Investigation of the Anti-Inflammatory Effects of Bauhinia purpurea L. Ethosome Suspension Gel Using Methanolic Leaf Extract: Optimization via Central Composite Design (CCD) and Evaluation In Silico, In Vitro, and In Vivo

The prime objective of the present study was to evaluate the probable in vivo antiinflammatory potential of various extracts of the leaves of Acacia auriculiformis Benth. using carrageenan and formalin-induced inflammation in the rats. Thereafter gas chromatography-mass spectrometry analysis of the bioactive extract was performed to identify the compounds responsible for the antiinflammatory activity. The in vitro antioxidant potential was determined through 2,2-diphenyl-1-picrylhydrazyl scavenging assay, hydrogen peroxide scavenging assay and reducing power assay. Overall butanol and methanol leaf extracts showed significant antioxidant free radical scavenging activity. The methanol and petroleum ether leaf extracts at a dose of 400 mg/kg showed the highest percent inhibition of 84.88 and 82.12, respectively in the carrageenan-induced rat paw edema model, whereas, in formalin-induced rat paw edema model the chloroform and methanol leaf extracts showed the highest percent inhibition of 65.68 and 63.34 at a dose of 400 mg/kg, respectively. In both the antiinflammatory models, indomethacin (standard, 40 mg/kg) was used for comparison with test extracts. The gas chromatography-mass spectrometry analysis of bioactive butanol and methanol extracts showed the presence of compounds like sterols (stigmasterol, β-sitosterol and γ-sitosterol) and phenolic compounds (phenol, 2,4-bis(1,1-dimethylethyl),2,4-ditert-butylphenol), which could be responsible for the pharmacological activities observed. These in vitro and in vivo studies indicated that the antiinflammatory and antioxidant potential of the methanol leaf extract of Acacia auriculiformis Benth. could due to the presence of high phenolic and sterol content.

This study determined the bio active substances in the physic nut plant, Jatropha curcas and further examined the larvicidal potentials of its hexane, methanol and aqueous leaf and stem extracts on locally reared larvae of the malaria vector, Anopheles gambiae in accordance with the World Health Organization’s guidelines for laboratory and field testing of mosquito larvicides. Various concentrations (25mg/mL, 50 mg/mL 100mg/mL and 200 mg/mL) of the plant extracts were tested against third instar larvae of Anopheles gambiae. Qualitative phytochemical analysis of the different portions of J. curcas leaf and stem extracts revealed the presence of active toxic compounds including alkaloids, saponins, flavonoids, glycoside and tannins. Methanolic extracts were found to be richer in phytochemicals than hexane and aqueous extracts. All plant extracts at the various concentrations showed significant larvicidal activity against Anopheles gambiae mosquito larvae between 30 minutes to 24 hours of exposure. Methanol leaf extract of J. curcas was most effective as it showed larval mortality of 75 to 100% on the test larvae after 30 minutes to 24 hours of exposure while the methanol stem extract showed 60 to 100% larval mortality. Hexane leaf extract showed larval mortality of 65 to 100% after 30 minutes to 24 hours of exposure whereas hexane stem extract had larval mortality of 60 to 100%. However, the aqueous leaf extract had 40 to 100% mortality as the aqueous stem extract showed 35 to 100% mortality after 30 minutes to 24 hours respectively. The methanol leaf extract showed highest toxicity against the test larvae with LC₅₀ value of 2.52 mg/ml; and LC₉₀ value of 218.15 mg/ml while the least toxicity was observed on aqueous stem extract with LC₅₀ value of 70.71 mg/ml; and LC₉₀ value of 1635.76 mg/ml after 30 minutes of exposure respectively. All the test larvae treated with various extracts exhibited 100% mortality after 24 hours of exposure with less concentrations of the extract required to kill the larvae as time of exposure increased. The toxicity of the various leaf extracts on the mosquito larvae were relatively greater than those of the stem. This is supported by the abundance of secondary metabolites. The findings suggest that the hexane, methanol and aqueous leaf and stem extracts of J. curcas have the potential to be used as an effective botanical larvicide.

The purpose of this study was to implement Quality by Design (QbD) concept to Solid Lipid Nanoparticles (SLN) containing Asenapine maleate (AM) in order to identify critical process and formulation variables which can affect product quality such as particle size (PS) and entrapment efficiency (EE). Initially, risk assessment using ishikawa diagram and preliminary investigation of critical variables was carried out. Two statistical designs were used to optimize critical variables which can affect product quality attributes i.e. PS and EE. Plackett Burman Design (PBD) was used to screen 8 variables and results showed that lipid concentration, surfactant concentration and sonication time had significant effect on PS and EE. These critical factors were further optimized using Central Composite Design (CCD), a type of response surface methodology, to assess its effect on PS and EE. Design space was identified and implementation of control strategy for responses generated quality of the desired product. Design space was generated for SLN for reducing intra-batch and inter-batch variability in formulation development process. Analysis of robustness of design space predicted that the formulation must be prepared in established design space to reduce batch variations. The results conclusively demonstrated the potential of QbD concept to build quality in SLN formulation.

Background: Discontinuation and poor compliance with long-term oral medicine are major therapeutic issues in psychosis treatment. Poorer long-term outcomes may result from non-compliance as well as a higher chance of relapse. In order to sustain therapeutic drug plasma levels, co-administration of oral antipsychotics is necessary for commercially available longacting injections of second-generation antipsychotics, as they have a lag period of approximately three weeks during the drug release process. Methods: Poly(lactic-co-glycolic acid) (PLGA) encapsulated microspheres loaded with risperidone were fabricated in the current research for intramuscular administration. The single emulsion solvent evaporation technique was applied for the fabrication of microspheres. Risperidone microspheres were prepared using PLGA grade 75:25. Particle size, drug content and entrapment efficiency with a central composite design were the main optimization parameters for the formulation. The microspheres were characterized by different techniques, namely Fourier transform infrared spectroscopy (FT-IR), differential scanning calorimetry (DSC), X-ray diffractometry (XRD), scanning electron microscopy (SEM), and transmission electron microscopy (TEM). The drug content, entrapment efficiency, morphology, particle size, and in vitro release profiles, along with release kinetics of the risperidone microspheres, were studied. Results and Discussion: The microspheres produced by the single emulsion solvent evaporation approach show smooth and spherical morphology, with particle size ranging from 3 μm to 6 μm, drug content of 99.7%, and entrapment efficiency of 98.2% with little burst release of 3% to 10%, providing drug release for 45 days and exhibiting zero order release kinetics and Korsmeyer- Peppas model for non-fickian drug release from the polymeric matrix. By applying the Quality by Design (QbD) approach and formulation parameters, microspheres with appropriate particle size, morphology, enhanced drug content, entrapment efficiency and desirable drug release profile for depot formulation can be obtained successfully. Conclusion: The optimized microspheres, in comparison to the marketed Risperdal Consta™, show enhanced potential for a better depot formulation, which can further improve patient compliance.

The purpose of present study was to optimize rizatriptan (RZT) chitosan (CS) nanoparticles using ionic gelation method by application of quality by design (QbD) approach. Based on risk assessment, effect of three variables, that is CS %, tripolyphosphate % and stirring speed were studied on critical quality attributes (CQAs); particle size and entrapment efficiency. Central composite design (CCD) was implemented for design of experimentation with 20 runs. RZT CS nanoparticles were characterized for particle size, polydispersity index, entrapment efficiency, in-vitro release study, differential scanning calorimetric, X-ray diffraction, scanning electron microscopy (SEM). Based on QbD approach, design space (DS) was optimized with a combination of selected variables with entrapment efficiency > 50% w/w and a particle size between 400 and 600 nm. Validation of model was performed with 3 representative formulations from DS for which standard error of − 0.70–3.29 was observed between experimental and predicted values. In-vitro drug release followed initial burst release 20.26 ± 2.34% in 3–4 h with sustained drug release of 98.43 ± 2.45% in 60 h. Lower magnitude of standard error for CQAs confirms the validation of selected CCD model for optimization of RZT CS nanoparticles. In-vitro drug release followed dual mechanism via, diffusion and polymer erosion. RZT CS nanoparticles were prepared successfully using QbD approach with the understanding of the high risk process and formulation parameters involved and optimized DS with a multifactorial combination of critical parameters to obtain predetermined RZT loaded CS nanoparticle specifications.

In vitro antibacterial activity of methanol and ethanol leaf extracts of Euphorbia heterophylla and Pterocarpus lucens were investigated against six bacterial clinical isolates using the tube dilution and agar diffusion methods. Salmonella typhi was the most susceptible to methanol leaf extracts of E. heterophylla with a zone of inhibition ranging from 16 to 24 mm for 12.5 to 100 mg/ml concentration. This was followed by Streptococcus lactis, Escherichia coli, Staphylococcus aureus, and Shigella species in that order with Proteus vulgaris not susceptible to the different test concentrations of both plant extracts. E. heterophylla had the least minimum inhibitory concentration (MIC) of 6.25 mg/ml against E. coli and S. typhi while P. lucens extract MIC of 25.00 mg/ml was the least against S. typhi. Since there is an inverse relationship between MIC value and susceptibility of the clinical test isolates, the MIC values also shows that E. heterophylla methanol leaf extracts were more potent to the susceptible test organisms having lower MIC values than the corresponding ethanol leaf extract MIC value. E. heterophylla extract minimum bactericidal concentration (MBC) was 25.00 mg/ml for the sensitive isolates except for methanol extract with 12.50 mg/ml against S. typhi and ethanol extract with 12.50 mg/ml against S. aureus. P. lucens extract MBC was 100.00 mg/ml for the sensitive test isolates except for ethanol leaf extracts with 50.00 mg/ml against S. typhi. The killing rate of E. heterophylla methanol leaf extract MBC shows that E. coli was most rapidly killed at a rate of 4.53 × 106 CFU/min with S. aureus as the least killed at a rate of 0.62 × 106 CFU/min. S. lactis and E. coli were the most rapidly killed by P. lucens leaf extract MBC at a rate of 1.90 × 106 CFU/min. The killing rate of the extracts showed a positive support in the potential use of these plants in curing some infections as done by the traditional herbal healers in Anyigba, Kogi State, Nigeria.       Key words: In vitro, antibacteria, extract, inhibitory.In vitro antibacterial activity of methanol and ethanol leaf extracts of Euphorbia heterophylla and Pterocarpus lucens were investigated against six bacterial clinical isolates using the tube dilution and agar diffusion methods. Salmonella typhi was the most susceptible to methanol leaf extracts of E. heterophylla with a zone of inhibition ranging from 16 to 24 mm for 12.5 to 100 mg/ml concentration. This was followed by Streptococcus lactis, Escherichia coli, Staphylococcus aureus, and Shigella species in that order with Proteus vulgaris not susceptible to the different test concentrations of both plant extracts. E. heterophylla had the least minimum inhibitory concentration (MIC) of 6.25 mg/ml against E. coli and S. typhi while P. lucens extract MIC of 25.00 mg/ml was the least against S. typhi. Since there is an inverse relationship between MIC value and susceptibility of the clinical test isolates, the MIC values also shows that E. heterophylla methanol leaf extracts were more potent to the susceptible test organisms having lower MIC values than the corresponding ethanol leaf extract MIC value. E. heterophylla extract minimum bactericidal concentration (MBC) was 25.00 mg/ml for the sensitive isolates except for methanol extract with 12.50 mg/ml against S. typhi and ethanol extract with 12.50 mg/ml against S. aureus. P. lucens extract MBC was 100.00 mg/ml for the sensitive test isolates except for ethanol leaf extracts with 50.00 mg/ml against S. typhi. The killing rate of E. heterophylla methanol leaf extract MBC shows that E. coli was most rapidly killed at a rate of 4.53 × 106 CFU/min with S. aureus as the least killed at a rate of 0.62 × 106 CFU/min. S. lactis and E. coli were the most rapidly killed by P. lucens leaf extract MBC at a rate of 1.90 × 106 CFU/min. The killing rate of the extracts showed a positive support in the potential use of these plants in curing some infections as done by the traditional herbal healers in Anyigba, Kogi State, Nigeria.       Key words: In vitro, antibacteria, extract, inhibitory.

As a result of increased interest in the production of plant-based drugs for the treatment of many diseases has become a significant reason why people have become more coversant in the use of traditional medicine for the treatment of mild and serious illness. Due to increase in the thrust for the production of plant-based drugs, this present study was carried out to compare the phytochemical constituents and antioxidant potencies of acetone, methanol and aqueous leaf extracts of Acalypha wilkesiana collected from Kaura Namoda Botanical Garden in Zamfara State-Nigeria. The antioxidant activities was evaluated using various assays; The total phenolic content of aqueous, methanol and acetone leaf extract were 15.58 0.66 mg GAE/g, 14.10 2.17 mg GAE/g and 8.70 0.01 mg GAE/g respectively. Total flavonol contents; 207.10 11.53 mg QE/g, 196.08 5.53 mg QE/g and 112.04 8.27 mg QE/g respectively. Total flavonoid contents; 240.99 9.50 mg QE/g, 252.52 3.73 mg QE/g and 123.88 5.58 mg QE/g respectively. FRAP values were 679.14 0.45 mmol/g, 611.90 7.09 mmol/g and 292.07 11.38mmol/g respectively. ABTS activity of aqueous, methanol and acetone leaf extract were 24.30 5.86 mg AAE/g, 14.49 1.02 mg AAE/g and 7.00 0.57 mg AAE/g respectively, methanol leaf extract had the highest percentage DPPH Inhibition value of 42.64 5.13, followed by aqueous (31.77 4.08) at 0.25mg/ml while aqueous had the highest (52.63 0.67), followed by methanol extract (44.80 2.80) at 0.50mg/ml. Aqueous extract had the highest percentage inhibition of Nitric Oxide with a value of 59.74 1.30, followed by methanol extract (46.11 2.54) at 0.25mg/ml. inhibition for aqueous was also highest at 0.5 mg/ml. Aqueous extract had the highest percentage lipid peroxidation inhibition value of 22.66 2.93, followed by methanol leaf extract with the value of 18.89 0.80 while at 0.50mg/ml methanol leaf extract had the highest percentage inhibition of lipid peroxidation (39.42 3.10), followed by aqueous leaf extract with the value of 31.48 1.61. The results showed that aqueous and methanol leaf extract of Acalypha wilkesiana displayed potent antioxidant effects with the aqueous having an edge. This present study therefore supports the view that Acalypha wilkesiana can be used in the management of oxidative stress and other related diseases.

Poeciloneuron indicum Bedd. is an endemic medicinal plant of Western Ghats belonging to the family Clusiaceae. Bark is used to treat dysentery, diarrhea and cholera and its root decoctionwas used as oral contraceptive. Phytochemical screening showed the presence of sterols, triterpenes, saponins, alkaloids, phenols, tannins, flavonoids, carbohydrates, resins, proteins and glycosides using extractsof hexane, chloroform, ethyl acetate, methanol and aqueous of stem, leaf and callus. Majority of were present in methanol extract and aqueous extract of leaf, whereas for stem in methanol extract and ethyl acetate extract and methanol extract of callus. Methanol extract of leaf, stem and callusshowed maximum phenols of 35.02 GAE/g, 32.05 GAE/g and 19.03 GAE/g, respectively, total flavonoid content wasmaximum with 628.57 mg/g in aqueous extract of stem followed by ethyl acetate extract of leaf with 585.71 mg/g and 342.62 mg/g in aqueous extract of callus extract. DPPH scavenging activity revealed highest scavenging activity of 89.53% inmethanol extract of stem and aqueous extract of leaf with 76.57%. Methanolic extracts of stem, leaf and callus have good reducing power with 0.461, 0.453 and 0.253 at 700 nm, respectively,the methanol extracts of stem, aqueous extract of leaf and methanolic extract of leaf derived callus extractsof P. indicum consists of active antioxidants.

Context: Several Polygonum species (Polygonaceae) are used in traditional medicine in Asia, Europe and Africa to treat inflammation and diabetes. Objective: Evaluate the in vitro antioxidant, anti-inflammatory and antidiabetic potential of methanol and dichloromethane extracts of leaves and roots of the halophyte Polygonum maritimum L. Material and methods: Antioxidant activity was determined (up to 1 mg/mL) as radical-scavenging activity (RSA) of 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS), copper (CCA) and iron (ICA) chelating activities and iron reducing power (FRAP). NO production was measured in lipopolysaccharide (LPS)-stimulated macrophages for 24 h at concentrations up to 100 μg/mL and antidiabetic potential was assessed by α-amylase and α-glucosidase inhibition (up to 10 mg/mL) assays. The phytochemical composition of the extracts was determined by gas chromatography-mass spectrometry (GC-MS). Results: The methanol leaf extract had the highest activity against DPPH • (IC50 = 26 μg/mL) and ABTS +• (IC50 = 140 μg/mL), FRAP (IC50 = 48 μg/mL) and CCA (IC50 = 770 μg/mL). Only the dichloromethane leaf extract (LDCM) showed anti-inflammatory activity (IC50 = 48 μg/mL). The methanol root (IC50 = 19 μg/mL) and leaf (IC50 = 29 μg/mL) extracts strongly inhibited baker’s yeast α-glucosidase, but LDCM had higher rat’s α-glucosidase inhibition (IC50 = 2527 μg/mL) than acarbose (IC50 = 4638 μg/mL). GC-MS analysis identified β-sitosterol, stigmasterol, 1-octacosanol and linolenic acid as possible molecules responsible for the observed bioactivities. Conclusions: Our findings suggest P. maritimum as a source of high-value health promoting commodities for alleviating symptoms associated with oxidative and inflammatory diseases, including diabetes.

Background: AKT inhibition presents a promising avenue for cancer treatment strategies. By exploring natural analogues using in-silico docking and ADMET profiles, this work aims to design effective anti-cancer therapies shown by binding affinities and pharmacokinetic assessments. Aims and Objectives: The aim of this research paper is to utilize in-silico ADMET profiling and molecular docking studies to investigate the potential of natural analogues as inhibitors of the AKT enzyme. By leveraging computational techniques, including Molegro Virtual Docker (MVD) 6.0 and computational techniques like pkCSM, we aim to identify promising compounds with strong binding affinities to the target protein (PDB ID: 3OCB) and favorable pharmacokinetic properties. Our objectives include identifying key molecular interactions, evaluating optimal molecular weight ranges, and prioritizing compounds based on their MolDock scores for cancer treatment. Through this approach, our goal is to contribute to the design and development of effective anti-cancer therapies targeting the AKT signaling pathway. Methods: The Protein Data Bank provided the target protein (PDB ID: 3OCB) for the molecular docking study, which was conducted using Molegro Virtual Docker (MVD) 6.0. The selection of ligands from PubChem was focused on natural analogues. ADMET profiling benefited from the use of computational techniques such as pkCSM. Results: A molecular docking study of selected natural compounds was performed, and the top three compounds with higher MolDock scores were considered to be the best among all sixteen natural analogues. The compounds [00]UNX_16, [01]UNX_13, and [00]UNX_11 showed the highest MolDock score of -111.09, -98.31, and -96.37, respectively, and can show great potential in treating cancer. Conclusion: The analysis primarily focuses on a docking study investigating the potential inhibition of the AKT enzyme by natural analogs. The study explores molecular interactions and ADMET properties, offering insights into their role in drug discovery. Key findings include strong binding affinities of selected analogs against the target 3OCB, with specific amino acid residues and steric/hydrogen bond interactions influencing binding success. Compounds within the 400-500 Da molecular weight range show favorable interactions, suggesting implications for future drug design. Additionally, ADMET analysis identifies compounds like [00]UNX_16, [01]UNX_13, and [00]UNX_11 with high MolDock scores, indicating potential as AKT inhibitors for cancer treatment.

  Antimicrobial activity of aqueous, methanol and chloroform leaf extracts of Cissusmultistriata were investigated against 8 bacterial and 2 fungal test organisms, using the tube dilution and agar ditch diffusion methods. Aqueous leaf extract had no activity against both the bacterial and fungal test organisms. Both the methanol and chloroform leaf extracts inhibited all the test organisms with chloroform leaf extract showing the highest zone of inhibition against Escherichia coli (diameter 25 mm) and least against Staphylococcus aureus  (diameter 13 mm). The methanol leaf extract was least inhibitory against Salmonella typhi (diameter 8 mm) and most inhibitory against S. aureus (diameter 15 mm). The methanol leaf extract of C. multistriata show more antifungal activity compared with chloroform leaf extract, with Candida albicans being more susceptible than Aspergillus niger to both methanol and chloroform leaf extracts. The minimum inhibitory concentration (MIC) of methanol leaf extract show least activity against Yersinia enterocolitica andPseudomonas aeruginosa (MIC = 100 mg/ml) and higher activity of MIC at 50 mg/ml against the other bacterial test organisms. The chloroform leaf extract MIC of 100 mg/ml had least activity against Proteus mirabilis and P. aeruginosa and MIC of 20 mg/ml most inhibitory against E. coli, Klebsiella pneumonia and S. typhi. The antimicrobial activity of the heated extracts persisted after exposure to various temperatures between 30oC to 121oC for 15 to 30 min. However, the extract activity decreased as the temperature increased. The killing rate of the MBC of chloroform extract on E. coli was 1 cfu/3 min while on S.  typhi was 1 cfu/3.8 min.   Key words:  Cissus multistriata, antimicrobial, extract, inhibition, susceptible.

Screening of anti-inflammatory phytocompounds from Crateva adansonii leaf extracts and its validation by in silico modeling.

Quality by design in pharmaceutical manufacturing: A systematic review of current status, challenges and future perspectives

The present study was conducted to evaluate the antimicrobial activity of various extracts (viz. methanol, acetone and ethyl acetate) of Thuja orientalis and its preliminary phytochemical screening. In vitro antimicrobial activity was tested by agar well diffusion assay against human pathogenic microbes which are Gram positive (viz. S. aureus andB. subtilis), Gram negative bacteria (viz. P. aeruginosa, A. faecalis and K. pneumoniae) and fungi (A. flavus and A. niger). Methanol and acetone extract of leaf exhibited maximum activity against B. subtilis with zones of inhibition of 20.33 mm and 17.83 mm, respectively. All the extracts of stem were found to be most effective against P. aeruginosa. S. aureus was found to be sensitive to leaf extracts prepared in methanol, acetone and ethyl acetate with 13.66, 14.03 and 15.00 mm zone of inhibition, respectively.A. flavus and A. niger, both were found to be sensitive towards the methanol extract of leaf with inhibition zones of 15.50 and 16.00 mm, respectively compared to fungicides (viz. ketoconazole and fluconozole). Methanol extract of leaf and ethyl acetate extract of stem exhibit marked antimicrobial activity against B. subtilis and P. aeruginosa, which was comparable to standard antibiotics (that is, penicillin, ampicillin, tetracycline and streptomycin). It was also observed that leaf extracts were more effective as compared tostem extracts of the plant. Thus, Thuja orientalis could be considered as potential source of natural antimicrobials used for the treatment of bacterial and fungal infections.   Key words: Plant extracts, antibacterial activity, minimal inhibitory concentration, phytochemical screening.

Neurological disorders (ND) pose a major global health challenge, in large part due to the restrictive nature of the blood-brain barrier (BBB), which prevents most therapeutic agents from reaching the central nervous system (CNS). Intranasal delivery (IN) offers a non-invasive and patient-friendly route to bypass the BBB via the olfactory and trigeminal pathways, but its success requires advanced nanocarrier systems capable of enhancing drug retention, stability, and controlled release. In this study, a Quality by Design (QbD) framework was applied to systematically develop and optimize 1,2-distearoyl-sn-glycero-3-phosphoethanolamine (DSPE-PEG2000)-functionalized nanostructured lipid carriers (NLCs) using Donepezil (DPZ) as a model drug. Through sequential risk assessment, Plackett-Burman screening design (PBD), and Central Composite Design (CCD) optimization, the Critical Formulation Variables (CFVs) and Critical Process Parameters (CPPs) influencing particle size (PS) and entrapment efficiency (%EE) were identified. The optimized DSPE-PEG2000-NLCs exhibited nanoscale size (133.4 ± 2.91nm), high %EE (89.5 ± 1.51%), strong mucin binding (84.6 ± 2.68%), and a distinct core-shell morphology. In vitro and ex vivo studies confirmed a biphasic and sustained drug release up to 60h & 67h, outperforming uncoated NLCs and conventional formulations. Stability studies demonstrated improved preservation under refrigerated conditions. Beyond DPZ, the QbD-guided strategy presented here provides a generalizable and regulatory-aligned platform for designing IN nanocarriers, paving the way for reproducible, scalable, and translational drug delivery systems targeting a wide range of ND.