No abstract

Amphiphilic macrocycles, such as p -sulfonatocalix[6]arenes ( p -SC6), have demonstrated great potential in designing synthetic nanovesicles based on self-assembly approaches. These supramolecular nanovesicles are capable of improving the solubility, stability, and biological activity of various drugs. In the present study, the biologically active harmala alkaloid-rich fraction (HARF) was extracted from Peganum harmala L . seeds. Ultraperformance liquid chromatography–electrospray ionization–tandem mass spectrometry (UPLC/ESI-MS) analysis of HARF revealed 15 alkaloids. The reversed-phase high-performance liquid chromatography (RP-HPLC) analysis revealed three peaks: peganine, harmol, and harmine. The HARF was then encapsulated in p -SC6 nanocapsules employing a thin-film hydration approach. The designed nanocapsules had an average particle size of 264.8 ± 10.6 nm, and a surface charge of −30.3 ± 2.2 mV. They were able to encapsulate 89.3 ± 1.4, 74.4 ± 1.3, and 76.1 ± 1.7% of the three harmala alkaloids; harmine, harmol, and peganine; respectively. The in vitro drug release experiments showed the potential of the designed nanocapsules to release their cargo at a pH of 5.5 (typical of cancerous tissue). The IC 50 values of HARF encapsulated in p -SC6 (H/ p -SC6 nanocapsules) were 5 and 2.7 μg/mL against ovarian cancer cells (SKOV-3) and breast adenocarcinoma cells (MCF-7), respectively. The prepared nanocapsules were found to be biocompatible when tested on human skin fibroblasts. Additionally, the antioxidant activity of the designed nanocapsules was 5 times that of the free powder fraction; the IC 50 of the H/ p -SC6 nanocapsules was 30.1 ± 1.3 μg/mL, and that of the HARF was 169.3 ± 7.2 μg/mL. In conclusion, encapsulation of P. harmala alkaloid-rich fraction into self-assembled p -SC6 significantly increases its antioxidant and cytotoxic activities.

Wound healing is a major healthcare concern, and complicated wounds may lead to severe outcomes such as septicemia and amputations. To date, management choices are limited, which warrants the search for new potent wound healing agents. Natural products loaded in poly (lactic-co-glycolic acid) (PLGA) coated with chitosan (CS) constitute a promising antibacterial wound healing formulation. In this work, harmala alkaloid-rich fraction (HARF) loaded into PLGA nanoparticles coated with chitosan (H/CS/PLGA NPs) were designed using the emulsion-solvent evaporation method. Optimization of the formulation variables (HARF: PLGA and CS: PLGA weight ratios, sonication time) was performed using the 33 Box–Behnken design (BBD). The optimal NPs were characterized using transmission electron microscopy (TEM) and Attenuated Total Reflection Fourier-Transformed Infrared Spectroscopy (ATR-FTIR). The prepared NPs had an average particle size of 202.27 ± 2.44 nm, a PDI of 0.23 ± 0.01, a zeta potential of 9.22 ± 0.94 mV, and an entrapment efficiency of 86.77 ± 4.18%. In vitro drug release experiments showed a biphasic pattern where an initial burst of 82.50 ± 0.20% took place in the first 2 h, which increased to 87.50 ± 0.50% over 72 h. The designed optimal H/CS/PLGA NPs exerted high antibacterial activity against Staphylococcus aureus and Escherichia coli (MIC of 0.125 and 0.06 mg/mL, respectively) compared to unloaded HARF (MIC of 0.50 mg/mL). The prepared nanoparticles were found to be biocompatible when tested on human skin fibroblasts. Moreover, the wound closure percentage after 24 h of applying H/CS/PLGA NPs was found to be 94.4 ± 8.0%, compared to free HARF and blank NPs (68.20 ± 5.10 and 50.50 ± 9.40%, respectively). In conclusion, the three components of the developed nanoformulation (PLGA, chitosan, and HARF) have synergistic antibacterial and wound healing properties for the management of infected wounds.

Rhei Rhizoma (rhubarb), called Daio in Japanese, is one of the important herbal drugs. To date, qualitative analysis of its typical laxative components, anthraquinone derivatives including sennoside A, has been used for the qualitative evaluation of rhubarb by means of several analytical methods. 1)However, such evaluation is incomplete because rhubarb has been used for the treatment of "Oketsu" (various syndromes caused by the obstruction of blood circulation such as dysmenorrhoea, hypermenorrhea, hematemesis, lower abdominal pain, etc.), jaundice, diarrhea and food poisoning, in addition to constipation. Although various individual pharmacological effects related to the above treatments, such as purgative activity, 2,3) anti-bacterial and anti-fungal activities, 4) anti-tumor activity, 5) anti-inflammatory and analgesic activities, 6,7) improvement of renal disorders, [8][9][10][11] improvement of nitrogen metabolism, 12,13) psychotropic activity, 14) anti-allergic effects, 15,16) anti-cholera toxin activity, 17,18) promoting blood circulation and removing blood stasis, 19) and the various involved active compounds have been reported, comprehensive chemical study of the composition of bioactive constituents of rhubarb has been rare. Kashiwada et al. 20) set up a HPLC method to analyze almost all the phenolic compounds simultaneously, and reported that the majority of the compounds could be separated by a 0.05 M H 3 PO 4 solution-acetonitrile gradient elution system. However, several compounds, i.e. procyanidin B-1 3-Ogallate and 1,2,6-trishowed overlapping peaks. In addition, polymeric procyanidins, i.e. RG-tannin and rhatannin, showed serious band broadening and tailing. Therefore, it is still necessary to develop a HPLC method which can be used for the effective separation and quantitative determination of the active components, which would then be used to evaluate the quality of rhubarb samples.In this study, we developed new HPLC methods to analyze 30 compounds (1-30, Fig. 1) in rhubarb, quantitatively, and the contents of the active components were compared in rhubarb samples of different botanical origins. ExperimentalMaterials Three Rhei Rhizoma samples derived from the following species were quantitatively examined: rhizomes of R. tanguticum (Huangnan County, Qinghai Prov., TMPW no. 20065), R. palmatum (Jiulong County, Sichuan Prov., TMPW no. 20216) and R. officinale (Wanyuan County, Sichuan Prov., TMPW no. 20267). The botanical origins of each sample were correctly identified by the molecular biological methods previously reported. 21,22) The rhubarb sample (Qinghai Prov., TMPW no. 19929) used for the isolation of standard compounds was purchased from Uchida Wakanyaku Co., Ltd. (Japan). Voucher specimens have been deposited in the Museum of Materia Medica, Institute of Natural Medicine, University of Toyama (TMPW).Chemicals and Reagents Sephadex LH-20 (Amersham Biosciences, Sweden) and reversed phase gel MCI CHP-20P (70-150 mesh, Mitsubishi Chemical Co., Japan) were used for column chromatography. A...

Ginseng drugs, the roots and/or rhizomes of Panax spp. (Araliaceae), are a group of the most important herbal medicines in the Orient. More than 10 Panax taxa reported worldwide are available as medicinal resources in traditional Chinese medicine as well as in folk medicine, such as Ginseng, American Ginseng, Notoginseng, Chikusetsu-ninjin (Japanese Ginseng), Vietnamese Ginseng, etc. The main bioactive constituents of Ginseng drugs are considered to be triterpene saponins, generally referred to as ginsenosides. To date, more than 80 ginsenosides have been isolated from these drugs and most of them possess four types of aglycones, i.e., protopanaxadiol, protopanaxatriol, ocotillol and oleanolic acid types. [1][2][3] Pharmacologic studies showed that the bioactivities of ginsenosides vary depending on the type of aglycone and the sugar moiety. [4][5][6] Phytochemical studies revealed that the composition and relative abundance of various ginsenosides in these drugs are different. [7][8][9] Comparative studies of the protopanaxadiol type [ginsenoside Rb 1 (1), Rb 2 , Rc (2), and Rd (3)] and protopanaxatriol type [ginsenoside Re (5), Rg 1 (6), and Rf] in Ginseng, American Ginseng and Notoginseng have been reported. [10][11][12] However, a systematic comparison of the four-type ginsenosides, including the ocotillol and oleanolic acid types, has not been investigated so far. Recently, majonoside R 2 (8), a typical ocotillol type saponin, has attracted much attention for its high medicinal value as an antistress and antitumor active agent. 13,14) The oleanolic acid saponins, ginsenoside Ro (9) and chikusetsusaponin IV (10) have also been reported to have an antiinflammatory effect and to protect against stress ulcers. 15)Therefore a convenient and efficient method that allows simultaneous determination of the four-type ginsenosides is desirable for chemical and biological evaluation of different Ginseng drugs.In the present study, 11 ginsenosides (1-11) (Fig. 1) representing the four types of saponins were chosen as standards based on extensive existence of relatively high concentration in different Ginseng drugs, which also have biological potential and phytotaxonomic significance. Here, we report the investigation and validation of a convenient HPLC method for the simultaneous determination of 11 ginsenosides with fourtype aglycones and its utilization in the quantitative analysis of three Ginseng drugs with completely different compositions of ginsenosides, 7,8,16) which are Ginseng, Ye-Sanchi, and Satsuma-ninjin derived from Panax ginseng, Panax vietnamensis var. fuscidiscus, and Panax japonicus (Japan), respectively. A HPLC method for the simultaneous determination of 11 triterpene saponins with four-type aglycones (protopanaxadiol, protopanaxatriol, ocotillol and oleanolic acid types) in Ginseng drugs was developed and validated. Using a gradient of acetonitrile and 10 mM K-phosphate buffer (pH 5.80) as the mobile phase and UV detection at 196 nm, more than 18 ginsenosides with different aglycones were separated ...