Rafael Gámez

Effects of Policosanol and Lovastatin in Patients with Intermittent Claudication: A Double-Blind Comparative Pilot Study

Policosanol is a cholesterol-lowering drug with concomitant antiplatelet effects. The present study was undertaken to compare the effects of policosanol and lovastatin on patients with moderately severe intermittent claudication. The study had a 4-week baseline step, followed by a 20-week double blinded, randomized treatment period. Twenty-eight patients who met study entry criteria were randomized to policosanol 10 mg or lovastatin 20 mg tablets once daily. Walking distances in a treadmill (constant speed 3.2 km/hr, slope 10°, temperature 25°C) were assessed before and after 20 weeks of treatment. Both groups were similar at random ization. Compared with baseline, policosanol increased significantly (p<0.01) the initial claudi cation distance (ICD) from 160.39 ±15.82 m to 211.31 ±21.48 m (+33.7%) and the absolute claudication distance (ACD) (p<0.001) from 236.39 ±25.44 m to 288.09 ±28.47 m (+24.3%); meanwhile both variables remained unchanged after lovastatin therapy. Changes in ICD and ACD were significantly larger in the policosanol than in the lovastatin group (p<0.01). Policosanol, but not lovastatin, significantly increased (p < 0.05) the ankle/arm index, although between-group differences were not significant. The frequency of patients reporting improve ment on quality of life domains was greater in the policosanol than in the lovastatin group. Policosanol significantly (p< 0.00 1 ) lowered total cholesterol (TC) and low-density lipoprotein- cholesterol (LDL-C) by 17.5% and 31.0%, respectively, and meanwhile increased (p<0.01) high- density lipoprotein-cholesterol (HDL-C) levels by 31.5%. Lovastatin reduced (p<0.01) TC (18.0%), LDL-C (22.6%), and (p<0.05) triglycerides (9.8%). In addition, policosanol, but not lovastatin, moderately, but significantly, reduced (p < 0.05) fibrinogen levels, so that final values and percent changes in both groups were different (p<0.01). Treatments were well tolerated. Only 1 lovastatin patient withdrew from the study because of a nonfatal myocardial infarction. Five lovastatin patients, but none from the policosanol group, experienced 6 adverse events (AE) (p<0.01). The present results indicate that policosanol, but not lovastatin, is a suitable alternative to manage patients with intermittent claudication because of pleiotropic properties beyond its cholesterol-lowering effects.

Toxicity of policosanol in beagle dogs: one-year study

Policosanol is a new chemical entity composed of 8 higher aliphatic alcohols obtained from sugar cane (Saccharum officinarum), L. wax, whose cholesterol-lowering effects have been demonstrated in experimental models, healthy volunteers and patients with type II hypercholesterolemia. This study investigated the oral toxicity of policosanol administered for 52 weeks to beagle dogs. Twenty-four beagle dogs (12 males and 12 females) were distributed randomly in 3 experimental groups (4 animals/group): a control and 2 treated groups receiving policosanol at 30 and 180 mg/kg daily (7 days/week) by gavage. No mortality was observed in any group. Overall, policosanol was well tolerated throughout the study and no toxic symptoms were observed. All groups showed similar weight gain and food consumption. Lipid profile determinations showed that policosanol decreased total cholesterol by 20% approximately from 8 to 52 weeks. Cholesterol-lowering effects did not wear off during the study, thus demonstrating the persistence of the effectiveness. Triglycerides and high density lipoprotein-cholesterol (HDL-C) were not changed significantly. No blood biochemistry or histopathological disturbances attributable to treatment were observed. This study has shown that no drug-related toxicity was induced by policosanol administered up to 180 mg/kg/day for 52 weeks to beagle dogs. Since this dose is approximately 620 times higher than the maximal recommended therapeutic dose (20 mg/day) it indicates a good safety margin of this product.

Dose-dependent cholesterol-lowering effects of d-003 on normocholesterolemic rabbits

Abstract Objectives The goal of the present study was to evaluate the effects of D-003 (5 to 200 mg/kg) administered orally for 30 days on the serum lipid profile of normocholesterolemic New Zealand rabbits as well as the effects of treatment withdrawal for the following 30 days. Background D-003 is a mixture of higher primary aliphatic acids; octacosanoic acid is its major component. Methods Thirty-five animals were adapted to experimental conditions for 15 days and then randomized to 5 study groups: a control group, which received only orally equivalent volumes of the vehicle, and 4 groups treated with the various doses of D-003 (5, 25, 100, or 200 mg/kg). Results After 30 days of treatment, D-003 significantly ( P P Conclusions Results of this study suggest that D-003 has reversible cholesterol-lowering effects characterized by a dosedependent reduction of TC and LDL-C values accompanied by a non-dose-dependent increase in HDL-C levels.

A long-term study of policosanol in the treatment of intermittent claudication

Policosanol is a cholesterol-lowering drug with concomitant antiplatelet effects. This study was undertaken to investigate the long-term effects of policosanol administered to patients with moderately severe intermittent claudication. The study consisted of a 6-week single-blind, placebo-controlled run in phase, followed by a 2-year double-blind, randomized treatment step. Fifty-six patients who met study entry criteria were randomized to receive placebo or poli cosanol 10 mg twice daily. Walking distances on a treadmill (constant speed 3.2 km/h, slope 10°, temperature 25°C) were assessed before and after 6, 12, 18, and 24 months of treatment. Both groups were similar at randomization. After 6 months of therapy, policosanol significantly increased (p < 0.0 1) the initial claudication distance from 125.9 ±8.7 m to 201.1 ±24.8 m and the absolute claudication distance from 219.5 ± 14.1 m to 380.7 ±50.2 m. Both variables remained unchanged in the placebo group (p<0.01). These effects did not wear off but improved after long-term therapy, so that final values were 333.5 ±28.6 m (initial claudication distance) and 648.9 ±54.1 m (absolute claudication distance); both significantly greater (p<0.0001) than those obtained in the placebo group, which showed values of 137.9 ±21.8 m (initial claudication distance) and 237.7 ±28.1 m (absolute claudication distance), respectively. At study completion, 21 policosanol and 5 placebo patients attained increases in claudication distance values > 50% (p < 0.001). Policosanol, but not placebo, significantly increased the ankle/arm pressure index. In addition, from month 6 up to study completion, the frequency of patients reporting improve ment of lower limb symptoms was greater in the policosanol group than in the placebo group. The treatment was tolerated well. There were 16 withdrawals (12 placebo, 4 policosanol) from the study. Eight patients in the placebo group experienced a total of 10 serious adverse events, 8 of which were vascular events, compared with none in the policosanol group (p<0.01). In addition, 3 patients in the policosanol group and 3 patients in the placebo group reported mild adverse events during the study. The present results demonstrate the long-term usefulness of policosanol therapy to treat patients with intermittent claudication.

Acute and oral subchronic toxicity of D-003 in rats

D-003 is a mixture of higher aliphatic primary acids purified from sugar cane wax (Saccharum officinarum) with cholesterol-lowering and antiplatelet effects experimentally proven. The present work reports the results of two studies investigating the acute and subchronic oral toxicity of D-003 in rats. Oral acute toxicity of D-003 (2000 mg/kg) was investigated according to the Acute Toxic Class (ATC) method (an alternative for the classical LD(50) test), which was performed in Wistar rats. The results obtained in this study defined D-003 oral acute toxicity as unclassified. In the subchronic study, rats of both sexes were orally treated with D-003 at 50, 200 and 1250 mg/kg for 90 days. At this time, animals were sacrificed. No evidence of treatment-related toxicity was detected during the study. Thus, data analysis of body weight gain, food consumption, clinical observations, blood biochemical, haematology, organ weight ratios and histopathological findings did not show significant differences between control and treated groups. It is concluded that D-003 orally administered to rats was safe and that no drug-related toxicity was detected even at the highest doses investigated in both acute (2000 mg/kg) and subchronic (1250 mg/kg) studies.

Comparison of the effects of D-003 and policosanol on lipid profile and endothelial cells in normocholesterolemic rabbits

Abstract Background: Policosanol is a cholesterol-lowering drug purified from sugar cane wax that consists of a mixture of higher aliphatic primary alcohols. D-003 is a mixture of long-chain aliphatic primary acids isolated from the same source with experimentally proven cholesterol-lowering effects. Objective: The present study was undertaken to compare the cholesterol-lowering effects of D-003 and policosanol in normocholesterolemic New Zealand rabbits. In addition, the effects of the 2 drugs on levels of plasma circulating endothelial cells (PCEC) before and after citrate-induced endothelial damage were compared. Methods: Animals were randomly distributed to 3 experimental groups: 1 control and 2 treatment groups. Rabbits in the treatment groups received orally administered policosanol or D-003 daily at 5 mg/kg, the lowest effective dose of policosanol in this model, for 30 days. Results: After 15 days of treatment, both D-003 and policosanol significantly ( P P P P P P P P P Conclusions: D-003 administered orally at 5 mg/kg for 30 days to normocholesterolemic rabbits induced more beneficial changes in serum LDL-C levels than policosanol at the same dose. In addition, a slightly greater beneficial effect on HDL-C levels and PCEC count was noted with D-003 versus policosanol. Although D-003 was more effective than policosanol in these experimental conditions, additional comparative studies that include a wide dose range of both drugs must be conducted before definitive conclusions can be made about the comparative cholesterol-lowering effects of D-003.

Assessment of the Effects of D-003, a New Antiplatelet and Lipid-Lowering Compound, in Healthy Volunteers

AbstractObjective: To investigate the effects of D-003 on the bleeding time (BT) and lipid profile of healthy human volunteers. Methods: This single-blind, randomised, placebo-controlled, parallel-group study was conducted in healthy volunteers. Step 1 investigated the effects of single doses of D-003 5, 25 or 50mg on BT in comparison with placebo. Step 2 investigated the effects of 30 days of D-003 5, 25 or 50 mg/day compared with placebo on lipid profile with an interim assessment at 14 days. BT, lipid profile, physical and haematological safety indicators were measured and adverse events (AEs) recorded.Both steps were followed by a 14- or 30-day washout period Results:Step 1: D-003 25 and 50mg significantly increased mean BT 2 hours after administration compared with baseline, but a significant difference versus placebo occurred only with the 50mg dose. Individual values from participants taking this dose, however, remained within normal limits. This effect was reversible. BT values obtained 2 hours after drug administration showed a moderate dose-dependent relationship. No drug-related changes in safety indicators were found with D-003. Step 2: After 7 days on D-003 50 mg/day, BT was significantly increased compared with baseline and placebo up to the end of the active treatment period. However, all individual values for participants taking this dosage remainedwithin the normal range. This effect was reversible by the end of the washout period. After 30 days, D-003 (5, 25 and 50 mg/day) significantly reduced serum TC (by 13.3 to 17.4%) and LDL-C (by 11.6 to 22.6%) levels, and raised HDL-C levels (by 14.6 to 29.7%), but did not affect triglyceride levels. The significant increase in HDL-Cwas observed after 14 days on treatment. The effects on the lipid profile were reversible by the end of the 30-day washout period, although after 14 days of washout the effects on HDL-C and LDL-C still remained significant, revealing a certain persistence of effect. Eight participants (four receiving placebo and four receiving D-003 5, 25 or 50 mg/day) reported a total of nine AEs, none of which were drug-related. Of these patients, only two treated with D-003 25 and 50 mg/day discontinued treatment. Conclusions: D-003 in single or repeated doses (50mg) induced significant and reversible increases in BT. In addition, repeated doses (5, 25 and 50 mg/day) significantly and reversibly lowered serum LDL-C and TC levels and significantly raised serum HDL-C levels. These effects were reversible by 30 days after the end of treatment.

In Vivo Genotoxic Evaluation of D-003, a Mixture of Very Long Chain Aliphatic Acids

D-003 is a mixture of very long chain aliphatic acids purified from sugar cane wax with cholesterol-lowering effects. The present study was undertaken to investigate the in vivo cytotoxic and genotoxic potential of D-003 using three established assays: bone marrow micronucleus, sperm morphology, and single cell gel electrophoresis (Comet) assay. In a first experimental series, CEN/NMRI mice (6-8 animals per sex per group) were administered D-003 by gastric gavage at 5, 50, or 500 mg/kg for 90 days, then sacrificed 24 hours after the last administration. The effects on bone marrow micronucleus were evaluated only in female mice. D-003 (5-500 mg/kg) did not increase the frequency of micronucleated polychromatic erythrocytes, nor the ratio of polychromatic to normochromatic erythrocytes, compared with the controls. The assessment of the effects on sperm morphology showed that D-003 did not change the sperm count or the frequency of all types of abnormal head shapes, compared with the controls. In a second series, the micronucleus assay was performed in mice of both sexes given 2,000 mg/kg for 6 days. Likewise, in this series, neither cytotoxic nor genotoxic effects were found. Finally, five male Sprague-Dawley rats were treated with D-003 (1,250 mg/kg) by oral gavage for 90 days, and Comet assay on liver cells was performed. No single-strand breaks or alkali-labile site induction on DNA was observed. These results indicate that D-003 does not show evidence of cytotoxic or genotoxic activity on either somatic or germ cells in rodents.

Effects of D-003, a New Compound Purified from Sugarcane Wax, on Platelet Aggregation in Healthy Volunteers: A Randomised, Double-Blind Clinical Study

BackgroundD-003 is a mixture of long-chain aliphatic primary acids purified from sugarcane wax with experimentally demonstrated antiplatelet effects. D-003 shows hypocholesterolaemic effects in rabbits and healthy volunteers, lowering serum total cholesterol (TC) and low-density lipoprotein-cholesterol (LDL-C), but increasing high-density lipoprotein-cholesterol (HDL-C).ObjectiveTo investigate the effects of D-003 administered for 10 days on platelet aggregation in healthy volunteers.ParticipantsHealthy men and women aged 20–55 years.MethodsThe present double-blind, randomised, placebo-controlled study investigated the effects of D-003 (5, 10 and 20 mg/day), on platelet aggregation induced by arachidonic acid (AA) [0.75 and 1.5 mmol/L], collagen (1 mg/L) and adenosine diphosphate (ADP) [1 and 2 μmol/L]. The reversibility of the effects was also investigated. Forty-one subjects were randomised to receive placebo or D-003 at 5, 10 and 20 mg/day for 10 days followed by a washout period of 7 days. At baseline and after active treatment and washout completion, platelet aggregation, lipid profile and safety indicators were assessed.ResultsD-003 significantly, markedly and reversibly inhibited platelet aggregation induced by AA 0.75 and 1.5 mmol/L in a dose-dependent manner. D-003 at 10 mg/day significantly inhibited (p < 0.05) aggregation induced by AA 0.75 mmol/L from 60.4-21.0% and that induced by AA 1.5 mmol/L from 61.5-26.8%. The dosage of 20 mg/day inhibited platelet aggregation induced by AA 0.75 mmol/L from 57.2-11.2%, and platelet aggregation induced by AA 1.5 mmol/L from 65.1-21.6%. D-003 at 10 and 20 mg/day also inhibited platelet aggregation induced by collagen 1 mg/L, the effect being moderate, not dose related and reversible after washout. Platelet aggregation induced by ADP and coagulation time were unaffected by the treatment. D-003 at 5 mg/day was ineffective in inhibiting platelet aggregation. TC and triglycerides remained unchanged after therapy, as was expected for such a short duration of administration. Nevertheless, D-003 (10 and 20 mg/day) significantly (p < 0.05 vs baseline and placebo ) raised HDL-C by 20%, an effect that was not dose dependent and was partially reversible after washout. As a consequence, final values of LDL-C were lower (p < 0.05) in the 20 mg/day group than in the placebo group. No significant changes in lipid profiles occurred with placebo. No drug-related changes in safety indicators were observed. Four subjects, one from each group, withdrew from the study, none because of adverse events (AEs). No patients reported AEs during the study.ConclusionsD-003 at 10 and 20 mg/day for 10 days significantly inhibited AA-and collagen-induced platelet aggregation, without changes in ADP-induced platelet aggregation or coagulation time. The effects on AA-induced aggregation were marked, dose dependent and reversible, whereas the effects on collagen-induced aggregation were moderate and not dose dependent. D-003 (10 and 20 mg/day) also significantly but not dose dependently raised HDL-C. D-003 was well tolerated, with no AEs being reported and no changes in safety indicators being observed. The effects described suggest that D-003 is a promising agent with concomitant antiplatelet and lipid-profile modifying effects, and is potentially useful for the treatment and/or prevention of atherothrombotic disorders. Clinical investigation of D-003 is justified and further studies are needed to demonstrate such a hypothesis.

Effects of Addition of Policosanol to Omega-3 Fatty Acid Therapy on the Lipid Profile of Patients with Type II Hypercholesterolaemia

AbstractBackground: Policosanol is a mixture of higher aliphatic primary alcohols purified from sugar-cane wax. The mixture has cholesterol-lowering efficacy, its specific effects being to reduce serum total (TC) and low-density lipoprotein cholesterol (LDL-C), and to increase high-density lipoprotein cholesterol (HDL-C). The effects of policosanol on triglycerides (TG) are modest and inconsistent. Omega-3 fatty acids (FA) from fish oil protect against coronary disease, mainly through antiarrhythmic and antiplatelet effects. Omega-3 FA also have lipid-modifying effects, mostly relating to TG reduction. Thus, potential benefits could be expected from combined therapy with omega-3 FA and policosanol. Objective: To investigate whether combined therapy with omega-3 FA + policosanol offers benefits compared with omega-3 FA + placebo with respect to the lipid profile of patients with type II hypercholesterolaemia. Methods: This randomised, double-blind study was conducted in 90 patients with type II hypercholesterolaemia. After 5 weeks on a cholesterol-lowering diet, patients were randomised to omega-3 FA + placebo, omega-3 FA + policosanol 5 mg/day or omega-3 FA + policosanol 10 mg/day for 8 weeks. Omega-3 FA was supplied as 1g capsules (two per day); placebo and policosanol were provided in tablet form. Physical signs and laboratory markers were assessed at baseline and after 4 and 8 weeks on therapy. Drug compliance and adverse experiences (AEs) were assessed at weeks 4 and 8. The primary efficacy variable was LDL-C reduction; other lipid profile markers were secondary variables. Results: After 8 weeks, omega-3 FA + policosanol 5 and 10 mg/day, but not omega-3 FA + placebo, significantly reduced LDL-C by 21.1% and 24.4%, respectively (both p < 0.0001). Omega-3 FA + policosanol 5 mg/day also significantly lowered TC (12.7%; p < 0.01) and TG (13.6%; p < 0.05), and significantly increased HDL-C (+14.4%; p < 0.001). Omega-3 FA + policosanol 10 mg/day significantly decreased TC (15.3%; p < 0.001) and TG (14.7%; p < 0.01), and significantly increased HDL-C (+15.5%; p < 0.0001). Omega-3 FA + placebo significantly reduced TG (14.2%; p < 0.05) but had no significant effect on other lipid profile variables. The proportion of randomised patients in the omega-3 FA + policosanol 5 or 10 mg/day groups that achieved LDL-C targets or reductions ≥15% was significantly greater than in the omega-3 FA + placebo group (p < 0.001). Combined therapy with omega-3 FA + policosanol 5 or 10 mg/day resulted in significantly greater changes in LDL-C, TC and HDL-C than treatment with omega-3 FA + placebo, but did not modify the TG response compared with the omega-3 FA + placebo group.Four patients (two in the omega-3 FA + placebo group and two in the omega-3 FA + policosanol 10 mg/day group) withdrew from the study; none of these withdrawals was due to AEs. Two patients reported mild AEs, namely nausea/headache (one in the omega-3 FA + placebo group) and heartburn (one in the omega-3 FA + policosanol 5 mg/day group). Conclusions: Policosanol 5 or 10 mg/day administered concomitantly with omega-3 FA 1 g/day improved LDL-C, TC and HDL-C, maintained the reduction in TG attributable to omega-3 FA monotherapy, and was well tolerated. Treatment with omega-3 FA + policosanol could be useful for regulating lipid profile in patients with type II hypercholesterolaemia, but further studies involving larger sample sizes are needed before definitive conclusions can be drawn.