In Vitro Effect on Plasmodium falciparum and In Vivo Effect on Plasmodium berghei of Annomaal, an Oily Fraction Obtained from the Seeds of Annona squamosa.

Malaria remains a life-threatening health problem and is responsible for the high rates of mortality and morbidity in the tropical and subtropical regions of the world. The increasing threat of drug resistance to available artemisinin-based therapy warrants an urgent need to develop new antimalarial drugs that are safer, more effective, and have a novel mode of action. Natural plants are an excellent source of inspiration in searching for a new antimalarial agent. This research reports a systematic investigation for determining the antimalarial potential of the seeds of A. squamosa. The study shows that the crude seed extract (CSE), protein, saponin, and the oily fractions of the seeds were nontoxic at a 2000 mg/kg body weight dose when tested in Wistar rats, thus revealing high safety is classified as class 5. The oily fraction, Annomaal, demonstrated pronounced antimalarial activity with low IC50 (1.25 ± 0.183 µg/mL) against P. falciparum in vitro. The CSE and Annomaal significantly inhibited the growth of P. berghei parasites in vivo with 58.47% and 61.11% chemo suppression, respectively, while the standard drug artemether showed chemo suppression of 66.75%. Furthermore, the study demonstrated that oral administration of Annomaal at a daily dose of 250 mg/kg/day for 3 days was adequate to provide a complete cure to the P. berghei-infected mice. Annomaal thus holds promise as being patient-compliant due to the shorter treatment schedule, eliminating the need for frequent dosing for extended time periods as required by several synthetic antimalarial drugs. Further studies are needed to determine the active compounds in the oily fraction responsible for antimalarial activity.

Potential of Phytomolecules in Alliance with Nanotechnology to Surmount the Limitations of Current Treatment Options in the Management of Osteoarthritis.

Osteoarthritis (OA), a chronic degenerative musculoskeletal disorder, progressively increases with age. It is characterized by progressive loss of hyaline cartilage followed by subchondral bone remodeling and inflammaging. To counteract the inflammation, synovium releases various inflammatory and immune mediators along with metabolic intermediates, which further worsens the condition. However, even after recognizing the key molecular and cellular factors involved in the progression of OA, only disease-modifying therapies are available such as oral and topical NSAIDs, opioids, SNRIs, etc., providing symptomatic treatment and functional improvement instead of suppressing OA progression. Long-term use of these therapies leads to various life-threatening complications. Interestingly, mother nature has numerous medicinal plants containing active phytochemicals that can act on various targets involved in the development and progression of OA. Phytochemicals have been used for millennia in traditional medicine and are promising alternatives to conventional drugs with a lower rate of adverse events and efficiency frequently comparable to synthetic molecules. Nevertheless, their mechanism of action in many cases is elusive and uncertain. Even though many in vitro and in vivo studies show promising results, clinical evidence is scarce. Studies suggest that the presence of carbonyl group in the 2nd position, chloro in the 6th and an electron- withdrawing group at the 7th position exhibit enhanced COX-2 inhibition activity in OA. On the other hand, the presence of a double bond at the C2-C3 position of C ring in flavonoids plays an important role in Nrf2 activation. Moreover, with the advancements in the understanding of OA progression, SARs (structure-activity relationships) of phytochemicals and integration with nanotechnology have provided great opportunities for developing phytopharmaceuticals. Therefore, in the present review, we have discussed various promising phytomolecules, SAR as well as their nano-based delivery systems for the treatment of OA to motivate the future investigation of phytochemical-based drug therapy.

Recent advances in lipid-engineered multifunctional nanophytomedicines for cancer targeting.

Cancer is a leading cause of death in many countries around the world. However, the efficacy of current treatments available for variety of cancers is considered to be suboptimal due to the pathophysiological challenges associated with the disease which limits the efficacy of the anticancer drugs. Moreover, the vulnerability towards off-target effects and high toxicity also limits the use of drugs for the treatment of cancers. Besides, the biopharmaceutical challenges like poor water solubility and permeability of the drugs, along with the absence of active targeting capability further decreases the utility of drugs in cancer therapy. As a result of these deficiencies, the current therapeutic strategies face noncompliance to patients for providing meaningful benefits after administration. With the advancements in nanotechnology, there has been a paradigm shift in the modalities for cancer treatment with the help of phytomedicine-based nanosized drug delivery systems coupled with variegated surface-engineering strategies for targeted drug delivery. Among these delivery systems, lipid-based nanoparticles are considered as one of the highly biocompatible, efficient and effective systems extensively explored for anticancer drug delivery. These include diverse range of systems including liposomes, nanoemulsions, solid lipid nanoparticles, nanostructured lipidic carriers and supramolecular carriers, which alters pharmacokinetic and biodistribution of the drugs for active targeting to the desired site of action by overcoming the biopharmaceutical challenges associated with anticancer drug delivery. The present review endeavours to provide a comprehensive account on the recent advances in the application of lipid-based nanostructured systems for improving the pharmacotherapeutic performance of phytomedicines for cancer targeting application.

In vitro anti-HIV activity of some Indian medicinal plant extracts.

BACKGROUND: Human Immunodeficiency Virus (HIV) persists to be a significant public health issue worldwide. The current strategy for the treatment of HIV infection, Highly Active Antiretroviral Therapy (HAART), has reduced deaths from AIDS related disease, but it can be an expensive regime for the underdeveloped and developing countries where the supply of drugs is scarce and often not well tolerated, especially in persons undergoing long term treatment. The present therapy also has limitations of development of multidrug resistance, thus there is a need for the discovery of novel anti-HIV compounds from plants as a potential alternative in combating HIV disease. METHODS: Ten Indian medicinal plants were tested for entry and replication inhibition against laboratory adapted strains HIV-1IIIB, HIV-1Ada5 and primary isolates HIV-1UG070, HIV-1VB59 in TZM-bl cell lines and primary isolates HIV-1UG070, HIV-1VB59 in PM1 cell lines. The plant extracts were further evaluated for toxicity in HEC-1A epithelial cell lines by transwell epithelial model. RESULTS: The methanolic extracts of Achyranthes aspera, Rosa centifolia and aqueous extract of Ficus benghalensis inhibited laboratory adapted HIV-1 strains (IC80 3.6-118 µg/ml) and primary isolates (IC80 4.8-156 µg/ml) in TZM-bl cells. Methanolic extract of Strychnos potatorum, aqueous extract of Ficus infectoria and hydroalcoholic extract of Annona squamosa inhibited laboratory adapted HIV-1 strains (IC80 4.24-125 µg/ml) and primary isolates (IC80 18-156 µg/ml) in TZM-bl cells. Methanolic extracts of Achyranthes aspera and Rosa centifolia, (IC801-9 µg/ml) further significantly inhibited HIV-1 primary isolates in PM1cells. Methanolic extracts of Tridax procumbens, Mallotus philippinensis, Annona reticulate, aqueous extract of Ficus benghalensis and hydroalcoholic extract of Albizzia lebbeck did not exhibit anti-HIV activity in all the tested strains. Methanolic extract of Rosa centifolia also demonstrated to be non-toxic to HEC-1A epithelial cells and maintained epithelial integrity (at 500 µg/ml) when tested in transwell dual-chamber. CONCLUSION: These active methanolic extracts of Achyranthes aspera and Rosa centifolia, could be further subjected to chemical analysis to investigate the active moiety responsible for the anti-HIV activity. Methanolic extract of Rosa centifolia was found to be well tolerated maintaining the epithelial integrity of HEC-1A cells in vitro and thus has potential for investigating it further as candidate microbicide.

Enhanced anti-psoriatic efficacy and regulation of oxidative stress of a novel topical babchi oil (Psoralea corylifolia) cyclodextrin-based nanogel in a mouse tail model.

Psoriasis is a proliferative inflammatory skin disorder with relapsing episodes. Herein, the efficacy of babchi oil (BO) loaded nanostructure gel was evaluated for antipsoriatic activity and oxidative stress biomarkers assessment using mouse tail model. BO was entrapped into cyclodextrin-based nanocarriers (360.9 ± 19.55 nm), followed by incorporation into Carbopol gel and characterised for viscosity, spreadability, and texture analysis. The gels were topically applied on mouse-tails once daily for fourteen days. Evaluation of antipsoriatic activity as determined by histopathological observations of orthokeratotic epidermis revealed two times higher efficacy of BO nanogel in comparison to the native BO gel. Further, significantly enhanced superoxide dismutase (SOD) and reduced glutathione (GSH) levels, and diminished malondialdehyde (MDA) and nitrite (NO) levels revealed that prepared nanogels played a major role in the management of reactive oxygen species (ROS) associated in psoriasis pathogenesis. Hence, this study provides strong evidence for use of cyclodextrin-based nanogels as a safe and better delivery carrier of BO for management of psoriasis.

Antipsoriatic potential of Annona squamosa seed oil: An in vitro and in vivo evaluation.

BACKGROUND: Corticosteroids remains compound of choice for topical treatment of psoriasis. Several side effects associated with chronic application of corticosteroids limit its uses. Hence, there is a need to find a safe and effective alternative agent for psoriasis treatment. PURPOSE: The study aimed to investigate the in vitro and in vivo efficacy of petroleum ether extract of Annona squamosa seeds (ASO) as an antipsoriatic agent. The toxicity profile of ASO and its effect on psoriasis-induced inflammation has also been determined. METHODS: Physicochemical characterization was performed to determine constituents of ASO. Anti-proliferative activity of ASO was studied by Sulforhodamine B (SRB) assay using HaCaT cell lines. Oxazolone-induced psoriasis in female Balb/C mice was used as an animal model for investigating in vivoefficacy of ASO. Inflammatory markers were analyzed by immunohistochemical staining of mice ears. Safety profile of ASO was confirmed by performing acute dose dermal toxicity and repeated dose dermal toxicity testing. RESULTS: Predominant presence of polyunsaturated fatty acids viz. linoleic acid and oleic acid in ASO was confirmed by 1H NMR, 13C NMR and GC-MS analysis. The petroleum ether extract of Annona squamosa seeds showed inhibition of cell proliferation of keratinocytes (HaCaT cells). The growth inhibitory property of ASO was significantly higher than that was observed in presence of corticosteroid, clobetasol propionaste (CP). Application of ASO to the ears of Balb/C mice with oxazolone induced psoriasis showed marked reduction in erythema and edema, which was comparable to treatment with 0.05% CP cream. The increased levels of cytokines IL6, IL17, TNF-α, INF-γ, GMCSF, and infiltration of CD4 T cells observed in psoriasis lesions were decreased upon application of ASO. Acute and repeated dermal toxicity studies of ASO did not reveal any adverse events affirming the safety of ASO. CONCLUSION: The present data has demonstrated that ASO is a safe and effective anti-psoriatic agent when tested in animal models. The efficacy of ASO in preclinical studies could further be exploited for the development of potential novel topical antipsoriatic agent for therapy in humans.

Exploring Nanotechnologies for the Effective Therapy of Malaria Using Plant-Based Medicines.

Malaria is a potentially lethal disease caused by species of the plasmodium parasite. Despite the advances in the interventions for its control and approaches to manage fatality, morbidity and mortality rates associated with malaria are still high. At present, artemisinin-based combination therapy is the first line of treatment. However, there is the need to explore newer approaches as reduced effectiveness and multi-drug resistance (even to artemisinin) has been reported in some regions and is expected to widen in scope. Phytomedicines have shown promise for the management of this debilitating disease and there are abundant resources in most of the areas where this disease is endemic. This report would systematically review the literature, emphasizing the challenges encountered in the control of malaria, active phytochemicals currently utilised in the management, drug delivery approaches as well as the nanotechnology-based systems that could be exploited in its treatment. These phytomedicines, either delivered conventionally or via the use of advanced delivery systems may suggest new strategies towards the better management of malaria.

Development and evaluation of colloidal modified nanolipid carrier: application to topical delivery of tacrolimus, Part II--in vivo assessment, drug targeting, efficacy, and safety in treatment for atopic dermatitis.

In atopic dermatitis (AD), topical anti-inflammatory therapy with skin barrier restoration to prevent repeated inflammatory episodes leads to long-term therapeutic success. Tacrolimus, although effective against AD, is a challenging molecule due to low solubility, low-penetration, poor-bioavailability, and toxicity. Part I of this paper, reported novel modified nanolipid carrier system for topical delivery of tacrolimus (T-MNLC), offering great opportunity to load low-solubility drug with improved entrapment efficiency, enhanced stability and improved skin deposition. Present investigation focused on restoration of skin barrier, site-specific delivery, therapeutic effectiveness, and safety of novel T-MNLC. T-MNLC greatly enhanced occlusive properties, skin hydration potential and reduced transepidermal water loss. This might help to reduce the number of flares and better control the disease. Cutaneous uptake and drug deposition in albino rats by HPLC and confocal laser scanning microscopy revealed prominently elevated drug levels in all skin strata with T-MNLC as compared to reference. T-MNLC demonstrated efficient suppression of inflammatory responses in BALB/c mice model of AD. Safety assessment by acute and repeated-dose dermal toxicity demonstrated mild keratosis and collagenous mass infiltration at the treatment area with repeated application of reference. Interestingly, T-MNLC showed no evident toxicity exhibiting safe drug delivery. Thus, novel T-MNLC would be a safe, effective, and esthetically appealing alternative to conventional vehicles for treatment for AD.

Contraceptive efficacy and safety of HerbOshield™ vaginal gel in rats.

BACKGROUND: Spermicides represent one of the methods of contraception. The synthetic agents available as spermicides produce severe side effects. Hence, there is a need to replace these agents with safe and effective agents such as plant-based contraceptive agents. STUDY DESIGN: The objective of the present study was to develop and evaluate a stable, safe, effective and easily acceptable contraceptive delivery system containing herbal drug. Efforts were made to evaluate the contraceptive potential of the hydroalcoholic extract from the seeds of Annona squamosa Linn. and the vaginal gel HerbOshield™ containing the extract. RESULTS: Spermicidal effect was evaluated in vitro using healthy human spermatozoa and in vivo in rats. The in vitro results demonstrated that HerbOshield™ vaginal gel is an effective spermicide. At a 100-mg/mL dose, complete immobilization of human spermatozoa was observed within 20 s. None of the treated animals conceived, indicating 100% contraceptive effect as compared to Gynol II, a nonoxynol-9-containing marketed formulation, which showed only 67% contraceptive effect in vivo. HerbOshield™ vaginal gel was found to be safe in animals during a 14-day toxicity study. CONCLUSIONS: HerbOshield™ vaginal gel was found to be safe and effective in rats and could be developed as a potential vaginal contraceptive for future use in humans.

Development and evaluation of colloidal modified nanolipid carrier: application to topical delivery of tacrolimus.

Low solubility of tacrolimus in carrier matrix and subsequent poor in vivo bioavailability was overcome by constructing modified nanolipid carrier (MNLC) as a novel approach. The aim of this study was to develop MNLC with enhanced drug solubility in carrier lipid matrix using lipophilic solubilizers for topical delivery. Comprehensive characterization of tacrolimus-loaded MNLC (T-MNLC) was carried out for particle size, morphology, and rheology. Lipid modification resulted in the formation of less perfect crystals offering space to accommodate the dissolved drug leading to high entrapment efficiency of 96.66%. Compatibility and mixing behavior of carrier constituents was evaluated using DSC, FT-IR, and (1)H NMR. T-MNLC displayed sufficient stability that could be attributed to possibility to reduce total lipid concentration in carrier. T-MNLC-enriched gels showed significantly higher in vitro drug release, skin permeation, and in vivo bioavailability with dermatopharmacokinetic approach in guinea pigs compared to commercial ointment, Protopic® as reference. Penetration-enhancing effect was confirmed using gamma scintigraphy in vivo in rats. Radioactivity remained localized in skin at the application site avoiding unnecessary biodisposition to other organs with prospective minimization of toxic effects. Skin irritation studies showed T-MNLC to be significantly less irritating than reference. Research work could be concluded as successful development of novel T-MNLC using lipophilic solubilizers to increase the encapsulation efficiency of colloidal lipid carriers with advantage of improved performance in terms of stability and skin localization.