Isolation, characterization and evaluation of oxypeucedanin and osthol from local endemic Prangos aricakensis Behçet and Yapar root as antioxidant, enzyme inhibitory, antibacterial and DNA protection: molecular docking and DFT approaches.

Prangos species were previously used against many disorders due to their chemical component. Prangos aricakensis Behçet & Yapar is a newly discovered local endemic species in Turkey's eastern region, and there is no research on P. aricakensis in the literature. In this work, oxypeucedanin and osthol molecules have been isolated from the root part of P. aricakensis for the first time. Oxypeucedanin and osthol structures were elucidated by 1D and 2D NMR analysis. For the bioactivities determination, antioxidant (DPPH· and ABTS·+ scavenging), enzyme inhibition (AChE, BChE, tyrosinase, and urease), antibacterial and DNA protection activity studies were applied for both molecules and compared with standard drug molecules, after applying enzyme kinetic assays and in silico approaches to clarify the mechanism of action for both molecules with enzymes, using molecular docking and density functional theory (DFT). Oxypeucedanin (2.19 ± 0.38 µg/mL) and osthol (4.57 ± 1.28 µg/mL) exhibited better activity than standards in DPPH∙ scavenging activity. Osthol (11.76 ± 0.59 µg/mL) showed a better tyrosinase inhibition effect than kojic acid (12.82 ± 0.91 µg/mL), and oxypeucedanin (3.03 ± 0.01 µg/mL) showed better urease inhibition effect than thiourea (5.37 ± 1.86 µg/mL). Our results showed that the osthol molecule was an excellent skin protective agent while the oxypeucedanin molecule could be a remarkable antiulcer agent. Therefore, although this study is the first in its field, it remained in the in vitro and in silico stages and is thought to pave the way for in vivo studies in the future.Communicated by Ramaswamy H. Sarma.

Design, Preparation, and Evaluation of Osthol Poly-Butyl-Cyanoacrylate Nanoparticles with Improved In Vitro Anticancer Activity in Neuroblastoma Treatment.

Osthol (osthole), known as a neuroprotective drug, has shown potent anticancer activity. However, the potential clinical application of osthol is limited due to its low water solubility and low bioavailability. Polybutyl cyanoacrylate (PBCA) has been widely used to improve the solubility of drugs with poor water solubility. In this study, an orthogonal experimental design (OED) was applied to design the preparation process of PBCA nanoparticles (NPs). Then, nanoparticles were prepared and evaluated in terms of physicochemical properties, in vitro release, and cellular uptake, etc. Further, the anti-cancer activity of osthol-PBCA NPs was demonstrated in SH-SY5Y cells. The pharmacokinetics and area under the curve (AUC) were investigated. The obtained osthol-NPs presented a spherical shape with a particle size of 110 ± 6.7 nm, a polydispersity index (PDI) of 0.126, and a zeta potential of −13 ± 0.32 mV. Compared with the free osthol, the drugs in osthol-NPs presented better stability and sustained release pattern activity. In vitro analysis using SH-SY5Y neuroblastoma cells showed that osthol-loaded nanoparticles displayed a significantly enhanced intracellular absorption process (three times) and cytotoxicity compared with free osthol (p < 0.05, increased 10−20%). The in vivo pharmacokinetic study revealed that the AUC of osthol-NPs was 3.3-fold higher than that of free osthol. In conclusion, osthol-PBCA NPs can enhance the bioactivity of osthol, being proposed as a novel, promising vehicle for drug delivery.

Synthesis, characterization, and antimicrobial evaluation of polyvinylalcohol-osthol composite films.

Although poly-vinyl alcohol (PVA) has certain mechanical drawbacks such as a weak barrier, it has widely been used in food packaging over the last many years. To increase the suitability of PVA (C2H4O)n and render it ideal for food packaging, a diversity of studies have already been carried out. In the below-mentioned script, we, for the first time, report the use of natural product osthol in making a new composite with PVA for enhancing thermal, physicochemical, and antimicrobial properties. The significant aim of the report is the insertion of osthol (C15H16O3) into PVA polymer, which is to be subsequently used for antimicrobial applications. The synthesis of the polymer composite film is done by solvent casting method and is characterized by SEM, XRD, FT-IR, and UV-Vis spectroscopy analysis. The manifestation of antimicrobial activity against (S. aureus) (ATCC8738P), E. coli (ATCC8739), Aspergillus niger, Alternaria alternata, and Fusarium solani by the film composite is remarkable. The addition of osthol molecule increases the tensile strength of PVA films from 18.73 ± 0.56 Mpa (PVA) to 24.58 ± 0.49 Mpa (15 mL). As a result, tensile strength increases by 23.79% in a film containing a higher concentration of osthol (15 mL). The barrier properties of PVA osthol composite films improve with the incorporation of osthol. OTR and WVTR decrease by 43.03% and 30.24%, respectively, on the addition of 15 mL osthol. Reduction in OTR and WVTR of the films could increase their applicability in the food sector. An increase in contact angle from 43° (pure PVA) to 66.7° increases the hydrophobic character of the composite films which is desirable for food packaging. This noticeable enhancement of the properties of the PVA film like hydrophobicity, mechanical, barrier, and antimicrobial is supporting the potential application of achieved material in packaging of easily perishable foods like fruits and vegetables by extending their shelf life.

[Pharmacokinetic study on five components in phthalide target areas of Chaxiong and its ß-CD inclusion compounds based on UPLC-MS/MS].

This study aims to establish a method for the determination of the concentration of five main components of phthalide target areas of Chaxiong(CPTA) and its inclusion of ß-CD in the plasma of rats, and determine the pharmacokinetic parameters, absolute bioavailability and relative bioavailability of CPTA/ß-CD inclusion compound in vivo. The plasma concentrations of senkyunolide A, N-butylphthalide, new osthol lactone, Z-ligustilide and butenyl phthalide were determined with UPLC-MS/MS. The content determination was conducted at the chromatographic conditions as follows: Shim-pack GIST C_(18)-AQ HP column(2.1 mm×100 mm, 3 µm), mobile phase of 0.1% formic acid solution(A)-acetonitrile(B), gradient elution, flow rate of 0.3 mL·min~(-1), column temperature of 35 ℃ and injection volume of 2 µL. The mass spectra were obtained with electrospray ion source(ESI), positive ion mode and multi reaction monitoring. CPTA/ß-CD inclusion compound was prepared by grinding method, DAS 2.0 software was used to model the data, and the absolute bioavailability of CPTA and relative bioavailability of inclusion compound were calculated. Finally, the methods for the determination of five components of senkyunolide A, N-butylphthalide, new osthol lactone, Z-ligustilide and butenyl phthalide in CPTA, were successfully established. The linear relationship among the five components was good within their respective ranges, r>0.99. The absolute bioavailability of the five components in rats was 22.30%, 16.32%, 21.90%, 10.16% and 12.43%, respectively. After CPTA/ß-CD inclusion was prepared, the relative bioavailability of the five components was 138.69%, 198.39%, 218.01%, 224.54% and 363.55%, respectively, significantly improved. This method is rapid, accurate and sensitive, so it is suitable for the pharmacokinetic study of extracts in traditional Chinese medicine and their preparations.

Pharmacokinetic study on five components in phthalide target areas of Chaxiong and its β-CD inclusion compounds based on UPLC-MS/MS / 中国中药杂志

This study aims to establish a method for the determination of the concentration of five main components of phthalide target areas of Chaxiong(CPTA) and its inclusion of β-CD in the plasma of rats, and determine the pharmacokinetic parameters, absolute bioavailability and relative bioavailability of CPTA/β-CD inclusion compound in vivo. The plasma concentrations of senkyunolide A, N-butylphthalide, new osthol lactone, Z-ligustilide and butenyl phthalide were determined with UPLC-MS/MS. The content determination was conducted at the chromatographic conditions as follows: Shim-pack GIST C_(18)-AQ HP column(2.1 mm×100 mm, 3 μm), mobile phase of 0.1% formic acid solution(A)-acetonitrile(B), gradient elution, flow rate of 0.3 mL·min~(-1), column temperature of 35 ℃ and injection volume of 2 μL. The mass spectra were obtained with electrospray ion source(ESI), positive ion mode and multi reaction monitoring. CPTA/β-CD inclusion compound was prepared by grinding method, DAS 2.0 software was used to model the data, and the absolute bioavailability of CPTA and relative bioavailability of inclusion compound were calculated. Finally, the methods for the determination of five components of senkyunolide A, N-butylphthalide, new osthol lactone, Z-ligustilide and butenyl phthalide in CPTA, were successfully established. The linear relationship among the five components was good within their respective ranges, r>0.99. The absolute bioavailability of the five components in rats was 22.30%, 16.32%, 21.90%, 10.16% and 12.43%, respectively. After CPTA/β-CD inclusion was prepared, the relative bioavailability of the five components was 138.69%, 198.39%, 218.01%, 224.54% and 363.55%, respectively, significantly improved. This method is rapid, accurate and sensitive, so it is suitable for the pharmacokinetic study of extracts in traditional Chinese medicine and their preparations.

Evaluation of the Physicochemical Properties, Pharmacokinetics, and In Vitro Anticancer Effects of Docetaxel and Osthol Encapsulated in Methoxy Poly(ethylene glycol)-b-Poly(caprolactone) Polymeric Micelles.

Docetaxel (DTX), a taxane-based anticancer drug, and osthol (OTH), a coumarin-derivative compound, have shown anticancer effects against different types of cancers through various mechanisms. However, these drugs have low solubility in water and low oral bioavailability, and thus their clinical application is difficult. To overcome these problems, we encapsulated DTX and OTH in methoxy poly(ethylene glycol)-b-poly(caprolactone) (mPEG-b-PCL) and conducted studies in vitro and in vivo. We selected a 1:4 ratio as the optimal ratio of DTX and OTH, through combination index analysis in A549 cancer cells, and prepared micelles to evaluate the encapsulation efficiency, drug loading, particle size, and zeta potential. The in vitro drug-release profile showed that DTX/OTH-loaded mPEG-b-PCL micelles could slowly release DTX and OTH. In the clonogenic assay, DTX/OTH-loaded mPEG-b-PCL micelles showed 3.7 times higher inhibitory effect than the DTX/OTH solution. Pharmacokinetic studies demonstrated that micelles in combination with DTX and OTH exhibited increased area under curve and decreased clearance values, as compared with single micelles.

Effects of osthol on activity, mRNA and protein expression of Cyp3a in rats in vivo.

Osthol (OST) has a wide range of pharmacological effects and has long been used in clinical medicine in China. Previous studies have indicated that osthol has weak inhibitory effects on CYP3A4 in human liver microsomes. The aim of the present study was to investigate the inhibition of Cyp3a by osthol in rats in vivo. A substrate assay was used to corroborate the inhibitory effect on Cyp3a by osthol in rats, and the substrate probe (midazolam) was detected by high-performance liquid chromatography (HPLC). Semi-quantitative RT-PCR (SqRT-PCR) analysis was used to study the effect of osthol on Cyp3a1 and Cyp3a2 mRNA expression and Western blot analysis was used to investigate the effect of OST on Cyp3a1 and Cyp3a2 protein expression. Our study confirmed the inhibitory effect of osthol on Cyp3a and indicated that the inhibitory effect on Cyp3a was stronger in the group receiving multiple doses compared with the single dose group. The SqRT-PCR analysis results showed that medium and high doses of osthol (20 and 40 mg/kg, respectively) had an inhibitory effect on Cyp3a1 mRNA expression but not on Cyp3a2 mRNA expression. Western blot analysis results indicated that the inhibitory effect of the medium and high osthol doses on Cyp3a1 and Cyp3a2 protein expression was significantly different. It was also demonstrated that the inhibitory effect of osthol on Cyp3a in rats resulted from the comprehensive effect of the direct inhibition of the Cyp3a enzyme, as well as the down-regulation of its mRNA and protein expression level.

HPLC fingerprint of Cnidii Fructus formula granule and determination of six main coumarin components / 中草药

Objective: To establish an HPLC fingerprint of Cnidii Fructus formula granule analysis method for simultaneous determination of six main coumarin components, including osthol, xanthotoxin, xanthotol, bergapten, imperatorin and isopimpinellin, in order to provide reference for the study of the material basis of Cnidii Fructus formula granule. Methods: The method was performed by high performance liquid chromatography with a Waters XBridge C18 (250 mm × 4.6 mm, 5 μm) column and methanol (A)-0.1% acetic acid (B) as the mobile phase for gradient elution. The flow rate was 0.5 mL/min, the injection volume was 10 μL and the column temperature was 40 ℃. The detection wavelength was set at 320 nm. The chromatographic fingerprint evaluation system published by the State Pharmacopoeia Commission (2012 Edition) was used to establish the fingerprint of Cnidii Fructus formula granule, and the content of six main coumarin components was simultaneously determined. Results: The research on the 18 batches of Cnidii Fructus formula granule showed that the fingerprint similarity was greater than 0.992 and 19 common peaks were calibrated with satisfied peak resolution. The content determination results showed that the content of both xanthotoxin and osthol were the main coumarin components in Cnidii Fructus formula granule. According to the methodological investigation, the precision RSD values were all less than 1.6%. The sample was stable within 48 h and this method had good repeatability. The average recovery rates of xanthotol, xanthotoxin, imperatorin, isopimpinellin, bergapten and osthol were 100.69%, 101.03%, 99.48%, 100.88%, 101.27% and 100.35%, respectively. All of these coumarin components’ RSD were less than 2.5%. The six components showed a good linear relationship within a certain concentration range. The results of the content determination of xanthotol, xanthotoxin, isopimpinellin, bergapten, imperatorin and osthol respectively were 8.01-8.29, 2.37-2.63, 4.30-4.61, 4.04-4.40, 3.45-3.90 and 6.02-6.80 mg/g among the 18 batches of the Cnidii Fructus formula granule. Conclusion: The fingerprint method and the determination method of six main coumarin components in the Cnidii Fructus formula granule established in this study are simple, stable, accurate and reliable. This method can be used for the quality control of the Cnidii Fructus formula granule.

Synthesis and Biological Evaluation of Novel Osthol Derivatives as Potent Cytotoxic Agents.

BACKGROUND: Natural product, osthol has been found to have important biological and pharmacological roles particularly having inhibitory effect on multiple types of cancer. OBJECTIVE: The unmet needs in cancer therapeutics make its derivatization an important and exciting field of research. Keeping this in view, a whole new series of diverse analogues of osthol (1) were synthesized. METHOD: All the newly synthesized compounds were made through modification in the lactone ring as well as in the side chain of the osthol molecule and were subjected to anti-proliferative screening through 3-(4,5-Dimethylthiazol-yl)-diphenyl tetrazoliumbromide (MTT) against four different human cancers of diverse origins viz. Colon (Colo-205), lung (A549), Leukemia (THP- 1) and breast (MCF-7) including SV40 transformed normal breast epithelial cell (fR-2). RESULTS: Interestingly, among the tested molecules, most of the analogs displayed better antiproliferative activity than the parent Osthol 1. However, among all the tested analogs, compound 28 exhibited the best results against leukemia (THP1) cell line with IC50 of 5µM.Compound 28 induced potent apoptotic effects and G1 phase arrest in leukemia cancer cells (THP1). The population of apoptotic cells increased from 13.8% in negative control to 26.9% at 8µM concentration of 28. Compound 28 also induced a remarkable decrease in mitochondrial membrane potential (ΛΨm) leading to apoptosis of the cancer cells. CONCLUSION: A novel series of molecules derived from natural product osthol were synthesized, wherein compound 28 was found to be most effective against leukemia and with 10 fold less toxicity against normal cells. The compound induced cancer inhibition mainly through apoptosis and thus has a potential in cancer therapeutics.

The ethnopharmacology, phytochemistry and pharmacology of Angelica biserrata - A review.

ETHNOPHARMACOLOGICAL RELEVANCE: Angelica biserrata (R.H. Shan & C.Q. Yuan) C.Q. Yuan & R.H. Shan (Angelica pubescens Maxim. f. biserrata Shan et Yuan) (A.biserrata) is a widely used traditional Chinese medicine; its roots known as 'Duhuo' in China. The herb is used for expelling wind, eliminating dampness, and terminating pain. Moreover, it is used for treating the onset of anemofrigid-damp arthralgia, pain of the waist and knee and headache caused by latent wind pathogenic factor or damp-cold pathogenic factor. A.biserrata is slightly warm, bitter and pungent in taste, and it is well distributed in regions such as Sichuan and Hubei Provinces. AIM OF THE STUDY: This review aims to provide critical summary of the current evidence on A.biserrata. In particular, the progress of studies in the fields of botany, ethnopharmacology, phytochemistry, pharmacology and toxicity are discussed. Possible directions for future research are also briefly proposed. MATERIALS AND METHODS: Information on A.biserrata was collected from the internet database PubMed, Elsevier, China Knowledge Resource Integrated databases, ResearchGate, Web of Science, Wiley Online Library and Europe PMC using a combination of various relevant keywords. Other published books providing an overview of extant literature studies were considered for reference if they are related to the taxonomy, traditional knowledge, phytochemistry, pharmacology and toxicity of the plant. RESULTS: A substantial proportion of the isolated and identified compounds of the herb were reported to be coumarins and volatile oils. Biological effects, such as neuroprotective, anti-tumor, anti-arthritis, anti-inflammatory, and sedative, were also validated in In vitro and in vivo studies. Therapeutic effects are attributed to the bioactivities of the naturally occurring compounds in this herb. CONCLUSIONS: A.biserrata has been proven as a valuable medicinal sources from traditional herb. Some conventional uses has been evaluated by pharmacological investigation. Although the crude extracts of A.biserrata has been emerged to possess more pharmacological activities, it is now time to isolate and identify more active chemical constituents by Bioactivity-Guided and elucidate their structure-activity relationship. More designed investigations are need to focus on understanding the multi-target network pharmacology, clarity the molecular mechanism of action and efficacy as well as identifying the effective doses of A.biserrata. In addition, A.biserrata is not fully assessed regarding its safety. Further studies are essential to investigate its toxicity on human. It's useful to provide identify its underlying therapeutic remedy and economic value of developing new medicine in the future.

Preparation of Chitosan-stearic Acid Nano-micelles Modified with Folic Acid and mPEG / 中国药房

OBJECTIVE： To prepare insoluble anti-tumor drug-loading polymer micelles， and to increase inhibitive effect of insoluble anti-tumor drug. METHODS： Chitosan （CSO） and stearic acid （SA） were used to prepare blank micelles （CSO-SA）， then modified with mPEG and folic acid （FA） to prepare PEG-CSO-SA and FA-PEG-CSO-SA. Characteristic functional groups of CSO-SA， PEG-CSO-SA and FA-PEG-CSO-SA were detected by infrared spectroscopy. The morphology of micelles was observed by transmission electron microscopy. The particle size and Zeta potential of micelles were measured by laser particle size analyzer. Osthole （OST） was used as the model drug and drug-loading micelles （FA-PEG-CSO-SA/OST） were prepared by dialysis. MTT assay was used to detect the inhibitory rate of FA-PEG-CSO-SA， OST solution and FA-PEG-CSO-SA/OST to human liver cancer cell HepG2. Half inhibitory concentration （IC50） was calculated. RESULTS： FA-PEG-CSO-SA was successfully prepared. CSO-SA， PEG-CSO-SA， FA-PEG-CSO-SA were oval in shape； particle sizes were （96.01±5.99）， （112.93±1.06）， （216.01±4.76） nm （n＝3） and Zeta potentials were （39.30±1.75）， （38.03±2.91）， （15.17±2.10） mV （n＝3）， respectively. Encapsulation efficiency and drug-loading amount of OST in FA-PEG-CSO-SA were （84.47±2.07）％ and （16.01±0.90）％ （n＝3）， respectively. The inhibition rates of FA-PEG-CSO-SA to HepG2 cells were＜20％. IC50 of OST solution and FA-PEG-CSO-SA/OST to HepG2 cells were （62.08±5.21）， （27.49±0.50） μg/mL （n＝3）， respectively. CONCLUSIONS： Prepared FA-PEG-CSO-SA can significantly increase inhibitive effect of insoluble drug OST to HepG2 cells， and it is expected to become a new anti-tumor drug carrier.

Effects of Osthol Isolated from Cnidium monnieri Fruit on Urate Transporter 1.

(1) Background: Crude drugs used in traditional Japanese Kampo medicine or folk medicine are major sources of new chemical entities for drug discovery. We screened the inhibitory potential of these crude drugs against urate transporter 1 (URAT1) to discover new drugs for hyperuricemia. (2) Methods: We prepared the MeOH extracts of 107 different crude drugs, and screened their inhibitory effects on URAT1 by measuring the uptake of uric acid by HEK293/PDZK1 cells transiently transfected with URAT1. (3) Results: We found that the extract of the dried mature fruit of Cnidium monnieri inhibited urate uptake via URAT1. We isolated and identified osthol as the active ingredient from this extract. Osthol noncompetitively inhibited URAT1 with an IC50 of 78.8 µM. We evaluated the effects of other coumarins and found that the prenyl group, which binds at the 8-position of coumarins, plays an important role in the inhibition of URAT1. (4) Conclusions: Cnidium monnieri fruit may be useful for the treatment of hyperuricemia or gout in traditional medicine, and its active ingredient, osthol, is expected to be a leading compound for the development of new drugs for hyperuricemia.

Potential Anticancer Properties of Osthol: A Comprehensive Mechanistic Review.

Cancer is caused by uncontrolled cell proliferation which has the potential to occur in different tissues and spread into surrounding and distant tissues. Despite the current advances in the field of anticancer agents, rapidly developing resistance against different chemotherapeutic drugs and significantly higher off-target effects cause millions of deaths every year. Osthol is a natural coumarin isolated from Apiaceaous plants which has demonstrated several pharmacological effects, such as antineoplastic, anti-inflammatory and antioxidant properties. We have attempted to summarize up-to-date information related to pharmacological effects and molecular mechanisms of osthol as a lead compound in managing malignancies. Electronic databases, including PubMed, Cochrane library, ScienceDirect and Scopus were searched for in vitro, in vivo and clinical studies on anticancer effects of osthol. Osthol exerts remarkable anticancer properties by suppressing cancer cell growth and induction of apoptosis. Osthol's protective and therapeutic effects have been observed in different cancers, including ovarian, cervical, colon and prostate cancers as well as chronic myeloid leukemia, lung adenocarcinoma, glioma, hepatocellular, glioblastoma, renal and invasive mammary carcinoma. A large body of evidence demonstrates that osthol regulates apoptosis, proliferation and invasion in different types of malignant cells which are mediated by multiple signal transduction cascades. In this review, we set spotlights on various pathways which are targeted by osthol in different cancers to inhibit cancer development and progression.

Antifungal activity of osthol in vitro and enhancement in vivo through Eudragit S100 nanocarriers.

In vitro interaction of osthol (Ost) and fluconazole (FLC) was investigated against 11 fluconazole-resistant clinical isolates of Candida albicans. Synergistic activities were determined using the checkerboard microdilution assay. The results of agar diffusion test confirmed the synergistic interaction. We used an enteric material Eudragit S100 for preparation of Ost nanoparticle (Ost-NP) to improve the oral bioavailability, biological activity of Ost. The physicochemical characteristics of Ost-S100-NP revealed Ost-S100-NP with mean particle size of 55.4±0.4 nm, encapsulation efficiency of 98.95±0.06%, drug loading efficiency of 23.89±0.25%, yield of 98.5±0.1% and a polydispersity index (PDI) of 0.165. As the Ost concentration-time curve showed, Ost-S100-NP can increase the plasma concentration and relative bioavailability of Ost compared with Ost-suspension by oral administration. In vivo, Ost-S100-NP enhanced the therapeutic efficacy of Ost against FLC-resistant C. albicans in immunosuppressed candidiasis mice model. The available information strongly suggests that Ost-S100-NP may be used as a promising compound against drug-resistant fungi.

Isolation and structural characterization of chemical constituents from fruits of Physalis pubescens / 中草药

Objective To study the chemical constituents from the fresh fruits of Physalis pubescens. Methods Compounds were isolated and purified by repeated column chromatography on silica gel column, Sephadex LH-20 gel column, and preparative HPLC methods. Physicochemical properties and spectroscopic methods were employed for the structure elucidation. Results Twenty compounds were obtained and identified as osthol (1), xanthotoxin (2), isopimpinellin (3), imperatorin (4), (-)-meranzin hydrate (5), auraptenol (6), 1-(4-hydroxy-3,5-dimethoxyphenyl) ethanone (7), 2,5-dimethoxybenzoquinone (8), 2-(4-hydroxyphenyl) acetic acid (9), (S)-methyl 2-hydroxy-2-(4-hydroxyphenyl) acetate (10), pyrocatechol 1-O-β-D-glucopyranoside (11), benzyl β-D-glucopyranosyl (1→6)-β-D-glucopyranoside (12), 2-phenylethyl-O-β-D-glucopyranoside (13), p-hydroxybenzene propanoic acid (14), 3,4-dihydroxybenzene propionic acid (15), (1-O-p-coumaroyl)-(6-O-β-glucosyl)-β-glucoside (16), 1-O-trans-cinnamoyl-β-D- glucopyranosyl-(1→6)-β-D-glucopyranoside (17), N-trans-feruloyltyramine (18), kaempferol 3-O-β-D-glucopyranoside (19), and bergapten (20), respectively. Conclusion Compounds 1-16 are obtained from Physalis genus for the first time.

Chemical constituents of Gelsemium elegans / 中草药

Objective: The chemical constituents were isolated and identified in order to find the bioactive natural products from Gelsemium elegans. Methods: Silica gel, sephadex LH-20 and HPLC column chromatographic techniques were used for separation and purification of the compounds and extensive spectral analysis spectrum were employed for structural elucidation. Results: Fifteen compounds were separated from G. elegans, identified by physicochemical properties, spectral analysis and other means. They are koumine (1), gelsemine (2), gelsevirine (3), gelsemicine (4), xanthotoxin (5), bergapten (6), isopimpinellin (7), imperatorin (8), osthol (9), p-hydroxybenzoic acid (10), vanilla acid (11), β-sitosterol (12), β-daucosterol (13), gelsemamide (14), and 16-epivoacarpine (15). Conclusion: Compounds 5-9 are first isolated from the plants in genus Gelsemium Juss.

Antileishmanial activity of prenylated coumarins isolated from Ferulago angulata and Prangos asperula.

Leishmaniasis has a wide spectrum of signs and symptoms due to infection to numbers of Leishmania species and makes enormous mortality and morbidity. There are clues of antileishmanial effects of prenylated coumarins. Apiaceae family is one of the most important sources of coumarins. Air-dried aerial parts of Ferulago angulata and fruits of Prangos asperula were extracted with n-hexane, using a soxhlet apparatus. The solvents were evaporated under reduced pressure. Column chromatography and crystallization process resulted to isolation of three prenylated coumarins. (1)H-nuclear magnetic resonance, electron ionization Mass and Infrared spectra were used for elucidation of isolated compounds. Leishmanicidal activity of isolated coumarins was assessed on Leishmania major strain (MRHO/IR/75/ER) for the first time. Suberosin epoxide and suberosin were isolated from aerial parts of F. angulata and osthol was extracted from grounded fruits of P. asperula. Osthol showed a significant antileishmanial effect on promastigotes in early hours of exposure with IC50 of 14.40 µg/mL but suberosin epoxide showed only a weak antileishmanial activity. IC50 of osthol and suberosin epoxide after 48 h were 10.79 and 54.0 µg/mL, respectively. Suberosin showed no remarkable effect in these concentrations. This is the first report on the pharmacological activity of suberosin epoxide. Substantial difference between efficacies of two isomers, osthol and suberosin remarks the importance of prenyl substituent location on C-8.

Osthol attenuates neutrophilic oxidative stress and hemorrhagic shock-induced lung injury via inhibition of phosphodiesterase 4.

Oxidative stress caused by neutrophils is an important pathogenic factor in trauma/hemorrhagic (T/H)-induced acute lung injury (ALI). Osthol, a natural coumarin found in traditional medicinal plants, has therapeutic potential in various diseases. However, the pharmacological effects of osthol in human neutrophils and its molecular mechanism of action remain elusive. In this study, our data showed that osthol potently inhibited the production of superoxide anion (O2(•-)) and reactive oxidants derived therefrom as well as expression of CD11b in N-formylmethionylleucylphenylalanine (FMLP)-activated human neutrophils. However, osthol inhibited neutrophil degranulation only slightly and it failed to inhibit the activity of subcellular NADPH oxidase. FMLP-induced phosphorylation of extracellular signal-regulated kinase (ERK) and protein kinase B (Akt) was inhibited by osthol. Notably, osthol increased the cAMP concentration and protein kinase A (PKA) activity in activated neutrophils. PKA inhibitors reversed the inhibitory effects of osthol, suggesting that these are mediated through cAMP/PKA-dependent inhibition of ERK and Akt activation. Furthermore, the activity of cAMP-specific phosphodiesterase (PDE) 4, but not PDE3 or PDE7, was significantly reduced by osthol. In addition, osthol reduced myeloperoxidase activity and pulmonary edema in rats subjected to T/H shock. In conclusion, our data suggest that osthol has effective anti-inflammatory activity in human neutrophils through the suppression of PDE4 and protects significantly against T/H shock-induced ALI in rats. Osthol may have potential for future clinical application as a novel adjunct therapy to treat lung inflammation caused by adverse circulatory conditions.

Optimization of Inclusion Compound for Osthol with Hydroxypropyl-β-Cyclodextrin by Orthogonal and Rotational Combination Design / 中国中医药信息杂志

Objective To study the process of hydroxypropyl-β-cyclodextrin (HP-β-CD) inclusion techniques for osthol. Methods The inclusion complex of osthol and HP-β-CD was prepared by unsaturated water solution and freeze-drying technique. Inclusion techniques were selected by screening on quadratic orthogonal rotation combination design method, and the entrapment efficiency was identified by HPLC. Results The optimal technical conditions were as follows:the ratio of HP-β-CD and drug was 4.5∶1;temperature for electric mixer was 35 ℃;the stirring time for thermostatic waterbath was 210 min. Conclusion This method is reasonable and it may have a prosperous future of development and application.

Preparation of Osthol Loaded Solid Lipid Nanoparticles / 医药导报

Objective To study the main factors affecting the preparation of osthol ( Ost ) loaded solid lipid nanoparticies ( SLN ) . Methods The SLN were prepared by melt-homogenization method. The optimum formulation and process were selected by orthogonai design. The shape, particle size, loading capacity were investigated. Results The obtained Ost-loaded SLN were sphere or oval at a range of 100-200 nm, and were well distributed without adhesion, the loading capacity was 59. 78%. Conclusion The melt-homogenization method is available for the preparation of Ost loaded SLN.