Natural compounds, such as citronellal (CTL), daidzin (DZN), and phytol (PHY), have demonstrated potential anticonvulsant properties, but their combined effects remain unexplored. This study aimed to evaluate the anticonvulsant efficacy of CTL, DZN, and PHY, individually and in combination, using in vivo and in silico approaches. In the in vivo study, pentylenetetrazole (PTZ)-induced seizures in chicks were used to assess seizure latency, frequency, and duration. CTL (250mg/kg), DZN (20mg/kg), and PHY (75mg/kg) were administered oral (p.o.), alone and in combination. Carbamazepine (CAR, 80mg/kg) and diazepam (DZP, 5mg/kg) served as standard controls. In silico molecular docking was performed against γ-aminobutyric acid A (GABAA) and voltage-gated sodium channel (VGSC), whereas pharmacokinetics and toxicity predictions were evaluated. Results showed that CTL, DZN, and PHY significantly increased seizure latency, reduced seizure frequency, and shortened seizure duration. The combination of CTL+DZN+PHY enhanced these effects by increasing latency and reducing seizure frequency and duration. When combined with DZP, the combination provided the highest protection (83.33%) and significantly reduced seizure duration. Docking studies revealed strong binding affinities (BAs) to GABAA (CTL: -7.0kcal/mol, DZN: -8.4kcal/mol, PHY: -5.4kcal/mol) and VGSC (DZN: -8.9kcal/mol). Pharmacokinetic analysis confirmed good absorption and blood-brain barrier (BBB) permeability, whereas toxicity profiling indicated a high safety margin. These findings suggest that CTL, DZN, and PHY exert significant anticonvulsant effects via GABAergic and sodium channel modulation, warranting further clinical investigation.

Citronellal (CTL), a natural monoterpenoid aldehyde, and phytol (PHY), a chlorophyll-derived diterpenoid, have been reported to exert neuropharmacological activities with possible GABAergic involvement. This study investigated their anxiolytic-like effects in Swiss albino mice, supported by molecular docking and pharmacokinetic evaluations. Animals received CTL (62.5, 125, and 250mg/kg, per orally (p.o.)) or PHY (25, 50, and 75mg/kg, p.o.) either alone or in combination with the gamma-aminobutyric acid (GABA) agonist diazepam (DZP, 2mg/kg, p.o.) or the GABA antagonist flumazenil (FLU, 0.1mg/kg, intraperitoneally (i.p.)). Behavioral analyses (open-field, hole-cross, swing, and light-dark tests) revealed that CTL significantly and dose-dependently reduced locomotor activity, while PHY produced variable responses, with higher doses enhancing exploratory behavior. Both compounds potentiated DZP-induced anxiolytic effects and counteracted FLU, suggesting GABAergic modulation. Molecular docking demonstrated stronger binding affinities of PHY (-4.9 and -4.2kcal/mol) than CTL (-4.7 and -4.0kcal/mol) at the α2 and α3 subunits of the GABAA receptor, with PHY forming hydrogen bonds (HBs) indicative of stable interactions. Pharmacokinetics and toxicity predictions confirmed favorable drug-likeness and higher LD50 values for CTL and PHY compared to DZP and FLU, with no major toxic liabilities. Collectively, these findings highlight that CTL may exert sedative-type anxiolytic-like effects, whereas PHY may show antidepressant-like anxiolytic properties, both mediated via the GABAergic pathway. These results support further investigation of CTL and PHY as potential natural anxiolytic candidates.

Phytol (PHY) is awater immiscible natural diterpene with interestingpharmacologic and antioxidant properties with great potential forapplication in foods, cosmetics, and pharmaceuticals. In this letter,a natural 1,3-distearyl-2-oleyl glycerol (TG1) extracted from Platonia insignis Mart was explored as a lipid matrix forpreparing uniformly sized and stable PHY-loaded solid lipid nanoparticles(SLN). Different tested compositions induced spherical (<300 nm),stable SLN with high encapsulation efficiency (>90%), able to induceslow release and a dose-dependent cytotoxic effect of PHY. These achievementscorroborated with a promising nanocarrier for PHY and are able tobe tested in different application fields.

The chlorophyll-derived diterpenoid phytol (PHY) is evident for its diverse biological effects, including neuroprotective activities in experimental animals. This study aims at the evaluation of anticonvulsant effects along with the possible mechanism of action of PHY through animal and in silico studies. For this, we performed pentylenetetrazole (PTZ)-induced convulsion tests in chicks and in silico studies against GABAA receptor subunits and voltage-gated sodium channel (VGSC) receptors. PHY was tested at 25, 50, and 75 mg/kg with or without the standard drugs diazepam (DZP: 5 mg/kg) and carbamazepine (CAR: 80 mg/kg) using a vehicle as a control. Acute oral toxicity was evaluated in chicks per OECD guidelines by administering PHY (500-2000 mg/kg, p.o.) and monitoring for 48 h for mortality, toxicological signs, and behavioral changes. PHY exhibited a dose-dependent anticonvulsant effect, significantly increasing latency and reducing convulsion frequency and duration at 75 mg/kg. PHY (75 mg/kg) combined with CAR and DZP showed the most potent reduction in convulsion frequency and duration, indicating a synergistic effect. Acute toxicity tests in chicks confirmed safety up to 2000 mg/kg. In silico studies demonstrated that PHY had good binding affinity with both the GABAA receptor (-6.5 kcal/mol) and VGSC (-7.0 kcal/mol), potentially contributing to its anticonvulsant action through GABAergic and sodium channel modulation. PHY showed significant anticonvulsant activity, likely via GABAA and VGSC modulation, warranting further studies to clarify its mechanisms and assess its adjunct potential in drug-resistant convulsion management.

A previous report suggests that phytol (PHY) may exert its antidepressant effects in mice, possibly through GABAA receptor interaction pathways. We aimed to check its antidepressant effect with possible molecular mechanisms through behavioral and in silico studies. For this, adult mice were randomly divided into different groups (n = 6), namely control (vehicle), standards (DZP: diazepam at 2 mg/kg, FLU: flumazenil at 0.1 mg/kg, FLUX: fluoxetine at 20 mg/kg), PHY (25, 50, and 75 mg/kg), and combined groups (PHY-75 with DZP-2 and/or FLU-0.1, and FLUX-20). Thirty minutes after treatment, each animal was subjected to tail suspension and forced swimming tests, and their immobility time (IMT) was counted for 5 min. In silico studies were performed with the GABAA receptor α1, α2, α3, α5, and γ2 subunits and 5HT1A to investigate possible molecular mechanisms. Additionally, invitro GABA activity of PHY and/or reference drugs was also performed by using the colorimetric method. The results demonstrated that PHY and/or DZP significantly (p < 0.05) and concentration-dependently inhibited GABA, while FLU alone or its combination with PHY reversed it. In mice, PHY dose-dependently reduced the IMT in both protocols, while FLUX-20 showed lower IMT compared to the control and DZP, indicating elevated locomotion in mice. It showed a reduced IMT value in male animals than in female animals. In both sexes, PHY at 75 mg/kg significantly (p < 0.05) increased the IMT values with DZP-2, while reducing this parameter with FLU-0.1. In silico studies demonstrated that PHY exhibited higher binding affinities with the α2 and α3 subunits of the GABAA and 5HT1A receptors by -6.5, -7.2 and 6.7 kcal/mol, respectively. Taken together, PHY exerted sedative-like antidepressant effects in mice and modulated the effects of GABAergic drugs DZP and FLU and serotonergic drug FLUX. PHY may be a potential candidate for the management of depression.

Phytol exerts sedative-like effects and modulates the diazepam and flumazenil’s action, possibly through the GABAA receptor interaction pathway

The current study is designed to evaluate the antiemetic effect of the diterpenoid phytol (PHY) using in vivo and in silico studies. For this, emesis was induced in 4-day-old chicks by the oral administration of copper sulfate (CuSO4.5H2O) at 50mg/kg. To see the possible antiemetic mechanism of PHY, we used a number of reference drugs such as domperidone (80mg/kg), ondansetron (24mg/kg) and hyoscine (100mg/kg) as positive controls, while the vehicle served as a negative control group. PHY was administered orally at the doses of 50 and 75mg/kg. Both PHY and reference drugs were given alone or in combined groups to evaluate their synergistic or antagonistic effects on the chicks. Molecular docking of PHY and reference drugs was carried out against 5HT3, D2, D3, H1, NK1,and mAChRs (M1-M5) receptors for estimating binding affinity to the receptors. Drug-receptor interactions and active sites of the receptors were observed with the aid of different computational tools. The drug-likeness and pharmacokinetics of all the drugs were predicted through the SwissADME online database. The results suggest that PHY reduces the mean number of retches and increases latency dose-dependently in the birds. In the combination groups, PHY75 showed better antiemetic effects with domperidone and ondansetron. In addition, PHY exhibited the highest binding affinity with the D2 receptor (6CM4) (-7.3kcal/mol). In conclusion, PHY showed an antiemetic activity in chicks, possibly through the D2 receptor interaction pathway.

Annona macroprophyllata Donn (A. macroprophyllata) is used in traditional Mexican medicine for the treatment of cancer, diabetes, inflammation, and pain. In this work, we evaluated the antitumor activity of three acyclic terpenoids obtained from A. macroprophyllata to assess their potential as antilymphoma agents. We identified the terpenoids farnesyl acetate (FA), phytol (PT) and geranylgeraniol (Gg) using gas chromatography–mass spectroscopy (GC-MS) and spectroscopic (1H, and 13C NMR) methods applied to petroleum ether extract of leaves from A. macroprophyllata (PEAm). We investigated antitumor potential in Balb/c mice inoculated with U-937 cells by assessing brine shrimp lethality (BSL), and cytotoxic activity in these cells. In addition, to assess the potential toxicity of PEAm, FA, PT and Gg in humans, we tested their acute oral toxicity in mice. Our results showed that the three terpenoids exhibited considerable antilymphoma and cytotoxic activity. In terms of lethality, we determined a median lethal dose (LD50) for thirteen isolated products of PEAm. Gg, PT and AF all exhibited a higher lethality with values of 1.41 ± 0.42, 3.03 ± 0.33 and 5.82 ± 0.58 µg mL−1, respectively. To assess cytotoxic activity against U-937 cells, we calculated the mean cytotoxic concentration (CC50) and found that FA and PT were closer in respect to the control drug methotrexate (MTX, 0.243 ± 0.007 µM). In terms of antilymphoma activity, we found that FA, PT and Gg considerably inhibited lymph node growth, with median effective doses (ED50) of 5.89 ± 0.39, 6.71 ± 0.31 and 7.22 ± 0.51 mg kg−1 in females and 5.09 ± 0.66, 5.83 ± 0.50 and 6.98 ± 0.57mg kg −1 in males, respectively. Regarding acute oral toxicity, we classified all three terpenoids as category IV, indicating a high safety margin for human administration. Finally, in a molecular docking study of 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase, we found binding of terpenoids to some amino acids of the catalytic site, suggesting an effect upon activity with a resulting decrease in the synthesis of intermediates involved in the prenylation of proteins involved in cancer progression. Our findings suggest that the acyclic terpenoids FA, PT, and Gg may serve as scaffolds for the development of new treatments for non-Hodgkin’s lymphoma.

Background and objective: Medicinal plants used in traditional medicine as prevention and treatment of disease and illness or use in foods, has a long history. Plants belonging to genera Allium have widely been acquired as food and medicine. In many countries, including Iran, a variety of species of the genus Allium such as garlic, onions, leeks, shallots, etc use for food and medicinal uses. Method: The leaves and bulbs of Allium atroviolaceum, collected from Borujerd (Lorestan Province, Iran) in May 2015 and their essential oils of were obtained by hydro-distillation. The oils were analyzed by gas chromatography coupled with mass spectrometry (GC/MS) and their chemical composition was identified. Results: The major constituents of A. atroviolaceum leaves oil were dimethyl trisulfide (59.0%), ethyl linolenate (12.4%), phytol (11.4%) and in bulb oil were methyl methyl thiomethyl disulfide (61.3%), dimethyl trisulfide (15.1%) and methyl allyl disulfide (4.3%). The major constituents of both essential oils are sulfur compounds. Conclusion: The results of the present study can help to increase of our information about composition of an edible herb in Iran. The major components of the oils can use as marker for standardization of the herbs.

BackgroundAntioxidants have protective capacity, and can be used combinedly with other substances. Both, ascorbic acid (AA) and phytol (PHY) have many important biological activities, including antioxidant, anti-inflammatory, and organ protective activity. Recently, PHY has been found to exert an anti-pyretic effect in a mouse model. This study aims to evaluate the combined effects of AA and/or PHY with paracetamol (PARA) in Brewer’s yeast-induced fever mice model.MethodsAA (125 mg/kg) and/or PHY (200 mg/kg) were orally co-treated with the PARA (100 mg/kg, p.o.) in Brewer’s yeast-induced fever Swiss mice. Data were analyzed by using GraphPadPrism software (version: 6.0), considering p < 0.05 at 95% confidence interval, and using one-way analysis of variance (ANOVA) through time, followed by Dunnett’s post hoc multiple comparison test.ResultsPARA alone and with PHY and/or AA significantly (p < 0.05) reduced rectal temperature at 1st h of observation. PHY reduced rectal temperature at 2nd h, then maintained basal temperature over the observation period (4 h). AA showed an insignificant anti-pyretic effect in experimental animals. However, in combination groups, AA (i.e., with PHY or PARA) did not found to interfere the PHY and PARA mediated reduction of rectal temperature in the animals. Furthermore, AA when co-treated with the PARA + PHY, it caused a slight hypothermic temperature at 1st h, which was then started to restablish from 2nd to 3rd h, and normalized at 4th h.ConclusionTaken together, AA did not interfere anti-pyretic effects of PARA and PHY, suggesting its possible use as a combination substance.

Abstract: Background: Black seed oil and Phytol (PHY) are evident for their promising biological effects in various test systems. Aim: This study evaluates anti-atherothrombosis activity of Nigella sativa oil (NSO) and Phytol (PHY). Materials and Methods: The clotlysis activity of the NSO and/or PHY has been investigated by taking Streptokinase (SK) as a standard clotlysis drug. Results: The results suggest that the NSO and PHY exhibited a concentration-dependent clotlysis activity in human clotted blood. The highest clotlysis of the NSO and PHY was seen by 57.83 ± 0.23% and 53.05 ± 2.93% with 80 μL and 150 μg per tube, respectively. NSO co-treated with PHY showed a better clotlysis capacity than the NSO and PHY. The vehicle showed negligible clotlysis (2.86 ± 4.33%) capacity, while the standard SK (100 I.U.) showed 78.96 ± 1.08% clotlysis activity. Conclusion: NSO and/or PHY exhibited clotlysis activity in human clotted blood in in vitro. Key words: Nigella sativa oil, Phytol, Antioxidant, Atherothrombosis, Cardiovascular diseases.

Plant Science For seed crops, such as maize and wheat, flowers need to develop at the right time to optimize harvest. Hormonal signals and metabolic status coordinate to set the transition from vegetative to reproductive growth. During acute nitrogen starvation, the plant flowers in a last-chance attempt to reproduce. Olas et al. studied what happens in the model plant Arabidopsis when nitrogen status is suboptimal, but not at starvation levels. MicroRNA regulators and nitrate-responsive gene promotor elements detect nitrogen status directly in the shoot apical meristem, from which flowers arise. With low nitrogen and short days, flowering depends more upon the availability of carbon in the form of sugar. With longer days, low nitrogen matters less. Thus, when faced with limited nitrogen supplies, the plant integrates photoperiod, hormonal, and metabolic signals to define the moment of flowering. New Phytol. 10.1111/nph.15812 (2019).

Phytol (PHY) (3,7,11,15-tetramethylhexadec-2-en-1-ol) exhibits various pharmacological properties including toxicity and cytotoxicity, and exerts antitumor activity. Owing to the urgent need of new pharmaceutical formulations for breast cancer therapy, this study aimed at the evaluation of antitumor activity of PHY in 7,12-dimethylbenzanthracene-cancer-induced animal model. Comet assay was employed to evaluate the cytogenetics, DNA repair, and antigenotoxic activities of PHY in neoplastic (breast) and non-neoplastic rodent cells (bone marrow, lymphocytes, and liver). Additionally, hematological, biochemical, histopathological, and immunohistochemical analyses were carried out in experimental animals. Thirty nonpregnant female mice (n = 5) underwent 7 weeks treatment with 6 mg/kg pro-carcinogen, PHY (4 mg/kg), and cyclophosphamide (25 mg/kg). Induction of cancer was confirmed by histopathology and immunohistochemistry for Ki-67. Results suggest that PHY exhibits low toxicity in comparison with other groups in hematological, biochemical, histopathological, and organ size parameters. Additionally, PHY showed modulatory effects on the pro-carcinogen, and induced genotoxicity and apoptosis in breast cancer cells. Furthermore, it showed a DNA damage repair capacity in mouse lymphocytes. These data indicate that PHY may have the potential as an anticancer candidate in pharmaceutical consumption. © 2018 IUBMB Life, 71(1):200-212, 2019.

Phytol: A review of biomedical activities

In the present investigation, phytochemical compounds and GC-MS analysis of Boerhavia diffusa whole plant was studied, and presence of phytochemical compounds was screened by qualitative method. The results confirmed the presence of phytochemicals namely terpenoids, saponin, flavonoids, cardiac glycosides, phlobatannins and tannin. In GC-MS analysis, 42 phytochemical compounds were identified in the methanol extract of Boerhavia diffusa, the components were identified by comparing their relation indices and mass spectra fragmentation patterns with those stored on the MS-Computer library, PubChem and also from the published literatures. Phytol, hexadecane, hexadecanoic acid, 2-hydroxy-1-(hydroxymethyl) ethyl ester, stigmast-5-en-3-ol, (3.BETA.) and stigmasterol seem to be the major constituents. The results supported the ethno-medicinal knowledge of the tribes of state of Jharkhand, India; is worth for the scientific validation. The results were also helpful in the development of new therapeutics for the treatment of various types of deadly disease including cancer. Keywords: Ethno-medicinal, phytochemical, GCMS, flavonoids, phytol Cite this Article Patnaik A, Sharma HP. GC-MS Analysis of Boerhavia diffusa Linn. Whole Plant Extracts: An Ethno-medicinal Plant. Research & Reviews: A Journal of Pharmacognosy . 2018; 5(1): 7–12p.

Glioblastoma (GBM) is the most common primary brain tumor. Genetic mutations may reprogram the metabolism of neoplastic cells. Particularly, alterations in cholesterol and fatty acid biosynthetic pathways may favor biomass synthesis and resistance to therapy. Therefore, compounds that interfere with those pathways, such as phytol (PHY) and retinol (RET), may be appropriate for cytotoxic approaches. We tested the effect of PHY or RET on the viability of human GBM cell lines (U87MG, A172 and T98G). Since the compounds showed a dose-dependent cytotoxic effect, additional analyses were performed with IC50 values. Transcriptome analyses of A172 cells treated with PHY IC50 or RET IC50 revealed down-regulated genes involved in cholesterol and/or fatty acid biosynthetic pathways. Thus, we investigated the expression of proteins required for cholesterol and/or fatty acid synthesis after treating all lineages with PHY IC50 or RET IC50 and comparing them with controls. Sterol regulatory element-binding protein 1 (SREBP-1) expression was reduced by PHY in U87 and T98G cells. However, fatty acid synthase (FAS) protein expression, which is regulated by SREBP-1, was down-regulated in all lineages after both treatments. Moreover, farnesyl-diphosphate farnesyltransferase (FDFT1) levels, a protein associated with cholesterol synthesis, were reduced in all lineages by PHY and in U87MG and A172 cells by RET. Our results suggest that SREBP-1, FAS and FDFT1 are potential target(s) for future in vivo approaches against GBM and support the use of inhibitors of their synthesis, including PHY and RET, for such approaches.

Background and Objective: Chlorophyllase catalyzes the hydrolysis of chlorophylls to chlorophyllide and phytol. Recently, several applications including removal of chlorophylls from vegetable oils, use in laundry detergents and production of chlorophyllides have been described for chlorophyllase. However, there is little information about the biochemical characteristics of chlorophyllases. Material and Methods: 35 chlorophyllase protein sequences were obtained from the National Centre for Biotechnology Information database. All of the sequences were analyzed using bioinformatics tools for their conserved domain, phylogenetic relationships and biochemical characteristics. Results and Conclusion: The overall domain architecture of chlorophyllases consisted of the esterases/lipases superfamily domain over their full length and the alpha/beta hydrolase family domain over the middle part of their sequences. Plant chlorophyllases could be classified into 4 clades. Molecular weight and pI of the chlorophyllases ranged 32.65-37.77 kDa and 4.80-8.97, respectively. The most stable chlorophyllase is probably obtained from Malus domestica. Chlorophyllases form Solanum pennellii, Triticum aestivum, Triticum urartu, Arabidopsis lyrata, Pachira macrocarpa, Prunus mume and Malus domestica were predicted to be soluble upon overexpression in Escherichia coli, Beta vulgaris and Chenopodium album chlorophyllases were predicted to form no disulfide bond. Chlorophyllases from Jatropha curcas, Amborella trichopod, Setaria italica, Piper betle, Triticum urartu and Arabidopsis thaliana were predicted to be in non-N-glycosylated form. Conflict of interest: The authors declare no conflict of interest.

Evaluation of toxic, cytotoxic and genotoxic effects of phytol and its nanoemulsion

An important chlorophyll-derived diterpenoid essential oil, phytol (PYT) bio-metabolite, phytanic acid (PA) has a number of pathophysiological contributions. The PA metabolism and its plasma levels associated phenomena are continuously being researched on. This study aims to look at the complete current scenario of PA. The findings suggest that PA has anti-diabetic, cytotoxic, anticancer and anti-teratogenic activities. Although PA-mediated Refsum&rsquo;s Diseases (RD), Sjogren-Larsson Syndrome (SLS) and prostate cancer are still controversial; Zellweger&#39;s Disease Hyperpipecolic Academia (ZDHA), Rhizomelic Chondrodysplasia Punctata (RCDP), Leber Disease (LD) and oxidative stress leading to mitochondrial and cardiac complications are also evident. In conclusion, PA may be a good biomarker of some pathophysiological phenomena and can be used for medico-pharmaceutical functions. Key words: Phytanic acid, phytol bio-metabolite, biomarkers, pathophysiological contributions.

This study aims at investigating a possible pathway of cytotoxicological status of the diterpenoid essential oil, phytol (PYL). For this brine shrimp lethality bioassay (BSLB) and hemolysis (HL) test systems were selected. In the BSLB, PYL either alone or co-treated with ethylenediaminetetraacetic acid (EDTA), potassium di-chromate (K2Cr2O7; KD), copper sulphate (CuSO4.5H2O; CS) and 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (Trolox, TRO) as membrane lyser, strong oxidizer, oxidizer-cytogenotoxicant and antioxidative-cell-protestant, respectively. The HL was carried out in rat erythrocytes (RBCs) taking TRO as a standard. In addition, to view a time-dependent cytotoxic activity of PYL, the mortality of the shrimps was counted at 24 and 48 h. Results suggest PYL is non-cytotoxic at low (40-160 µM) but toxic at high concentration (2-8 mM) to the shrimps and RBCs. An increased cytotoxicity was observed for 24 h to 48 h in brine shrimps. In both cases groups co-treated with cytotoxicants/protestant suggest that PYL is cytoprotective in the presence of oxidizer. The cytoprotectivityof PYL may be connected to its antioxidant potential and cytotoxicity for antioxidant-mediated pro-oxidative effects. In conclusion, PYL is cytoprotective at low concentration but toxic at high, activities found, however, may be linked to the radical scavenging pathway