Daphnetin inhibits proliferation and inflammatory response in human HaCaT keratinocytes and ameliorates imiquimod-induced psoriasis-like skin lesion in mice

BACKGROUND: Psoriasis is a common chronic inflammatory skin disease. Keratinocytes hyperproliferation and excessive inflammatory response contribute to psoriasis pathogenesis. The agents able to attenuate keratinocytes hyper-proliferation and excessive inflammatory response are considered to be potentially useful for psoriasis treatment. Daphnetin exhibits broad bioactivities including anti-proliferation and anti-inflammatory. This study aims to evaluate the anti-psoriatic potential of daphnetin in vitro and in vivo, and explore underlying mechanisms. METHODS: HaCaT keratinocytes was stimulated with the mixture of IL-17A, IL-22, oncostatin M, IL-1α, and TNF-α (M5) to establish psoriatic keratinocyte model in vitro. Cell viability was measured using Cell Counting Kit-8 (CCK-8). Quantitative Real-Time PCR (qRT-PCR) was performed to measure the mRNA levels of hyperproliferative marker gene keratin 6 (KRT6), differentiation marker gene keratin 1 (KRT1) and inflammatory factors IL-1ß, IL-6, IL-8, TNF-α, IL-23A and MCP-1. Western blotting was used to detect the protein levels of p65 and p-p65. Indirect immunofluorescence assay (IFA) was carried out to detect p65 nuclear translocation. Imiquimod (IMQ) was used to construct psoriasis-like mouse model. Psoriasis severity (erythema, scaling) was scored based on Psoriasis Area Severity Index (PASI). Hematoxylin and eosin (H&E) staining was performed to examine histological change in skin lesion. The expression of inflammatory factors including IL-6, TNF-α, IL-23A and IL-17A in skin lesion was measured by qRT-PCR. RESULTS: Daphnetin attenuated M5-induced hyperproliferation in HaCaT keratinocytes. M5 stimulation significantly upregulated mRNA levels of IL-1ß, IL-6, IL-8, TNF-α, IL-23A and MCP-1. However, daphnetin treatment partially attenuated the upregulation of those inflammatory cytokines. Daphnetin was found to be able to inhibit p65 phosphorylation and nuclear translocation in HaCaT keratinocytes. In addition, daphnetin significantly ameliorate the severity of skin lesion (erythema, scaling and epidermal thickness, inflammatory cell infiltration) in IMQ-induced psoriasis-like mouse model. Daphnetin treatment attenuated IMQ-induced upregulation of inflammatory cytokines including IL-6, IL-23A and IL-17A in skin lesion of mice. CONCLUSIONS: Daphnetin was able to attenuate proliferation and inflammatory response induced by M5 in HaCaT keratinocytes through suppression of NF-κB signaling pathway. Daphnetin could ameliorate the severity of skin lesion and improve inflammation status in IMQ-induced psoriasis-like mouse model. Daphnetin could be an attractive candidate for future development as an anti-psoriatic agent.

Dual role of daphnetin in suppressing HMGB1 release and HMGB1-induced inflammation in murine macrophage RAW264.7 cells and human monocytic THP-1 cells in vitro / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the dual role of daphnetin in suppressing high mobility group box-1 protein (HMGB1) release and blocking HMGB1-induced inflammatory response.</p><p><b>METHODS</b>Murine macrophage RAW264.7 cells were cultured in the presence of daphnetin, lipopolysaccharide (LPS), or both. HMGB1 release from the cells was determined using ELISA, and phosphorylations of JAK1/2 and of STAT1 were detected by Western blotting. Human monocytic THP-1 cells exposed to daphnetin, rhHMGB1, or both were examined for NO production using a NO detection kit, for the release of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and prostaglandin E2 (PGE2) using ELISA, and for expressions of iNOS, COX-2 and phosphorylated p38, ERK, and JNK with Western blotting.</p><p><b>RESULTS</b>Daphnetin dose-dependently reduced the release of HMGB1 in RAW264.7 cells and suppressed rhHMGB1-induced iNOS and COX-2 expressions and release of TNF-α, IL-6, PGE2, and NO in THP-1 cells. Western blotting revealed that daphnetin significantly down-regulated the phosphorylations of JAK-STAT1 pathway in LPS-stimulated RAW264.7 cells but did not suppress the phosphorylations of MAPKs signaling pathway induced by rhHMGB1 in THP-1 cells.</p><p><b>CONCLUSION</b>Daphnetin can reduce the release of HMGB1 and suppress HMGB1-induced inflammatory response. In RAW264.7 cells, daphnetin inhibited LPS induced HMGB1 release is at least partly mediated by suppressing JAK-STAT1 signaling pathway activation.</p>

Influencia del tiempo de cosecha sobre el contenido de cumarina y de umbeliferon hallado en los extractos de Justicia pectoralis Jacq. (tilo) / Harvest time influences on coumarin and umbelliferone contents in extracts of Justicia pectoralis Jacq. (tilo)

INTRODUCTION: Justicia pectoralis Jacq. (Acanthaceae) is medicinal plant species commonly used in Cuba for the treatment of nervous disorders because of its sedative effect. Coumarin is one of its main active phytochemicals present in the extracts obtained from this plant and used as analytic marker in quality control. On the other hand, this compound contributes to the sedative effect attributed to this plant. OBJECTIVE: to evaluate the influence of harvest time on the coumarin and umbelliferone (7-hydroxycoumarin) in Justicia pectoralis extracts. METHODS: the experiment lasted two years. The harvest was performed at 4, 6 and 8 months after planting. Aqueous and hydroalcohol extracts were produced and the coumarin and umbelliferone contents were determined by high resolution liquid chromatography. RESULTS: the achieved results showed the presence of coumarin and umbelliferone in both extracts. Both methods can be used for the extraction of these components from the plant, although in the case of umbelliferone, the best extraction results were achieved by using aqueous extract. In both cases, the recovery percentages were more than 98 percent. This study confirmed that the harvest time significantly influences on the coumarin and umbelliferone contents. CONCLUSIONS: the best results are observed in the first two harvests (4 and 6 months at summer time), which indicates that the industry should process the vegetal material in these two periods of the year(AU)

INTRODUCCIÓN: Justicia pectoralis Jacq. (Acanthaceae) es una planta medicinal comúnmente usada en Cuba para el tratamiento de enfermedades nerviosas por su efecto sedante. La cumarina es uno de los fitocomponentes mayoritarios en los extractos obtenidos con esta planta y empleado como marcador analítico en los controles de calidad. Por otro lado, este componente contribuye con el efecto sedante atribuido a esta planta. OBJETIVO: evaluar la influencia del tiempo de cosecha sobre el contenido de cumarina y umbelliferona (7 hidroxicumarina) en extractos de Justicia pectoralis. MÉTODOS: se desarrolló el experimento durante 2 años. Se realizaron las cosechas a los 4, 6 y 8 meses de plantada. Se elaboraron extractos acuosos e hidroalcohólicos y se determinó el contenido de cumarina y umbelliferone por cromatografía líquida de alta resolución. RESULTADOS: se mostró la presencia de cumarina y umbelliferona en ambos extractos. Además, en el caso de la umbelliferona, los mejores resultados se alcanzaron al aplicar extracción acuosa. En ambos casos, los por cientos de recobrados fueron superiores al 98 por ciento. Se confirmó que el tiempo de cosecha influyó significativamente sobre el contenido de cumarina y umbelliferona. CONCLUSIONES: los mejores resultados se obtienen en la primeras dos cosechas (4 y 6 meses que coincide con el verano), lo que sugiere que el material de la planta debe procesarse por la industria en esos periodos del año(AU)

Determination of skimmin, scopolin and umbelliferone in Tibetan medicine Saussurea hieracioides by HPLC / 中国中药杂志

This study is aimed to establish a high-performance liquid chromatography (HPLC) method for simultaneous determination of skimmin, scopolin and umbelliferone in Saussurea hieracioides. Samples were analyzed on a Wondasil C18-WR column (4.6 mm x 250 mm, 5 microm) with methanol (A) and water containing 0.1% phosphate (B) as mobile phases for gradient elution at a flow rate of 1.0 mL x min(-1). The detection wavelength and column temperature were set at 325 nm and 35 degrees C, respectively, and the sample size was 10 microL. The results showed that skimmin, scopolin and umbelliferone were simultaneously achieved within 40 min under the above conditions. A good linearity was observed in the range of 0.18-5.6 microg (r = 1.000 0), 0.060-1.8 microg (r = 0.999 9), 0.032-0.97 microg (r = 0.999 8) for skimmin, scopolin and umbelliferone, respectively, with the average recoveries of 99.16% (RSD = 0.41%), 100.3% (RSD = 0.79%), 102.2% (RSD = 0.87%). The method is simple, accurate and reproducible and can be used for the quality control of S. hieracioides.

[Purification and structured termination of compounds in chloroform extract of the aerial parts of the plant Ferula hirtella]

Ferula is a genus of about 130 species of flowering plants in the family Umbelliferae, of which 30 species are found in Iran. Many species of this family possess medicinal qualities and have long been used in traditional medicine. This research aims to isolate, separate and determine the structure of the compounds found in the chloroform extract of the aerial parts of Ferula hirtella. We collect F. hirtella from the site of its natural habitat in Abade, Fars Province. The chloroform extract of the aerial parts is derived and then mixture separation using Column chromatography is performed. Using Column chromatography purification methods, we were able to separate, for the first time, two coumarins, Farneciferol C and Umbelliprenin, and a steroid compound, beta-stigmasterol from the chloroform extract of the F. hirtella which were identified using C NMR and H NMR spectral data. A majority of the compounds found in the chloroform extract of the aerial parts of F. hirtella are coumarins. Separation of beta-stigmasterol from the chloroform extract of this genus was achieved for the first time

Phytochemical and biological study of vangueria edulis cultivated in Egypt

Fractionation and purification of the alcoholic extract of the leaves and stem -bark of Vangueria edulis cultivated in Egypt afforded thirteen compounds identified as: p-sitosterol acetate [1], stigmasterol [2], palmitic acid [3], scopoletin [4], p-coumaric acid [5], protocatechuic acid [6], esculetin [7], ethyl I-O-glucosyi-4-O-[E] caffeoyl quinate [8], kaempferol 3-O-rhamnoside, 7-O-rutinoside [9], p-sitosterol [10], ceryl alcohol [11], vanillic acid [12] and p-sitosterol-3-Q-beta-D-glucopyranoside [13]. Identification of these compounds has been established by physical, chemical and spectral data as well as comparison with authentic samples. Compounds 8 and 9 are firstly reported from a natural source while the rest of the compounds have been isolated for the first time from the genus Vangueria. The LD[50] antibacterial antifungal anti-inflammatory, antipyretic and the effect on CNS were studied

Chemical constituents of Eupatorium lindleyanum / 中国中药杂志

To study chemical constituents of Eupatorium lindleyanum. Ethyl acetate extractive fractions were separated with silica gel and Sephadex LH-20 by column chromatography, and their structures were identified on the basis of spectroscopic analysis and chemical evidence. Sixteen compounds were separated and identified as scopoletin (1), 6, 7-dimethylesculetin (2), nepetin (3), eupatrin (4), luteolin (5), isoquerecitrin (6), jaceosidin (7), quceritin (8), kaempferol (9), rutin (10), cirsiliol (11), taraxasterylacetate (12), pseudotaraxasteryl acetate (13), pseudotaraxasterol (14), butanoic acid (15) and n-hexadecanoic acid (16). Of them, compounds 1-6 and 11, 13 and 15 were separated from this plant for the first time.

Analysis of metabolites of daphnetin in the intestinal wall of rats by liquid chromatography and quatrupole-time of flight mass spectrometry / 药学学报

In this study, daphnetin and its major metabolites in the intestinal wall of rats were identified by liquid chromatography and quatrupole-time of flight mass spectrometry. Perfusion fluid of duodenum, jejunum, ileum and colon were collected separately for 2 hours from the rat intestine following perfusion with daphnetin. The metabolites of daphnetin in the perfusion fluid of different intestine segments were analyzed by the liquid chromatography and quatrupole-time of flight mass spectrometry. It is shown that the parent drug daphnetin and four metabolites were found in the perfusion fluid of duodenum, jejunum and ileum. However, no metabolites were found in the colon. Among the four metabolites, two daphnetin sulfates (m/z 257) were first discovered as the phase II metabolites of daphnetin in rats, which revealed a new way of daphnetin metabolism in rats.

Effects of glycyrrhiza extract on pharmacokinetics property of daphnetin in rats / 中国中药杂志

<p><b>OBJECTIVE</b>To research the influence of glycyrrhiza extract on the pharmacokinetics characteristic parameters of daphnetin, which was aimed to explore the rationality of concert application of drugs.</p><p><b>METHOD</b>The rats received intragastric administration of daphnetin and glycyrrhiza extract containing the same daphnetin respectively. The blood concentration of daphnetin was assayed by LC-MS. The data was processed by program DAS2.1.1.</p><p><b>RESULT</b>Glycyrrhiza extract can reduce the t(1/2), tmax and Ke of daphnetin, while increased the Ka and AUC(0-infinity).</p><p><b>CONCLUSION</b>Glycyrrhiza extract promoted the oral absorption of daphnetin, slowed down the elimination and increased the biological availability.</p>

Biotransformation of daphnetin by suspension transgenic hairy roots of Polygonum multiflorum / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the biotransformation of daphnetin by suspension transgenic hairy root of Polygonum multiflorum and provide a biotechnological method for large-scale production of the daphnetin-8-O-beta-D-glucoside using this new culture system.</p><p><b>METHOD</b>Daphnetin was added into the media of suspension to culture 36 h. The biotransformation product was detected with TLC and HPLC, and isolated by various chromatographic methods. The influence of co-cultured time on conversion ratio, content of degradation product and the reason for the degradation of product II were investigated using HPLC.</p><p><b>RESULT</b>One biotransformation product, daphnetin-8-O-beta-D-glucoside, was obtained, the optimal co-cultured time in suspension hairy root of P. multiflorum was 36 h with the highest biotransformation molar ratio of 32.11%, the sucrose medium (sucrose-only) can increase the biotransformation molar ratio to 72.44%. The result demonstrated that the degradation products of the product II was induced by the MS medium.</p><p><b>CONCLUSION</b>The potential application of suspension transgenic hairy root of P. multflorum in the sucrose-only medium on generating daphnetin-8-Obeta-D- glucoside could be prospective.</p>

Esculetin inhibits N-methyl-D-aspartate neurotoxicity via glutathione preservation in primary cortical cultures / 한국실험동물학회지

Recently, loss of endogenous glutathione during N-methyl-D-aspartate (NMDA) receptor-mediated excitotoxic injury, and the resultant overproduction of reactive oxygen species (ROS) through an arachidonic acid cascade process in brain, have been implicated in neuronal damage in various neurodegenerative diseases. Glutathione depletion induced by L-buthionine-(S,R)-sulfoximine (BSO), an inhibitor of glutathione synthesis, is known to cause arachidonic acid-mediated excitotoxicity in primary mixed cortical cultures. The aim of this study was to investigate whether esculetin (6,7-dihydroxycoumarin), an inhibitor of lipoxygenase, protects against neurotoxicity induced by NMDA or BSO. We observed that neurotoxicity induced by NMDA but not kainic acid was attenuated by esculetin. At the same concentration (100 microM), esculetin attenuated the 45Ca2+ uptake elevation induced by NMDA. Free radical-mediated neuronal injury induced by H2O2 and xanthine/xanthine oxidase was concentration-dependently blocked by esculetin. Esculetin (1-30 microM) dose-dependently inhibited BSO-induced neuronal injury. In addition, arachidonate-induced neurotoxicity was completely blocked by esculetin. BSO also reduced glutathione peroxidase (GPx) activity, but did not change glutathione reductase (GR) activity 24 h after treatment. Esculetin dose-dependently increased GR activity, but did not alter GPx activity. These findings suggest that esculetin can contribute to the rescue of neuronal cells from NMDA neurotoxicity and that this protective effect occurs partly through NMDA receptor modulation and the sparing of glutathione depletion.

Synthesis and purification of 7-prenyloxycoumarins and herniarin as bioactive natural coumarins

7-prenyloxycoumarins including 7-isopentenyloxycoumarin, auraptene and umbelliprenin, and herniarin have been widely recognized as bioactive coumarins. This paper presents the ways to synthesis these compounds. 7-prenyloxycoumarins were synthesized by reaction between 7-hydroxycoumarin [1 M] and relevant prenyl bromides [1.5 M] in acetone at room temperature. The reaction was carried out in the presence of DBU [1, 8-diazabicyclo [5.4.0] undec-7-ene] [2 M]. After 24 hr. the mixture was concentrated under reduced pressure. The compounds were purified by column chromatography. Three bioactive 7-prenyloxycoumarins, namely, umbelliprenin, auraptene and 7-isopentenyloxycoumarin, together with herniarin were synthesized from 7-hydroxycoumarin under alkaline conditions [DBU] and then purified by column chromatography. The structures of the products were characterized by NMR spectroscopic method including [1]H- and [13]C-NMR experiments. The method of synthesis for 7-prenyloxycoumarins and herniarin which is presented here has not been reported yet. Moreover, for the first time, umbelliprenin was chemically prepared in this work

Intestinal absorption of daphnetin by rats single pass perfusion in situ / 药学学报

This study investigated the effects of concentration, intestinal section, pH and P-gp on the absorption of daphnetin. The absorptions of three concentrations (10, 20, 40 microg x mL(-1)) of daphnetin in different intestinal segments were studied with phenol red as the marker by in situ rats single pass perfusion model. The results showed that daphnetin was stable under pH 6.0 condition and little affected by metabolism enzyme. There was upgrade tendency between the Peff of duodenum, jejunum, ileum and colon in different concentration of daphnetin, and it has obvious difference between the high concentration and low concentration in jejunum and colon, which indicated that the absorption of daphnetin was passive diffusion and no difference in different segments of rat intestine. However, compared with colon, the absorption of small intestine was better significantly (P < 0.05). Daphnetin may be not a substrate of P-gp as verapamil had not significantly affected the absorption of daphnetin in different intestinal segments of rats.

Coumarins from Peucedanum harry-smithii var. subglabrum / 中国中药杂志

The root of Peucedanum harry-smithii var. subglabrum was extracted with methanol, then separated with solvents at different polarity into four fractions: aqueous (H2O), ethyl acetate (AcOEt), chloroform (CHCl3) and petroleum ether (DAB-6). From AcOEt psoralen, bargapten, xanthotoxin, marmesin, umbelliferone, scopoletin, (+/-) peuformosin, Pd-I b, (+/-) selinidin, praeruptorin D were isolated by column chromatography on silica gel, using petroleum ether/ethyl acetate as eluent. The structures of the coumarins were identified by 1H-NMR and 13C-NMR.

Studies on chemical constituents from stem barks of Fraxinus paxiana / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the chemical constituents of Fraxinus paxiana.</p><p><b>METHOD</b>The chemical constituents were isolated and purified by chromatographic techniques and the structures of the compounds were identified with or by spectroscopic methods.</p><p><b>RESULT</b>Fifteen compounds were obtained from the methanol extract of F. paxiana and their structures were elucidated as esculin (1), esculetin (2), fraxin (3), fraxetin (4), salidroside (5), osmanthuside H (6), liriodendrin (7), 3-(4-beta-D-glucopyranosyloxy-3-methoxy)-phenyl-2E-propenol (8), threo-syringylglycerol (9), euscaphic acid (10), 3-hydroxy-1-(4-hydroxy-3, 5-dimethoxyphenyl)-1-propanone (11), omega-hydroxypropioguaiacone (12), sinapyladehyde (13), betulinic acid (14) and mannitol (15).</p><p><b>CONCLUSION</b>All compounds were obtained from this plant for the first time.</p>

Coumarins from branch of Fraxinus sieboldiana and their antioxidative activity / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the chemical constituents from the branch of Fraxinus sieboldiana, and evaluate their antioxidative activity.</p><p><b>METHOD</b>The chemical constituents were isolated and purified by chromatographic techniques over silica gel, macroporous adsorbent resin, Sephadex LH-20, and preparative HPLC. Structures of the compounds were identified by spectroscopic methods. The antioxidant activity was evaluated by Fe(+2)-cystine induced rat liver microsomal lipid peroxidation.</p><p><b>RESULT</b>Eight coumarins were obtained and their structures were elucidated as esculin (1) , esculetin (2), fraxin (3), fraxetin (4), 6, 7-di-O-beta-D-glucopyranosylesculetin (5), scopoletin (6), cleomiscosin D (7) and cleomiscosin B (8). At a concentration of 10(-6) mol x L(-1), compound 4 showed antioxidative activity inhibiting Fe(+2)-cystine induced rat liver microsomal lipid peroxidation with inhibitory rate of 60%.</p><p><b>CONCLUSION</b>Compounds 5, 7 and 8 were obtained from the genus Fraxinus for the first time. Compound 4 showed remarkable antioxidative activity, which was higher than that of VE (35%).</p>

Studies on chemical constituents from leaves of Vaccinium bracteatum / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the chemical constituents from the leaves of Vaccinium bracteatum.</p><p><b>METHOD</b>Many column chromatographic techniques were used for the isolation and separation of chemical constituents. Their structures were elucidated on the basis of spectral analysis and chemical evidences.</p><p><b>RESULT</b>Twelve compounds were isolated from the plant, and they were identified as chrysoeriol (1), scopoletin (2), trans-p-hydroxycinnamic acid (3), trans-p-hydroxycinnamic acid ethyl ester (4), cafeic acid ethyl ester (5), beta-sitosterol (6), iuteolin (7), quercetin (8), esculetin (9), cafeic acid (10), isolariciresinol-9-O-beta-D-xyloside (11), 10-O-trans-p-coumaroylsandoside (12).</p><p><b>CONCLUSION</b>Compounds 4, 5, 11, 12 were isolated from the genus Vaccinium for the first time, and compounds 1, 2, 9, 10 were isolated from this plant for the first time.</p>

Studies on chemical constituents of Saussurea laniceps / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the chemical constituents of Saussurea laniceps.</p><p><b>METHOD</b>The ethanol extract of S. laniceps was separated by means of silica gel chromatography. The compounds isolated from the plant were identified by their spectral evidence.</p><p><b>RESULT</b>Fifteen compounds were isolated and identified as beta-stiosterol (1), umbelliferone (2), 4-hydroxyacetophenone (3), scopoletin (4), isoscopoletin (5), xuelianlactone (6), methyl 3-(2', 4'-dihydroxyphenyl) propanoate (7), apigenin (8), neoechinulin A (9), daucosterol (10), scopolin (11), xuelianlactone 8-O-beta-D-glcuoside (12), apigenin 7-glcuoside (13), apigenin 7-lutinoside (14) and syringin (15).</p><p><b>CONCLUSION</b>Compounds 5-15 were isolated from S. laniceps, and among them, 7 and 9 were isolated from genus Saussurea for the first time.</p>

Studies on constitutes from Taraxacum mongolicum / 中国中药杂志

<p><b>OBJECTIVE</b>To study the chemical constituents of Taraxacum mongolicum.</p><p><b>METHOD</b>Repeated column chromatography over silica gel, C-18 and Sephadex LH-20 were used to isolate chemical constitutes and their structures were elucidated by spectroscopic methods MS, 1H-NMR, 13C-NMR including 2D-NMR techniques and by direct comparing spectral data with those reported in literatures.</p><p><b>RESULT</b>Eight compounds were identified as artemetin (1), quercetin (2), luteolin (3), luteolin-7-O-beta-D-glucopyranoside (4), caffeic acid (5), esculetin (6), stigmasterol (7), taraxasteryl acetate (8), respectively.</p><p><b>CONCLUSION</b>Compound 1 was obtained from this genus and compound 6, 8 from this species for the first time. Furthermore, the 13 carbon data of compound 6 from the literature were corrected with 2D-NMR techniques in this paper.</p>

Chemical constituents from roots of Ferula sinkiangensis / 中国中药杂志

<p><b>OBJECTIVE</b>To study the chemical constituents of the roots of Ferula sinkiangensis.</p><p><b>METHOD</b>Compounds were isolated by repeated chromatography on silica gel. Their structures were elucidated by chemical and spectroscopic methods.</p><p><b>RESULT</b>Seven compounds were identified as fekrynol (1), fekolone (2), farnesiferol B (3), isosamarcandin (4), episamarcandin (5), farnesiferol C (6), umbelliferone (7).</p><p><b>CONCLUSION</b>All the compounds were obtained from this plant for the first time.</p>