Synthesis of Andrographolide glycosides and evaluation of their antibacterial activity in vitro and in vivo.

Andrographis paniculata, a medicinal plant widely found in Asia, contains andrographolide as its main active compound, known for its wide-ranging pharmacological effects, including anti-inflammatory, anti-cancer, anti-obesity, and anti-diabetic properties. Recent investigations have highlighted the anti-infective potential of andrographolide and its derivatives, with demonstrated antiviral, antibacterial, and antimalarial activities. This review summarizes progress in andrographolide’s anti-infective applications, focusing on its structure–activity relationship (SAR) and mechanisms of action. Researchers have used semi-synthetic methods, such as esterification, oxidation, Michael addition, salification, and hybrid design, to enhance andrographolide’s physicochemical properties and biological activity. These derivatives show potent antiviral activity against RNA and DNA viruses, antibacterial activity against Gram-positive and Gram-negative bacteria, antifungal effects, and antiparasitic activity against Plasmodium spp. and Leishmania spp. Nevertheless, poor solubility and limited bioavailability still hinder their clinical translation. Strategies such as nano delivery systems and β-cyclodextrin complexes are discussed to improve bioavailability. Although andrographolide itself has not received regulatory approval as a stand-alone drug, several andrographolide-containing preparations have been clinically used in certain countries. Overall, this review brings together evidence on antiviral, antibacterial, antifungal, and antiparasitic activities, linking them with structure–activity trends and pharmacokinetic insights, thereby providing a consolidated foundation for future development and clinical translation.

Objective: To enhance solubility of efavirenz by using chitosan-graft-HPβCD copolymer synthesized by novel one pot technique. Methodology : Poor aqueous solubility and limited bioavailability is the major problem for more than 40% NEC (new chemical entities). Efavirenz (EFV), widely used non nucleoside reverse transcriptase inhibitor (NNRTI) belongs to BCS class II drug is having very poor intrinsic water solubility and limited oral bioavailability. Chitosan –graft-HPβCD (CS-g-HPβCD) copolymer was synthesized by tosylation of HPβCD followed by grafting on chitosan (CS) backbone. CS-g-HPβCD was prepared by varying CS: HPβCD weight ratios (1:1, 1:2, 1:3 and 1:4). The copolymer was characterized by FT-IR, NMR and DSC. The molecular weight of grafted copolymer was determined and copolymer was further evaluated for its solubility enhancement capability. Result: Tosylated HPβCD was grafted on CS backbone by simple one pot synthesis method. FT-IR, NMR and DSC results avowed the synthesis of grafted copolymer. The molecular weight of grafted polymer was greater than molecular weight of CS also support confirmation of HPβCD grafting on CS. The copolymer was found to tremendously enhance aqueous solubility of EFV (380 times the solubility of drug in water). These results conclusively demonstrated synthesis of grafted copolymer possessing good potential in the solubility enhancement of the poorly water soluble drug like EFV. Conclusion: Grafted copolymer is an excellent strategy to conquer the solubility issues of hydrophobic drugs like EFV owing to solubility enhancement capability and mucoadhesivity. Key words: Chitosan, Chitosan-g-HPβCD, Efavirenz, Hydroxypropyl-β- cyclodextrin, Grafting, Solubility enhancement.

Current developments in Nano/Micro-formulations for enhanced delivery and bioactivity of andrographolide

The study was designed to evaluate the antibacterial potential of three plants which are Andrographis paniculata, Durio zibethinus and Psidium guajava. Andrographis paniculata leaves (30mg/ml) and roots (30 mg/ml), Durio zibethinus wood bark (10mg/ml), and Psidium guajava leaves (15mg/ml) extract was obtained through the process called maceration, filtration, evaporation and the paste form was freshly reconstitute in dimethyl sulfoxide (DMSO) and tested against Staphylococcus aureus for Andrographis paniculata, Psidium guajava. Streptococcus agalactiae for Durio zibethinus and Psidium guajava and Escherichia coli for Durio zibethinus using Kirby Baur technique and the plates were incubated at 37 oC. The zone of inhibition was measured after 24 hours and recorded in millimeters. The combination study was conducted using extract in combination with Penicillin G (6.25 µg/ ml) and erythromycin (15 µg/ ml; Andrographis paniculata) with the propotion of 1:1 in homogenous condition and incubated at 37 oC for 24 hours. The zone of inhibition was measured and recorded. Mean and standard deviation was calculated. Andrographis paniculata do possesses some antibacterial activity against Staphylococcus aureus. Leaves (17.33 mm), roots (10.67 mm), erythromycin (24.00 mm), leaves and erythromycin (20.67 mm), roots and erythromycin (21.67 mm), leaves and roots (17.33 mm). Wood bark against Streptococcus agalactiae (14.67 mm), Penicillin G (14.00 mm), and combination (16.67 mm). Durio zibethinus showed antibacterial activity against Escherichia coli (11.00mm) and Penicillin G (13.33 mm). Psidium guajava leaves extract were having slightly higher activity than Penicillin G and in combination activity, leaves were having a slightly higher activity than Penicillin G.

NATURAL RESISTANCE OF HORSES OF GUTSUL BREED FROM THE POKUTTYA CARPATHIANS

Andrographis paniculata is a well-known Asian medicinal plant with a major phytoconstituent of diterpene lactones, such as andrographolide, 14-deoxyandrographolide, and neoandrographolide. A World Health Organization (WHO) monograph on selected medicinal plants showed that A. paniculata extracts and its major diterpene lactones have promising anti-inflammatory, antidiabetic, antimalarial, anticancer, antifungal, antibacterial, antioxidant, and hypoglycemic activities. However, these active phytochemicals have poor water solubility and bioavailability when delivered in a conventional dosage form. These biological barriers can be mitigated if the extract or isolated compound are delivered as nanoparticles. This review discusses existing studies and marketed products of A. paniculata in solid, liquid, semi-solid, and gaseous dosage forms, either as an extract or isolated pure compounds, as well as their deficits in reaching maximum bioavailability. The pharmaceutics and pharmacological activity of A. paniculata as a nano-delivery system are also discussed.

Endophytic fungi of medicinal plants are abundant, and their metabolites often have antioxidant, antibacterial, and antitumor effects and can produce secondary metabolites identical or similar to those of their hosts, which can mitigate the problem of insufficient supply of medicinal plants. In this study, we screened endophytic fungi for strains that produce the same diterpene lactones as Andrographis paniculata based on their biological activity. Firstly, the dominant group of endophytic fungi of Andrographis paniculata was screened and pathogenicity was studied using Koch’s rule. Secondly, DPPH, ABTS, OH, PTIO radical scavenging, and FRAP assays were used to detect the antioxidant activity of the extracellular extracts of the strains, and total phenol and total flavonoid contents of the strains with high antioxidant capacity were determined. S. aureus, B. subtilis, E. coli, and P. aeruginosa were used to determine the antibacterial activity of the mycelial extracts of the strains. Finally, the secondary metabolites of the mycelial extracts of the strains were examined by high-performance liquid chromatography. The results showed that 32 strains of Andrographis paniculata were relatively isolated > 70% and non-pathogenic. Extracellular extracts of strains AP-1 and AP-4 showed vigorous antioxidant activity, and AP-4, AP-12, AP-47, and AP-48 showed antibacterial activity against four strains of bacteria. The HPLC results indicated that the mycelial extracts of AP-4 and AP-12 contained diterpene lactones. The two endophytic fungi were recognized as Colletotrichum sp. The study successfully obtained diterpene lactones from the endophytic fungus of Andrographis paniculata and confirmed the feasibility of using endophytic fungal strains to produce active substances consistent with the host. It was also useful for exploring endophytic fungi and medicinal plants. The relationship provides theoretical guidance.

<abstract> <p>Quality of dried <italic>Andrographis paniculata</italic> (Burm.f.) Nees materials is important to determine its effectiveness in traditional medicine. The present study aimed to investigate an effect of harvesting age and drying condition on andrographolide content and its consequences on antioxidant and antibacterial activities. The plants were cultivated and harvested at 90,100,115, and 127 days after sowing (DAS) prior to drying under the sun or using hot air oven at 50, 65, and 80 ℃. The results indicated that drying condition significantly influenced andrographolide content, antioxidant capacities, and antibacterial activity of <italic>A. paniculata</italic>, whereas the harvesting age had no significant impact on those parameters. The andrographolide contents ranged from 0.74–4.11% (w/w) dry weight. The highest andrographolide contents were obtained at 90 DAS/65 ℃, 127 DAS/65 ℃, and 127 DAS/65 ℃. <italic>A. paniculata</italic> extracts were found to exhibit antibacterial activity against gram-positive bacterial strains (<italic>B. cereus</italic>, <italic>M. luteus</italic>, <italic>S. epidermidis</italic>, and <italic>S. aureus</italic>), which the highest antibacterial activity was observed at 90 DAS/65℃. The used of hot air oven at 65 ℃ effectively preserved andrographolide content and antibacterial activity of <italic>A. paniculata</italic>. In contrast, drying at 50 ℃ was the preferable drying condition for antioxidant capacity. The antioxidant activities of <italic>A. paniculata</italic> extracts ranged from 3.43–26.73 and 1.93–17.28 mg Trolox/g dry weight for DPPH• scavenging activity and FRAP reducing power activity, respectively. Overall, <italic>A. paniculata</italic> is suggested to dry using hot air oven at 65 ℃ to maintain high levels of andrograhpolide and antibacterial activity. Drying using hot air oven at 50 ℃ is advised, if antioxidant activity is the main focus. Even though the harvesting age is not a key parameter, <italic>A. paniculata</italic> is suggested to harvest at 50% flowering stage onward for a better total herbage and andrographolide yield.</p> </abstract>

To identify the features of development of a necrotic and inflammatory process in different forms of nonalcoholic fatty liver disease (NAFLD), by comparatively analyzing a full set of clinical and laboratory parameters, including the cytokine status and the expression level of enzyme genes controlling the apoptosis of peripheral leukocytes. 86 patients with NAFLD, including 8 (9.3%) with hepatic steatosis (HS), 70 (81.4%) with nonalcoholic steatohepatitis (NASH), 40, 19, and 11 with mild, moderate, and high disease activity, respectively, and 8 (9.3%) with liver cirrhosis (LC), were examined. A control group consisted of 34 healthy donors. Clinical and biochemical blood indices, cytokine profile, and the level of caspase gene transcripts in the peripheral blood leukocytes (PBL) were estimated. As compared to the controls, the patients with HS had higher tumor necrosis factor-α (TNF-α) and interleukin 6 (IL-6) levels and lower caspase 3, 6, and 8 mRNA in PBL. The concentration of IL-10 in NASH was higher than that in steatosis and positively correlated with the level of proinflammatory cytokines. The levels of TNF-α and IL6 were higher in the patients with NASH than in the controls. Those of C-reactive protein, γ-globulin, IL-6, and cytokeratin-18 fragment increased with the progression of NASH. In the latter, the transcriptional activity of caspase-3 gene decreased relative to the reference value and negatively correlated with the level of proinflammatory cytokines. In the patients with LC, the gene expression profile of caspases in PBL was similar to that in the control group; the level of IL-6 was higher than that in steatosis and NASH, that of IL-1β was higher than in HS and positively correlated the concentration of IL-6 and the activity of alanine aminotransferase and aspartate aminotransferase. The features of a necrotic and inflammatory process were identified in different forms of NAFLD. When the latter progressed, the cytokine profile and gene expression levels of caspases in PBL altered along with a change in the general clinical picture.

Introduction: Andrographis paniculata, Tinospora crispa and Centella asiatica are known to have various pharmacological functions. This research was carried out to investigate the antibacterial activities of protein extracts from A. paniculata, T. crispa and C. asiatica. Methods: Total soluble proteins from these herbs were extracted using a modified TCA/acetone method. The protein extracts were then quantified using the Bradford assay and separated using SDS-PAGE. The antibacterial activities were determined by disc diffusion method. Results: T. crispa had a significantly higher amount of proteins (83.86 ± 0.4 µg/µl) compared to A. paniculata (81.57 ± 0.4 µg/µl) and C. asiatica (78.93 ± 0.5 µg/µl). The proteins separated by SDS-PAGE were ranged from 30kDa to 260kDa, 25kDa to 110kDa and 25kDa to 160kDa for A. paniculata, T. crispa and C. Asiatic, respectively. The high abundance proteins were observed in A. paniculata and T. crispa but not in C. asitica. Protein extracts from C. asiatica have demonstrated antibacterial activity against all tested bacteria with the diameter of inhibition zone of 11.0 ± 0.5 mm, 12.3 ± 0.6 mm, 10.7 ± 0.7 mm and 20.0 ± 0.8 mm against B. cereus, S. aureus, K. pneumonia and S. typhimurium respectively. Meanwhile, protein extracts of A. paniculata showed a positive antibacterial activity only against B.cereus (13.7 ± 0.4 mm), S. aureus (7.0 ± 0.8 mm) and S. typhimurium (11.5 ± 0.3 mm). Protein extracts from T. crispa only showed a positive antibacterial activity against B. cereus (9.7 ± 0.5 mm). Conclusions: There is a constant need in the discovery of new antibiotics for the treatment of infectious diseases.

BackgroundHydrogen-rich saline has been reported to have antioxidant and anti-inflammatory effects and effectively protect against organ damage. Oxidative stress and inflammation contribute to the pathogenesis and/or development of pulmonary hypertension. In this study, we investigated the effects of hydrogen-rich saline on the prevention of pulmonary hypertension induced by monocrotaline in a rat model.MethodsIn male Sprague-Dawley rats, pulmonary hypertension was induced by subcutaneous administration of monocrotaline at a concentration of 6 mg/100 g body weight. Hydrogen-rich saline (5 ml/kg) or saline was administred intraperitoneally once daily for 2 or 3 weeks. Severity of pulmonary hypertension was assessed by hemodynamic index and histologic analysis. Malondialdehyde and 8-hydroxy-desoxyguanosine level, and superoxide dismutase activity were measured in the lung tissue and serum. Levels of pro-inflammatory cytokines (tumor necrosis factor-α, interleukin-6) in serum were determined with enzyme-linked immunosorbent assay.ResultsHydrogen-rich saline treatment improved hemodynamics and reversed right ventricular hypertrophy. It also decreased malondialdehyde and 8-hydroxy-desoxyguanosine levels, and increased superoxide dismutase activity in the lung tissue and serum, accompanied by a decrease in pro-inflammatory cytokines.ConclusionsThese results suggest that hydrogen-rich saline ameliorates the progression of pulmonary hypertension induced by monocrotaline in rats, which may be associated with its antioxidant and anti-inflammatory effects.

Owing to the disseminating resistance among pathogenic bacteria, especially Klebsiella pneumoniae, there is a high need for alternate compounds with antibacterial activity. Herein, lycopene was isolated from Lycopersicon esculentum L. Molecular docking approach was employed to explore lycopene binding affinity to selected vital proteins of K. pneumoniae with the binding mechanisms being investigated. This proposed a promising antibacterial activity of lycopene. However, the pharmacological use of lycopene is hampered by its poor solubility and limited oral bioavailability. Accordingly, bilosomes were fabricated for oral lycopene delivery. The computed entrapment efficiency, mean vesicular size, and zeta potential values for the optimized formulation were 93.2 ± 0.6%, 485.8 ± 35.3 nm, and −38.3 ± 4, respectively. In vitro drug release studies revealed controlled lycopene release from constructed bilosomes, with the drug liberation being based on the Higuchi kinetics model. Transmission electron microscopic evaluation of bilosomes revealed spherical nanovesicles free from aggregates. Moreover, the in vitro and in vivo antibacterial activity of lycopene and its constructed formulations against multidrug-resistant K. pneumoniae isolates were explored. The optimized bilosomes exhibited the lowest minimum inhibitory concentrations ranging from 8 to 32 µg/mL. In addition, scanning electron microscopy revealed remarkable deformation and lysis of the bilosomes-treated bacterial cells. Regarding in vivo investigation, a lung infection model in mice was employed. The tested bilosomes reduced the inflammation and congestion in the treated mice’s lung tissues, resulting in normal-sized bronchioles and alveoli with very few congested vessels. In addition, it resulted in a significant reduction in pulmonary fibrosis. In conclusion, this study investigated the potential activity of the naturally isolated lycopene in controlling infections triggered by multidrug-resistant K. pneumoniae isolates. Furthermore, it introduced bilosomes as a promising biocompatible nanocarrier for modulation of oral lycopene delivery and in vivo antimicrobial activity.

New drug delivery technologies are transforming drug discovery and development, as well as establishing research and development-focused pharmaceutical firms that are accelerating global progress. The bioactive rutin molecule is used in a wide range of food and medicinal goods. Its limited bioavailability and poor water solubility are major issues. Rutin is a polyphenolic natural compound with antibacterial, anticancer, antioxidant, chemopreventive, and anti-inflammatory activities. However, no research has been published to yet to improve its bioavailability and efficacy. As a result, an attempt was made in this study to load rutin into a nanoparticlulate system in order to improve its bioavailability and efficacy. Six formulations (F1-F6) of nanoparticles were prepared by solvent evaporation technique and were evaluated for particle size and shape using Zeta Sizer, Scanning Electron Microscopy (SEM) and Fourier Transform Infra-Red (FT-IR) Spectroscopy. The optimized formulation was further subjected to in vitro evaluation. Practical percent yield, drug entrapment efficiency and In vitro drug release were evaluated. Out of various formulations F1 have shown best results in particle size 80.71 (1-100 nm), particle shape (spherical nanoparticles with a smoothed surface), average size distribution (105.0), zeta potential (-20.6) percentage yield (70.83), drug entrapment efficiency (83.6%) and drug loading (95%). Pure rutin showed incomplete dissolution of 47.69% in 330 min while Rutin loaded nanoparticles gave 94.75% release in 330 min. It is obvious from the foregoing that rutin chitosan nanoparticles were used as a novel drug delivery technology to improve therapeutic efficacy and sustained release features while overcoming issues such as poor solubility and limited bioavailability.

Nano-emulsification of Osmanthus essential oil: Characterizations, stability and molecular interactions explaining antibacterial activity

Background: Septic shock, the most serious complication of sepsis, is a life-threatening disease that is mainly characterized by hypoperfusion and multiple organ failure. Various aberrantly expressed microRNAs (miRNAs) have been reported to be related to septic shock. We explored the regulatory effect of microRNA-203 (miR-203) on lung injury in septic shock mice. Methods: Microarray-based gene expression profiling related to septic shock identified the differentially expressed gene vanin-1 (VNN1) and potential regulatory miR-203. miR-203 was predicted to mediate VNN1 expression, thus affecting septic shock, which was investigated by treatment with miR-203 mimic, miR-203 inhibitor, and siRNA-VNN1 in septic shock mouse models. Polymorphonuclear neutrophils (PMNs) and pulmonary alveolar macrophages in bronchoalveolar lavage fluid (BALF) as well as the wet/dry ratio of the lung were also measured to assess lung injury. Additionally, the effects of miR-203 on inflammatory cytokines, oxidative stress indexes, blood biochemical indexes, serine-threonine protein kinase (AKT) signaling pathway-related factors, and apoptosis-related factors were determined. Results: VNN1 was verified to be targeted and negatively regulated by miR-203. In mouse models of septic shock, weak expression of miR-203, high expression of VNN1, and inhibition of AKT signaling pathway were identified. In response to miR-203 mimic and VNN1 gene silencing, mouse models of septic shock displayed reduced apoptosis, MDA, ALT, and AST in lung tissues, decreased levels of TNF-α, IL-1β, IFN-γ, IL-10, and IL-6, in serum, and reduced PMN and PAM levels in BALF, in addition to elevated SOD activity. Notably, the presence of miR-203 mimic led to AKT signaling pathway activation. Conclusion:This study shows that upregulating miR-203 can alleviate lung injury through activation of the AKT signaling pathway by downregulating VNN1 in septic shock.