Two triterpenoids, arjunolic acid (1), belleric acid (2), five phenylethanoids, martynoside (3), orobanchoside (4), 3,4-dihydroxyphenethylalcohol-6-O-caffeoyl-β-D-glucoside (5), leucosceptoside B (6), lunariifolioside (7), four phenolic acids, ferulic acid (8), syringic acid (9), vanillic acid (10), 4-hydroxybenzoic acid (11), and one lignan, (+)-syringaresinol-β-D-glucoside (12), were isolated from the roots of P. umbrosa. All isolated compounds were explored for their antioxidant potential in the DPPH and ABTS assays. In DPPH assay, compound 5 showed high antioxidant capacity. Compounds 3, 4, 6, and 7 displayed considerable antioxidant activities. In addition, compounds 5–7 exhibited potential antioxidant capacities in the ABTS assay.
Phenol ; Phlomis ; Vanillic Acid

The roots of Phlomis umbrosa (Turcz.) (Phlomidis Radix) have been traditionally used to treat cold, reduce swelling and staunch bleeding. Four iridoids (1 – 3 and 5) and six phenylethanoid derivatives (4, and 6 – 10) were isolated from the roots of P. umbrosa. A simple, sensitive, and reliable analytical HPLC/PDA method was developed, validated, and applied to determine 10 marker compounds in Phlomidis Radix. Furthermore, the isolates were evaluated for cytotoxic and anti-oxidant activities as well as DPPH-HPLC method. Among them, compounds 4 and 6 – 9 displayed potent anti-oxidant capacities using DPPH assay with IC50 values of 27.7 ± 2.4, 10.2 ± 1.1, 18.0 ± 0.8, 19.1 ± 0.3, and 19.9 ± 0.6 µM, and compounds 6, 8, and 9 displayed significant cytotoxic activity against HL-60 with IC50 values of 35.4 ± 3.1, 18.6 ± 2.0, and 42.9 ± 3.0 µM, respectively.
Hemorrhage ; Inhibitory Concentration 50 ; Iridoids ; Methods ; Phlomis

OBJECTIVETo study the chemical constituents of Phlomis umbrosa.

METHODChemical constituents were isolated by repeated column chromatography (Toyopearl HW-40C and preparative HPLC). The structures were elucidated on the basis of spectral data analysis.

RESULTThirteen compounds were identified as oleanolic acid (1), corosolic acid (2), hederagenin (3), arjunolic acid (4), belleric acid (5), 3beta-hydroxy-29-al-12-en-28-oleanoic acid (6), butyrospermol (7), beta-sitosterol (8), daucosterol (9), caffeic acid (10), vanillic acid (11), p-hydroxy-benzoic acid (12), 3, 4-dihydroxy-benzoic acid (13).

CONCLUSIONCompounds 3 9, 11-13 were isolated from the plant for the first time.

Organic Chemicals ; analysis ; chemistry ; isolation & purification ; Phlomis ; chemistry ; Plant Roots ; chemistry

OBJECTIVETo develop an RP-HPLC method for the determination of five iridoid glycosides in Phlomis younghusbandii.

METHODHPLC analysis was performed on a Symmetry C18 (4.6 mm x 150 mm, 5 microm, Waters) column eluted with acetonitrile (A) and water (B) in gradient elution. The gradient program was as follows: 0-5 min kept 7% A; 5-10 min changed to 12% A; 10-40 min kept 12% A. The flow rate was 1.0 mL x min(-1). The column temperature was 20 degrees and the detection wavelength was 235 nm.

RESULTThe linear ranges of sesamoside, shanzhiside methyl ester, 7, 8-dehydropenstemoside, penstemoside and 8-O-acetylshanzhiside methyl ester were 0.050-0.650 (r = 0.999 3), 0.050-0.350 (r = 0.999 5), 0.040-0.280 (r = 0.999 4), 0.010-0.070 (r = 0.999 6), 0.040-0. 280 (r = 0.999 7) g x L(-1), respectively. The average recoveries (n = 6) of them were all between 96% and 104%, RSD < 5.0%.

CONCLUSIONThe method is simple, accurate, repeatable and stable, which can be used for quality control of P. younghusbandii.

Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; Iridoid Glycosides ; analysis ; Phlomis ; chemistry ; Quality Control ; Reproducibility of Results

OBJECTIVETo study the chemical constituents of the roots of Phlomis medicinalis.

METHODThe compounds were isolated and repeatedly purified by macroporous resin, silica gel column chromatography, TLC and PREP-HPLC. Their structures were elucidated by physical and chemical properties and NMR spectra.

RESULTTwo iridoid glucosides were obtained and elucidated as 7-epilamalbide (1), chlorotuberoside (2).

CONCLUSIONCompound 1 was a new compound, compound 2 was isolated from the plant for the first time.

Glucosides ; analysis ; isolation & purification ; Iridoid Glucosides ; Iridoids ; analysis ; isolation & purification ; Magnetic Resonance Spectroscopy ; Phlomis ; chemistry ; Plant Roots ; chemistry

In order to find the compound basis of Phlomis younghusbandii Mukerjee that related to pharmacodynamic action, various chromatographic techniques were used to separate and purify the constituents of this plant, and physicochemical and spectral data were used to identify the structures of obtained compounds. A new furanolabdane diterpene glycoside, named as phlomisoside F, was isolated and identified, which was 15,16-epoxy-8(9),13(16), 14-labdatrien-7-ketone-19-oic acid-beta-D-glucopyranosyl ester.
Diterpenes ; chemistry ; isolation & purification ; Glucosides ; chemistry ; isolation & purification ; Glycosides ; chemistry ; isolation & purification ; Molecular Structure ; Phlomis ; chemistry ; Plant Tubers ; chemistry ; Plants, Medicinal ; chemistry

A new isobenzofuranone derivative has been isolated from Phlomis betonicoides by using various chromatographic techniques, including silica gel, Sephadex LH-20, MCI-gel resin and RP-HPLC. This compound was determined as 5-(3-hydroxypropyl)-2,2-dimethyl-2-furo[3,4-]chromen-7(9)-one (1) by NMR, MS, IR and UV spectroscopic data. Compound 1 showed potent antibacterial activity with an MIC₉₀ value of (58.4 ± 4.2) mg·L⁻¹ for methicillin resistant Staphylococcus aureus (MRSA) strain [levofloxacin as a control with MIC₉₀ value of (52.8±4.6) mg·L⁻¹].
Anti-Bacterial Agents ; isolation & purification ; pharmacology ; Benzofurans ; isolation & purification ; pharmacology ; Methicillin-Resistant Staphylococcus aureus ; drug effects ; Microbial Sensitivity Tests ; Phlomis ; chemistry ; Phytochemicals ; isolation & purification ; pharmacology

To study the antitumor effect of phlomiol extracted from Phlomis younghusbandii in vivo and in vitro. The inhibitory effect of phlomiol on two kinds of human tumor cells proliferation was assayed by MTT method. Transplant tumor models of S180 and H22 were used. After transplantation, different doses of phlomiol were given to the mice for 14 days. The inhibitory rates were calculated. MTT method was used to assess the proliferation of T spleen lymphocyte cells and the activity of NK cells in tumor-bearing mice with S180. Phlomiol (50-100 mg x L(-1)) inhibited the proliferation of three kinds of tumor cells in vitro, antitumor effect of phlomiol was in a dose-dependent manner (r = 0.989, P < 0.05). The inhibitory rates of phlomiol (2.5, 5, 10 mg x kg(-1)) were 28.5%-65.0% and 35.0%-74.5% in tumor-bearing mice with S180 and H22 respectively, It could stimulate the spleen T-cells in tumor-bearing mice with S180 and increase the activity of the NK cells. Phlomiol could inhibit the proliferation of three kinds of tumor cells in vitro, present antitumor effect on the tumor-bearing mice, and improve the immunological function.
Animals ; Antineoplastic Agents, Phytogenic ; administration & dosage ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Disease Models, Animal ; Humans ; Iridoid Glucosides ; Iridoids ; administration & dosage ; Lymphocytes ; immunology ; Male ; Mice ; Mice, Inbred ICR ; Neoplasms ; drug therapy ; immunology ; physiopathology ; Phlomis ; chemistry

A total of 52 endophytic fungi were isolated from roots and stems of Tibetan medicinal plant Phlomis younghusbandii Mukerjee. These fungal isolates were molecularly identified based on ITS sequnces and 28S sequences distributed to 12 genera, including Phoma, Chaetosphaeronema, Fusarium and Leptosphaeria, etc. Among them, the dominant genus was Phoma. Extracts of all strains were evaluated for anti-HIV-1 integrase activity by using soluable integrase expressed in E. coli BL21 (DE3). The results showed that seven samples from five fungal endophytes PHY-24, PHY-38, PHY-40, PHY-51, PHY-53, which belonged to genus Chaetosphaeronema, inhibited strand transfer reaction catalyzed by HIV-1 integrase with IC50 values, of 6.60, 5.20, 2.86, 7.86, 4.47, 4.56 and 3.23 microg x mL(-1) respectively. In conclusion, the endophytic fungi of Phlomis younghusbandii Mukerjee are valuable for further screening anti-HIV-1 integrase agents.
Ascomycota ; enzymology ; isolation & purification ; Chaetomium ; enzymology ; isolation & purification ; Endophytes ; enzymology ; isolation & purification ; Escherichia coli ; enzymology ; HIV Integrase ; genetics ; metabolism ; HIV Integrase Inhibitors ; pharmacology ; Phlomis ; microbiology ; Phylogeny ; Plant Roots ; microbiology ; Plant Stems ; microbiology ; Plants, Medicinal ; microbiology ; Plasmids ; Recombinant Proteins ; genetics ; metabolism

Many bone-related diseases such as osteoporosis, rheumatoid arthritis are occurred by excessive bone resorbing activity of osteoclasts. Recently, many studies have been proceeded to find out the new therapeutic materials from natural products of plants. Phlomis umbrosa Turcz, one of the natural products of plants has been known to improve bone health. However, the precise effects and treatment mechanisms of Phlomis umbrosa Turcz about bone diseases has been unknown. So, we examined the effects of Phlomis umbrosa Turcz on expression of receptor activator of nuclear factor-kappaB ligand (RANKL)-induced osteoclast differentiation in bone marrow-derived macrophages (BMMs) and bone resorption. Also, we investigated the treatment mechanisms of Phlomis umbrosa Turcz relating to osteoclast differentiation. Here, we showed that Phlomis umbrosa Turcz significantly suppressed RANKL-induced osteoclast differentiation and bone resportion. Furthermore, Phlomis umbrosa Turcz suppressed the activation of NF-kappaB in bone marrow macrophage treated RANKL and M-CSF. The mRNA expression of c-Fos, nuclear factor of activated T-cells (NFAT)c1, osteoclast-associated receptor (OSCAR), tartrate-resistant acid phosphatase (TRAP) in BMMs was inhibited by Phlomis umbrosa Turcz. Integrin alphanu, beta3 relating to cell adhesion and dendritic cell-specific transmembrane protein (DC-STAMP) relating to the structure of filamentous actin (F-actin) ring and cathepsin K relating to bone resorbing activity are disrupted too. These results suggest that Phlomis umbrosa Turcz will be a good materials to treat bone diseases like osteoporosis.
Acid Phosphatase ; Actins ; Arthritis, Rheumatoid ; Biological Agents ; Bone Diseases ; Bone Marrow ; Bone Resorption ; Cathepsin K ; Cell Adhesion ; Cytokines ; Isoenzymes ; Macrophage Colony-Stimulating Factor ; Macrophages ; NF-kappa B ; Osteoclasts ; Osteoporosis ; Phlomis ; RANK Ligand ; RNA, Messenger ; T-Lymphocytes