Antitumor activity of solamargine in mouse melanoma model: relevance to clinical safety.

Melanoma is the most aggressive type of skin cancer, and thus it is important to develop new drugs for its treatment. The present study aimed to examine the antitumor effects of solamargine a major alkaloid heteroside present in Solanum lycocarpum fruit. In addition solamargine was incorporated into nanoparticles (NP) of yttrium vanadate functionalized with 3-chloropropyltrimethoxysilane (YVO4:Eu3+:CPTES:SM) to determine antitumor activity. The anti-melanoma assessment was performed using a syngeneic mouse melanoma model B16F10 cell line. In addition, systemic toxicity, nephrotoxic, and genotoxic parameters were assessed. Solamargine, at doses of 5 or 10 mg/kg/day administered subcutaneously to male C57BL/6 mice for 5 days, decreased tumor size and frequency of mitoses in tumor tissue, indicative of a decrease in cell proliferation. Treatments with YVO4:Eu3+:CPTES:SM significantly reduced the number of mitoses in tumor tissue, associated with no change in tumor size. There were no apparent signs of systemic toxicity, nephrotoxicity, and genotoxicity initiated by treatments either with solamargine alone or plant alkaloid incorporated into NP. The animals treated with YVO4:Eu3+:CPTES:SM exhibited significant increase in spleen weight accompanied by no apparent histological changes in all tissues examined. In addition, animals treated with solamargine (10 mg/kg/day) and YVO4:Eu3+:CPTES:SM demonstrated significant reduction in hepatic DNA damage which was induced by tumor growth. Therefore, data suggest that solamargine may be considered a promising candidate in cancer therapy with no apparent toxic effects.

Common anticholinergic solanaceaous plants of temperate Europe - A review of intoxications from the literature (1966-2018).

Datura stramonium, Atropa belladonna, Hyoscyamus niger, and Scopolia carniolica are all temperate plants from the family Solanaceae, which as a result of their anticholinergic tropane alkaloids, hyoscyamine/atropine and scopolamine, have caused many cases of poisoning around the world. Despite the danger these nightshade plants represent, the literature often presents incomplete cases lacking in details and filled with ambiguity, and reviews on the topic tend to be limited in scope. Many also point to a gap in knowledge of these plants among physicians. To address this, the following review focuses on intoxications involving these plants as reported in the literature between 1966 and 2018, with brief mention to pertinent related plants to contextualise and provide a fuller picture of the situation surrounding the presently discussed temperate plants. Analysis of the literature displays that D. stramonium is largely associated with drug use among teens while A. belladonna is primarily ingested as a result of the berries being mistaken for edible fruits. H. niger was found to be largely ingested when mistaken for other plants, and S. carniolica was the cause of incredibly few intoxications.

In Vivo Toxicity of Solasonine and Its Effects on cyp450 Family Gene Expression in the Livers of Male Mice from Four Strains.

Solasonine was reported to inhibit tumour cell growth in several different models. The in vivo toxicity of solasonine, the effects of genetic background on its toxicity, and its possible roles in regulating the expression of cyp450 family genes were still unclear and required characterisation. Here, Horn's assays were performed on male mice from four different strains, and the expression of cyp450 family genes in their livers was examined by RT-PCR and ELISA. Mice treated by intraperitoneal injection with high levels of solasonine showed immediate post-excitatory depression, intraperitoneal tissue adhesion, and dissolving of cells in the liver. Furthermore, these four mouse strains showed different toxicological sensitivity to solasonine. The strains, in decreasing order of LD50 value, rescuing speed of body weight, and more severe pathological symptoms, were KM, ICR, C57BL/6, and BALB/c. Interestingly, more cyp450 genes were downregulated at the mRNA and/or protein level in the livers of male mice from C57BL/6 or BALB/c strains than those from KM or ICR strains. These results suggest that (1) Solasonine has hepatic toxicity and downregulates cyp450 genes expression at transcriptional and/or post-transcriptional levels; (2) Genetic background is an important factor which can affect the in vivo toxicity; (3) Downregulation of cyp450 gene expression in the liver may be a clue to help understand whether or not a given strain is sensitive to solasonine; (4) Influences on the expression of cyp450 genes should be considered when using solasonine alone, or in combination with other drugs.

Combination of Solamargine and Metformin Strengthens IGFBP1 Gene Expression Through Inactivation of Stat3 and Reciprocal Interaction Between FOXO3a and SP1.

BACKGROUND/AIMS: Solamargine, one natural photochemical component from traditional plants, has been shown to have anti-cancers properties. We previously showed that solamargine inhibited the growth of non-small-cell lung cancer (NSCLC) cells through suppression of prostaglandin E2 (PGE2) receptor EP4 gene and regulation of downstream signaling pathways. However, the detailed mechanism underlying this, especially in combination of metformin, a known AMPK activator, still remained to be determined. METHODS: Cell viability was measured using a 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) and colorimetric 5-bromo-2-deoxyuridine (BrdU) ELISA methods, respectively. Western blot analysis and immunohistochemistry were performed to examine the phosphorylation and protein expressions of signal transducer and activator of transcription 3 (Stat3), SP1, forkhead box O3a (FOXO3a), and insulin-like growth factor (IGF)-IGF binding protein 1 (IGFBP1). The expression of IGFBP1 mRNA was measured by quantitative real time PCR (qRT-PCR). Silencing of FOXO3a and IGFBP1 were examined by siRNA procedures. Exogenously expression of SP1, FOXO3a, and IGFBP1 were carried out by transient transfection assays. The promoter activity of IGFBP1 was tested using Secrete-PairTM Dual Luminescence Assay Kit. A xenografted tumor model was used to further test the effect of solamargine in combining with metformin in vivo. RESULTS: We further demonstrated that solamargine inhibited growth and induced cell cycle arrest in other NSCLC cell lines. Through mechanism-based approaches, we showed that solamargine decreased the phosphorylation of Stat3; In addition, solamargine induced FOXO3a, whereas reduced SP1 protein levels; all of which were abrogated in cells with overexpressed Stat3 gene. Interestingly, there is interaction between FOXO3a and SP1. Moreover, solamargine increased mRNA, protein expression and promoter activity of IGFBP1, which was not observed in cells with overexpressed SP1 or with silenced FOXO3a genes. Finally, ablation of IGFBP1 expression by siRNA blocked the effect of solamargine on cell growth inhibition. More importantly, there was a synergy of combination of solamargine and metformin. Similar findings were also observed in vivo. CONCLUSION: Our results show that solamargine increases IGFBP1 gene expression through inactivation of Stat3, followed by regulation and reciprocal interaction of FOXO3a and SP1 in vitro and in vivo. This ultimately leads to suppression of human lung cancer cell growth. Moreover, this is a synergy of solamargine in combination with metformin in this process. This study unravels a novel mechanism underlying the anti-lung cancer effects of solamargine in combination of metformin, and suggests a potential new lung cancer associated therapy.

The Steroidal Glycoalkaloids from Solanaceae: Toxic Effect, Antitumour Activity and Mechanism of Action.

Steroidal glycoalkaloids present in Solanaceae are toxic compounds biosynthesised for the protection of the plants. However, many health benefits of these compounds have been reported so far. One of their promising targets might be cancer, as demonstrated in a large number of studies. However, the main mechanism of action seems to be unclear. It could include the induction of apoptosis or trigger a necrosis with a subsequent inflammatory response. The relatively high systemic toxicity of steroidal compounds is another effect that must be taken into account in anticancer research. The main aim of this work was to summarise the recent progress in the investigation of the mechanisms of their antitumour action and to discuss their potential.

Biosynthesis of antinutritional alkaloids in solanaceous crops is mediated by clustered genes.

Steroidal glycoalkaloids (SGAs) such as α-solanine found in solanaceous food plants--as, for example, potato--are antinutritional factors for humans. Comparative coexpression analysis between tomato and potato coupled with chemical profiling revealed an array of 10 genes that partake in SGA biosynthesis. We discovered that six of them exist as a cluster on chromosome 7, whereas an additional two are adjacent in a duplicated genomic region on chromosome 12. Following systematic functional analysis, we suggest a revised SGA biosynthetic pathway starting from cholesterol up to the tetrasaccharide moiety linked to the tomato SGA aglycone. Silencing GLYCOALKALOID METABOLISM 4 prevented accumulation of SGAs in potato tubers and tomato fruit. This may provide a means for removal of unsafe, antinutritional substances present in these widely used food crops.

In vitro leishmanicidal and cytotoxic activities of the glycoalkaloids from Solanum lycocarpum (Solanaceae) fruits.

Leishmaniasis is an infection caused by a protozoan parasite of the genus Leishmania and is the second most prevalent parasitic protozoal disease after malaria in the world. We report the in vitro leishmanicidal activity on promastigote forms of Leishmania amazonensis and cytotoxicity, using LLCMK2 cells, of the glycoalkaloids from the fruits of Solanum lycocarpum, determined by colorimetric methods. The alkaloidic extract was obtained by acid-base extraction; solamargine and solasonine were isolated by silica-gel chromatography, followed by reversed-phase HPLC final purification. The alkaloidic extract, solamargine, solasonine, as well as the equimolar mixture of the glycoalkaloids solamargine and solasonine displayed leishmanicidal activity against promastigote forms of L. amazonensis, whereas the aglycone solasodine was inactive. After 24 and 72 h of incubation, most of the samples showed lower cytotoxicities (IC50 6.5 to 124 µM) as compared to leishmanicidal activity (IC50 1.1 to 23.6 µM). The equimolar mixture solamargine/solasonine was the most active with an IC50 value of 1.1 µM, after 72 h. Likewise, solamargine was the most active after 24 h with an IC50 value of 14.4 µM, both in comparison with the positive control amphotericin B.

Prenatal exposure to tobacco extract containing nicotinic alkaloids produces morphological and behavioral changes in newborn rats.

Tobacco exposure is not only a health concern for adults but has also been shown to exert deleterious effects on the health of the fetus, newborn, child, and adolescent. Decreased cognitive function, lower Intellectual Quotient (IQ) and deficits in learning and memory in children have been associated with maternal smoking during pregnancy. In this study, we have studied the effect of a tobacco plant extract on the growth and development in the rat. The extract contained relative proportions of alkaloids, including nicotine, purified by chemical separation. Pregnant rats received oral doses of either control (NaCl) or tobacco extract during the entire gestational period. Offspring length and body weight were measured. Each day, the offspring were observed for the following physical parameters: hair growth, incisor eruption and eye opening. The day of appearance of these developments was recorded. Before weaning, the offspring were examined to test their cliff avoidance response (6 postnatal day (PN)), surface righting reflex (05, 07, 13 postnatal day), swimming development (10, 12 postnatal day), negative geotaxis response (7,9,13 and 17 postnatal day) and jumping down choice cage (15, 17 postnatal day). Administration of tobacco extract to dams during the entire gestation period affects behavior and development in pups. The observed effects were a delay in opening eyes, incisor eruption and hair appearance, behavioral developments and an alteration in the rate of success behavior. However, in the jumping down choice cage test there was no difference compared to control animals. The results suggest that tobacco extract has a significant effect on the development of behavioral patterns, orientation and motor coordination and function. They also suggest significant growth retardation and teratogenic effects.

In vivo antimalarial activities of glycoalkaloids isolated from Solanaceae plants.

CONTEXT: Malaria is one of the most common and serious protozoan tropical diseases. Multi-drug resistance remains pervasive, necessitating the continuous development of new antimalarial agents. OBJECTIVE: Many glycosides, such as triterpenoid saponins, were shown to have antimalarial activity against Plasmodium falciparum in vitro. This study was to elucidate the ability of five glycoalkaloids against Plasmodium yoelii and develop new antimalarial lead compounds. MATERIALS AND METHODS: Glycoalkaloids were isolated from three kinds of Solanaceae plants: chaconine and solanine were isolated from Solanum tuberosum L. sprouts, solamargine and solasonine from Solanum nigrum L. fruit, tomatine from Lycopersicon esculentum Mill. fruit. The five isolated glycoalkaloids were evaluated against Plasmodium yoelii 17XL in mice with 4-day parasitemia suppression test in different concentrations. RESULTS: Chaconine showed a dose-dependent suppression of malaria infection, ED50, 4.49 mg/kg; therapeutic index (TI), approximately 9. At a dose of 7.50 mg/kg, the parasitemia suppressions of chaconine, tomatine, solamargine, solasonine and solanine were 71.38, 65.25, 64.89, 57.47 and 41.30%, respectively. At 3.75 mg/kg, the parasitemia suppression of chaconine was 42.66%, but the derivative, chaconine-6-O-sulfate, appeared to show no antimalarial activity. Simultaneous administration of chaconine and solanine in 1:1 did not show any synergistic effects. DISCUSSION AND CONCLUSION: The results showed that the glycoalkaloids with chacotriose (chaconine and solamargine) were more active than those with solatriose (solanine and solasonine). Chaconine was the most active among the five glycoalkaloids. We propose that the activity is dependent upon non-specific carbohydrate interactions. The 6-OH of chaconine is important for antimalarial activity.

Application of high performance capillary electrophoresis on toxic alkaloids analysis.

We employed CE to identify mixtures of the toxic alkaloids lappaconitine, bullatine A, atropine sulfate, atropine methobromide, scopolamine hydrobromide, anisodamine hydrobromide, brucine, strychnine, quinine sulfate, and chloroquine in human blood and urine, using procaine hydrochloride as an internal standard. The separation employed a fused-silica capillary of 75 microm id x 60 cm length (effective length: 50.2 cm) and a buffer containing 100 mM phosphate and 5% ACN (pH 4.0). The sample was injected in a pressure mode and the separation was performed at a voltage of 16 kV and a temperature of 25 degrees C. The compounds were detected by UV absorbance at wavelengths of 195 and 235 nm. All the ten alkaloids were separated within 16 min. The method was validated with regard to precision (RSD), accuracy, sensitivity, linear range, LOD, and LOQ. In blood and urine samples, the detection limits were 5-40 ng/mL and linear calibration curves were obtained over the range of 0.02-10 microg/mL. The precision of intra- and interday measurements was less than 15%. Electrophoretic peaks could be identified either by the relative migration time or by their UV spectrum.

Toxicological effects of alpha-solamargine in experimental animals.

Alpha-solamargine isolated from the fresh fruits of Solanum americanum Miller was studied for its toxicity. Lethality studies in rats showed a dose-mortality relationship with a LD(50) of 42 mg/kg body weight intraperitoneally. The chronic and subchronic toxicity investigations indicated that the size of the glycoalkaloid dose was more important than the total glycoalkaloid intake. No appreciable toxic effects were observed at doses below 35 mg/kg body weight as indicated by blood parameters, enzyme levels and histological sections of kidney, liver and cardiac muscle. Alpha-solamargine did not affect the weight of the testes and epididymis or the number of spermatozoa but produced a slight irritation and congestion in the epididymis and testis at doses up to 50 mg/kg body weight.

Toxicological evaluations of long-term consumption of Solanum lycocarpum St. Hill fruits in male and female adult rats.

Solanum lycocarpum St. Hill is a common plant in the Brazilian savanna. This plant contains an alkaloid with stereospecific configuration to the synthesis of steroid hormones. Because the plant may be consumed long-term, the present study was undertaken to determine the possible toxic effects of S. lycocarpum fruit ingestion (3% added to the diet) on male (60 days of administration) and female (37 days) adult rats. Few significant differences in body weight and consumption of food and water, no significant differences in male and female weight gain or estrous cycle were detected. Female treated rats showed a significant reduction in uterus and liver weights; however, no significant differences were observed in other organ (adrenal, liver, seminal vesicle, testicle and ovary) weights in either sex. Additionally blood enzymes and proteins evaluated were not affected by treatment with 3% S. lycocarpum added to the diet. The present data, however, show sex-related differences in S. lycocarpum toxicity. Thus, other studies have to be conducted to better investigate female toxicity and other toxic effects of higher levels of exposure to this plant.

Assessment of the perinatal effects of maternal ingestion of Solanum malacoxylon in rats.

A perinatal study was performed to verify the toxic effects of Solanum malacoxylon, which contains a glycoside conjugated to Vitamin D(3). In the gestational study, female rats received S. malacoxylon leaves in the diet at 0, 0.1, 0.2, 0.5, and 1% from days 6 to 21 of pregnancy. At 21 days of gestation, blood samples were taken from the dams for evaluation of serum Ca and P. A laparotomy was performed and the rats were examined for standard parameters of reproductive performance. Fetuses were examined for skeletal changes and histopathologic evaluation. In the second trial, dams were fed diets containing 0 or 0.1% S. malacoxylon leaves during the gestation and lactation periods. After weaning, all animals were euthanized and biochemical and histopathologic evaluations were performed. The biochemical evaluation showed increase in Ca and P levels in females from all experimental groups; however, this effect did not occurred in a dose-related manner. Pups from dams exposed during gestation and lactationi also showed increased Ca and P levels. Fetal data suggested a delay of fetal development manifested by decreased body weight and skeletal alterations. There was also a reduction in live fetuses. Histopathologic study revealed alterations of the soft tissue in litters from dams given 1% dietary S. malacoxylon during pregnancy and 0.1% during pregnancy and lactation. These findings support our hypothesis that Vitamin D(3) glycoside crosses the placenta and suggests milk transfer of this substance.

Effect of feeding solanidine, solasodine and tomatidine to non-pregnant and pregnant mice.

The aglycone forms of three steroidal glycoalkaloids-solanidine (derived by hydrolytic removal of the carbohydrate side chain from the potato glycoalkaloids alpha-chaconine and alpha-solanine), solasodine (derived from solasonine in eggplants) and tomatidine (derived from alpha-tomatine in tomatoes)-were evaluated for their effects on liver weight increase (hepatomegaly) in non-pregnant and pregnant mice and on fecundity in pregnant mice fed for 14 days on a diet containing 2.4 mmol/kg of aglycone. In non-pregnant mice, observed ratios of % liver weights to body weights (%LW/BWs) were significantly greater than those of the control values as follows (all values in % vs matched controls+/-S.D.): solanidine, 25.5+/-13.2; solasodine 16.8+/-12.0; and tomatidine, 6.0+/-7.1. The corresponding increases in pregnant mice were: solanidine, 5.3+/-10.7; solasodine, 33.1+/-15.1; tomatidine, 8.4+/-9.1. For pregnant mice (a) body weight gains were less with the algycones than with controls: solanidine, -36.1+/-14.5; solasodine, -17.9+/-14.3; tomatidine, -11.9+/-18.1; (b) litter weights were less than controls: solanidine, -27.0+/-17.1; solasodine, -15.5+/-16.8; tomatidine, no difference; (c) the %LTW/BW ratio was less than that of the controls and was significant only for solasodine, -8.7+/-13.7; and (d) the average weight of the fetuses was less than the controls: solanidine, -11.2+/-15.2; solasodine, -11.4+/-9.4; tomatidine, no difference. Abortion of fetuses occurred in five of 24 pregnant mice on the solanidine and none on the other diets. To obtain evidence for possible mechanisms of the observed in vivo effects, the four glycoalkaloids (alpha-chaconine, alpha-solanine, solasonine and alpha-tomatine) mentioned above and the aglycones solanidine and tomatidine were also evaluated in in vitro assays for estrogenic activity. Only solanidine at 10 microM concentration exhibited an increase in the MCF-7 human breast cancer cell proliferation assay. Generally, the biological effects of solanidine differ from those of the parent potato glycoalkaloids. Possible mechanisms of these effects and the implication of the results for food safety and plant physiology are discussed.

Absence of interceptive effect in rats treated with Solanum lycocarpum (St. Hil).

The interceptive effect of the powder of Solanum lycocarpum (St. Hil) (Solanaceae) fruit, used as a hypoglycemic agent by diabetic patients in Minas Gerais state (Brazil), was evaluated to observe possible effects upon zygote and pre-embryo transport in rats, since it contains solamargine and solasonine from which a 3beta-acetoxipregna-5,16-dien-20-one is obtained as well as an alkaloid with stereospecific configuration to the synthesis of steroid hormones. Inseminated rats received an aqueous suspension of 100 mg of the lobeira powder/kg of body weight, by oral gavage, from the 1st to the 4th day of pregnancy. A control group received 5 mL of distilled water in the same schedule. The pregnant rats were weighed at the beginning of treatment and on sacrifice day. Animals were killed on the 5th day of pregnancy. The oviducts and uterine horns were removed and flushed with saline solution to count expanded blastocysts. It was concluded that administration of lobeira did not cause maternal toxicity, alteration of the pre-embryo transport or reduction of the number of expanded blastocysts.

Effects of plant-induced hypervitaminosis D on cutaneous structure, cell differentiation and cell proliferation in cattle.

Solanum glaucophyllum (Sg) (synonym S. malacoxylon) is a plant toxic to cattle due to its high levels of 1,25-dihydroxyvitamin D3 as glycoside derivatives. Sg causes a disease characterized by wasting and calcification of soft tissues. The effects of vitamin D are not only important in calcium homeostasis, but also in immune regulation, cell growth and cell differentiation. Skin samples in Sg-intoxicated and control heifers were studied histologically. Cellular differentiation and proliferation were analysed by immunohistochemical expression of cytokeratins, involucrin and proliferating cell nuclear antigen (PCNA). The results were obtained by image processing and analysis and were statistically evaluated. Sg-intoxicated cattle showed atrophy of epidermis and severe involution of hair follicles and of sebaceous and sweat glands. As judged by PCNA expression, cellular proliferation was reduced, even though the reduction was not statistically significant. The analysed markers of differentiation, e.g. involucrin and cytokeratins 10 and 11, changed in relation to Sg-poisoning. The possible pathogenesis of the skin lesions is discussed.

Evaluation of the toxicity of Solanum lycocarpum in the reproductive system of male mice and rats.

Powdered Solanum lycocarpum fruit is commonly used to treat diabetes, but apparently no studies have been conducted to evaluate potential adverse side effects. In the present paper the toxic effect of S. lycocarpum was evaluated in adult male Wistar rats and Swiss mice. The administration of an aqueous extract prepared using a powder obtained from the S. lycocarpum fruit at two different dose levels (60 mg/15 ml and 120 mg/15 ml distilled water for rats and 30 mg/15 ml and 60 mg/15 ml distilled water for mice, twice daily for 5 days in each case) did not produce body weight variations in either species although a significant weight change was observed in some organs. Significant weight loss was observed only in the ventral prostate of mice receiving the high dose treatment. These results suggest a toxic effect of S. lycocarpum on the male reproductive system of the Swiss mouse, with possible antiandrogenic activity, but there was no apparent antifertility activity in rats at the doses given.

Toxic effect of Solanum Lycocarpum in the male Wistar rats and swiss mice reproductive organs

Uma suspensäo de Solanum Lycocarpum foi administrada a ratos Wistar e camundongos suiços adultos em duas doses diferentes (60 mg/15ml e 120 mg/15ml de água destilada para ratos e 30 mg/15ml e 60 mg/15ml de água destilada para camundongos) para avaliar seu efeito no peso corporal, de testículos, de epidídimo esquerdo, de glândulas sexuais acessórias, de hipófise e na concentraçäo de espermatozóides. O extrato foi administrado duas vezes ao dia durante 5 dias, com sacrifício dos animais 3 dias após o término do tratamento. Os resultados indicaram que, a princípio, Solanum Lycocarpum näo tem efeito tóxico no rato Wistar adulto mas causou perda significativa de peso corporal, de próstata ventral, vesícula seminal e hipófise de camundongo suiço adulto. A reduçäo de peso de hipófise poderia estar indicando alto nível de testosterona ou ausência de estímulo para sua secreçäo. Quanto ao baixo peso de próstata ventral e vesícula seminal, mais estudos säo necessários para explicar as causas dessa reduçäo.

Evaluation in rabbits of the fetal effects of maternal ingestion of Solanum malacoxylon.

A study was performed to determine the possible toxic effects on the young of does that had been fed during the gestational period on a ration containing Solanum malacoxylon (Sm), a calcinogenic plant that contains a vitamin D3-glycoside conjugate. Experimental animals received a ration containing 0.03% or 0.04% of Sm leaves on days 6 to 30 of gestation. The levels of calcium, phosphorus and alkaline phosphatase in their sera, as well as their feed intake and body weight, were evaluated weekly. The does were euthanized 3 days after parturition and paraffin sections stained with haematoxylin and eosin were prepared from their heart, lungs, kidneys and aorta for histopathological examination. The young from does in the Sm 0.03% group were euthanized 3 days after birth and biochemical and histopathological determinations were performed, as described for the does. The does in both experimental groups showed decreased feed consumption and those in the Sm 0.04% group showed lower body weights throughout their gestation. Animals treated with Sm 0.04% presented a high incidence of abortion and stillbirth. There were biochemical and histopathological alterations in both experimental groups, which were more prominent in the does in the Sm 0.04% group. Litters from does treated with Sm 0.03% showed mineralization of soft tissue and an increase in phosphorus and calcium levels. These findings indicate that the vitamin D3-glycoside passes through the placental barrier to the fetus.