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Introduction Nimbolide (Nim), a limonoid obtained from the neem tree, Azadirachta indica, has several pharmacological properties, including anticancer effects in different type of cancers. No drug-delivery system has been reported for enhancing the therapeutic application of this novel hydrophobic molecule. Methods In the present research, poly(lactic-co-glycolic acid) (PLGA) nanoparticles of Nim (Nim-nano) were formulated by nanoprecipitation, characterized for physicochemical properties, and screened for anticancer potential in breast (MCF-7 and MDA-MB-231) and pancreatic (AsPC-1) cancer cell lines. Results The Nim-nano had a particle size of 183.73±2.22 nm and 221.20±11.03 nm before and after lyophilization, respectively. Cryoprotectants (mannitol and sucrose) significantly inhibited growth in particle size due to lyophilization. The ζ-potential of the Nim-nano was -22.40±4.40 mV. Drug loading and encapsulation efficiency of Nim-nano were 5.25%±1.12% and 55.67%±12.42%, respectively. The Nim-nano exhibited sustained release of Nim for more than 6 days in PBS (pH 7.4) and showed two- to three-fold enhanced cytotoxicity in breast and pancreatic cancer cell lines compared with free Nim. Conclusion The Nim-nano formulation has great potential for treatment of cancers, such as pancreatic and breast cancer. Further, the PLGA-polymer surface can be modified by conjugation with polyethylene glycol, receptor-binding ligands (eg, folic acid), and other that which may lead to targeted delivery of Nim in the treatment of cancer.

Pueraria tuberosa(P. tuberosa), known as Indian Kudzu belongs to family Fabaceae and it is solicited as “Rasayana” drugs in Ayurveda. In the present study, we analyzed the efficacy an antioxidant enriched fraction (AEF) from the tuber extract ofP. tuberosaagainst menopausal osteoporosis and breast and ovarian cancer cell lines. The AEF fromP. tuberosawas identified by determining phenolic composition (total phenolic and flavonoid amount). Antioxidant property (in vitroassays) was also carried out followed by analysis of the AEF for its antiosteoporotic and anticancer potentials. Antiosteoporotic activity of AEF was investigated in ovariectomy-induced osteoporosis in rats andin vitroanticancer activity by MTT assay. Also, the GC/MS analysis of AEF was performed to determine various phytoconstituents. A docking analysis was performed to verify the interaction of bioactive molecules with estrogen receptors (ERs). Ethyl acetate fraction of the mother extract was proved as the AEF. AEF significantly improved various biomechanical and biochemical parameters in a dose dependent manner in the ovariectomized animals. AEF also controlled the increased body weight and decreased uterus weight following ovariectomy. Histopathology of femur revealed the restoration of typical bone structure and trabecular width in ovariectomized animals after AEF and raloxifene treatment. AEF also exhibitedin vitrocytotoxicity in breast (MCF-7 and MDA-MB-231) and ovarian (SKOV-3) cancer cells. Further, genistein and daidzein exhibited a high affinity towards both estrogen receptors (α and β) in docking study revealing the probable mechanism of the antiosteoporotic activity. GC/MS analysis confirmed the presence of bioactive molecules such as stigmasterol, β-sitosterol, and stigmasta-3,5-dien-7-one. The observations of this study vindicate the potency of AEF fromP. tuberosain the treatment of menopausal osteoporosis and cancer.

The present study was undertaken to develop and validate a novel high-performance thin-layer chromatography (HPTLC) method for the quantification of apigenin and luteolin in Hygrophila spinosa. Separation was carried out on HPTLC plates using the mobile phase toluene—ethyl acetate—formic acid (6.0:4.0:1.0, v/v), and detection was achieved at 349 nm. The method was validated by the International Conference on Harmonization (ICH) guidelines. The calibration range was 80–560 ng per band (R2 = 0.997) for apigenin and 40–280 ng per band (R2 = 0.998) for luteolin. The method was accurate in triplicate results at different standard addition levels with average recoveries of 99.94 and 100.01% for apigenin and luteolin, respectively. The limits of detection and quantification were 6.25 and 18.95 ng, respectively, for apigenin, and 2.36 and 7.55 ng, respectively, for luteolin. The quantity of apigenin was 8.84 and 15.67 mg g−1 in the alcoholic extract and its ethyl acetate fraction, respectively. The luteolin contents in the alcoholic extract and ethyl acetate fraction were 0.172 and 0.464 mg g−1, respectively. Further, luteolin was found to be present only in the flower (6.87 mg g−1) extract, but apigenin was detected in the husk (2.38 mg g−1), flower (7.97 mg g−1), spine (1.01 mg g−1), fruits (0.60 mg g−1), and leaf (4.28 mg g−1). The developed method could be used for the quality control analysis and quantification of apigenin and luteolin in herbal preparations containing H. spinosa.

In the current study, we have explored an antioxidant enriched fraction (AEF) from whole plant of H. spinosa for management of menopausal complications like osteoporosis and cancer. The AEF was identified by determining the total phenolic content, total flavonoid content and antioxidant potential (DPPH method and total antioxidant capacity assay). Ethyl acetate fraction of the crude ethanol extract comprised of the highest antioxidant potential and considered as AEF. HPTLC analysis of AEF confirmed the presence of apigenin and luteolin, which were further investigated for their estrogen receptors modulator effect by docking study. GC/MS analysis also demonstrated the presence of β-sitosterol, and lupeol in AEF. Antiosteoporotic activity was studied by ovariectomized animal model and in vitro anticancer potential by MTT assay. AEF significantly improved biomechanical parameters, body and uterine weight, phosphorous and calcium level in serum and urine. AEF also restored bone turnover markers like alkaline phosphatase, tartrate resistant acid phosphatase and hydroxyproline along with serum cholesterol and triglycerides (biochemical parameters). Histopathology showed restoration of normal bone architecture after AEF and raloxifene treatment. Docking study revealed the high affinity of apigenin and luteolin towards both estrogen receptors (α and β). AEF exerted dose dependent anticancer potential against breast and ovarian cancer cell lines with lowest IC50 (43 μg/ml) against SKOV-3 cell line. Our findings conclude that AEF from H. spinosa could be used as an alternative and economical botanical drug for treatment of menopausal osteoporosis and cancer.

Background Quercetin (QCT), a naturally occurring flavonoid has a wide array of pharmacological properties such as anticancer, antioxidant and anti-inflammatory activities. QCT has low solubility in water and poor bioavailability, which limited its use as a therapeutic molecule. Polymeric micelles (PMs) is a novel drug delivery system having characteristics like smaller particle size, higher drug loading, sustained drug release, high stability, increased cellular uptake and improved therapeutic potential. In the present study, we have formulated and characterized mixed PMs (MPMs) containing QCT for increasing its anticancer potential. Methods The MPMs were prepared by thin film hydration method, and their physicochemical properties were characterized. The in vitro anticancer activity of the MPMs were tested in breast (MCF-7 and MDA-MB-231, epithelial and metastatic cancer cell lines, respectively), and ovarian (SKOV-3 and NCI/ADR, epithelial and multi-drug resistant cell lines, respectively) cancer. Results The optimal MPM formulations were obtained from Pluronic polymers, P123 and P407 with molar ratio of 7:3 (A16); and P123, P407 and TPGS in the molar ratio of 7:2:1 (A22). The size of the particles before lyophilization (24.83±0.44 nm) and after lyophilisation (37.10±4.23 nm), drug loading (8.75±0.41%), and encapsulation efficiency (87.48±4.15%) for formulation A16 were determined. For formulation A22, the particle size before lyophilization, after lyophilization, drug loading and encapsulation efficiency were 26.37±2.19 nm, 45.88±13.80 nm, 9.01±0.11% and 90.07±1.09%, respectively. The MPMs exhibited sustained release of QCT compared to free QCT as demonstrated from in vitro release experiments. The solubility of QCT was markedly improved compared to pure QCT. The MPMs were highly stable in aqueous media as demonstrated by their low critical micelle concentration. The concentration which inhibited 50% growth (IC50) values of both micellar preparations in all the cancer cell lines were significantly less compared to free QCT. Conclusion Both the MPMs containing QCT could be used for effective delivery to different type of cancer and may be considered for further development., Video abstract

The present study was performed to estimate the concentration of genistein and daidzein in ethanol extract of tubers of Pueraria tuberosa (Indian kudzu or Vidarikanda) and its various fractions (n-hexane, ethyl acetate, n-butanol, and aqueous) by high-performance thin-layer chromatography (HPTLC). The separation of bioactive compounds was performed using mobile phase, toluene:ethyl acetate:acetone:formic acid (20.0:4.0:2.0:1.0) and detected at wavelength 269 nm. The method was validated for linearity, accuracy, precision, limit of detection (LOD), limit of quantification (LOQ), etc. by International Conference on Harmonization guidelines. The calibration range was found to be 100–600 ng/band for both the bioactive compounds. Daidzein was separated with an Rf value of 0.39 ± 0.02 and genistein with an Rf value of 0.54 ± 0.02. Average recovery was 99.96 and 99.90% for genistein and daidzein, respectively. The LOD and LOQ were 14.786 and 44.805 ng, respectively, for genistein, and 9.607 and 29.114 ng...

Plant extract-mediated green synthesis of gold nanoparticles (AuNPs) and their biomedical application is currently gaining huge interest in the field of nanotechnology. In the present study, AuNPs were synthesized using Hygrophila spinosa aqueous extract (HSAE). The formation of AuNPs was confirmed by observing color change and characteristic absorbance peak for surface plasmon resonance by UV–Vis spectroscopy. Different reaction parameters such as HSAE concentration, salt concentration, pH, reaction time and temperature were optimized for biosynthesis of AuNPs. The NPs were characterized for particle size, surface morphology, crystallinity, elemental composition and surface functionalities. Cytotoxicity of the AuNPs was studied against different cancer cell lines by MTT assay and antioxidant potential as total antioxidant capacity was also evaluated. The biosynthesized AuNPs were small in size and negatively charged. Phytochemical screening of HSAE, and FTIR spectrum of synthesized AuNPs demonstrated participation of various groups of chemical compounds as capping and stabilizing agents in biosynthesis of AuNPs. The green synthesized AuNPs exhibited significantly higher anticancer activity compared to HSAE against breast, ovarian, glioblastoma/brain and multi-drug resistant ovarian cancer cell lines. Antioxidant activity of the synthesized AuNPs was less than HSAE. The biosynthesized AuNPs could be studied further to access their potency in the treatment and imaging of various cancers.

In the present study, we have synthesized silver nanoparticles (AgNPs) using Pueraria tuberosa aqueous extract (PTAE) in a straightforward manner without involvement of toxic chemicals. Various reaction parameters (reaction time, AgNO

The purpose of the present study is to investigate the pharmacognostical characteristics of the roots of Saccharum officinarum Linn. (poaceae) to establish standardized parameters. Macroscopic, microscopic, and physicochemical evaluation; preliminary phytochemical analyses and high performance thin layer chromatography (HPTLC) profile were done for qualitative phytochemical evaluation. Roots are grey to blackish brown, fibrous, solid, jointed, odour and taste not distinct. Transverse section of both thin and fairly thick root showed presence of epidermal, hypodermal layers, aerenchymatous cortex and a wide stele. Powder microscopy revealed presence of long thick walled fibers with pointed ends, vessels and tracheids with reticulate, scalariform and bordered pitted walls and thin walled parenchyma cells. Physicochemical parameters viz. percentage of moisture content (9.3% w/w), total ash (7.5% w/w), acid insoluble ash (5.0% w/w), water soluble extractive (6.9% w/w) and ethanol soluble extractive (3.8% w/w) were also determined. Various groups of phytoconstituents found in the plant are carbohydrate, steroids, flavonoids, saponins, tannins and phenolic compound. HPTLC profile of hydroalcoholic extract showed presence of twelve different components in the solvent system, chloroform: methanol (9:1, v/v). The different salient diagnostic features established in this study will help for proper identification and standardization of the drug in crude form, and then the plant material can be used for further studies.

Postmenopausal osteoporosis, a silent epidemic, has become a major health hazard, afflicting about 50% of postmenopausal women worldwide and is thought to be a disease with one of the highest incidences in senile people. It is a chronic, progressive condition associated with micro-architectural deterioration of bone tissue that results in low bone mass, decreased bone strength that predisposes to an increased risk of fracture. Women are more likely to develop osteoporosis than men due to reduction in estrogen during menopause which leads to decline in bone formation and increase in bone resorption activity. Estrogen is able to suppress the production of proinflammatory cytokines like interleukin (IL)-1, IL-6, IL-7 and tumor necrosis factor (TNF-α). This is why these cytokines are elevated in postmenopausal women. In this review article we have made an attempt to collate the various methods and parameters most frequently used for screening of antiosteoporotic activity in postmenopausal osteoporosis. Pertaining to ovariectomized animal model, this is the most appropriate model for studying the efficacy of different drugs to prevent bone loss in postmenopausal osteoporosis.

The plant Polyalthia longifolia (Annonaceae) is an ornamental tree that finds its reference in Indian medicinal literature owing to its popular Hindi name Ashoka i.e, Saraca indica. However, P. longifolia is equated with the name Asoka and often used as an adulterant or substitute of the genuine Asoka bark. The present investigation was carried out with an object to separate and isolate active phytomolecule(s) from stem bark of P. longifolia and to screen their antibacterial and antioxidant potential. Column chromatography of the butanol fraction of the hydroalcoholic extract (methanol:water, 1:1) has led to the isolation of a phenolic compound. Structural elucidation was done by IR, 1H NMR, 13C NMR, DEPT, COSY, HSQC, HMBC and mass spectroscopy techniques, and purity was checked by HPTLC and HPLC. Butanol fraction and the isolated compound were screened for antibacterial activity (against facultative aerobic and fastidious aerobic bacterial strains) and antioxidant potential (DPPH method). The compound was revealed to be 3-O-methyl ellagic acid 4'-rhamnoside (1), and the purity of the compound was 99.2%. The isolated compound comprises promising antibacterial and antioxidant activities.

Hygrophila spinosa T. Anders (Acanthaceae) is traditionally used in Indian medicine for the treatment of microbial infections, liver diseases, cancer, inflammation, rheumatism, diabetes, pain, fever etc. The aim of the present study is to evaluate the anti-inflammatory activity of chloroform and alcoholic extracts of the leaves of H. spinosa in chronic inflammation models in rats as our previous study revealed that these two extracts had anti-inflammatory activity in carrageenan induced paw oedema model. Anti-inflammatory activity was evaluated by cotton pellet-induced granuloma and Freund’s adjuvant-induced arthritis in rats. Antioxidant activity of the extracts was revealed by their scavenging activity of the stable 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical and a flavonoid compound (apigenin) was also isolated and characterized from the alcoholic extract of the plant. Chloroform and alcoholic extracts showed anti-inflammatory activity both in cotton pellet-induced granuloma and Freund’s adjuvant-induced arthritis in a dose dependent manner. The decrease in body weight due to injection of CFA was improved significantly by the above two extracts also. Both the extracts also exhibited antioxidant activity. The results demonstrated that H. spinosa has anti-inflammatory activity in chronic models of inflammation which support the traditional use of H. spinosa in the treatment of rheumatism. Introduction Hygrophila spinosa T. Anders (Acanthaceae) commonly known as ‘Talmakhana’ in Hindi is found in water-logged areas throughout India, contains a number of phytoconstituents viz. lupeol (Shailajan and Abhishek, 2008), βsitosterol (Tiwari et al., 1967), stigmasterol (Khare, 2007), ascorbic acid, βcarotene (Sahoo and Acharya, 2005), nicotinic acid (Sharma et al., 2002), hentriacontane (Chopra et al., 1958), luteolin, luteolin-7-rutinoside (Balraj and Nagarajan, 1982), syringic acid, vannilic acid (Daniel, 2005) etc. In our earlier studies we have reported the other constituents, traditional uses, pharmacological activities of the plant (Patra et al., 2009a); various groups of phytoconstituents in petroleum ether, chloroform, alcoholic and aqueous extracts of the leaves of the plant (Patra et al., 2009b) and also few pharmacological studies of the above extracts (Patra et al., 2008, 2009c). The aim of this study was to determine the anti-inflammatory activity of chloroform and aqueous extracts of the leaves of H. spinosa in chronic inflammation models in rats. Materials and Methods Plant material H. spinosa plants were collected from Berhampur, Orissa, India and botanical identification was done through Department of Pharmaceutical Sciences, Birla Institute of Technology, Mesra, Ranchi (Voucher no. BITPcog. 463/07-08). Voucher specimen was preserved in the department for further verification. Preparation of different extracts The leaves were washed thoroughly, dried under shade and pulverized. The coarse powder was extracted successively with petroleum ether, chloroform and alcohol using soxhlet apparatus. The extracts were dried using a rotary vacuum evaporator and stored in a desiccator until further use. Anti-inflammatory activity Anti-inflammatory activity was evaluated by cotton pellet-induced granuloma and Freund’s adjuvant-induced arthritis in rats. Antioxidant activity Antioxidant activity of the extracts was revealed by their scavenging activity of the stable 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical. Isolation of compound 1 from alcoholic extract 50 gm of alcoholic extract was column chromatographed on a silica gel column (mesh 60-120) and eluted with solvent mixtures of increasing polarity: chloroform (300 ml), chloroform-acetone [(80:20, 300 ml), (60:40, 300 ml), (40:60, 300 ml), (20:80, 300 ml)], acetone (300 ml), acetonemethanol [(80:20, 300 ml), (60:40, 300 ml), (40:60, 300 ml), (20:80, 300 ml)] and methanol (300 ml). Fractions (20ml) were collected and monitored on TLC. Fractions (151 to 165) collected were pulled together as these fractions showed a single spot of same Rf value in TLC. It was evaporated in a water bath (70-800C) to afford a solid residue. The residue was dissolved in EtOH with little warming on a water bath. It was left undisturbed in refrigerator when crystals of compound 1 was obtained (yield: 0.0015% w/w). Characterization of compound 1 The various instruments used for recording the data for compound 1 are: FTIR spectroscopy (Shimadzu, IRPrestige-21), elemental analysis (Elementar, Vario EL III), 1H and 13C NMR spectra (PABBO BB NMR spectrophotometer), mass spectroscopy etc. Other physicochemical characters as melting point, solubility and physical appearance were also recorded. Statistical analysis The results were expressed as mean ± standard error mean (SEM). Statistical analysis of the data was carried out using one way analysis of variance (ANOVA) followed by Student’s t-test to determine the significant difference between the control and the treated groups. P < 0.05 was considered significant. Results Fig. 1. Effect of different extracts of H. spinosa leaf on body weight of animals in CFA induced arthritic rats. HSCH, chloroform extract; HSAL, alcoholic extract Treatment Dose (mg/kg, p.o.) Weight of dry granuloma (mg) Percentage inhibition of granuloma Vehicle (Control) 122.36 ± 2.32 Indomethacin 10 65.42 ± 1.52* 46.53* Chloroform 200 109.65 ± 1.05* 10.38* 400 92. 88 ± 0.98* 24.09* Alcoholic 200 102.28 ± 1.06* 16.41* 400 78.68 ± 0.86* 35.69* Table 1. Effect of different extracts of H. spinosa leaf on cotton pellet-induced granuloma