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EvaluatingtheEfficacyofDiaMetrixinaRandomized,DoubleBlind PlaceboControlledHumanClinicalTrial KennethR.Hampshire1,StevenMiles1,RickMiles1,RobertT.Streeper2,JoelMichalek3, ChristopherLouden3,ElzbietaIzbicka2&

1

SyntratechCorporation,Denver,Colorado,Colorado,USA,80237,8007380650,

www.syntratech.com,
2 3

CPCLTD,SanAntonio,Texas,USA UniversityofTexasHealthScienceCenteratSanAntonio,SanAntonio,Texas,USA

&correspondingauthor

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ABSTRACT Background:Peoplewithdiabetesareathighriskofcardiaccomplications,whichare exacerbatedbyelevatedbloodpressureanddysfunctionalsugarandfatmetabolism. Theaimofthisstudywastoevaluateeffectsofanherbalsupplement,DiaMetrix,on bloodglucoselevels,triglycerides,cholesterol,andbloodpressureinsubjectswith chronicuncontrolledbloodglucose. Methods:Wedidarandomizedprospectivedoubleblindedstudyin100subjects(47 menand53women)withfastingbloodglucoselevelsbetween160and240mg/dL.The participantswererandomlyassignedtooralDiaMetrixat6tabletsdaily(n=50)or placebo(n=50)for90days.Theendpointsincludedfastingbloodglucose,bodymass, oralglucosetolerancetest,totalcholesterol,HDL,triglycerides,glycosylated hemoglobinandbloodpressure. Results:Allstudyparticipantscompletedthestudy.DiaMetrixreducedHbA1c,blood glucose(fastingandafterchallenge),triglycerides,systolicanddiastolicbloodpressure, totalcholesterol,andbodymassovertime.Meanfastingglucosewassignificantly decreasedinsubjectstakingDiaMetrixat30,60and90daysafterbaselinerelativeto thosetakingplacebo.HbA1cmeanchangefrombaselineto90dayswassignificantly decreasedinsubjectstakingDiaMetrix.TheDiaMetrixmeanserumglucosewas significantlydecreasedrelativetoplaceboat30,120and180minutesafterthe challenge.Triglyceridesmeanchangesfrombaselineweresignificantlydecreasedby

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DiaMetrixat30,60and90days.Fortotalcholesterol,DiaMetrixmeanchangefrom baselineto90dayswassignificantlydecreasedrelativetoplacebo.Highandlow densitylipoproteinswerenotsignificantlyaffected.Systolicanddiastolicblood pressurewassignificantlydecreasedat6090days,andmeanbodymasswas significantlydecreasedrelativetoplaceboat90days. Conclusions:Thisstudydemonstratedareductioninthemeanfastingbloodsugar, bloodpressureandbodymassinsubjectsrandomizedtoDiaMetrix.Thesefindingsof suggestthatDiaMetrix,anoverthecounterpreparation,deservesfurtherevaluationof itsactivityinlargerclinicalstudies.

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Introduction Basedonthedatafromthe2007NationalDiabetesFactSheetpublishedbythe AmericanDiabetesAssociation,23.6millionchildrenandadultsintheUnitedStates (7.8%ofthepopulation)havediabetesand1.6millionnewcasesofdiabetesare diagnosedinpeopleaged20yearsandoldereachyear(1).In20032004,therewerean estimated5.7millionpeoplewithundiagnoseddiabetesand57millioninaprediabetes stateintheUnitedStates(2).Inthelatter,bloodglucoselevelsknownasimpaired fastingglucose(IFG)orimpairedglucosetolerance(IGT)areintherangeof100125 mg/dL,whilehigherlevelsareassociatedtype2diabetesmellitus,nowincreasingly recognizedasanautoimmuneinflammatorydisease(2).Notably,peoplewithpre diabetesareatincreasedriskofdevelopingtype2diabetes,aconditioncharacterized byimpairedinsulinsecretionfrompancreaticLangerhansislandscells,increasedhepatic glucoseproductionandinconsequencehighbloodglucoselevels,anddecreaseduseof glucoseinmuscletissue.Thesedefectsaremainlyresponsibleforthedevelopmentand progressionoftype2diabetes(3).Furthermore,insulinresistanceisassociatedwith decreasedratesofglycolysis,glycogenesis,lipogenesis,andproteinsynthesis.People withtype2diabetesarealsoathighriskoffatalandnonfatalvascularevents.In 20032004,75%ofadultswithselfreporteddiabeteshadelevatedbloodpressureof> 130/80mmHgorusedprescriptionmedicationsforhypertension.Intensivecontrolof glycemiadecreasessomevascularcomplicationssuchasretinopathyandnephropathy (4).

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DiaMetrixisanherbaldietarysupplementcomposedofvitaminC(ascorbicacid), biotinUSP,chromium(chelate),vanadium(chelate),garciniacambogiaextract(50% hydroxycitricacid),gymnemasylvestreextract(25%),cinnamonextract(4:1),bitter melonextract(10:1),betaineHCL,banabaextract(1%corosolicacid),fennugreek, dicalciumphosphate,cellulose,croscarmellosesodium,stearicacid,silicondioxide, magnesiumstearate,andhydroxypropylmethylcellulose. AnotherrecentlycompletedstudycomparedtheactivityofDiaMetrixtothatofthree antidiabeticdrugs;Metformin,Actos(pioglitazonehydrochloride),andByetta (exenatide)inanobesediabeticmousemodelusingBKS.Cgm+/+Leprdb/BomTacfemale micefedeithernormaldietorhighfatdiet(HFD)+/drugs.DiaMetrixsprotection againstorgandamagewascomparabletothatofByettaandbetterthanActosand Metforminintheanimalsonnormaldiet.Themeanvaluesofmostinflammatory plasmabiomarkerswereelevatedinhighfatdietrelativetonormaldiet.Biomarker meansvariedsignificantlybytreatmentgroupanddiet.Onnormaldiet,DiaMetrix decreasedlevelsofanumberofproinflammatorycytokines,chemokinesandgrowth factorssuchaseotaxin,MCP1,MCP3,MCSF,andincreasedantiinflammatory cytokineIL4relativetountreated.DiaMetrixtreatmentdecreasedGCSF,GMCSF,and TGFrelativetountreatedinhighfatanimals.PyruvatekinaseandAGEincreased, whileinsulinwasdecreasedinanimalstreatedwithDiaMetrixrelativetountreatedon normaldiet.DiaMetrixdemonstratedsuperiorantiinflammatoryactivityrelativetothe commonlyusedantidiabeticdrugsagainstabackgroundofgeneticobesity,supporting

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thecontentionthatDiaMetrixmaybeaneffectiveinterventionfortype2diabetes (Hampshireetal.,submitted). ThegoalofthisclinicalstudywastoassesstheefficacyofDiaMetrixtreatmentin diabeticsubjectswithfastingbloodglucoselevelsbetween160and240mg/dLandto evaluateeffectsoforalDiaMetrixonclinicalendpointsusedintheroutineevaluationof diabeticsubjects.Safetydatawasnotcollected. METHODS Subjects SyntraTechcontractedtheclinicalstudywithacontractresearchorganization,Fenestra ResearchLaboratories(Fenestra),LasVegas,NV.Fenestraalsoconductedthetestsas describedbelow.In20062007,Fenestrarecruited47menand53womenages23to50 (33Black,32Caucasian,25Asianand10Hispanic)usingparticipantsrecruitedfroma largediversepopulationofpeoplelivinginorneartheLasVegas,NVarea,withchronic uncontrolledbloodglucose(peoplewithfastingbloodglucoselevelsbetween160 mg/dLand240mg/dLwithameanof197mg/dL). Excludedweresubjectswithahistoryofheadtrauma,seriousdiseasesorillness diagnosedatthistime,knownmoderatetosevererenalinsufficiency,recenthistory(<6 monthspriortoVisit1)ofmyocardialinfarction.Alsoexcludedwerethosewho regularlyusedoxygentherapy,thosewithknownactivetuberculosis,withtreatedbasal

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cellcarcinoma,historyofcancerwithinthelast5years,thoracotomywithpulmonary resectionwithin1yearpriortothetrial,thoseinapulmonaryrehabilitationprogramor whocompletedapulmonaryrehabilitationprograminthe6weekspriortothe screeningvisit(Visit1),takingprescribeddiureticmedications,cardiacstimulants,or anyotherprescribedornonprescribedmedicationthatmightaltertestingresults, thosetakingopiateanalgesicsprescribedorotherwiseobtainedforanytreatment reasonincludingmigrainetreatment,orforrecreationandhistoryofdrugaddictionor alcoholaddictionwithinsix(5)monthsofthisstudyperiod,femaleswhowerepregnant, lactating,ornursingorwhomaybecomepregnantduringthecourseofthestudy,and thosediagnosedasHIVpositive,diagnosedwithAIDS,orwithanyneuromuscular conditionincludingCP,MS,ALS,orHuntington'sChorea.Alsoexcludedwerepeople withuncontrolledhypertension(e.g.BP>140/90),oranyconditionnotpreviously namedthat,intheopinionofFenestrainvestigatorsorintakestaff,wouldjeopardize safetyoraffectthevalidityoftheresultscollectedinthisstudy. Thestudyincludedfemalesormales18yearsofageorolder,whosignedawritten informedconsent.Tobeeligibleforinclusion,potentialsubjectswererequiredto documentbloodsugarimbalanceforaminimumofthepreviousconsecutive6months ofthescreeningvisitatafrequencyofatleasttwenty(20)timeseachmonthbasedon dailyglucometerreadings.Eligibleparticipantswererandomizedintotwogroupsof50 participants.Thestudywasdoubleblinded.

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Compliancetoinclusionandexclusioncriteriawasmonitoredandmaintainedthrough biweeklyphonecallswithFenestrapersonnelandinpersonofficevisits.All participantswereinstructedtocontacttheirregularhealthcareprofessionaliftheyhad anyunusualoruncomfortablesymptomsduringthecourseofthisstudy.All participantswereinstructedtomakenochangestotheirdailyactivityorconsumption offoodorliquidrelatingtotheamount,volume,ortypeconsumed. Procedures Participantsinbothtreatmentgroups,DiaMetrixandplacebo(vegetariantablet, magnesiumcitrate,silicondioxide),wereinstructedtotaketwotabletsthreetimes daily,orthreetabletstwotimesdailyifonlytwomeals,approximately15minutes beforemeals.Participantswereinstructedtotakeproductonlyiftheyateameal. Participantswhometallinclusioncriteriaandnoneoftheexclusioncriteriaatthefirst visit(day0)andgavesignedinformedconsentwerethenprovidedeithertheplaceboor DiaMetrixalongwithinstructionsdescribingdailydosingtofollowforthedurationof thestudy.Baselinemeasurementsweremadeatday1(visit2).Followingscreening (day0),randomization,andsignedinformedconsent,participantsentereda1week baselineperiod.Inthattime,participantswereaskedtorefrainfromtakingany unnecessaryoverthecounterorprescriptionmedications,ornaturalproductsthatthey werenotalreadytakingfortheremainderofthestudy.Ibuprofen,acetaminophen,and aspirinwereallowed.Atthethirdandfourthvisits(days14and30)evaluationswere performedfollowingstandardproceduresandtheinclusionandexclusioncriteriawere

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againreviewedwitheachparticipantonanindividualbasis.Thefifthvisittookplaceon day60andthesixthandfinalvisitonday90. Atthescreenandpriortorandomization,14oftheparticipantsreportedmild headachesand"notfeelingwell".All100studyparticipantswereattemptingtocontrol theirglucoselevelswithdietandexercise.Dietandexercisewerenotmonitoredduring thestudy,butallparticipantswereinstructedtomakenochangesintheirdiet, activities,orwaterintakeduringthedurationofthestudyatthescreeningandat subsequentvisits. Tests ThetestsbrieflydescribedbelowwereperformedbyFenestrausingstandardclinical procedures. FastingBloodGlucose:Approximately7mLofvenousbloodwascollectedfromeach participantateachblooddrawandfastingbloodglucosewasmeasured. BodyMass:Bodymass(lbs)wasrecordedonallparticipantsatall6visitsusinga monthlycalibratedweightscale. GlucoseChallenge:Eachparticipant'sfastingbloodglucose(FBG)levelwasmeasured atbaseline(day2).Fortheoralglucosetolerancetest(OGTT),allparticipantswere givenatesttablet(halfoftheparticipantsweregivenactiveproductandhalfofthe

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participantsweregivenplacebo),followedbyastandard75gcarbohydrateloaddrink. Bloodplasmareadingsweretakenat30minute,120minute,and180minuteintervals. TotalCholesterol:Participantsbloodwastakenaftera1214hourfastingperiod. Participantswereaskedtoabstainfromalcoholforatleast24hoursbefore519mlof arterialbloodwasdrawn. Lipoproteins:LDLlevelswerecalculatedbysubtractingHDLreadingsplusonefifthof thetriglyceridesfromthetotalcholesterol.LDL=Totalcholesterol(HDL+Triglycerides/5). Triglycerides:Thebloodtestwasperformedonfasting(1214hour)participants. Participantswereaskedtoabstainfromanyalcoholfor24hoursbeforethetest.Fiveto 10mLofvenousbloodwasdrawnfromeachparticipantateachtestinterval. BloodPressure:Bloodpressurewastakenbymedicalprofessionalswithaminimumof 20yearsexperienceusingasphygmomanometerandstethoscope.Bloodpressure measurementsweretakenanaverageofthreetimesafteraminimumrestofatleast15 minutes. HemoglobinA1c(HbA1c,Glycohemoglobin,Glycatedhemoglobin,Glycosylated hemoglobin):BloodwasdrawnonDay1andDay90.Anarterialbloodsamplewas obtainedbyinsertinganeedleintoanarteryoftheparticipantsarmandastandard HbA1cvialofbloodwascollectedbyahealthcareprofessional. Statisticalmethods Continuouslydistributedoutcomesweresummarizedwiththemeanonestandard deviation.Treatmentgroups(DiaMetrix,Placebo)werecontrastedonthemeanof continuouslydistributedoutcomesusingrepeatedmeasureslinearmodelsofthe

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outcomeintermsoftime,treatmentgroup,andthetimebytreatmentgroup interactionwithanautoregressiveorder1autocorrelationmatrix.Allstatisticaltesting was2sidedwithasignificancelevelof5%.RVersion2.11.1(RFoundation)wasused throughout;tableswerecreatedusingSASVersion9.2forWindows(SASInstitute,Cary, NC). RESULTS Allparticipantscompletedthestudy;therewerenomissingdata. DemographicCharacteristics:Treatmentgroupsweresimilarwithregardtothe proportionmale[DiaMetrix22(44%),Placebo25(50%)]andmeanage(DiaMetrix 43.512.7,Placebo45.810.8,p=0.32)andthepercentageofsubjectsofthewhiterace wassignificantlydecreasedamongthoserandomizedtoDiaMetrix(DiaMetrix24(48%), Placebo37(74%),p=0.01);seeTable1. FastingSerumGlucose:Treatmentgroupfastingglucosemeans(ng/dL),showninTable 2andFigure2,weresimilaratbaseline(DiaMetrix196.617.5,Placebo196.917.4, p=0.93)whereastheDiaMetrixmean(14days:DiaMetrix96.610.3,Placebo 205.530.7,p<0.001;30days:DiaMetrix96.511.0,Placebo204.921.6,p<0.001;60 days:DiaMetrix94.810.1,Placebo209.422.5,p<0.001;90days:DiaMetrix89.49.2, Placebo205.220.2,p<0.001)andmeanchangesfrombaseline(DiaMetrix107.214.5, Placebo8.314.8,p<0.001)weresignificantlydecreasedat90days. HbA1c(%):TreatmentgroupHbA1c(%)means(Table2)weresimilaratbaseline (DiaMetrix7.70.5,Placebo7.70.5;p=0.93)whereasthemeanat90days(DiaMetrix

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4.70.3,Placebo80.6,p<0.001)andthemeanchangefrombaselineto90days (DiaMetrix3.00.5,Placebo0.30.4,p<0.001)weresignificantlydecreasedinsubjects takingDiaMetrix.At90days,nosubjectwhoreceivedDiaMetrixhadanHbA1cabove 5%.Incontrast,at90daysnosubjectwhoreceivedPlacebohadanHbA1cbelow6.9%. GlucoseChallengeTest(ng/dL):TheDiaMetrixmeanserumglucose(Table2)was significantlydecreasedrelativetoplaceboat30,120and180minutesafterthe challenge(30minutes:DiaMetrix110.124.1,Placebo205.118.5,p<0.001;120 minutes:DiaMetrix110.023.0,Placebo221.914.6,p<0.001;180minutes:DiaMetrix 109.523.3,Placebo225.812.6,p<0.001). Triglycerides:Treatmentgrouptriglyceridesmeans(mg/dl),seeTable2andFigure1, weresimilaratbaseline(DiaMetrix255.248.1,Placebo254.546.5,p=0.95)whereas theDiaMetrixmean(30days:DiaMetrix234.650.2,Placebo259.345.4,p=0.01;60 days:DiaMetrix213.944.4,Placebo269.648.1,p<0.001;90days:DiaMetrix 203.242.4,Placebo271.348.3,p<0.001)andmeanchangesfrombaseline(90days: DiaMetrix52.023.7,Placebo16.828.6,p<0.001)weresignificantlydecreasedat90 days. TotalCholesterol:TheDiaMetrixmeanCholesterol(g/dL),seeTable2,was significantlyincreasedatbaselinerelativetoplacebo(DiaMetrix:338.596.9,Placebo: 301.389.4,p=0.01).ThetreatmentgroupCholesterolmeansdidnotdiffersignificantly at90days(90days:DiaMetrix240.032.8,Placebo261.229.2,p=0.13)whereasthe DiaMetrixmeanchangefrombaselinewassignificantlydecreasedrelativetoplacebo

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(changesfrombaselineto90days:DiaMetrix98.5110.0,Placebo40.199.6, p=0.004). HighDensityLipoproteins(HDL):TheDiaMetrixmeanHLDL(g/dL),seeTable2,did notdiffersignificantlyfromtheplacebomeanatbaseline(DiaMetrix:56.829.8, Placebo:57.329.8,p=0.93)orat90days(90days:DiaMetrix47.79.3,Placebo 52.610.5,p=0.28;changesfrombaselineto90days:DiaMetrix9.033.6,Placebo 4.735.0,p=0.45). LowDensityLipoproteins(LDL):TheDiaMetrixmeanLDL(mg/dL),showninTable2, wassignificantlyincreasedatbaselinerelativetoplacebo(DiaMetrix:230.7106.9, Placebo:193.287.2,p=0.01)whereasthetreatmentgroupLDLmeansandmean changesinLDLdidnotdiffersignificantlyat90days(90days:DiaMetrix151.733.5, Placebo154.430.3,p=0.86;changesfrombaselineto90days:DiaMetrix79.0120.8, Placebo3894.9,p=0.11). RatioOfTotalCholesterolToHighDensityLipoproteins:TheDiaMetrixmeanratioof totalcholesteroltohighdensitylipoproteins(Table2)didnotdiffersignificantlyfrom theplacebomeanatbaseline(DiaMetrix:8.55.9,Placebo:7.25.1,p=0.10)orat90 days(90days:DiaMetrix5.31.5,Placebo5.11.1,p=0.88). RatioOfLowDensityLipoproteinsToHighDensityLipoproteins:TheDiaMetrixmean ratiooftotalcholesteroltohighdensitylipoproteins(Table2)didnotdiffersignificantly fromtheplacebomeanatbaseline(DiaMetrix:6.25.2,Placebo:4.94.3,p=0.06)orat 90days(90days:DiaMetrix3.41.3,Placebo3.10.9,p=0.65).

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SystolicBloodPressure:TheDiaMetrixsystolicbloodpressuremean(Table2)was significantlyincreasedrelativetoplaceboatbaseline(Baseline:DiaMetrix171.319.9, Placebo155.225.1,p<0.001),whereasthetreatmentgroupmeanswerenot significantlydifferentat30and60days(30days:DiaMetrix157.925.9,Placebo 154.221.4,p=0.37;60days:DiaMetrix149.716.3,Placebo155.419.4,p=0.16)and theDiaMetrixmeanwassignificantlydecreasedat90days(90days:DiaMetrix 142.914.6,Placebo155.218.9,p=0.003).Analysesonchangesfrombaselineshowed significantlydecreasedmeanchangesfrombaselineto90(p<0.001)daysinsubjects randomizedtoDiaMetrix. DiastolicBloodPressure:Treatmentgroupdiastolicbloodpressuremeans(mmHg),see Table2andFigure3,weresimilaratbaselineandat30days(Baseline:DiaMetrix 75.29.9,Placebo76.08.7,p=0.55;30days:DiaMetrix74.56.4,Placebo75.56.9, p=0.44),whereastheDiaMetrixmeanwassignificantlydecreaseat60and90days(60 days:DiaMetrix71.94.3,Placebo76.75.7,p=0.001;90days:DiaMetrix70.33.2, Placebo76.65.2,p<0.001).Themeanchangefrombaselineto90dayswas significantlydecreasedinsubjectsrandomizedtoDiaMetrix(p<0.001). BodyMass:TheDiaMetrixmeanbodymass(lb),seeTable2,wasnotsignificantly differentfromtheplacebomeanatbaseline(DiaMetrix:211.454.7,Placebo: 211.653.6,p=0.99).Thetreatmentgroupweightmeansdidnotdiffersignificantlyat 90days(90days:DiaMetrix202.154.0,Placebo211.652.8,p=0.38)whereasthe DiaMetrixmeanchangefrombaselinewassignificantlydecreasedrelativetoplacebo (changesfrombaselineto90days:DiaMetrix9.33.1,Placebo0.02.9,p<0.001).

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Discussion Thisstudyinsubjectswithprediabetesanddiabetesdemonstratedstatistically significanteffectsofDiaMetrixonfastingglucoselevelandtheglucosechallengetest, HbA1c,andbloodpressure.TotalcholesterolwasdecreasedbutLDLandHDLwerenot significantlyaffected.Withrespecttobodymass,DiaMetrixtreatmentgroupmean changefrombaselinewassignificantlydecreasedrelativetoplacebo. Basedonthemagnitudeofclinicalresponses,DiaMetrixcomparedfavorablyto standardtherapeuticsusedintreatmentofdiabetes.Metforminisoneoftheoldest antidiabetesdrugsandwasclaimedto"lowerthebloodsugartominimum physiologicallimit"intreatedpatients.Basedonitsfavorabletoxicityprofileitwas widelyusedbutwasapprovedbytheU.S.FoodandDrugAdministrationfortype2 diabetesin1994(6).MedianHbA1cwas7.4%intheMetformingroupcomparedwith 8.0%intheconventionalgroup,andintensiveglucosecontrolwithMetformin decreasedtheriskofdiabetesrelatedendpointsinoverweightdiabeticpatients,and wasassociatedwithlessweightgain(6).Pioglitazone(Actos),adruginthe thiazolidinedionefamily,selectivelystimulatesthenuclearreceptorperoxisome proliferatoractivatedreceptorgamma(PPAR)andmodulatesthetranscriptionof

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insulinresponsivegenesinvolvedinthecontrolofglucoseandlipidmetabolism(7). Actossignificantlydecreasedfastingplasmaglucoselevelfrom11.0+/2.0mmol/literto 8.9+/1.1mmol/literwithasignificantimprovementinthehemoglobinHbA1clevel from9.2+/1.8%to8.3+/1.5%(11).Rosiglitazone,anotherthiazolidinedioneoral antidiabeticagent,hasbeenshowntosignificantlyreduceHbA1candfastingplasma glucoseunderdifferentdosingregimens(8).Exenatideisasyntheticversionof exendin4,ahormonefoundinthesalivaoftheGilamonster.Itdisplaysbiological propertiessimilartohumanglucagonlikepeptide1throughregulationofglucose metabolismandinsulinsecretionbutitsmechanismofactionisstillunderstudy[2].A 52weektreatmentwithexenatidereducedHBA1Cby0.80.1%(9). Atpresent,themanagementoftype2diabetesfocusesonglucosecontrolthrough loweringofbloodglucoseandbyextensionHbA1candotheradvancedglycationend products(3).Inouropinion,theseclinicalendpointsaresymptomsofdisease,not diseasemechanisms.Persistenthighbloodsugarinpeopleisaproblem;however,even withperfectglucosecontrolpeoplestillsufferfromsignificantorganpathologies, morbidityandmortality.Drugsthataredesignedtocontrolbloodsugarhavemany liabilitiessuchasinductionofhypoglycemia.Thusthecommonlyacceptedemphasison controllingbloodsugarasasoleendpointmaybesomewhatmisguidedinthatblood sugarlevelisonlyoneofthemultiplefacetsofdiabetes.Wefeelthatmoreemphasis shouldbeplacedoncontroloftheimmunedysfunctionthatisamajorbiochemical driverofthepathologyofdiabetes.Therefore,understandinghowdrugsimpact

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inflammatorybiomarkerprofileswillbeextremelyimportantinunderstandingand treatingdiabetes.Weproposethattheoptimaltherapeuticstrategyfordiabetesshould addressnotonlybloodsugarcontrol,butalsobedirectedtocontrolofimmunesystem imbalanceinaefforttodelaydiseaseprogression(3). Giventhattype2diabetesisfundamentallyaninflammatorydisease(2),inourrecentin vivostudyinamodelofhumantype2diabetesinobesemice,weselectedabroad rangeofendpointstoevaluatemechanismofactionofDiaMetrix.Inadditionto commonlytestedmetabolites(glucose,advancedglycationendproduct,insulin, cholesterol,triglycerides),weincludedpyruvatekinase,hexokinaseII,andcitrateas potentiallyrelevantendpointsalongwithalargepanelofcytokines,chemokines, endocrinemarkersandgrowthfactorsandhaveshownthatofDiaMetrixsignificantly downregulatedseveralproinflammatorychemokines,cytokines,andgrowthfactors andupregulatedlevelsoftheantiinflammatorycytokineIL4.Wespeculatethat DiaMetrixmaybreaktheselfreinforcinginflammatorycycleandthusslowdownthe naturalcourseofthedisease(Hampshireetal.,submitted). Inconclusion,thisstudydemonstratedareductioninthemeanfastingbloodsugar, bloodpressureandbodymassinsubjectsrandomizedtoDiaMetrix.Thesefindings suggestthatDiaMetrix,anoverthecounterpreparation,deservesfurtherevaluationof itsactivityinlargerclinicalstudies.

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Competinginterests KRH,SM,andRMareownersofSyntraTech. EI,RTS,JM,andCLareconsultantsforSyntraTech. Authors'contributions KRH,SM,andRMfundedthestudyandanalysisoftheclinicaldata. JMandCLperformedstatisticalanalysisoftheclinicaldata. EI,JMandRTSreviewedtheresultsandwrotethemanuscript.

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References 1. Steinbrook,R.Facingthediabetesepidemicmandatoryreportingof

glycosylatedhemoglobinvaluesinNewYorkCity,NEnglJMed2006;354(6):545548. 2. Kohn,L.D.,Wallace,B.,Schwartz,F.,andMcCall,K.(2005)Istype2diabetesan

autoimmuneinflammatorydisorderoftheinnateimmunesystem?,Endocrinology 2005;146(10):41924199. 3. DeFronzo,R.A.Currentissuesinthetreatmentoftype2diabetes.Overviewof

neweragents:wheretreatmentisgoing,AmJMed2010;123(3suppl):S3848. 4. Dormandy,J.A.,Charbonnel,B.,Eckland,D.J.,Erdmann,E.,MassiBenedetti,M.,

Moules,I.K.,Skene,A.M.,Tan,M.H.,Lefebvre,P.J.,Murray,G.D.,Standl,E.,Wilcox,R. G.,Wilhelmsen,L.,Betteridge,J.,Birkeland,K.,Golay,A.,Heine,R.J.,Koranyi,L.,Laakso, M.,Mokan,M.,Norkus,A.,Pirags,V.,Podar,T.,Scheen,A.,Scherbaum,W., Schernthaner,G.,Schmitz,O.,Skrha,J.,Smith,U.,andTaton,J.(2005)Secondary preventionofmacrovasculareventsinpatientswithtype2diabetesinthePROactive Study(PROspectivepioglitAzoneClinicalTrialInmacroVascularEvents):arandomised controlledtrial,Lancet2005;366(9493):12791289. 5. http://diabetes.niddk.nih.gov/dm/pubs/insulinresistance/.NIHPublicationNo.

094893;2008.

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6.

EffectofintensivebloodglucosecontrolwithMetforminoncomplicationsin

overweightpatientswithtype2diabetes(UKPDS34).UKProspectiveDiabetesStudy (UKPDS)Group,Lancet1998;352(9131):854865. 7. Yamasaki,Y.,Kawamori,R.,Wasada,T.,Sato,A.,Omori,Y.,Eguchi,H.,Tominaga,

M.,Sasaki,H.,Ikeda,M.,Kubota,M.,Ishida,Y.,Hozumi,T.,Baba,S.,Uehara,M.,Shichiri, M.,andKaneko,T.Pioglitazone(AD4833)amelioratesinsulinresistanceinpatientswith NIDDM.AD4833GlucoseClampStudyGroup,Japan,TohokuJExpMed1997;183:173 183. 8. 9. Goldstein,B.J.Rosiglitazone,IntJClinPract2000;54(5):333337. Bunck,M.C.,Diamant,M.,Corner,A.,Eliasson,B.,Malloy,J.L.,Shaginian,R.M.,

Deng,W.,Kendall,D.M.,Taskinen,M.R.,Smith,U.,YkiJarvinen,H.,andHeine,R.J. Oneyeartreatmentwithexenatideimprovesbetacellfunction,comparedwithinsulin glargine,inMetformintreatedtype2diabeticpatients:arandomized,controlledtrial, DiabetesCare2009;32(5):762768.

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DiaMetrix Human Study (14 October 2010) Table 1. Demographics Syntra-5 (N=50) Male [N (%)] White [N (%)] Age (Mean SD) 22 (44) 24 (48) 43.4812.68 Placebo (N=50) 25 (50) 37 (74) 45.8210.77 Total 47 (47) 61 (61) 44.6512 P-Value 0.691 0.011 0.322

1 2

Pearson's Chi Square Test F Test for treatment

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DiaMetrix Human Study (14 October 2010) Table 2. Blood Chemistry Outcome (Mean SD) High Density Lipoprotein (? g/? L) Baseline Day 90 Change from Baseline to Day 90 Low Density Lipoprotein (? g/? L) Baseline Day 90 Change from Baseline to Day 90 Ratio of Low Density Lipoprotein to High Density Lipoproteins Baseline Day 90 Systolic Blood Pressure (mm Hg) Baseline Day 30 Day 60 Day 90 Change from Baseline to Day 90 Diastolic Blood Pressure (mm Hg) Baseline Day 30 Day 60 Day 90 Change from Baseline to Day 90 Weight (lbs) Baseline Day 90 Change from Baseline to Day 90 Time DiaMetrix - (N=50) 56.829.8 47.79.3 -933.6 230.7106.9 151.733.5 -79120.8 6.25.2 3.41.3 171.319.9 157.925.9 149.716.3 142.914.6 -28.417.1 75.29.9 74.56.4 71.94.3 70.33.2 -4.99 211.454.7 202.154 -9.33.1 Placebo (N=50) 57.329.8 52.610.5 -4.735 193.287.2 154.430.3 -38.894.9 4.94.3 3.10.9 155.225.1 154.221.4 155.419.4 155.218.9 010.5 768.7 75.56.9 76.75.7 76.65.2 0.76.5 211.653.6 211.652.8 02.9 Total 5730 50.110 -6.934 21299 15332 -58.9110 5.65 3.21 163.224 15624 152.618 14918 -14.220 75.69 757 74.36 73.55 -2.18 211.554 206.853 -4.66 P-Value 0.931 0.282 0.452 0.011 0.862 0.112 0.061 0.652 < 0.0011 0.372 0.162 0.0032 < 0.0012 0.551 0.442 0.0012 < 0.0012 < 0.0012 0.991 0.382 < 0.0012

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F Test for treatment F Test for treatment based on a repeated measures ANOVA in terms of treatment, time and treatment by time and an AR(1) correlation structure (treatment by time interaction: p-value < 0.001).

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DiaMetrix Human Study (14 October 2010) Table 2. Blood Chemistry Outcome (Mean SD) Fasting Serum Glucose (mg/dL) Baseline Day 14 Day 30 Day 60 Day 90 Change from Baseline to Day 90 A1c (%) Baseline Day 90 Change from Baseline to Day 90 Serum Glucose (mg/dL) Minute 30 Minute 120 Minute 180 Triglycerides (mg/dL) Baseline Day 30 Day 60 Day 90 Change from Baseline to Day 90 Cholesterol (? g/? L) Baseline Day 90 Change from Baseline to Day 90 Ratio of Cholesterols to High Density Lipoproteins Baseline Day 90 Time DiaMetrix (N=50) 196.617.5 96.610.3 96.511 94.810.1 89.49.2 -107.214.5 7.70.5 4.70.3 -30.5 110.124.1 11023 109.523.3 255.248.1 234.650.2 213.944.4 203.242.4 -5223.7 338.596.9 24032.8 -98.5110 8.55.9 5.31.5 Placebo (N=50) 196.917.4 205.530.7 204.921.6 209.422.5 205.220.2 8.314.8 7.70.5 80.6 0.30.4 205.118.5 221.914.6 225.812.6 254.546.5 259.345.4 269.648.1 271.348.3 16.828.6 301.389.4 261.229.2 -40.199.6 7.25.1 5.11.1 Total 196.817 151.159 150.757 152.160 147.360 -49.560 7.70.01 6.32 -1.42 157.652 165.959 167.661 254.947 24749 241.754 237.257 -17.643 319.995 250.633 -69.3108 7.86 5.21 P-Value 0.931 < 0.0012 < 0.0012 < 0.0012 < 0.0012 < 0.0012 0.931 < 0.0012 < 0.0012 < 0.0011 < 0.0012 < 0.0012 0.951 0.012 < 0.0012 < 0.0012 < 0.0012 0.011 0.132 0.0042 0.101 0.882

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F Test for treatment F Test for treatment based on a repeated measures ANOVA in terms of treatment, time and treatment by time and an AR(1) correlation structure (treatment by time interaction: p-value < 0.001).

Figure 1 Mean triglycerides (mM) by treatment group (Blue: DiaMetrix - , Black: Placebo); whiskers extend to the mean 1 standard deviation. The treatment by time interaction was significant (p< 0.001); treatment group contrasts on the mean were significant at 30 (p=0.01), 60 (p<0.001) and 90 days (p<0.001) based on a repeated measures linear model with an autoregressive order 1 covariance assumption.

Figure 2 Mean fasting serum glucose(ng/dl) by treatment group (Blue: DiaMetrix, Black: Placebo); 1 standard deviation. The treatment by time interaction was significant whiskers extend to the mean (p<0.001); treatment group contrasts on the mean were significant at 14 (p<0.001), 30 (p<0.001 ), 60 (p<0.001) and 90 days (p<0.001) based on a repeated measures linear model with an autoregressive order 1 covariance assumption.

Figure 3 Mean diastolic blood pressure (mmHg) by treatment group (Blue: DiaMetrix - , Black: Placebo); whiskers extend to the mean1 standard deviation. The treatment by time interaction was significant (p<0.001); treatment group contrasts on the mean were significant at 6 0 (p<0.001) and 90 days (p<0.001) based on a repeated measures linear model with an autoregressive order 1 covariance assumption.