CHCl3-soluble Fraction Research Articles

Isolation of a Unique Monoterpene Diperoxy Dimer From Ziziphora clinopodioides subsp. bungeana Together With Triterpenes With Antidiabetic Properties.

Ziziphora clinopodioides subsp. bungeana (Juz.) Rech.f. is used in traditional medicine for various purposes. Previous phytochemical studies focused on phenolic compounds, but triterpenoids were almost overlooked. The study focused on the isolation of compounds with dual antidiabetic activity from the aerial parts of Z. clinopodioides subsp. bungeana. Separation of CHCl3-soluble fraction by silica gel column chromatography using different mobile phases and purification of compounds by semi-preparative HPLC or preparative TLC. The structures of pure compounds were elucidated by 1D and 2D NMR experiments along with HRMS. Compound 1 was additionally identified by the single crystal X-ray diffraction method. α-Glucosidase inhibitory assay and GLUT4 expression and translocation in C2C12 myotubes were conducted to evaluate antidiabetic potential of isolated compounds. This phytochemical study led to the isolation of 20 compounds, including a unique monoterpene diperoxy dimer (1). Compounds 7 and 9-11 displayed more potent α-glucosidase inhibitory activity (IC50 45.3-135.3 μM) than acarbose used as a positive control (IC50 264.7 μM), while only pomolic acid (5) increased GLUT4 translocation in C2C12 myotubes in a significant manner. Extensive chromatographic separation led to the isolation and identification of a unique monoterpene diperoxy dimer (1) from aerial parts of Z. clinopodioides subsp. bungeana. Some triterpenes inhibited α-glucosidase, another increased GLUT4 translocation. Although none of the isolated compounds demonstrated dual antidiabetic activity, selected triterpenes proved to be potent antidiabetic agents in vitro.

Chemical diversity of the herbal decoction of Plectranthus ornatus and its anti-nociceptive and anti-inflammatory activities in zebrafish models.

Plectranthus ornatus is a medicinal plant originally from Africa but adapted to Brazil's climate conditions. It is recognized for its therapeutic properties, particularly for treating liver and stomach diseases, gastritis control, heartburn, and hangover. Therefore, studies on its chemical composition and pharmacological evaluation are important for the safe use of the plant. To investigate the chemical composition and pharmacological activity of leaf decoction of P. ornatus. The lyophilized herbal decoction from P. ornatus leaves was extracted with MeOH (methodology A), and CHCl3 and n-BuOH (methodology B). The compounds were isolated using chromatographic techniques. Their structures were determined using spectroscopic methods (1H and 13C NMR, IR, and HRESIMS) and comparison with published data. The lyophilized herbal decoctions DPO A (decoction from methodology A) and DPO B (methodology B), and compounds 8, 9, 15, and 16 (4, 20, and 40 mg/kg) were evaluated for their toxicity, anti-nociceptive, and anti-inflammatory effects in experimental adult zebrafish (Danio rerio) models. The present study focused on the CHCl3 and MeOH soluble fractions from the lyophilized leaf decoction leading to the isolation of five new diterpenes (1, 4-7) and two new glucuronide flavonoid derivatives (2 and 3). In addition, four diterpenes (14-17), two glucuronide flavonoid derivatives (10 and 11), three phenolics (8, 12, and 13), and one disaccharide (9) previously reported were also isolated. In zebrafish essays, all samples showed no toxicity and exhibited an anti-nociceptive effect in at least one of the tested doses: 9, 15, 16, and DPO B (4 mg/kg), 8 (20 mg/kg), and DPO A (40 mg/kg). Moreover, the compounds 15 (4, 20, and 40 mg/kg) and 16 (4, 20, and 40 mg/kg) exhibited anti-inflammatory effects. The lyophilized decoctions (DPO A and DPO B) including the compounds 8, 9, 15, and 16 exhibited significant anti-nociceptive effects in adult zebrafish and showed no toxicity. Since pain can be a symptom of liver, stomach, and gastrointestinal disorders, and all the samples proved to be non-toxic, the herbal decoction of P. ornatus leaf could be considered a potential therapeutic option in pain management, supporting the ethnopharmacological use of the plant.

A new bicyclo [3.3.1]nonadienemethanol-type lignan from the leaves of Taxus wallichiana Zucc.

Following an α-glucosidase inhibitory activity-guided phytochemical study, a rare new bicyclo[3.3.1]nonadienemethanol-type lignan named 9,9′-oxidolanceolatanin B (1) and four known lignans, lanceolatanin B (2), pinoresinol (3), syringaresinol (4), and medioresinol (5), were isolated from a CHCl3-soluble fraction of Taxus wallichiana Zucc. leaves. The absolute configuration of 9,9′-oxidolanceolatanin B was elucidated based on NMR spectroscopic interpretation and ECD data. All isolated compounds 1–5 showed stronger α-glucosidase inhibitory potency than the positive control, acarbose. This data could support the conceit that Taxus wallichiana leaves could be a promising source for plant-based anti-diabetic drug discovery.

Cytotoxic and antibacterial compounds from Schinus terebinthifolia Raddi fruits

Chromatographic procedures of extracts of Schinus terebinthifolia Raddi fruits afforded (Z)-masticadienoic (1) and 3β-masticadienolic (2) acids, tetrahydroamentoflavone (3), and 4-O-methyl gallic acid (4). Addicionally, the derivative 6-oxo masticadienoic acid (1a) was prepared by an allylic oxidation. The chemical structures of obtained compounds were elucidated by spectrometric data analyses. Furthermore, both the semi-synthetic derivative and the metabolites were subjected to in vitro cytotoxicity against the A549 human lung cancer cell line, as well as antimicrobial activity tests. Compounds 2 and 1a exhibited cytotoxicity towards A549 cells with IC50 values of 20.13 and 6.11 µM, respectively. In the tests against pathogens, the CHCl3 and EtOAc soluble fractions of MeOH extract along with the pure compounds, exhibited antibacterial activity against Escherichia coli, Staphylococcus aureus, and Pseudomonas aeruginosa. Except for 4-O-methyl gallic acid, the other pure compounds showed inhibitory microbial activities with MIC values ranging from 0.25 μg/mL to 25 μg/mL doses.

Microwave extraction and molecular imprinted polymer isolation of bergenin applied to the dendrochronological chemical study of Peltophorum dubium

This study describes methodologies for extracting and isolating bergenin, a C-glucoside of 4-O-methylgallic acid found in some plants and it presents various in vitro and in vivo biological activities. Bergenin was previously obtained from the Pelthophorum dubim (Fabaceae) roots with a good yield. Conventional chromatographic procedures of the CHCl3 soluble fraction of the MeOH extract gave 3.62% of this glucoside. An HPLC/DAD method was also developed and validated for bergenin and its precursor, gallic acid quantifications. Microwave extractions with different solvents were tested to optimize the extraction of bergenin, varying the temperature and time. MAE (Microwave Assisted Extraction) was more efficient than conventional extraction procedures, giving a higher yield of bergenin per root mass (0.45% vs. 0.0839%). Molecularly imprinted polymer (MIP) and non-imprinted polymer (NIP) based on bergenin as the template molecule, methacrylic acid, and ethylene glycol dimethacrylate were synthesized and characterized by FTIR and SEM (Scanning Electron Microscopy). Bergenin adsorption experiments using MIP and NIP followed by molecular imprinted solid phase extraction (MISPE) showed that MIP had a higher selectivity for bergenin than NIP. A dendrochronological study using the proposed method for detection and quantification of gallic acid and bergenin in five P. dubium growth rings of a 31-year-old heartwood and in the phelloderm and barks indicated that bergenin was more abundant in the 11–14th growth rings of the heartwood and decreased from the heartwood to the barks.

CHEMICAL CONSTITUENTS AND ANTIBACTERIAL ACTIVITY OF THREE TYPES OF AMAZONIAN Melipona spp. GEOPROPOLIS

This work describes the first chemical study on the metabolic profile of three geopropolis samples from Melipona fulva, M. compressipes, and M. paraensis collected in the Amazonian region. The samples were submitted to solvent extractions. The silylated hexane extracts were submitted to GC-MS analysis and tentatively identified mainly the presence of triterpenes, steroids, fatty acids, and alcohols. The chemical profile of M. fulva of and M. compressipes geopropolis are similar. However, cholesterol, triacontanol, and palmitic acid are the main compounds in M. fulva; in M. compressipes, palmitic acid, linoleic acid, and oleic acid are predominant. The chemical profile of the M. paraensis sample is entirely different once more than 60% of the extract comprises lupenone and triacontanol. The CHCl3 soluble fraction of the MeOH extract of the M. paraensis geopropolis was submitted to chromatographic techniques, and it afforded cycloartenol, 24-methylene-cycloartenol, lupeol, α- and β-amirins, and 7-O-methylaromadendrin. The MeOH extract of this sample presents antimicrobial activities against strains of Streptococcus mutans, S. sobrinus, and S. aureus (MIC of 15.6-62.5 µg mL-1 and MBC of 62.5-1000 µg mL-1). The M. fulva and M. compressipes geopropolis were not active in this test.

In silico and in vitro anti-inflammatory study of phenolic compounds isolated from Eucalyptus maculata resin

Plant resins are rich in bioactive compounds with high medicinal values. However, the chemistry and anti-inflammatory activity of the resins produced by trees of the genus Eucalyptus were scarcely investigated. The inflammatory targets cyclooxygenase-1 (COX-1), COX-2, TNF-, NF-B, and NO were significantly inhibited by the methanolic extract of Eucalyptus maculata kino resin (EME) and its CH2Cl2 soluble fraction (MCF). Sakuranetin (C1), (E)-cinnamic acid (C2), kaempferol 7- methyl ether (C3), 7-O-methyl aromadendrin (C4), and 1,6- dicinnamoyl-O-α-D-glucopyranoside (C5) were isolated from MCF. Three compounds (C1, C2, and C4) showed potent in vitro COX-1 inhibition, while C5 inhibited COX-2, TNF-α, NF-κB, and NO significantly. An in-silico study revealed that C5 had the highest binding affinity to the active site in COX-2 with binding energy score (S) of -14.85 kcal/mol, better than celecoxib (COX-2 inhibitor). In conclusion, 1,6-dicinnamoyl-O-α-D-glucopyranoside (C5) could be investigated further in the search for anti-inflammatory agents.

Molecular Networking-Guided Isolation of Cycloartane-type Triterpenoids from Curculigo orchioides and Their Inhibitory Effect on Nitric Oxide Production.

The MolNetEnhancer workflow was applied to molecular networking analysis of the CH2Cl2-soluble fraction of the rhizomes of Curculigo orchioides, which showed a potent inhibitory effect on the lipopolysaccharide (LPS)-induced nitric oxide production. Among the molecular network, clusters of cycloartane-type triterpenoids were classified using the ClassyFire module of MolNetEnhancer, and their structures were predicted by the in silico fragment analysis tool, Network Annotation Propagation (NAP). Using mass spectrometry (MS)-guided isolation methods, six cycloartane-type triterpenoids (1–6) were isolated, and their structures were elucidated based on the interpretation of NMR, HRESIMS, and single-crystal X-ray diffraction. Among the isolates, compounds 1 and 4, which have an α,β-unsaturated carbonyl moiety on the A-ring, exhibited significant inhibitory effects on LPS-induced nitric oxide production in RAW264.7 cells with IC50 values of 12.4 and 11.8 μM, respectively.

Naphthoquinones from Catalpa bungei “Jinsi” as potent antiproliferation agents inducing DNA damage

Structure-guided isolation of a CH2Cl2-soluble fraction of the heartwood of Catalpa bungei “Jinsi” provided two new naphthoquinones, 9-hydroxy-4-oxo-α-lapachone (1) and 6-hydroxy-4-oxo-α-lapachone (2), together with three undescribed ones (3–5) and six known ones (6–11). The structures were elucidated on the basis of spectroscopic methods including electronic circular dichroism calculation. The antiproliferative effects of these isolates were evaluated in human breast adenocarcinoma cells MCF7. (4R)-4,9-dihydroxy-α-lapachone (5) and (4S)-4,9-dihydroxy-α-lapachone (6) exhibited the significant activities with IC50 values of 2.19 and 2.41 μM, respectively. The structure-activity relationship of 1–11 in the antiproliferative assay was then discussed. The most potent 5 and 6 were found to induce cell arrest in G1 phage through DNA damage. The findings provided some valuable insights for the discovery and structural modification of α-lapachone as antiproliferative lead compounds against human breast adenocarcinoma cells.

Chemical constituents of the aerial parts Helicteres hirsuta Lour (Malvaceae)

This paper reported the isolation of chemical constituents from the chloroform extract of aerial parts of Helicteres hirsuta Lour (Malvaceae) and isolated compounds were evaluated for their α-glucosidase inhibitory. This work aimed to discover effective α-glucosidase inhibitors for scientific evidence in using medicinal plants for diabetic treatment in Vietnamese traditional medicines as well as developing drugs for diabetic treatment. From the CHCl3-soluble fraction of the aerial parts of Helicteres hirsuta Lour, collected in An Giang province, four compounds, including (+)-pinoresinol (1), (3,4-dihydroxyphenyl)acetic acid ethyl ester (2), 7-O-methylisoscutellarein (3) and ethyl 3-(3,4-dihydroxyphenyl)lactate (4) were isolated. Their chemical structures were elucidated based on the NMR spectroscopic analysis and comparison with data in the literature. All four isolated compounds (1‒4) showed good α-glucosidase inhibitory assay activity with the IC50¬ of 37.9, 69.1, 12.5, 9.4 µM, respectively, better than that of the positive control acarbose (IC50¬ 168.0)

The Anti-Oxidative and Anti-Neuroinflammatory Effects of Sargassum horneri by Heme Oxygenase-1 Induction in BV2 and HT22 Cells.

Sargassum horneri is used as a traditional medicinal agent and exhibits various pharmacological effects. In this study, we found that the 70% EtOH extract contained 34.37 ± 0.75 μg/mg fucosterol. We tested the antioxidant activities of the 70% EtOH extracts and their fractions. The CH2Cl2-soluble fraction showed the strongest DPPH and ABTS radical scavenging activities. Next, we evaluated the anti-neuroinflammatory effects of S. horneri on lipopolysaccharide (LPS)-stimulated BV2 cells. Pretreatment with the extract and fractions suppressed LPS-induced production of nitric oxide (NO) in BV2 cells. The 70% EtOH, CH2Cl2-soluble fraction, and water-soluble fraction inhibited the production of prostaglandin E2, interleukin-6, and tumor necrosis factor-α, as well as markedly blocking LPS-induced expression of inducible NO synthase and cyclooxygenase-2 via inactivation of the nuclear factor-kappa B pathway. In addition, the CH2Cl2-soluble fraction showed the most remarkable heme oxygenase (HO)-1 expression effects and increased nuclear erythroid 2-related factor translocation in the nucleus. In HT22 cells, the CH2Cl2-soluble fraction inhibited cell damage and ROS production caused by glutamate via the regulation of HO-1. Therefore, CH2Cl2-soluble fractions of S. horneri can attenuate oxidative action and neuroinflammatory responses via HO-1 induction, demonstrating their potential in the treatment of neuroinflammatory diseases.

Selective Extraction of Bioactive Phenylethanoids from Digitalis obscura.

Cardenolide-free extracts from Digitalis obscura showed significant antifeedant effects against the aphid Myzus persicae and this activity correlated with their phenylethanoid content. The content in phenylethanoids of Digitalis obscura has been studied. Maceration of the aerial parts of D. obscura was used for the selective extraction of the natural compound rengyolone (1) and the aglycone of cornoside (compound 3). Pure rengyolone (1) can be obtained from D. obscura in approximately 90% purity from fresh plant from the CHCl3 soluble fraction of the ethanolic extract (0.8% yield). The ethanol extraction of freshly collected D. obscura showed the presence of compound 3 as the only phenylethanoid. Compound 3 was proven to easily evolve to rengyolone. Due to this instability, and although its presence in plants has been previously reported, the spectroscopical data of 3 are reported herein for the first time. Selective mono-acetylation of compound of 3 led to the active natural compound hallerone (5). The aphid antifeedant (against Myzus persicae) and nematicidal (against root-knot nematode Meloidogyne javanica) activities of these compounds have been evaluated. Here we report for the first time on the aphid antifeedant effects of 1, 3, and 5. Additionally, the nematicidal activity of hallerone (5) is described here for the first time.

A new cytotoxic cardenolide from the roots of Calotropis gigantea

Bioactivity-guided isolation of the CHCl3-soluble fraction of the roots of Calotropis gigantea was carried out to obtain a new cardenolide glycoside, caloside G. Its absolute structure was elucidated based on NMR and ECD spectroscopic data interpretation. Caloside G showed noteworthy cytotoxicity against the PANC-1 human pancreatic and HeLa human cervical carcinoma cell lines, with the submicromolar IC50 values of 0.038 and 0.09 µM, respectively.

The potential aphrodisiac effect of Ferula drudeana korovin extracts and isolated sesquiterpene coumarins in male rats

Background: Many members of the genus Ferula are used in traditional medicine as aphrodisiac. Objectives: The aim of this study is to confirm the aphrodisiac potential of Ferula drudeana Korovin as listed in the Turkish traditional medicine and to isolate the active metabolites using male rats. Materials and Methods: The CHCl3soluble fraction showed promising activity. Chromatographic purification resulted in the isolation of three sesquiterpene coumarins. Isolated compounds structures were determined as feselol (1), samarcandin (2), and 3'-O-acetyl samarcandin (3) based on the physical and spectral characters. Single doses of F. drudeana CHCl3soluble fraction, aqueous fraction (200 mg/kg BW), 1 and 2 (10 mg/kg BW), and sildenafil citrate (10 mg/kg BW) were orally administered to male Wistar albino rats by gavages. Mount latency, mount frequency (MF), intromission latency, intromission frequency (IF), ejaculation latency, and postejaculatory interval (PEI) were studied. In addition, copulatory efficiency and intercopulatory efficiency were calculated. Results: Oral administration of F. drudeana roots extracts, 1 and 2 significantly increased MF and IF. The latencies of mount and intromission were reduced significantly and ELs were prolonged. Treatment with the extracts, 1 and 2 resulted in the reduction of the PEI. The highest aphrodisiac activity in male rats was exhibited by 2. Conclusion: The present findings provide experimental evidence that F. drudeana roots, 1 and 2 possess aphrodisiac activities by enhancing the sexual behavior of male rats. The obtained results supported the traditional claims about the use of Ferula species for male sexual dysfunction.

New cytotoxic polyacetylene alcohols from the Egyptian marine sponge Siphonochalina siphonella.

Three new polyacetylenic alcohols, siphonellanols A-C (1-3), together with two known polyacetylenic alcohols (4-5), were isolated from the CHCl3-soluble fraction of the methanolic extract of the marine sponge Siphonochalina siphonella, collected in Egypt. The structures of 1-3 were determined by spectroscopic analyses of their 1D-, 2D-NMR, and MS spectra and by comparisons with reported data. The cytotoxicity assay revealed that 1-3 exhibited moderate cytotoxic activities against a human cervical cancer cell line (HeLa), a human breast cancer cell line (MCF-7), and a human lung cancer cell line (A549) with IC50 values ranging from 25.9 to 69.2μM.

Chemical constituents from methanolic extract of Solanum procumbens Lour (Solanaceae)

Solanum procumbens L. (Solanaceae) were extracted respectively with n-hexane, ethyl acetate, and methanol to obtain the corresponding extracts. The methanol-soluble extract was acidified by adding HCl 0.5M; this was followed by liquid-liquid partition with CHCl3 to afford CHCl3-soluble fraction. From this fraction, four compounds were isolated and identified as ergosterol peroxide (1), 9,11-dehydroergosterol peroxide (2), N-(N-benzoyl-L-phenylalanyl)-L-phenylalanol (3), and moupinamide (4). Their structures were elucidated based on the NMR spectroscopic analysis and comparison with the literature. All isolated compounds were first reported from S. procumbens.

Minor Nortriterpenoids from the Twigs and Leaves of Rhododendron latoucheae.

A hyphenated NMR technique (analytical HPLC with a DAD connected to MS, SPE, and NMR) has proven effective for the full structural analysis and identification of minor natural products in complex mixtures. Application of this hyphenated technique to the CH2Cl2-soluble fraction of Rhododendron latoucheae led to the identification of 15 new minor ursane-type 28-nortriterpenoids (1-15). Compounds 1 and 12 inhibited HSV-1 with IC50 values of 6.4 and 0.4 μM, respectively.

Cytotoxic garcimultiflorones K–Q, lavandulyl benzophenones from Garcinia multiflora branches

Seven undescribed lavandulyl benzophenones garcimultiflorones K–Q, and fourteen known compounds were isolated from the CHCl3 soluble fraction of 95% EtOH extract of Garcinia multiflora branches. Their structures and absolute configurations were determined by spectroscopic techniques including NMR spectroscopy, MS analysis, and ECD calculations. Seven isolated compounds expect for garcimultiflorone L and garcimultiflorone O exhibited cytotoxic activities in vitro against five cancer cell lines (HL-60, A549, SMMC-7721, MCF-7, and SW480). It is worth mentioning that garcimultiflorone Q exhibited most significant cytotoxicities against five cancer cell lines with IC50 values ranging from 3.07–12.56 μM.

Mangostanaxanthone VII, a new cytotoxic xanthone from Garcinia mangostana.

Garcinia mangostana L. (the queen of fruits, mangosteen, family Guttiferae) is a wealthy source of xanthones. The CHCl3 soluble fraction of the air-dried pericarps of G. mangostana provided a new xanthone: mangostanaxanthone VII (5), along with four known xanthones: mangostanaxanthones I (1) and II (2), gartanin (3) and γ-mangostin (4). The structural verification of these metabolites was achieved by different spectral techniques, including UV, IR, 1D and 2D NMR and HRESIMS. The new metabolite was assessed for cytotoxic potential, using sulforhodamine B (SRB) assay towards the A549 and MCF-7 cancer cell lines. Moreover, its antimicrobial effects were evaluated against various bacterial and fungal strains, using agar disc diffusion assay. Mangostanaxanthone VII showed moderate cytotoxic activity against the A549 and MCF7 cell lines with IC50s 26.1 and 34.8 μM, respectively, compared with doxorubicin (0.74 and 0.41 μM, respectively).

A new bischromanone from the stems of Semecarpus caudata

From an CHCl3-soluble fraction of the stems of Semecarpus caudata, one new bischromanone named semecarpanone (1), together with 5 known flavonoids (2–6) were isolated. Their structures were elucidated based on interpretation of spectroscopic data. The stereo-configuration of 1 was identified based on the calculated and experimental coupling constants. Compounds 4–6 showed potent tyrosinase inhibitory activity with the IC50 values ranging from 15.0 to 76.3 μM.