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Medchemexpress/Chloroquine diphosphate/HY-17589/10mM*1mL in Water
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Medchemexpress/Chloroquine diphosphate/HY-17589/10mM*1mL in Water
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Chloroquine(diphosphate)isanantimalarialdrugandautophagy/lysosomeinhibitor,inhibitingToll-likereceptor(TLR)signalling.

CustomerValidation

  • SciRep.2017Jun7;7(1):2929.
  • TumourBiol.2016Jul;37(7):8811-24.
  • IntJMolSci.2017May19;18(5).pii:E1063.
  • ActaBiochimBiophysSin(Shanghai).2016Dec;48(12):1075-1084.
  • EBioMedicine.2017May;19:49-59.
Description

Chloroquine(diphosphate)isanantimalarialdrugandautophagy/lysosomeinhibitor,inhibitingToll-likereceptor(TLR)signalling.

InVitro

Chloroquine(CHQ,20μM)inhibitsIL-12p70releaseandreducesTh1-primingcapacityofactivatedhumanmonocyte-derivedLangerhans-likecells(MoLC).Chloroquine(CHQ,20μM)enhancesIL-1–inducedIL-23secretioninMoLCandsubsequentlyincreasesIL-17AreleasebyprimedCD4+Tcells[1].Chloroquine(25μM)suppressesMMP-9mRNAexpressioninnormoxiaandhypoxiainparentalMDA-MB-231cells.Chloroquinehascell-,dose-andhypoxia-dependenteffectsonMMP-2,MMP-9andMMP-13mRNAexpression[2].TLR7andTLR9inhibitionusingIRS-954orchloroquinesignificantlyreducesHuH7cellproliferationinvitro[3].

InVivo

Chloroquine(80mg/kg,i.p.)doesnotpreventthegrowthofthetriple-negativeMDA-MB-231cellswithhighorlowTLR9expressionlevelsintheorthotopicmousemodel[2].TLR7andTLR9inhibitionusingIRS-954orchloroquinesignificantlyinhibitstumourgrowthinthemousexenograftmodel.HCCdevelopmentintheDEN/NMORratmodelisalsosignificantlyinhibitedbychloroquine[3].

ClinicalTrial
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References
  • [1].SaidA,etal.ChloroquinepromotesIL-17productionbyCD4+Tcellsviap38-dependentIL-23releasebymonocyte-derivedLangerhans-likecells.JImmunol.2014Dec15;193(12):6135-43.

    [2].TuomelaJ,etal.Chloroquinehastumor-inhibitoryandtumor-promotingeffectsintriple-negativebreastcancer.OncolLett.2013Dec;6(6):1665-1672.

    [3].MohamedFE,etal.Effectoftoll-likereceptor7and9targetedtherapytopreventthedevelopmentofhepatocellularcarcinoma.LiverInt.2014Jul2.doi:10.1111/liv.12626.

PreparingStockSolutions
ConcentrationVolumeMass1mg5mg10mg
1mM1.9385mL9.6926mL19.3851mL
5mM0.3877mL1.9385mL3.8770mL
10mM0.1939mL0.9693mL1.9385mL
Pleaserefertothesolubilityinformationtoselecttheappropriatesolvent.
CellAssay
[2]

Thecellsareculturedin6-wellplateswithnormalculturemediuminthepresenceofvehicleor25or50μMchloroquine,untilnearconfluency,afterwhichtheyarerinsedwithsterilephosphate-bufferedsaline(PBS)andculturedfurtherfortheindicatedtimesinserum-freeculturemedium.Atthedesiredtime-points,theculturemediumisdiscardedandthecellsarequicklyharvestedinlysisbufferandclarifiedbycentrifugation.Subsequenttoboilingthesupernatantsinreducingsodiumdodecylsulphate(SDS)samplebuffer,equalamountsofprotein(100μg)areloadedperlaneandthesamplesareelectrophoresedinto10or4-20%grADIentpolyacrylamideSDSgels,thentransferredtoanitrocellulosemembrane.TodetectTLR9,theblotsareincubatedovernightat4°Cwithanti-TLR9antibodies,diluted1:500inTris-bufferedsalinewith0.1%(v/v)Tween-20(TBST).Equalloadingisconfirmedwithpolyclonalrabbitanti-actin.Secondarydetectionisperformedwithhorseradishperoxidase-linkedsecondaryantibodies.TheproteinbandsarevisualizedbychemiluminescenceusinganECLkit.MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.

AnimalAdmiNISTration
[2]

ChloroquinediphosphateisdissolvedinPBS.

ControlandTLR9siRNAMDA-MB-231cells(5×105 cellsin100μL)areinoculatedintothemammaryfatpadsoffour-week-old,immune-deficientmice(athymicnude/nuFoxn1).Treatmentsarestartedsevendaysaftertumorcellinoculation.Themicearetreateddailyeitherwithintraperitoneal(i.p.)chloroquine(80mg/kg)orvehicle(PBS).Theanimalsaremonitoreddailyforclinicalsigns.TumormeasurementsareperformedtwiceaweekandtumorvolumeiscalculatedaccordingtotheformulaV=(π/6)(d1×d2)3/2,whered1 andd2 areperpendiculartumordiameters.Thetumorsareallowedtogrowfor22days,atwhichpointthemicearesacrificedandthetumorsaredissectedforafinalmeasurement.Throughouttheexperiments,theanimalsaremaintainedundercontrolledpathogen-freeenvironmentalconditions(20-21ºC,30-60%relativehumidityanda12-hlightingcycle).Themicearefedwithsmall-animalfoodpelletsandsuppliedwithsterilewater adlibitum. MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.

References
  • [1].SaidA,etal.ChloroquinepromotesIL-17productionbyCD4+Tcellsviap38-dependentIL-23releasebymonocyte-derivedLangerhans-likecells.JImmunol.2014Dec15;193(12):6135-43.

    [2].TuomelaJ,etal.Chloroquinehastumor-inhibitoryandtumor-promotingeffectsintriple-negativebreastcancer.OncolLett.2013Dec;6(6):1665-1672.

    [3].MohamedFE,etal.Effectoftoll-likereceptor7and9targetedtherapytopreventthedevelopmentofhepatocellularcarcinoma.LiverInt.2014Jul2.doi:10.1111/liv.12626.

MolecularWeight

515.86

Formula

C₁₈H₃₂ClN₃O₈P₂

CASNo.

50-63-5

Storage
Powder-20°C3years
 4°C2years
Insolvent-80°C6months
 -20°C1month
Shipping

RoomtemperatureincontinentalUS;mayvaryelsewhere

Solvent&Solubility

H2O:≥33mg/mL;DMSO:<0.1="">

*"<1 mg/ml"="" means="" slightly="" soluble="" or="" insoluble.="" "≥"="" means="" soluble,="" but="" saturation="">

Purity:99.93%