W16-P-059 Antiatherogenic effects of phytoestrogens in postmenopausal women

Workshops W16 Treatment of atherosclerosis risk W16-P-056

] PLASMA A L D O S T E R O N E AND OXIDATIVE STRESS IN PATIENTS W I T H ESSENTIAL HYPERTENSION

B. Monge, J. Marquez, E. Beraal, E. Fernandez, O. Sanchez, R. Fabregate, M. Fabregate, J. Villafruela, J. Saban-Ruiz. Endothelial Pathology Uni~

Ramon y Cajal Hospital, Madrid, Spain Introduction: Although the "circulating" Renin Angiotensin System (RAS) is not directly responsible for the rise in blood pressure in essential hypertension, angiotensin All is a potent vasoconstrictor and thus could participate in this process. On the other hand, the "tissue" RAS plays an outstanding role in the genesis of vascular damage. Although aldosterone has been related to hypertensive cardiopathy, its influence on oxidative stress, in patients with hypertension, remains unknown. Objectives: 1.To detemaine plasma aldosterone levels in patients with essential hypertension under non-diuretic pharmacological treatment.2.To correlate plasma aldosterone levels with the control of blood pressure, urinary sodium, and plasma oxidized-LDL. Methods: N= 127 patients with hypertension under RAS-blockers and/or calcium antagonist treatment were studied, aged 35 to 85 (55±17), of which 66M, 54 had hyperglycemia and 27 were smokers, from a cardiovascular risk population; BMI (kg/m2); waist circumference (cm); 24-hour ambulatory blood pressure monitoring (ABPM): Spacelabs 90207, creatinine, glycemia, HbAlc, triglycerides, LDL, HDL, total cholesterol with HITACHI; Urinary sodium: mmol/1 (Synchron). Plasma aldosterone levels(ng/dl): Inmunotech RIA; PRA (plasma renin activity) (ng/ml/h): Inmunotech RIA; Oxidized-LDL (U/L): Mercodia ELISA. Statistical analysis: t-student, chi-square. Results: 1. N=31 patients (24.4%) had plasma aldosterone levels > 13.2. 2. In this group, PRA levels were higher (1.63 ± 0.43 vs 1.14 ± 0.15) and the urinary sodium levels were lower (p=0.005). 3. At similar ages, control of blood pressure, anthropometric and metabolic parameters, patients with higher aldosterone levels had a higher oxidative stress evaluated by plasma oxidized-LDL levels (p=0.02). Conclusions: 1. Plasma aldosterone levels in patients under non-diuretic pharmacologic treatment act as mediators in the retention of sodium at the kidney but do not influence the control of blood pressure, probably due to the vasodilating action of the antihypertensive agents. 2. Studying comparable populations, the higher the aldosterone plasmatic levels found, the greater the oxidative stress. This correlation had not been described previously in patients with hypertension.

W16-P-057 ] INFLUENCE O F EXTENDED-RELEASE FLUVASTATIN ON SERUM LIPID LEVEL C H A N G E S IN PATIENTS W I T H TYPE 2 DIABETES M E L L I T U S AND C O M B I N E D HYPERLIPIDAEMIA G. Naumov, S. Kozlov, A. Lyakishev. Department of Atherosclerosis,

Cardiology Research Centre, Moscow, Russia Objective: There're data that extended-release fluvastatin is effective in patients (pts) with combined hyperlipidaemia (HLP). Such type of HLP is common in pts with type 2 diabetes mellitus (DM). Aim of the study was to assess lipid-lowering efficacy of fluvastatin in patients with type 2 DM and combined HLP. Methods: 20 male and female pts (mean age 58±6 years) with type 2 DM, low-density lipoprotein (LDL) cholesterol _>4.1 mmol/1, and triglycerides (TG) 2.4-4.5 1 mmol/1 were enrolled in this study. Changes in total cholesterol, LDL cholesterol, high-density lipoprotein (HDL) cholesterol, TG, and LDL cholesterol/HDL cholesterol ratio were monitored after 4, 8 and 12-week treatment. Safety and tolerability of therapy was assessed by clinical symptoms and changes in AST, ALT and CPK. Results: Therapy with extended-release fluvastatin 80 mg/day for 12 weeks decreased median LDL cholesterol from 4.24±1.09 mmol/1 to 2.77±0.78 mmol/l (-35%), decreased triglycerides from 2.94-0.67 to 2.05±0.4 mmol/1 (-29%), decreased LDL cholesterol/HDL cholesterol ratio from 4.03±1.52 to 2.29±0.81 (-43%), and increased HDL cholesterol from 1.11±0.23 to 1.25±0.19 (+13%) (p<0.001 in all cases). Fluvastatin provided the same lipid profile changes at 4, 8 and 12 weeks There weren't blood glucose changes and adverse events with fluvastatin therapy. Conclusions: Therapy with extended-release fluvastatin is effective, tolerable and safe in pts with type 2 DM and combined HLP.

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COMPARATIVE SAFETY OF ATORVASTATIN 80 M G VS l 0 M G D E R I V E D FROM ANALYSIS OF 49 C O M P L E T E D TRIALS

C. Newman, J. Tsai, M. Szarek, D. Luo, E. Gibson. Medical, Pfizer

Pharmaceuticals, New York, USA Objective and Methods: Clinicians continue to associate high-dose statin use with increased adverse events (AEs). We thus compared the safety of atorvastatin (Atv) 80 mg, Atv 10 nag, and placebo (Pbo) in 14,236 patients with varying CV risk enrolled in 49 trials completed as of September 15, 2004. Results: An overview of safety parameters is presented in the Table. Overall, the most frequent treatment-associated AEs were related to the digestive system (_<6.2% in all groups). No cases of rhabdomyolysis were reported. Incidence of myalgia was low and similar for both Atv groups. Incidence of CPK elevations > 10xULN was 0%, 0.2%, and 0.5% in the Pbo, Atv 10-mg, and Atv 80-mg groups, respectively.* One patient (with a stress fracture) in the 80-mg group and 2 in the 10-mg group also had muscle symptoms. Persistent CPK elevations > 10xULN were observed in 2 patients in the 80-mg group, but without muscle symptoms. Incidence o f L F T (ALT/AST) elevations >3xULN was 1.0%, 0.8%, and 3.7% in the Pbo, Atv 10-mg, and Atv 80-mg groups, respectively.* Population Pt # (pt-yrs exposure) Pt experiencing _>1 AE* Withdrawals due to AEs* Serious AEs* Myalgia* persistent CPK > 10xULN* persistent LFTs >3xULN*

Pbo n(%) 2180 (907) 270 (12.4) 27 (1.2) 92 (4.2) 15 (0.7) 0 4 (0.2)

Atv 10 mgn(%) 7258 (4925) 983 (13.5) 171 (2.4) 12 (0.2) 99 (1.4) 0 11 (0.2)

Atv 80 mg n(%) 4798 (4681) 699 (14.6) 84 (1.8) 25 (0.5) 72 (1.5)* 2 (0.06) 28 (0.6)

*Trealment-associated events; *One case of reported myopathy with CPK 1.4xULN; *Denominators for incidences are the #ofpts with lab values during treatment. Conclusions: Across 49 completed trials, Atv 80 mg had an overall AE profile similar to Atv 10 nag, although the incidence of LFF elevations >3xULN was higher in the 80-mg group. Contrary to the perception that the incidence of muscle-related AEs increases with statin dose, no such relationship was evident in this analysis. *Denominators for incidences are the # of patients with lab values during treatment.

W16-P-059 J A N T I A T H E R O G E N I C EFFECTS OF P H Y T O E S T R O G E N S IN POSTMENOPAUSAL WOMEN N. Nikitina 1, V. Korennaya 2 , V. Myasoedova 3 , E. Khalilov 1, I. Sobenin 4, A. Orekhov 2. 1Dept. of Biochemistry, Institute of Physical and Chemical

Medicine, Moscow, Russia; 2lnstitute for Atherosclerosis Research, Moscow, Russia; 3Institute of General Pathology and Pathophysiology, Moscow, Russia; 4Institute of Experimental Cardiology, Moscow, Russia Objective: The aim of this investigation was to study the effects of phytoestrogen-rich natural substances on blood serum atherogenicity in postmenopausal women in ex vivo model. Methods: Blood serum atherogenicity was estimated as the ability of serum to induce cholesterol accumulation in cultured human blood-derived monocytes-macrophages. Blood of apparently healthy postmenopausal women aged 47-61 years was taken before single dose oral intake of compound and 2, 4 and 6 hours after it. The following plant compounds was compared: extract of grapeseeds, grape stems, hop cones, sage and green tea leaves in dosages 50, 100, 250 500 and 1000 mg. Additionally, dietary supplement Phytosny (Arcnpharma, France) was used in single dosage of 35 nag of soy isoflavones. The content of total pelyphenols, procyanidins, genistein, daidzein, resveratrol and flavone was measured by thin-layer and high performance liquid chromatography. Results: At the baseline, serum taken from women was able to induce 1.3-1.5-fold increase in intracelhilar cholesterol content in cultured cells. The amount of accumulated intracellular cholesterol was taken for 100%. Blood serum atherogenicity was lowered by 68±2%, 36±4%, 48±5%, 36±8% and 85±6% (average reduction for 6-hour interval) after administration of grape seeds extract, grape stems, hop cones, sage and green tea leaves, respectively (p<0.05 for all). Soy isoflavones decreased serum atherogenicity by 324-3% (p<0.05). The antiatherogenic effects of plant compounds positively correlated with the content of polyphenols (r=0.975,

75th EAS Congress, 23-26 April 2005, Prague, Czech Republic

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Workshops Treatment of atherosclerosis risk

p<0.001), procyanidins (r=-0.971, p<0.001) and the sum of phytoestrogens (genistein, daidzein, resveratrol and flavone), (r=0.764, p<0.001). Conclusions: Single dose oral administration of phytoestrogen-rich botanicals suppressed serum-induced cholesterol accumulation in cultured cells, thus providing anti-atherogenic effect. These plants may be considered as the effective and promising compounds for the development of natural drug designed for atherosclerosis prevention in postmenopausal women.

The most apparent reduction of ET-1 mRNA expression, as well as ET-1 production, was seen after DHA supplementation (Fig. 1), and DPA had a similar effect. All three fatty acids tested caused an increase in NO production.

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N OF A STUDY COMPARING IW16-P-0601 DTOE SRCI GETRA P I B/ATO RVASTAT IN W I T H ATORVASTATIN ALONE ON ATHEROMA VOLUME IN PATIENTS W I T H CORONARY HEART DISEASE S.E. Nissen 1, J.-C. Tafdif 2, T. Crowe 1, T. Thuren 3, C.L. Shear 3, J.H. Revkin 3 . i Cleveland Clinic Foundation, Cleveland, OH, USA;

eMontreal Heart Institute, Montreal, Quebec, Canada; 3Pfizer Global Research and Development, New London, C~, USA Background: Elevated levels of LDL-C and low levels of HDL-C are strong, independent risk factors for coronary heart disease (CHD). Coronary intravascular ultrasound (IVUS) measurement of atheroma progression has been shown to correlate with CHD risk. Studies have shown that statins lower LDL-C and reduce cardiovascular risk by 25% to 40% and that intensive therapy with atorvastatin (A) can halt the progression of atherosclerosis measured by IVUS. Torcetrapib (T) is a novel cholesteryl ester transfer protein (CETP) inhibitor that raises HDL-C, decreases LDL-C, and increases lipid particle size. Combination therapy with T/A may provide additive benefits when treating various dyslipidemias that may slow the progression of atherosclerosis and reduce cardiovascular risk. Rationale and Objective: Measurement of coronary atheroma volume using IVUS is an accurate and increasingly used method for evaluating the progression of atherosclerosis in patients on drug treatment for cardiovascular risk factors. The objective of this trial is to show that the combination of T/A can alter the progression of atherosclerosis, as measured by the change in atheroma volume after 2 years, more than A alone in subjects with established CHD. Design: This Phase 3, multi-center, double-blind, randomized, parallelgroup study is being conducted at 141 sites in North America and Europe. Approximately 1200 men and women aged 18 to 75 years with angiographic evidence of CHD were randomized. Subjects underwent a clinically indicated cardiac catheterization and baseline IVUS assessment. They were treated with A (10, 20, 40, or 80 mg once daily) titrated to achieve a target LDL-C level of <100 mg/dL (2.6 mmol/L). Subjects achieving the target LDL-C level were randomized to receive (i) fixed combination T/A (T 60 mg combined with A once daily) or (ii) A alone. In both treatment arms, the dose of A was that established during the ran-in period. Following randomization, patients are evaluated at Months 1 and 3, and then every 3 months until the final IVUS study is performed at Month 24. Clinical safety and/or lipid efficacy assessments are performed at each visit. The primary efficacy variable is the nominal change in percent atheroma volume measured by IVUS. Enrollment has been completed and results axe expected in 2007.

Iw16-P-0611

I

THE IMPACT OF FISH OIL FATTY ACIDS ON ENDOTHELIAL FUNCTION

E. Olano-Ma_rtln, M. Vassiliadou, A.M. Minihane. The Hugh Sinclair Unit

of Nutrition, The University of Reading Reading UK Objective: To investigate in vitro the mechanism(s) through which fish oil fatty acids act to improve vascular tone. Methods: EPA, DHA and DPA were supplemented to EAHy926 endothelial cells at 10uM, 25uM and 100uM for 8, 24 and 48 h. Fatty acid enrichment after incubation was confirmed by Gas Chromatography. The effect of these treatments on key modulators of endothelial derived vasodilation (caused by nitric oxide, NO) and vasoconstriction (caused by endothelin-l, ET-1) was studied both at gene and protein level. Gene expression of eNOS and ET-1 was measured using real time RT-PCR. ET-1 protein concentration was determined using and Enzyme Immunometric Assay, and NO production was quantified using a Nitric Oxide Quantification Kit. Results: As expected, the amount of these fatty acids in the cells was higher with increasing concentrations of the treatments and incubation time. EPA supplementation resulted in an increase in both EPA and DPA.

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Conclusions: Overall, among the three fatty acids tested DHA seemed to he the most beneficial with respect to vascular tone, with EPA being the least beneficial and DPA being somewhere in-between the two. However, since our results showed clearly that EPA was elongated to DPA inside the cell the effects seen could have been due to the presence of EPA, DPA, or due to a synergistic effect between the two. EFFECTS OF PHYTOSTEROLS ON LIPOPROTEIN IW16-P-0621 METABOLISM IN SUBJECTS W I T H THE i

METABOLIC SYNDROME E.M.M. Ooi 1, G.F. Watts 1, P.H.R. Barrett 1, P.M. Clifton 2 , P.J. Nestel 3 .

1School of Medicine and Pharmacology (RPH), University of Western Australia, Perth 2 CSIRO, Health Sciences and Nutrition, Adelaide; 3Baker Heart Research Institute (Wynn Domain), Melbourne, Australia Objective: To investigate the effects of dietary plant sterols supplementation on lipoprotein metabolism in men with the metabolic syndrome. Subjects: Nine men with the metabolic syndrome as defined by the following: BMI > 30kg/m 2 or waist:hip ratio > 0.9, triglycerides (TG) > 1.7retool/L, high-density lipoprotein (HDL)-cholesterol <1.10 mmol/L and fasting glucose > 6.1 mmol/L, while consuming an ad llbitum weight-maintenance diet. Study Designs: Randomised, double-blind, cross-over study of 2 x 4 weeks therapeutic periods with placebo or plant sterols (2g/day), and 2 weeks placebo wash-out between therapeutic periods. Methods: VLDL-, IDL- and LDL-apoB and HDL apoA-I kinetics were measured using intravenous, bolus D3-1eucine (4mg/kg), GCMS and compartmental modelling (SAAMII). Results: Plant sterols did not have a significant effect on total cholesterol, TG, LDL-cholesterol, HDL-cholesterol and plasma concentrations of apoB, apoA-I or apoA-II. There were no significant changes to VLDL-, IDL-, LDL-apoB or HDL apoA-I fractional catabolic rate (F'CR) or production rate (PR) between treatment and placebo phases. Campesterol was significantly increased on plant sterols treatment (2.53 =k 0.35 vs 4.64 4- 0.59 I~l/ml, p<0.05). Lathosterol, a marker of endogenous cholesterol synthesis did not increase significantly during plant sterols treatment. Conclusions: There appears to be no significant advantage gained by addition of phytosterols to the daily diet of subjects with the metabolic syndrome with respect to cholesterol lowering or improvement in lipoprotein profile or metabolism. The lack of response may he attributable to the high rates of endogenous cholesterol synthesis relative to cholesterol absorption observed in subjects with the metabolic syndrome. Further studies with a higher dose of phytosterols or other therapeutic agents that reduce endogenous cholesterol synthesis may confer benefits in managing the dyslipidaernia observed in these individuals.

I W16-P-063 1 THE LIPID-ALTERING EFFECTS OF INEGY-TM i

ARE CONSISTENT ACROSS GENDER, RACE, AGE, AND BASELINE LDL-C LEVELS

L. Ose 1, S. Stender 2, J. Rotonda 3 , A. Shah 3, D. Maccubbin 3 , D. Tribble 3, E. Veltri 3, Y. Mitchel 3. 1Lipid Clinic, Medical Departmen~ Rikshospitale~

Oslo, Norway; 2Gentofle University Hospita~ Hellerup, Denmark; SMerck/Schering-Plough Pharmaceuticals, North Wales, PA, USA Background: INEGYTM, the combination tablet containing ezetimihe and simvastatin (EZE/SIM), inhibits both the intestinal absorption and endogenous production of cholesterol. We examined the consistency of LDL-C

75th EAS Congress, 23-26 April 2005, Prague, Czech Republic