecithin insult to the clinical course (Zhu et al . , 1998 ) . spinal cord produces by 24 h to 72 h a larger volume of [0188 ] Clomipramine has been used previously in mice in demyelination and axonal loss in 8 - 10 months old mice different dosages to study conditions such as anti -nocicep compared to young 6 weeks old animals (FIG . 12 , 13 ) . tion ( 0 . 5 mg/ kg ) (Schreiber et al . , 2015 ) , Chagas disease (7 . 5 [0192 ] FIG . 14 shows RNAseq data of 3 day laser -micro mg/ kg ) (Garcia et al. , 2016 ) and neurotransmitter and his dissected lesions that homed onto NADPH oxidase . a ) Heat tone deacetylase expression (50 mg/kg ) (Ookubo et al . , map ( 3 samples/ group , where each sample is a pool of 5 2013 ) . In humans taking clomipramine as an anti -depres mice ) after lysolecithin (LPC ) lesion in young and aging sant, mean serum levels after a mean daily intake of 127 + 91 mice . b ) Upregulation of canonical immune- associated path mg/ d have been reported to be 122 ng/ ml ( 387 nM , consid ways in aging vs young mice that converge, through Inge ering a molecular weight of 314 . 9 ) (Rodriguez de la Torre et nuity Pathway Analysis , into NADPH oxidase 2 subunits . d ) al ., 2001 ) . Of note , clomipramine levels after oral intake in The RNAseq levels of the catalytic subunit of NADPH humans have a wide range , leading to plasma concentrations oxidase 2 , gp91 phox ( also called CYBB ) are selected for of more than 600 nM in some individuals ( Thoren et al. , display. * p < 0 .05 . 1980 ), which is in the range of neuroprotection against iron [0193 ] FIG . 15 shows higher expression of gp91phox (an in our in vitro experiments . The injection of 20 mg/ kg IP in NADPH oxidase subunit ) and malondialdehyde in aging CD1 mice leads to peak plasma concentrations of 438 ng / ml lesions . a , b ) The catalytic subunit of NOX2, gp91phox , is ( 1 . 4 uM ) with a half - life of 165 min (Marty et al. , 1992 ) , and readily found within CD45 + cells in aging but not young in our experiments animals ( sacrificed 1 h after the last demyelinated lesions (d3 ) . ( c , d ) Similarly , malondialdehyde injection ) had mean serum clomipramine concentrations of as a marker of oxidative damage is in aging lesion associated 236 . 5 ngéml (751 nM ) . These plasma levels are close to the with MBP + myelin breakdown . ones measured in humans ( average of 387 nM , and up to 600 [0194 ] Since we found oxidative stress more prevalent nM ( Thoren et al. , 1980 ) ) , especially keeping in mind that within the lysolecithin lesion of the aging mice , we tested plasma levels drop faster in mice due to the relatively bigger indapamide , a well- tolerated angiotensin converting enzyme liver :body mass and that the half -life of clomipramine in inhibitor used as an anti - hypertensive , as it has strong humans is between 17 . 7 and 84 hours (Balant -Gorgia et al. , anti - oxidant properties as described in the appended manu 1991 ) compared to about 2 . 5 h in mice . We found that script. Also , indapamide limits the neurotoxicity of the clomipramine levels in the spinal cord of the EAE -afflicted MS- relevant insult iron in culture . We thus treated aging mice averaged 28 uM ; levels achieved in the brains of 8 - 10 months old mice with intraperitoneal indapamide ( 20 humans are not known . Thus , the dosage of 25 mg/ kg mg /kg ) immediately after lysolecithin demyelination , and clomipramine tested in our EAE study reflects standard dose once per day at 20 mg/kg for the next 2 days . Spinal cord used in humans in that both attain similar plasma levels . tissues were taken for histology . We found that indapamide [0189 ] In summary , we discovered several generic com treated mice have a smaller volume of demyelination , less pounds in this systematic screening approach that exhibit axonal loss , and reduced lesional malondialdehyde (a neuroprotective properties against iron -mediated neurotox marker of oxidant- mediated injury ) level ( FIG . 16 ) than their icity . Additionally , some of those compounds preventmito vehicle -administered controls . These results suggest the chondrial damage to neurons, inhibit immune cell prolifera potential of indapamide as a medication for progressive MS. tion and show antioxidative capacities . Tricyclic antidepressants , antipsychotics and indapamide may be use LIST OF ABBREVIATIONS ful for further development in progressive multiple sclerosis [0195 ] BDNF: Brain - derived neurotrophic factor due to their manifold properties . Clomipramine showed 10196 ] DMSO : Dimethyl sulfoxide particular promise due to its capacity to reduce iron -medi [0197 ] EAE : Experimental autoimmune encephalomy ated neurotoxicity and T - and B - cell proliferation , its anti elitis oxidative effect, and its complete suppression of disease in [0198 ] FBS : Fetal bovine serum acute - EAE and positive effects in chronic EAE . 01991 GAEs : Gallic acid equivalents [0200 ] HORAC : Hydroxyl radical antioxidant capacity Example 2 [ 0201 ] INN : International nonproprietary name [ 0190 ] Indapamide Reduces Myelin and Axon Loss in an [ 0202 ] IP : Intraperitoneal MS Model: [0203 ] JAN : Japanese Accepted Name [0191 ] Active demyelinating lesions can be found in MS [0204 ] MAP -2 : Microtubule -associated protein -2 specimens of all ages sampled , including late in life . Indeed , [0205 ] MAPK : Mitogen -activated protein kinases age has been identified to be a factor in the dreaded [ 0206 ] MEM : Minimal essential medium conversion from relapsing - remitting into secondary progres 102071 PFA : Paraformaldehyde sive MS. Contributing causes for aging- associated worsen 0208 ] PI: Propidium iodide ing in MS that drives progression include the steady loss of axons with longevity of disease , or the deficient repair of [ 0209 ] PPMS: Primary - progressive multiple sclerosis myelin in older compared to younger patients . We tested the [0210 ] RRMS : Relapsing - remitting multiple sclerosis hypothesis that the same demyelinating injury is more [0211 ] USAN : United States Adopted Names devastating to axons and myelin as the individual ages. 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[0241 ] Lauterbach E C . Neuroprotective effects of psy cardiovascular effects of tricyclic antidepressants and chotropic drugs in Huntington ' s disease . International selective serotonin reuptake inhibitors in depressed journal of molecular sciences 2013 ; 14 (11 ) : 22558 - 603 patients . Therapeutic drug monitoring 2001; 23 ( 4 ) : 435 [ 0242] Li R , Rezk A , Miyazaki Y , Hilgenberg E , Touil H , 40 . Shen P , et al. Proinflammatory GM -CSF -producing B [0254 ] Romme Christensen J , Bornsen L , Ratzer R , Piehl cells in multiple sclerosis and B cell depletion therapy . F , Khademi M , Olsson T , et al. Systemic inflammation in Science translational medicine 2015 ; 7 (310 ) : 310ra166 . progressive multiple sclerosis involves follicular T - helper , [ 0243 ] Lin HY, Yeh W L , Huang B R , Lin C , Lai C H , Lin Th17 - and activated B - cells and correlates with progres H , et al. Desipramine protects neuronal cell death and sion . 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Alterations , modifications and variations ropsychopharmacology: the journal of the European can be effected to the particular embodiments by those of College of Neuropsychopharmacology 2008 ; 18 ( 2 ): 128 skill in the art . The scope of the claimsshould not be limited 40 . by the particular embodiments set forth herein , but should be [0251 ] Prineas J W , Kwon E E , Cho E S , Sharer L R , construed in a manner consistent with the specification as a Barnett M H , Oleszak E L , et al. Immunopathology of whole . secondary - progressive multiple sclerosis . Ann Neurol [0264 ] All publications , patents and patent applications 2001 ; 50 ( 5 ) : 646 - 57 . mentioned in this Specification are indicative of the level of [0252 ] Ransohoff R M , Hafler D A , Lucchinetti C F . skill those skilled in the art to which this invention pertains Multiple sclerosis - a quiet revolution . 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[0265 ] The invention being thus described , it will be 13. Use of one or more of dipyridamole , clopidogrel, obvious that the same may be varied in many ways. Such cefaclor, clarithromycin , erythromycin , rifampin , loper variations are not to be regarded as a departure from the amide , ketoconazole , labetalol, methyldopa , metoprolol, spirit and scope of the invention , and all such modification atenolol, carvedilol, indapamide, mefloquine , primaquine , as would be obvious to one skilled in the art are intended to mitoxanthrone , levodopa , trimeprazine , chlorpromazine , be included within the scope of the following claims. clozapine , periciazine , flunarizine , dimenhydrinate , diphen What is claimed is : hydramine , promethazine , phenazopyridine , yohimbine , 1 . A method of treating progressive multiple sclerosis memantine , liothyronine , clomipramine , desipramine , dox comprising administering to a subject in need thereof, a epin , imipramine, trimipramine, or functional derivative therapeutically effective amount of one or more of dipyrida thereof, for the treatment of progressive multiple sclerosis in mole , clopidogrel, cefaclor, clarithromycin , erythromycin , a subject. rifampin , loperamide, ketoconazole , labetalol, methyldopa, 14 . Use of one or more of dipyridamole , clopidogrel , metoprolol, atenolol, carvedilol, indapamide, mefloquine , cefaclor, clarithromycin , erythromycin , rifampin , loper primaquine , mitoxanthrone , levodopa , trimeprazine , chlo amide , ketoconazole , labetalol, methyldopa , metoprolol, rpromazine, clozapine , periciazine , flunarizine , dimenhydri atenolol , carvedilol, indapamide, mefloquine , primaquine , nate , diphenhydramine , promethazine, phenazopyridine , mitoxanthrone, levodopa , trimeprazine , chlorpromazine , yohimbine , memantine , liothyronine , clomipramine , desip clozapine, periciazine , flunarizine , dimenhydrinate, diphen ramine , doxepin , imipramine , trimipramine , or functional hydramine , promethazine , phenazopyridine , yohimbine , derivative thereof . memantine , liothyronine, clomipramine , desipramine , dox 2 . A method of treating progressive multiple sclerosis epin , imipramine , trimipramine , or functional derivative comprising administering to a subject in need thereof, a thereof, in the manufacture of a medicament for the treat therapeutically effective amountof clomipramine , or a func ment of progressive multiple sclerosis in a subject . tional derivative thereof. 15 . A use of clomipramine , or a functional derivative 3 . A method of treating progressive multiple sclerosis thereof, for treating progressive multiple sclerosis in a comprising administering to a subject in need thereof, a subject in need thereof. therapeutically effective amount of imipramine, or a func 16 . A use of clomipramine, or a functional derivative tional derivative thereof. thereof, in the manufacture of a medicament for treating 4 . A method of treating progressive multiple sclerosis progressive multiple sclerosis in a subject in need thereof. comprising administering to a subject in need thereof, a 17 . A use of imipramine , or a functional derivative therapeutically effective amount of trimipramine, or a func thereof, for treating progressive multiple sclerosis in a tional derivative thereof. subject in need thereof. 5 . A method of treating progressive multiple sclerosis 18 . A use of imipramine , or a functional derivative comprising administering to a subject in need thereof, a thereof, in the manufacture of a medicament for treating therapeutically effective amount of clomipramine , or a func progressive multiple sclerosis in a subject in need thereof. tional derivative thereof, and a therapeutically effective amount of indapamide , or a functional derivative thereof. 19 . A use of trimipramine , or a functional derivative 6 . A method of treating progressive multiple sclerosis thereof, for treating progressive multiple sclerosis in a comprising administering to a subject in need thereof, a subject in need thereof. therapeutically effective amount of indapamide , or a func 20 . A use of a therapeutically effective amount of trim tional derivative thereof. ipramine , or a functional derivative thereof, in the manu 7 . A method of treating progressive multiple sclerosis facture of a medicament for treating progressive multiple comprising administering to a subject in need thereof, a sclerosis in a subject in need thereof. therapeutically effective amount of indapamide, or a func 21. A use of clomipramine , or a functional derivative tional derivative thereof, and one or more of hydroxychlo thereof, and a use of indapamide , or a functional derivative roquine , minocycline, or clomipramine or a functional thereof , for treating progressive multiple sclerosis in subject derivative thereof. in need thereof. 8 . The method of any one of claims 1 to 7 , wherein said 22 . A use of clomipramine , or a functional derivative multiple sclerosis is primary progressive multiple sclerosis . thereof, and a use of indapamide , or a functional derivative 9 . The method of any one of claims 1 to 7 , wherein said thereof , in the manufacture of a medicament for treating multiple sclerosis is secondary progressive multiple sclero progressive multiple sclerosis in subject in need thereof. sis . 23 . A use of indapamide, or a functional derivative 10 . The method of any one of claims 1 to 7 , wherein said thereof , for treating progressive multiple sclerosis in subject multiple sclerosis is progressive relapsing multiple sclerosis . in need thereof. 11 . The method of any one of claims 1 to 10 , wherein said 24 . A use of indapamide, or a functional derivative treatment further comprises administering a therapeutically thereof, in the manufacture of a medicament for treating effective amount of Laquinimod , Fingolimod , Masitinib , progressive multiple sclerosis in subject in need thereof. Ocrelizumab , Ibudilast , Anti -LINGO - 1 ,MD1003 (high con 25 . A use of indapamide , or a functional derivative centration Biotin ) , Natalizumab , Siponimod , Tcelna ( imile thereof, and one or more of hydroxychloroquine , minocy cleucel - T ) , Simvastatin , Dimethyl fumarate , Autologous cline , and clomipramine , or a functional derivative thereof, haematopoietic stem cell transplantation , Amiloride , Rilu for treating progressive multiple sclerosis in subject in need zole , Fluoxetine, Glatiramer Acetate , Interferon Beta , or a thereof . functional derivative thereof. 26 . A use of indapamide , or a functional derivative 12 . The method of any one of claims 1 to 9 , wherein said thereof, and one or more of hydroxychloroquine , minocy subject is a human . cline , and clomipramine , or a functional derivative thereof, US 2019 / 0262353 A1 Aug . 29 , 2019 36

in the manufacture of a medicament for treating progressive atenolol, carvedilol, indapamide , mefloquine, prima multiple sclerosis in subject in need thereof quine, mitoxanthrone , levodopa , trimeprazine , chlo 27 . The use of any one of claims 13 to 26 , wherein said rpromazine , clozapine , periciazine , flunarizine , dimen multiple sclerosis is primary progressive multiple sclerosis . hydrinate , diphenhydramine , promethazine , 28 . The use of any one of claims 13 to 26 , wherein said phenazopyridine , yohimbine , memantine, liothyronine , multiple sclerosis is secondary progressive multiple sclero clomipramine , desipramine , doxepin , imipramine , sis . trimipramine , or functional derivative thereofl ; and 29 . The use of any one of claims 13 to 26 , wherein said multiple sclerosis is progressive relapsing multiple sclerosis . b . Instructions for the use thereof. 30 . The use of any one of claims 13 to 29 , further 35 . A kit for the treatment of progressive multiple scle comprising a use of a therapeutically effective amount of rosis comprising : a therapeutically effective amount of clo Laquinimod , Fingolimod , Masitinib , Ocrelizumab , Ibudi mipramine , or a functional derivative thereof, and instruc last, Anti -LINGO - 1 , MD1003 (high concentration Biotin ) , tions for use . Natalizumab , Siponimod , Tcelna ( imilecleucel - T ) , Simvas 36 . A kit for the treatment of progressive multiple scle tatin , Dimethyl fumarate , Autologous haematopoietic stem rosis comprising : a therapeutically effective amount of imi cell transplantation , Amiloride , Riluzole , Fluoxetine , Glati pramine , or a functional derivative thereof, and instructions ramer Acetate , Interferon Beta , or a functional derivative for use . thereof, for the treatment of progressive multiple sclerosis , 37 . A kit for the treatment of progressive multiple scle primary progressive multiple sclerosis , or secondary mul rosis comprising : a therapeutically effective amount of trim tiple sclerosis . ipramine , or a functional derivative thereof, and instructions 31. The use of any one of claims 13 to 29 , further for use . comprising a use of a therapeutically effective amount of 38 . A kit for the treatment of progressive multiple scle Laquinimod , Fingolimod , Masitinib , Ocrelizumab , Ibudi rosis comprising : a therapeutically effective amount of clo last, Anti - LINGO - 1 , MD1003 (high concentration Biotin ) , mipramine, or a functional derivative thereof, a therapeuti Natalizumab , Siponimod , Tcelna (imilecleucel - T ) , Simvas cally effective amount indapamide, or a functional tatin , Dimethyl fumarate , Autologous haematopoietic stem derivative thereof, and instructions for use . cell transplantation , Amiloride , Riluzole , Fluoxetine, Glati 39 . A kit for the treatment of progressive multiple scle ramer Acetate , Interferon Beta , or a functional derivative rosis comprising : a therapeutically effective amount of inda thereof, in the manufacture of a medicament for the treat ment of progressive multiple sclerosis , primary progressive pamide , or a functional derivative thereof, and instructions multiple sclerosis , or secondary multiple sclerosis . for use . 32 . The use according to any one of claims 13 to 31 , 40 . A kit for the treatment of progressive multiple scle wherein the subject is a human . rosis comprising: a therapeutically effective amount of inda 33 . A method of identifying a compound for the treatment pamide , or a functional derivative thereof, and one or more of progressive multiple sclerosis , comprising : of hydroxychloroquine, minocycline , or clomipramine , or a ( a ) selecting one or more compounds from a library of functional derivative thereof , and instructions for use . compounds that prevent or reduce iron -mediated neu 41. The kit of any one of claims 34 to 40, wherein said rotoxicity in vitro , multiple sclerosis is primary progressive multiple sclerosis . (b ) selecting one or more compounds from step ( a ) that 42 . The kit of any one of claims 34 to 40 , wherein said prevent or reduce mitochondrial damage in vitro ; multiple sclerosis is secondary progressive multiple sclero ( c ) selecting one or more compounds from step ( a ) for sis . antioxidative properties , ( d ) selecting one or more compound from step (a ) for 43 . The kit of any one of claims 34 to 40 , wherein said ability to reduce T- cell proliferation in vitro , multiple sclerosis is progressive relapsing multiple sclerosis . ( e ) optionally, after step ( a ) , selecting a compound from 44 . The kit of any one of claims 34 to 43 , further step ( a ) which is predicted or know to be able to cross comprising one or more of Laquinimod , Fingolimod , Masi the blood brain barrier , or having a suitable side effect tinib , Ocrelizumab , Ibudilast, Anti -LINGO - 1 , MD1003 profile , or having a suitable tolerability. (high concentration Biotin ), Natalizumab , Siponimod , 34 . A kit for the treatment of progressive multiple scle Tcelna (imilecleucel - T ) , Simvastatin , Dimethyl fumarate , rosis, comprising: Autologous haematopoietic stem cell transplantation , a . one or more of dipyridamole , clopidogrel , cefaclor, Amiloride, Riluzole , Fluoxetine , Glatiramer Acetate , Inter clarithromycin , erythromycin , rifampin , loperamide , feron Beta , or a functional derivative thereof. ketoconazole , labetalol, methyldopa , metoprolol,