Evaluation of packaging, labels, and some physicochemical properties of herbal antimalarial products on the Ghanaian market.

Introduction: Malaria is a parasitic disease that is endemic in tropical areas and can be life-threatening. There has been a decrease in the prevalence of malaria in Ghana but the burden of the disease is still high in the country. Many Ghanaians depend on herbal products for malaria treatment. This study aimed to survey and evaluate commercial herbal antimalarials in the Volta Region of Ghana. Methods: A survey of finished herbal antimalarials was done at herbal shops, pharmacies, and over-the-counter medicine seller shops. Products available on shelves were purchased and their details were recorded, after which they were examined using a visual inspection tool. The density, pH, and extract weight per dose of each sample were also determined. Results: Thirty-four liquid formulations (A-1-34) containing 1-9 different herbs were found. The majority of the product labels had errors in consumer age classifications. Unconventional ways of stating doses were found on two products (A-13, "tot"; A-19, cupful). Six products did not have dosing devices. No duration of treatment was indicated on 24 products. Dose errors were found on A-14 and A-22. Samples A-17 and A-28 did not have registration or batch numbers. Product A-28 did not have its herbs listed on it and was indicated for persons aged 3-8 years at a dose of 45 mL. The relative density range for the products was 0.997-1.015. From the pH investigation, no product was extremely erosive; however, 10 samples were deemed erosive (pH, 3.0-3.99), whereas 24 were minimally erosive (pH, ≥4.0). The extract weight per dose volume (20-90 mL) was 0.048-1.766 g, indicating that unit dose capsules or tablets could be formulated from the products. Conclusion: The findings clearly show that Ghanaian authorities responsible for regulating herbal products must enforce guidelines for the formulation, label details, and sale of antimalarial products. Additionally, the unpleasant taste of liquid herbal mixtures can affect patient compliance and dosing convenience; therefore, it is recommended that oral solid dosage forms of herbal antimalarials are produced as alternatives to the liquid mixtures.

Novel indoloquinoline alkaloid glycosides from Cryptolepis sanguinolenta (Lindl.) Schltr.

To explore the phytochemistry of the African ethnomedicinal plant Cryptolepis sanguinolenta (Lindl.) Schltr. (Apocynaceae) for rare indoloquinoline alkaloids, two novel indoloquinoline alkaloid glycosides, namely Cryptospirosanguine A (1) and B (2), were isolated from an ethanolic extract of the root. Their structures were elucidated based on spectral evidence. Furthermore, two known terpenoids were isolated from this plant for the first time.

Metabolite profiling, antifungal, biofilm formation prevention and disruption of mature biofilm activities of Erythrina senegalensis stem bark extract against Candida albicans and Candida glabrata.

The antifungal activity of the 70% ethanol stem bark extract of Erythrina senegalensis (ESB) against different strains and drug resistant clinical isolates of Candida albicans and Candida glabrata were evaluated in the study. The effect of ESB on biofilms as well as its activity in combination with fluconazole, nystatin or caspofungin against the Candida strains were also evaluated. We then evaluated the antifungal activity of a microemulsion formulation of ESB against planktonic and biofilms of the Candida species. UPLC-QTOF-MS2 analysis was then undertaken to identify the phytoconstituents of the extract and UPLC fingerprints developed for the routine authentication as part of quality control measures. ESB exerted strong antifungal activities against C. albicans ATCC 10231 and SC5314 strains, and C. glabrata ATCC 2001 strain with minimum inhibitory concentration (MIC) values from 3.91 to 31.25 µg/mL and minimum fungicidal concentrations (MFCs) that ranged from 62.5 to 250 µg/mL. It also exhibited potent antifungal activities (MIC = 4-64 µg/mL) against a collection of C. albicans and C. glabrata clinical isolates that were resistant to either nystatin or azole antifungals. The formulated ESB demonstrated higher antifungal potency against the C. albicans and C. glabrata strains with MIC values of 3.91-31.25 µg/mL which was the same as the MFC values. The extract and its microemulsion formulation were active against biofilms of the strains of the Candida species inhibiting their biofilm formations (SMIC50 = 16-64 µg/mL) and their preformed biofilms (SMIC50 = 128 ->512 µg/mL). ESB also exhibited synergistic antifungal action with fluconazole and nystatin against C. albicans ATCC 10231 and C. glabrata ATCC 2001 strains in the checkerboard assay. Chemical characterization of the extract revealed the presence of phenolic compounds such as flavonoids and their prenylated derivatives, anthracene glycosides and alkaloids. UPLC Fingerprints of the extract was also developed and validated for routine identification and authentication of the stem bark of E. senegalensis. The study findings have demonstrated that the stem bark of E. senegalensis is as a potential source of bioactive compounds that could be developed as novel antifungal agents.

Antifungal Activities of Phytochemically Characterized Hydroethanolic Extracts of Sclerocarya birrea Leaves and Stem Bark against Fluconazole-Resistant Candida albicans Strains.

The study evaluated the antifungal activities of the 70% ethanol extracts of Sclerocarya birrea leaves (SBL) and stem bark (SBB) against C. albicans strains and fluconazole-resistant isolates, their antifungal effects in combination with conventional antifungals as well as their effects on the biofilms of the C. albicans strains and isolates. UPLC-QTOF-MS/MS analysis was then carried out to investigate the metabolite profile of the extracts and UPLC fingerprints developed for their routine identification as part of quality control measures. The extracts exhibited considerable antifungal activity with MIC ranging from 12.21 to 97.66 µg/mL and MFC from 12.21 to 390.63 µg/mL against the C. albicans strains and isolates. The antifungal activity of the stem bark extract was higher than the leaf extract. SBL and SBB also significantly inhibited biofilm formation (IC50 = 12.49 to 164.42 µg/mL) and the mature biofilms (IC50 = 91.50 to 685.20 µg/mL) of the strains and isolates of the C. albicans and demonstrated potential for their use in combination therapies with currently used antifungals especially the stem bark extract with nystatin. Metabolite profiling identified the presence of polyphenolic compounds in both leaves and stem bark mostly flavonoids, their derivatives, and proanthocyanidins, which contribute in part to the bioactivity of the plant. Whereas flavonoids like quercetin, myricetin, and their derivatives were abundant in the leaves, epicatechin monomers with their condensed tannins, including procyanidin B2 and procyanidin C, were abundant in the stem bark. Fingerprints of SBL and SBB were developed and validated and could be used as qualitative tools to authenticate the plant. The outcomes of the study show the promise of the leaf and stem bark extracts of S. birrea to be studied further and developed as antifungal agents.

Indole alkaloid from Nauclea latifolia promotes LDL uptake in HepG2 cells by inhibiting PCSK9.

BACKGROUND: Proprotein convertase subtilisin/kexin type 9 (PCSK9) has been found to play a major role in atherosclerotic cardiovascular disease (ASCVD) by promoting hyperlipidemia. Its inhibition has therefore emerged as a viable drug target for improving the outcome of ASCVD. However, current monoclonal antibody PCSK9 inhibitors are considered cost ineffective and there is the need to discover new effective and cheaper small molecule alternatives. PURPOSE: The methanolic and ethanolic crude extracts of Nauclea latifolia have been shown to possess anti-hyperlipidemic activity, but the chemical component(s) responsible for this activity and the mechanism of action have remained unknown. The objective of this study was therefore to identify N. latifolia constituents with anti-hyperlipidemic activity and to investigate the inhibition of PCSK9 as a probable mechanism of action. METHOD: In the present study, compounds were isolated from the ethanolic extract of the stem of N. latifolia. The alkaloids were evaluated for their DiI-LDL uptake promoting activity in HepG2 cell. The most active compound was further assessed for its effect on low density lipoprotein receptor (LDLR) and PCSK9 protein expressions by western blot. RESULTS: 3R-3,14-dihydroangustoline (5), showed a relatively good activity in promoting LDL uptake (1.26-fold). It further increased LDLR protein expression and decreased the protein expression of PCSK9 in a dose dependent manner (1-50  µM). CONCLUSION: Alkaloids from N. latifolia may serve as a source of new PCSK9 inhibitors.

Isocryptolepine, an indoloquinoline alkaloid from Cryptolepis sanguinolenta promotes LDL uptake in HepG2 cells

ABSTRACT About 31 percent of deaths worldwide result from atherosclerotic cardiovascular disease. Hyperlipidemia remains the major risk factor for this disease and therefore, it is necessary to identify antihyperlipidemic compounds for drug development. The crude ethanolic extract of Cryptolepis sanguinolenta (Lindl.) Schltr., Apocynaceae, has demonstrated antihyperlipidemic properties. However, the chemical constituents responsible for this action are unknown. Hence, to identify chemical constituent(s) of C. sanguinolenta with anti-hyperlipidemic effect, five indoloquinoline alkaloids were isolated and evaluated in 1,1′-dioctadecyl-3,3,3′,3′-tetramethyl-indocarbocyanine perchlorate labeled low density lipoprotein uptake assay using HepG2 cells. The minor alkaloid, isocryptolepine, showed strong activity in promoting low lipid lipoprotein uptake by 1.85-fold. Isocryptolepine may, therefore, serve as a lead compound for future studies in the development of novel antihyperlipidemic drugs.

Nanotoxicity: the toxicity research progress of metal and metal-containing nanoparticles.

Along with the exuberant development of nanotechnology, a large number of nanoformulations or non materials are successfully applied in the clinics, biomedicine, cosmetics and industry. Despite some unique advantages of nanoformulations, there exist potentially worrying toxic effects, particularly those related to metal and metal-containing nanoparticles (NPs). Although various researches have been conducted to assess the metallic and metal-containing nanoparticles toxic effects, only little is known about the toxicity expressive types and evaluation, reasons and mechanisms, influencing factors and research methods of metal and metal-containing nanotoxicity. Therefore, it is of importance to acquire a better understanding of metal and metal-containing nanoparticles toxicity for medical application. This review presents a summary on the metal and metal-containing nanoparticles toxicity research progress consulting relevant literature.

Mixed polyethylene glycol-modified breviscapine-loaded solid lipid nanoparticles for improved brain bioavailability: preparation, characterization, and in vivo cerebral microdialysis evaluation in adult Sprague Dawley rats.

Breviscapine is used in the treatment of ischemic cerebrovascular diseases, but it has a low bioavailability in the brain due to its poor physicochemical properties and the activity of P-glycoprotein efflux pumps located at the blood-brain barrier. In the present study, breviscapine-loaded solid lipid nanoparticles (SLN) coated with polyethylene glycol (PEG) derivatives were formulated and evaluated for their ability to enhance brain bioavailability. The SLNs were either coated with polyethylene glycol (40) (PEG-40) stearate alone (Bre-GBSLN-PS) or a mixture of PEG-40 stearate and 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-PEG2000 (DSPE-PEG2000) (Bre-GBSLN-PS-DSPE) and were characterized both in vitro and in vivo. The mean particle size, polydispersity index, and entrapment efficiency for Bre-GBSLN-PS and Bre-GBSLN-PS-DSPE were 21.60 ± 0.10 and 22.60 ± 0.70 nm, 0.27 ± 0.01 and 0.26 ± 0.04, and 46.89 ± 0.73% and 47.62 ± 1.86%, respectively. The brain pharmacokinetic parameters revealed that the brain bioavailability of breviscapine from the Bre-GBSLN-PS and Bre-GBSLN-PS-DSPE was significantly enhanced (p < 0.01) with the area under concentration-time curve (AUC) of 1.59 ± 0.39 and 1.42 ± 0.58 µg h/mL of breviscapine, respectively, in comparison to 0.11 ± 0.02 µg h/mL from the commercial breviscapine injection. The ratios of the brain AUC for scutellarin in comparison with the plasma scutellarin AUC for commercial breviscapine injection, Bre-GBSLN-PS, and Bre-GBSLN-PS-DSPE were 0.66%, 2.82%, and 4.51%, respectively. These results showed that though both SLN formulations increased brain uptake of breviscapine, Bre-GBSLN-PS-DSPE which was coated with a binary combination of PEG-40 stearate and DSPE-PEG2000 had a better brain bioavailability than Bre-GBSLN-PS. Thus, the coating of SLNs with the appropriate PEG derivative combination could improve brain bioavailability of breviscapine and can be a promising tool for brain drug delivery.

Preparation and evaluation of charged solid lipid nanoparticles of tetrandrine for ocular drug delivery system: pharmacokinetics, cytotoxicity and cellular uptake studies.

In this study, tetrandrine-loaded cationic solid lipid nanoparticles (TET-CNP) and solid lipid nanoparticles (TET-NP) were prepared by the emulsion evaporation-solidification at low temperature method. The particle size, zeta potential, and entrapment efficiency of TET-CNP and TET-NP were characterized. The results showed that the TET-CNP and TET-NP had average diameters of (15.29 ± 1.34) nm and (18.77 ± 1.23) nm with zeta potentials of (5.11 ± 1.03) mV and (-8.71 ± -1.23) mV and entrapment efficiencies of (94.1 ± 2.37)% and (95.6 ± 2.43)%, respectively. In vitro release studies indicated that the TET-CNP and TET-NP retained the drug entity better than tetrandrine ophthalmic solutions (TET-SOL). In the pharmacokinetics studies, the AUC values of TET-CNP and TET-NP were 1.96-fold and 2.00-fold higher than that of TET-SOL ( p < 0.05); the Cmax values of TET-CNP and TET-NP were 2.45-fold and 2.53-fold higher than that of the TET-SOL (p < 0.05), respectively. Cytotoxicity study showed that TET-CNP and TET-NP had no significant toxicity at low concentrations. Flow cytometry studies and confocal microscopy analysis demonstrated that calcein labeled NP (CA-NP) uptake by SRA 01/04 cells was much higher than those of calcein labeled CNP (CA-CNP) and calcein solution (CA-SOL).