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Pharmacological studies on coleonol, a hypotensive diterpene from Coleus forskohlii.

Coleus spp. have been used in Aurvedic medicine for heart diseases, spasmodic pain, painful micturition and convulsions. The pharmacological properties of coleonol, a diterpene, isolated from Coleus forskohlii were investigated. Its predominant effect is to lower the blood pressure of anaesthetised cat and rat as well as of the spontaneously hypertensive rat due to relaxation of the vascular smooth muscle. In small doses it has a positive inotropic effect on isolated rabbit heart as well as on cat heart in vivo. Coleonol also exhibits nonspecific spasmolytic activity on smooth muscle of the gastrointestinal tract in various species but not on bronchial musculature of guinea pig. Large doses of coleonol have a depressant action on the central nervous system. These results provide the rationale for the use of this plant in Aurvedic medicine.

Animals↗

Effects of Coleus forskohlii on fat storage in ovariectomized rats.

The antiobesity effects of Coleus forskohlii were investigated in ovariectomized (ovx) rats. Eight-week-old female Wistar rats were assigned to four groups: a sham-operated group fed the control diet (MF, sham-m) ; an ovx-m group fed the control diet; a sham-operated group fed the control diet containing 50 g/kg of Coleus forskohlii extract (sham-c) ; and an ovx-c group fed the control diet containing 50 g/kg of Coleus forskohlii extract. The body weight, adipose tissues, and cell diameter were investigated in ovx rats after Coleus forskohlii extract treatment. Administration of Coleus forskohlii extracts reduced body weight, food intake, and fat accumulation in ovx rats. Our results suggest that Coleus forskohlii may be useful in the treatment of obesity.

Adipocytes↗

A 12-week, double-blind, quasi-randomized, placebo-controlled study to evaluate the efficacy and safety of Coleus forskohlii (Forcslim) on body weight loss.

BACKGROUND: Overweight and obesity have emerged as a global epidemic, significantly impacting human health. Traditional usage and growing scientific evidence suggest that Coleus forskohlii extract (Forcslim) may aid in reducing excess body weight and fat. This study aimed to evaluate the efficacy and safety of Forcslim supplementation in overweight individuals. METHODS: A quasi-randomized, double-blind, placebo-controlled clinical trial was conducted in 60 overweight subjects aged 20-70&#x2009;years over a period of 12&#x2009;weeks. The participants were assigned to receive either Forcslim or placebo. The key outcome measures included body weight, body mass index (BMI), body composition, and anthropometric parameters. Additionally, lipid profile parameters and safety markers (including metabolic, hepatic, and cardiovascular indicators) were assessed throughout the study duration. RESULTS: Compared to the placebo group, the Forcslim group showed significant reductions in waist circumference (-1.83&#x2009;cm; p&#x2009;<&#x2009;0.01) and body weight (-1.93&#x2009;kg; p&#x2009;<&#x2009;0.001). Significant improvements in anthropometric parameters were observed exclusively in the Forcslim group. Furthermore, triglyceride (TG) levels were significantly reduced (p&#x2009;<&#x2009;0.01), while high-density lipoprotein (HDL) levels showed a significant increase (p&#x2009;=&#x2009;0.001). No clinically significant changes were observed in metabolic markers, liver and muscle enzyme levels, heart rate, blood pressure, or reported adverse effects, indicating a favorable safety profile. CONCLUSIONS: Forcslim demonstrated significant anti-obesity effects, including reductions in body weight, waist circumference, and improvements in the lipid profile. These findings suggest that C. forskohlii extract supplementation may serve as a safe and effective alternative to synthetic anti-obesity drugs.

Humans↗

Production of Forskolin by Axenic Coleus forskohlii Roots Cultivated in Shake Flasks and 20-l Glass Jar Bioreactors*.

Root cultures of COLEUS FORSKOHLII Briq. were initiated from primary callus or IBA-treated suspension cultures and maintained on Gamborg's B5 medium containing 1 mg/l IBA. Transformed root cultures were established by infecting surface-sterilized leaves with AGROBACTERIUM RHIZOGENES strain 15834. Transformation was confirmed by mannopine detection. These cultures displayed the typical characteristics of hairy root cultures, with the sole exceptions of slow growth in hormone-free medium and accelerated growth on medium containing phytohormones. All root cultures examined formed forskolin and its derivatives in amounts ranging from 500 to 1300 mg/kg dry weight, corresponding to about 4 to 5 mg/l. During cultivation roots could be cut into small pieces without affecting growth and forskolin production. Scale-ups of the cultivation procedure were performed in 20-l glass jars with a working volume of 10 to 13l. Forskolin production in bioreactors was better than in shake flasks. Levels of almost 14 mg/l could be reached after 21 d of cultivation. As in the shake flask experiments cutting the roots did not affect growth or productivity in a negative way.

Journal Article↗

Quantitative analysis of forskolin in Coleus forskohlii (Lamiaceae) by reversed-phase liquid chromatography.

A rapid method was developed for the evaluation of forskolin in Coleus forskohlii Briq. (Lamiaceae). Forskolin was quantitated in the root and stem of dried C. forskohlii and in 17 market products by reversed-phase liquid chromatography (LC) with a photodiode array detector at 210 nm. The temperature was held constant at 30 degrees C, and the retention time of forskolin was approximately 6.8 min. The samples were extracted with acetonitrile by sonication. The precision of the method was confirmed by a standard deviation < 5.0% (n = 3), and forskolin recovery was 99.1%. Limit of detection was 1.5 microg/mL, and the response was linear through zero from 6.3 to 630 microg/mL with a correlation coefficient (R2) of 0.9998. Identity of the marker compound was confirmed by an LC/mass spectrometry experiment. The method was successful in the qualitative and quantitative evaluation of the marker compound in C. forskohlii plant material and in market products claiming to contain C. forskohlii.

Chromatography, Liquid↗

Two new diterpenoids from Coleus forskohlii.

Two new diterpenoids, forskolins I and J, have been isolated in our further investigation on Coleus forskohlii (Willd.) Briq. collected in Yunnan Province. Their structures have been determined as 1alpha,6beta-diacetoxy-7beta,9alpha-dihydroxy-8,13-epoxylabd-14-en-11-one (1) and 1alpha,9alpha-dihydroxy-6beta,7beta-diacetoxy-8,13-epoxylabd-14-en-11-one (2) by spectral methods (including 1D and 2D NMR techniques).

Coleus↗

Three new diterpenoids from Coleus forskohlii Briq.

Three new diterpenoids, forskolin G(2), forskolin H(3), forskolin I(4), were isolated from the whole plant of the Coleus forskohlii Briq., and their structures were elucidated as 1alpha,6beta-diacetoxy-8,13-epoxylabd-14-en-11-one, 1alpha-hydroxy-6beta,7beta-diacetoxy-8,13-epoxylabd-14-en-11-one, and 1alpha,9alpha-dihydroxy-6beta,7alpha-diacetoxy-8,13-epoxylabd-14-en-11-one on the basis of spectral data.

Coleus↗

Simple and rapid method for the isolation of forskolin from Coleus forskohlii by charcoal column chromatography.

A simple, safe, rapid and economical method was developed for the isolation of high-purity forskolin from Coleus forskohlii roots using activated charcoal as an adsorbent in a column. The elution was carried out under reduced pressure to make the process rapid. Activated charcoal acted as a reversed phase adsorbent and allowed elution of forskolin without much impurities. The residue, obtained from the eluate was purified and crystallized using different solvent mixtures to obtain pure forskolin. The forskolin isolated was analyzed and characterized by UV, IR, RP-HPLC, electrospray ionization MS, 1H NMR and 13C NMR. The yield was 0.097% w/w (RSD 5.6%). The purity was 96.9% w/w (RSD 0.3%) as determined by RP-HPLC. The present method enables researchers to produce high-purity forskolin in their labs by using common chemicals.

Adsorption↗

Establishment of forskolin yielding transformed cell suspension cultures of Coleus forskohlii as controlled by different factors.

Suspension cultures derived from gall calli which were obtained following infection with Agrobacterium tumefaciens (C58) were established in Coleus forskohlii. Cell line selection following single cell cloning or cell aggregate cloning was carried out to select cell lines capable of fast growth and for producing high level of forskolin. A fast growing cell line (GSO-5/7) thus selected was found to accumulate 0.021% forskolin in 42 days. The effect of cultural conditions on cell growth was studied to identify factors influencing biomass yield. Cell growth in suspension was found to be influenced significantly by carbon source, initial cell density and light or dark condition. Optimal cell growth (20 fold increase in biomass in a 42 day period) was obtained when the cells were grown in dark condition in B5O media containing 3% sucrose as sole carbon source with an initial cell density of 1.5 x 10(5) cells per ml. Forskolin accumulation was maximum (0.021%) in the stationary phase of cell growth. These suspension cultures showed continuous and stable production of forskolin.

Cell Culture Techniques↗

Molecular cloning and functional expression of geranylgeranyl pyrophosphate synthase from Coleus forskohlii Briq.

BACKGROUND: Isopentenyl diphosphate (IPP), a common biosynthetic precursor to the labdane diterpene forskolin, has been biosynthesised via a non-mevalonate pathway. Geranylgeranyl diphosphate (GGPP) synthase is an important branch point enzyme in terpenoid biosynthesis. Therefore, GGPP synthase is thought to be a key enzyme in biosynthesis of forskolin. Herein we report the first confirmation of the GGPP synthase gene in Coleus forskohlii Briq. RESULTS: The open reading frame for full-length GGPP synthase encodes a protein of 359 amino acids, in which 1,077 nucleotides long with calculated molecular mass of 39.3 kDa. Alignments of C. forskohlii GGPP synthase amino acid sequences revealed high homologies with other plant GGPP synthases. Several highly conserved regions, including two aspartate-rich motifs were identified. Transient expression of the N-terminal region of C. forskohlii GGPP synthase-GFP fusion protein in tobacco cells demonstrated subcellular localization in the chloroplast. Carotenoid production was observed in Escherichia coli harboring pACCAR25DeltacrtE from Erwinia uredovora and plasmid carrying C. forskohlii GGPP synthase. These results suggested that cDNA encoded functional GGPP synthase. Furthermore, C. forskohlii GGPP synthase expression was strong in leaves, decreased in stems and very little expression was observed in roots. CONCLUSION: This investigation proposed that forskolin was synthesised via a non-mevalonate pathway. GGPP synthase is thought to be involved in the biosynthesis of forskolin, which is primarily synthesised in the leaves and subsequently accumulates in the stems and roots.

Alkyl and Aryl Transferases↗

Influence of different AM-fungi on the growth, nutrition and forskolin content of Coleus forskohlii.

A glasshouse investigation was conducted to study the effectiveness of 11 arbuscular mycorrhizal (AM) fungi on the medicinal plant Coleusforskohlii. Coleus plants raised in presence of most of the AM fungi in polythene bags showed an increase in plant growth (height, number of branches and biomass), P, and forskolin contents over those grown in the absence of soil inoculation with AM fungi. The extent of growth, P, and forskolin status varied with the AM fungi used. Based on the plant biomass, P uptake and forskolin content per plant, Glomus bagyarajii was found to be the best AM symbiont for inoculating C. forskohlii, the next being Scutellospora calospora.

Coleus↗

[Studies on the chemical constituents in root of Coleus forskohlii].

OBJECTIVE: To study the chemical constituents in root of Coleusforskohlii. METHOD: The chemical constituents were isolated by column chromatography. The structures were elucidated on the basis of IR, MS, 1H-NMR, 13C-NMR and 2D-NMR experiments. RESULT: Six compounds were obtained and the structures were identified as 14-deoxycoleon U (1), demethylcryptojaponol (2), alpha-amyrin (3), betulic acid (4), alpha-cedrol (5) and beta-sitosterol (6). CONCLUSION: Compounds 1 and 2 were isolated from the genus Coleus for the first time. Compounds 3 and 4 were isolated from C. forskohlii for the first time.

Coleus↗

Production of Forskolin in in vitro Cultures of Coleus forskohlii.

Shoot tip culture, root organ suspension, and callus culture producing roots were established from shoot tip, root tip, and hypocotyl segments, respectively, on Murashige and Skoog's and White's basal media with or without growth regulators. Forskolin was identified by TLC and HPLC in 60 days old shoot differentiating culture, 30 days old micropropagated plants, and root organ suspension but not in rhizogenic callus. Shoot differentiating culture established in the presence or absence of BAP revealed more or less similar amounts of forskolin as that of the micropropagated plants. Root organ suspension showed traces of forskolin production.

Journal Article↗

Forskolin: a potential antimetastatic agent.

Forskolin, a diterpene from the roots of an Indian plant, Coleus forskohlii, is a potent platelet aggregation inhibitor and has been examined for its effects on (a) tumor-induced human platelet aggregation and (b) pulmonary tumor colonization in mice. These studies employed a subline of B16 murine melanoma, B16-F10 (highly metastatic to lungs). Forskolin (2 microM) strongly inhibits the melanoma cell-induced human platelet aggregation. A single dose of forskolin (82 micrograms/mouse) administered intraperitoneally 30 or 60 min prior to tail vein injection of cultured B16-F10 cells (2 or 3 X 10(5) cells/mouse) reduced tumor colonization in the lungs by more than 70%. Similar results were obtained in three separate experiments. These findings raise the possibility that forskolin could prove of value in the clinic for the prevention of cancer metastasis.

Animals↗

The effect of adenylate cyclase stimulation on endocochlear potential in the guinea pig.

Forskolin, a diterpene extracted from Coleus forskohlii, is potentially an important tool for studying the modulation of ionic currents by cAMP because it stimulates adenylate cyclase in a variety of cells. We studied the effect of forskolin on cochlear potentials and found that its perfusion of the scala vestibuli (SV) to a concentration more than 10(-5) M and the scala tympani (ST) to more than 10(-4) M produced a reversible elevation of the endocochlear potential (EP) in a dose-dependent manner. The cochlear microphonics recorded simultaneously with the EP was not depressed during the EP elevation. A large negative EP was induced by anoxia following the SV perfusion with forskolin (2 X 10(-4) M). The results suggest that the EP elevation produced by forskolin does not result from the decrease in the negative component of EP but from the increase in the positive component of EP.

Adenylyl Cyclases↗