EUGIS

European Union Garlic Inflammation Study in Humans

Background

• No clear indications that ‘garlic’ influences lipid metabolism

• Indications from cell and animal studies that ‘garlic’ may influence inflammation parameters

• Cardiovascular disease (atherosclerosis) shares common features with inflammation

• Current treatment strategies for atherosclerosis also focus on inflammation

AimAim

StudyStudy the effects of garlic on the effects of garlic on inflammatoryinflammatory biomarkers of biomarkers of atherosclerotic disease atherosclerotic disease and on and on

cancer cancer biomarkersbiomarkers in man in man

• Primary endpoint:: changes in C-reactive protein ( CRP) changes in C-reactive protein ( CRP)

• Secondary Secondary endpoints:endpoints:

– Indicators of lipid metabolism Indicators of lipid metabolism

– Biochemical markers of inflammationBiochemical markers of inflammation

– BloodBlood pressurepressure / heart rate variability / heart rate variability

– BBiomarkers for anti-carcinogenic effects iomarkers for anti-carcinogenic effects

InclusionInclusion criteria criteria

• Able and willing to give informed consentAble and willing to give informed consent

• 9090 subjects of either gender subjects of either gender**

• AAged between 40-75 years ged between 40-75 years

• SmokingSmoking≥≥ 10 cigarettes/day and 10 cigarettes/day and BMI≥ 24.5 BMI≥ 24.5 kg/mkg/m22

* * Power calculation to find a reduction of 30% Power calculation to find a reduction of 30% in plasma CRP levels with in plasma CRP levels with = 0.05 and a power = 0.05 and a power of 80% about 30 subjects per group are neededof 80% about 30 subjects per group are needed

EExclusion criteriaxclusion criteria

• Any major clinicalAny major clinical significant significant abnormality detected by abnormality detected by a general health a general health questionnairequestionnaire or by or by blood samplingblood sampling

• Chronic drug treatment /Chronic drug treatment / use of medicationuse of medication

• Participation in prior studParticipation in prior studyy in the last 3 months in the last 3 months or or bblood donationlood donation

• Inadequate use of contraceptives Inadequate use of contraceptives / p/ pregnanregnancy / cy / lactating womenlactating women

• History of alcoholHistory of alcohol / / drug abuse drug abuse

• Positive test results for HepBPositive test results for HepB//C or HIVC or HIV

• Dislike of garlicDislike of garlic

Experimental design

I.C. / screening Intervention period

0 4 - 5 11-12 weeks

Blood and urine collection

-2

• Double dummy placebo-controlled trial

• 3 parallel groups (n=30)

12-weeks treatment12-weeks treatment

1. Garlic (from EU-sponsored Printanor 2001)

• Lichtwer capsule preparation (300 mg)

• 2.1g daily (3 cap - breakfast & 4 cap at dinner) (3 cap - breakfast & 4 cap at dinner) 2. Atorvastatin:

• 40 mg/day daily at dinnerat dinner

3. Placebo:

• garlic and statin placebo daily

MeasurementsMeasurements

• SafetySafety::

– Parameters of muscle and liver functionParameters of muscle and liver function– HaematologyHaematology

• ClinicalClinical::

– Blood pressureBlood pressure– 12 lead12 lead ECG (HR, conduction) ECG (HR, conduction)– Heart Rate Variability (HRV)Heart Rate Variability (HRV)

MeasurementsMeasurements

• BiochemicalBiochemical::

–Cholesterol, HDL, and triglycerides; Cholesterol, HDL, and triglycerides;

–CRP, vWF, Fbg, and cytokines after whole blood stimulation CRP, vWF, Fbg, and cytokines after whole blood stimulation without or with LPS (TNF-α, IL-10);without or with LPS (TNF-α, IL-10);

–Sensitivity of leukocytes to inflammatory stimulus;Sensitivity of leukocytes to inflammatory stimulus;

–Cancer biomarkers: Cancer biomarkers: anti-oxidant enzymes (GPX, SOD), DNA damage (Comet assay), anti-mutagenic properties of urine (Ames test);

–Metabolites of garlic compounds in plasma and urine;

Additional measurements suggested in the EU proposal

• SAA, and PAI-1

• s-VCAM / s-ICAM / s-Selectine

• MCP-1 and other chemokines

• Endothelin 1 and 3

• Plasma ox-LDL

• Urine isoprostanes

Current study statusCurrent study status

Subjects at information sessions: >148Subjects at information sessions: >148

Subjects signing IC: 142Subjects signing IC: 142

Subjects randomised : 92Subjects randomised : 92 Subjects completed: 84Subjects completed: 84

Subjects withdrawn: 8 (6 non-related AEs)Subjects withdrawn: 8 (6 non-related AEs)

• Entire clinical study period: < 5 months!Entire clinical study period: < 5 months!– last samples at Jan 15last samples at Jan 15thth

Study population - baselineStudy population - baseline

• FemaleFemale– n = 47n = 47

– age: 40-67 yrsage: 40-67 yrs

– BMI: 29.6 (SD: 4.1) kg/mBMI: 29.6 (SD: 4.1) kg/m2 2

– BP: 124 (18) / 76 (12) mm HgBP: 124 (18) / 76 (12) mm Hg

• MaleMale– n = 37n = 37

– age: 40-63 yrsage: 40-63 yrs

– BMI: 29.3 (SD: 3.2) kg/mBMI: 29.3 (SD: 3.2) kg/m22

– BP: 130 (11) / 82 (10) mm HgBP: 130 (11) / 82 (10) mm Hg

Baseline lipidsBaseline lipids

• FemaleFemale– Total Cholesterol:Total Cholesterol: 5.60 (1.01) mmol/L 5.60 (1.01) mmol/L (3.9-7.3)(3.9-7.3)

– HDL-Cholesterol:HDL-Cholesterol: 1.35 (0.32) mmol/L 1.35 (0.32) mmol/L (>1.16)(>1.16)

– LDL-Cholesterol:LDL-Cholesterol: 3.52 (0.95) mmol/L3.52 (0.95) mmol/L

– Triglycerides:Triglycerides: 1.63 (0.95) mmol/L1.63 (0.95) mmol/L (0.8-1.94)(0.8-1.94)

• MaleMale– Total Cholesterol:Total Cholesterol: 5.71 (1.04) mmol/L 5.71 (1.04) mmol/L (3.9-7.3)(3.9-7.3)

– HDL-Cholesterol:HDL-Cholesterol: 1.11 (0.32) mmol/L 1.11 (0.32) mmol/L (>0.90)(>0.90)

– LDL-Cholesterol:LDL-Cholesterol: 3.54 (0.99) mmol/L3.54 (0.99) mmol/L

– Triglycerides:Triglycerides: 2.35 (1.32) mmol/L2.35 (1.32) mmol/L (0.8-1.94)(0.8-1.94)

Adverse EventsAdverse Events

• AE’sAE’s– Headache:Headache:

• 52 episodes in 41 subjects52 episodes in 41 subjects

– Abdominal discomfortAbdominal discomfort• 36 episodes in 26 subjects36 episodes in 26 subjects

– OtherOther• 60 various events (URT infection, ‘pains’, 60 various events (URT infection, ‘pains’,

intercurrents)intercurrents)

PLANPLAN

• All samples have been / are being shippedAll samples have been / are being shipped

• Unblinding next week Unblinding next week

• Primary parameter laboratory analysis Primary parameter laboratory analysis completed at Jan 31completed at Jan 31stst

• Stats analysis time allocatedStats analysis time allocated

• Main report around March 1Main report around March 1stst 2004 2004

• Additional reports: to be discussedAdditional reports: to be discussed

THANKSTHANKS

• TNO-PGTNO-PG• P Meijer

• R Kret

• R Streunding

• CHDRCHDR• M van Doorn

• I Kamerling

• Nursing staff

• LUMC pharmacyLUMC pharmacy• R Press

• LichtwerLichtwer• S Striegl

• T Haffner

• Participants who tolerated 2.1 gram garlic/dayParticipants who tolerated 2.1 gram garlic/day