One year oral Toxicity of D-004, a lipid extract from Roystonea regia fruits, in Sprague Dawley rats.

D-004, a lipid extract of royal palm (Roystonea regia) fruits that contains a reproducible mixture of fatty acids, has been shown to prevent testosterone and phenylephrine-induced prostate hyperplasia in rodents. This study investigated the long-term oral toxicity of D-004 in rats. Rats from both sexes were randomized into four groups (20 rats sex/group): a control and three treated with D-004 (800, 1500 or 2000 mg/kg/day, respectively). At study completion, rats were sacrificed under anaesthesia. Determinations of blood biochemical and haematological parameters and organ weight were done. Also, necropsy and histopathological studies were performed. Four of 160 rats died before study completion. No clinical signs of toxicity were observed throughout the study. Food and water consumption, bodyweight, blood biochemical and haematological parameters, organ weight ratios and histopathological findings were similar in control and treated groups. The histological lesions found in treated animals are commonly present in this specie and strain according to literature and our historical data. In conclusion, long-term (12 months) oral treatment of rats with D-004 (800-2000 mg/kg/day) did not show evidences of D-004-related toxicity under our conditions. The highest dose tested (2000 mg/kg) was a no-observed adverse effect level in this study.

Study of the long-term carcinogenicity potential of D-003, a mixture of high molecular weight sugarcane wax acids, in mice.

D-003, a mixture of high aliphatic primary acids purified from sugar cane wax, has shown cholesterol-lowering, anti-platelet and antioxidant effects. Previous data demonstrated that D-003 was not toxic or carcinogenic when given orally to Sprague-Dawley rats up to 1500 mg/kg. This study investigated the potential long-term oral carcinogenicity of D-003 in a second rodent species. OF1 mice of both sexes were randomized into 4 groups treated for 18 months: a vehicle control group and three groups treated with D-003 at 50, 500 and 1500 mg/kg, respectively, orally gavaged 6 days per week. Mortality, clinical symptoms, weight gain, food consumption, organ weight, blood indicators and tumour incidence did not show significant differences between control and treated groups. D-003 did not increase the frequency of neoplastic or non-neoplastic lesions with respect to the controls. Lesions observed in the study were consistent with spontaneous lesions reported for this specie. It can be concluded that D-003 did not result toxic or carcinogenic when given orally to OF1 mice for 18 months and that the highest dose was a NOAEL, consistent with results of the oral carcinogenicity study of D-003 in rats.

Long-term effects of D-003, a mixture of high molecular weight acids from sugarcane wax, on bones of ovariectomized rats: a one year study.

This study was done to determine the long-term effect of D-003 on bones of ovariectomized (ovx) rats distrib-uted in 4 groups: a false-operated and three groups of ovx rats: one treated with the vehicle and two with D-003 (5 and 250 mg/kg). D-003 significantly prevented, in a dose-dependent fashion, the trabecular bone volume (TBV), trabecular number (TbN) and trabecular thickness (TbTh) reduction induced in ovx rats and the increase of trabecular separation (TbSp) osteoclast number (OcN) and osteoclast surface (OcS/BS) increased in the positive controls versus the sham group. It is concluded that D-003 administered for 12 months prevented bone loss and decreased bone resorption in ovx rats, without evidences of impaired bone quality.

Long-term carcinogenicity of D-003, a mixture of high molecular weight acids from sugarcane wax, in Sprague Dawley rats: a 24 months study.

D-003 is a mixture of high molecular weight sugarcane wax aliphatic primary acids with cholesterol-lowering, anti-platelet and antioxidant effects. This study investigated the long-term oral toxicity and carcinogenicity of D-003 in Sprague Dawley rats of both sexes, randomly distributed into four groups: a control group, treated only with the vehicle, and three treated with D-003 (50, 500 and 1500 mg/kg). All treatments were given orally for 24 months. Mortality (survival analysis), clinical symptoms, weight gain, food consumption, organ weights, time-to-tumour or tumour incidence data were not shown between group differences or trends. With the exception of serum cholesterol levels, lower in D-003-treated groups (500 and 1500 mg/kg) than in the controls, no other difference in blood indicators was found. D-003 did not increase the frequency of neoplastic and non-neoplastic lesions compared with the controls. The occurrence of all malignant and mammary tumours in D-003-treated females was lower than in the controls. The lesions observed were consistent with spontaneous lesions reported in this species. In conclusion, D-003 is not toxic or carcinogenic when given orally to Sprague Dawley rats up to 1500 mg/kg for 2 years, and 1500 mg/kg was a not-observable effect dose.

Lack of in vivo embryotoxic and genotoxic activities of orally administered stem bark aqueous extract of Mangifera indica L. (Vimang).

Mango (Mangifera indica L.) stem bark aqueous extract (MSBE) is a new natural product with antioxidant, anti-inflammatory and immunomodulatory effects known by the brand name of its formulations as Vimang. Previously, the oral toxicity studies of the extract showed a low toxicity potential up to 2000 mg/kg. This work reports the results about teratogenic and genotoxicologic studies of MSBE. For embryotoxicity study, MSBE (20, 200, or 2000 mg/kg/day) was given to Sprague-Dawley rats by gavage on days 6-15 of gestation. For genotoxicity, MSBE was administered three times during 48 h to NMRI mice. Cyclophosphamide (50 mg/kg) was used as a positive control. No maternal or developmental toxicities were observed when the rats were killed on day 20th. The maternal body-weight gain was not affected. No dose-related effects were observed in implantations, fetal viability or external fetal development. Skeletal and visceral development was similar among fetuses from all groups. No genotoxicity was observed in bone marrow erythrocytes and liver cells after administration. MSBE appears to be neither embryotoxic nor genotoxic as measured by bone marrow cytogenetics in rodents.

Perinatal/postnatal study of D-003, a mixture of long-chain fatty acids, in rats.

D-003 is a mixture of long-chain fatty acids isolated and purified from sugar cane wax with cholesterol-lowering and antiplatelet effects. In order to further characterize the developmental toxicity during the treatment period from late gestation up to weaning of the offspring, pregnant females received 0 (control), 500, and 1,000 mg/kg/day D-003 daily by oral gavage beginning at day 15 of pregnancy and through gestation until day 21 postpartum. Maternal clinical signs, body weight, and food intake were measured at regular intervals during gestation and lactation. Live pups were weighed, sexed, and examined for developmental signs. One female and male of each litter were randomly selected to evaluate the reproductive potential. There were no spontaneous or dose-related maternal deaths during the course of this study. The general health and behavioral condition of offspring was good in all groups. No significant differences among groups were found in comparisons of litter size, survival through the weaning period, sex ratio, and male and female weights. This peri- and postnatal study conducted with D-003 in rats indicated that treatment of the dam during late gestation and lactation did not show adversely effects on reproductive performance or fetal development over two generations.

Oral acute and subchronic toxicity of D-004, a lipid extract from Roystonea regia fruits, in rats.

D-004 is a lipid extract obtained from Cuban royal palm (Rosytonea regia) fruits, consisting of a mixture of fatty acids and esters. D-004 has shown protective effects on prostate hyperplasia induced by testosterone in rodents. We report the results of two studies investigating the acute and subchronic oral toxicity of D004 in rats. Oral acute toxicity of D-004 (2,000 mg/kg) was investigated in Sprague Dawley rats according to the acute toxic class method, and the results showed that D-004 oral acute toxicity was practically absent, being defined as unclassified. In the subchronic study, rats were orally treated with D-004 at 500, 1,000 and 2,000 mg/kg for 90 days. No evidence of treatment-related toxicity was detected. Thus, analysis of body weight gain, clinical observations, blood biochemistry, hematology, organ weight ratios and histopathological data did not show significant differences between control and treated groups. We conclude that D-004 orally administered to rats was safe and that no drug-related toxicity was detected even at the highest dose investigated in both acute and subchronic (2,000 mg/kg) studies. Thus, this dose can be considered as a nonobservable-effect dose in rats.

Evaluation of the reproductive and developmental toxicity of the D-003, a mixture of long-chain fatty acids, in rats and rabbits.

D-003 is a mixture of long-chain fatty acids isolated and purified from sugar cane wax with cholesterol-lowering properties. D-003 given orally (500 and 1000 mg/kg/day) to female rats for 15 days prior to mating, through mating and gestation to day 21 of lactation and male rats for 4 weeks prior and during mating did not induce toxic effects on reproduction. There were no significant reductions in the number of animals that conceived, in the numbers of pups born to those that did conceive, in the numbers of pups that survived until weaning, and in their body weights at weaning. Drug-treated and control groups' offspring were comparable in growth, physical and behavioral development, spontaneous activity and reproductive performance. Pregnant New Zealand rabbits were given D-003 as oral doses of 500 and 1000 mg/kg/day on days 6 through 18 of gestation without any evidence of embryotoxicity or teratogenicity. The no-observed-effect dose in these two experimental studies was 1000 mg/kg/day. After assessment of the potential of high doses of D-003 to act on developing embryo and reproduction process, no evidence supports the conclusion that D-003 is a reproductive and developmental toxicant/teratogen.

Effects of chronic administration of D-003, a mixture of sugar cane wax high molecular acids, in beagle dogs.

D-003 is a mixture of high molecular weight aliphatic primary acids purified from sugar cane wax (Saccharum officinarum, L) with cholesterol-lowering and antiplatelet effects. Previous studies, including a 6-month study conducted in rats, have shown no D-003-related toxicity. The present study was undertaken to investigate the effects of D-003 orally administered for 9 months in beagle dogs. The animals were randomly distributed in three groups: a control group receiving the vehicle only and two groups orally administered D-003 (200 and 400 mg/kg). Body weight gain, food consumption and clinical signs were controlled throughout the study. The effects of D-003 on collagen-induced platelet aggregation, bleeding time (BT) and coagulation parameters (prothrombin time and kaolin-activated thromboplastin time) were also investigated. Most blood biochemistry and hematological parameters were assessed at baseline and after 6 and 9 months of treatment, while total cholesterol (TC), triglycerides, platelet aggregation, BT and coagulation parameters were determined at baseline and after 9 months of treatment. At study completion, the animals were sacrificed. D-003 at a dose of 200 and 400 mg/kg significantly reduced TC (p < 0.05), significantly inhibited platelet aggregation and increased BT compared with levels in controls. Data analyses of body weight gain, food consumption, clinical observations, the remaining blood biochemistry and hematology indicators (including coagulation parameters, organ weight ratios and histopathological findings) showed no trends with D-003 doses or significant differences between control animals and treated groups. In conclusion, D-003 administered for 9 months to beagle dogs induced the expected effects with no evidence of drug-related toxicity.

Lack of developmental toxicity of D-003: a mixture of long-chain fatty acids in rats.

D-003 is a mixture of long-chain fatty acids isolated and purified from sugar cane wax, the major component of which is 1-octacosanoic acid and which possesses effective antiplatelet, antithrombotic and cholesterol-lowering effects. D-003 was suspended in 1% acacia gum solution, and given daily by gavage to rats at dose levels of 5, 100 and 1000 mg/kg/day on days 6 through 15 of gestation. Cyclophosphamide, serving as a positive control, was given at the dose of 50 mg/kg/day on day 15 of gestation. Evidence of maternal or developmental toxicity was not observed in the groups treated with D-003. Maternal clinical signs of toxicity were not observed and the analysis of initial body weight and the body weight gain during the treatment period was comparable among the groups treated with D-003 and control. As expected, cyclophosphamide caused both embryotoxic and teratogenic effects in rats. Meanwhile, no adverse effects on reproductive performance, or on embryonic or fetal development, including visceral and skeletal examination, were seen in any of the groups administered D-003. It is concluded that D-003 administered up to 1000 mg/kg/day did not induce any evidence of developmental toxicity.

Effects of D-003, a mixture of long-chain aliphatic primary acids, fluvastatin and the combined therapy of D-003 plus fluvastatin on the lipid profile of normocholesterolemic rabbits.

D-003 is a mixture of long-chain aliphatic primary acids isolated from sugar cane wax with cholesterol-lowering effects proven in animals and healthy human volunteers. D-003 reduced serum total cholesterol (TC) and low-density lipoprotein cholesterol (LDL-C) in rabbits, while it increased high-density lipoprotein cholesterol (HDL-C) and did not affect triglycerides. D-003 inhibits cholesterol synthesis by regulating, instead of directly inhibiting, hydroxamethylglutaryl-CoA (HMGCoA) reductase activity. Although the ways in which D-003 and statins inhibit cholesterol biosynthesis are not identical, the strong competitive inhibition of cholesterol biosynthesis induced by statins suggests that an enhanced decrease of LDL-C and TC caused by the combined therapy D-003 plus statins is not expected. Nevertheless, taking into account the differential effects of D-003 and statins in HDL-C and triglycerides in rabbits, potential benefits of such combined therapy on other lipid variables cannot been discarded. Fluvastatin is a statin that inhibits competitively HMGCoA reductase, like other members of this class. This study was undertaken to compare the cholesterol-lowering effects of D-003, fluvastatin and the combined therapy of D-003 plus fluvastatin in normocholesterolemic rabbits. Animals were randomly distributed into four groups of eight. One control group received the vehicle, two groups were treated with D-003 or fluvastatin at 5 mg/kg/day each, and the fourth group received the combined therapy of both drugs at 5 mg/kg/day each. Treatments were orally administered for 30 days. Body weight, food consumption and overall animal behavior were recorded to detect any warning sign resulting from combined therapy. After treatment, it was found that both D-003 and fluvastatin had significantly lowered LDL-C - D-003 by 81.5% (p < 0.01) and fluvastatin by 61.4% (p < 0.05). Combined therapy reduced LDL-C values (75.9%). Final values and percent changes reached in all groups were different from the control (p < 0.01). The reductions of TC were consistent with LDL-C decreases, so that D-003, fluvastatin and combined therapy significantly lowered TC by 48.4% (p < 0.01), 39.7% (p < 0.05) and 45.3%, respectively, values being different from those of the control (p < 0.01). The responses of LDL-C and TC to combined therapy were statistically similar, but less pronounced than those reached by D-003 alone. D-003 and combined therapy, but not fluvastatin alone, increased HDL-C (+21.5% and + 19.0%, respectively), these changes being significant versus the control (p < 0.05). In turn, fluvastatin and combined therapy, but not D-003 alone, lowered triglycerides (13.6% and 13.0%, respectively, p < 0.05 versus control). The effects of combined therapy on HDL-C were similar to those of D-003 alone, and the effects of combined therapy on triglycerides were similar to those of fluvastatin alone. The only advantage of combined therapy appears to be that it shows better effects on HDL-C than those of fluvastatin alone and better effects on triglycerides than D-003 alone. No significant changes in lipid profile were observed in the control group. All groups showed similar food consumption and body weight gain, health status being unaffected by the treatments. It is concluded that D-003 and fluvastatin at 5 mg/kg/day administered orally for 30 days to normocholesterolemic rabbits lowered LDL-C and TC, D-003 being more effective in increasing HDL-C and fluvastatin in lowering triglycerides. Combined therapy did not improve the response of LDL-C and TC with respect to monotherapies, but induced better responses of HDL-C and triglycerides than fluvastatin alone had on HDL-C or D-003 alone had on triglycerides.

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.