A novel series of 3,6,6-trimethyl-4-oxo-4,5,6,7-tetrahydroindazole-1-acetic acid derivatives was designed and synthesized by a new one-step pathway. Structure elucidation of the synthesized compounds was confirmed by various spectral and elemental analyses. The prepared compounds were evaluated for their ability to inhibit cyclooxygenase-2 (COX-2) and cyclooxygenase-1 (COX-1) enzymes in-vitro. Among the synthesized compounds, the 2-(3,6,6-trimethyl-4-oxo-4,5,6,7-tetrahydroindazol-1-yl)acetic acid 4 emerged as the most potent COX-2 inhibitor (IC50 value: 150 nM) with the highest selectivity index (COX-1 / COX-2 inhibition ratio: 570.6). Docking studies of compound 4 in the active site of COX-2 recognized its potential binding mode to the enzyme. Based on the preliminary results, compound 4 was considered as a lead compound for further optimization.

The delivery of biologically active agents to the desired site in the body and intracellular organelles is still a big challenge despite efforts made for more than five decades. With the elaboration of synthetic methodologies to branched and hyperbranched macromolecules such as miktoarm stars and dendrimers, the focus has shifted to nanocarriers able to release and direct drug molecules to a desired location in a controlled manner. We present here recent developments in the field of targeted drug delivery with a focus on two specific macromolecular nanocarriers, dendrimers and miktoarm stars, and provide examples of these nanocarriers tested in different biological systems. A particular attraction of miktoarm stars is their versatility in achieving superior drug loading within their self-assembled structures. Advantages of dendrimers over linear polymers are that the former provide a platform for development of multivalent and multifunctional nanoconjugates, in addition to their ability to accommodate a large number of molecules inside, or at their surfaces.

In order to better understand nanoparticle uptake and elimination mechanisms, we designed a controlled set of small, highly fluorescent quantum dots (QDs) with nearly identical hydrodynamic size (8-10 nm) but with varied short ligand surface functionalization. The properties of functionalized QDs and their modes of uptake and elimination were investigated systematically by asymmetrical flow field-flow fractionation (AF4), confocal fluorescence microscopy, flow cytometry (FACS), and flame atomic absorption (FAA). Using specific inhibitors of cellular uptake and elimination machinery in human embryonic kidney cells (Hek 293) and human hepatocellular carcinoma cells (Hep G2), we showed that QDs of the same size but with different surface properties were predominantly taken up through lipid raft-mediated endocytosis, however, to significantly different extents. The latter observation infers the contribution of additional modes of QD internalization, which include X-AG cysteine transporter for cysteine-functionalized QDs (QD-CYS). We also investigated putative modes of QD elimination and established the contribution of P-glycoprotein (P-gp) transporter in QD efflux. Results from these studies show a strong dependence between the properties of QD-associated small ligands and modes of uptake/elimination in human cells.

Impairments of mitochondrial functions have been associated with failure of cellular functions in different tissues, leading to various pathologies. We report here a mitochondria-targeted nanodelivery system for coenzyme Q10 (CoQ10) that can reach mitochondria and deliver CoQ10 in adequate quantities. Multifunctional nanocarriers based on ABC miktoarm polymers (A = poly(ethylene glycol (PEG), B = polycaprolactone (PCL), and C = triphenylphosphonium bromide (TPPBr)) were synthesized using a combination of click chemistry with ring-opening polymerization, self-assembled into nanosized micelles, and were employed for CoQ10 loading. Drug loading capacity (60 wt%), micelle size (25-60 nm), and stability were determined using a variety of techniques. The micelles had a small critical association concentration and were colloidally stable in solution for more than 3 months. The extraordinarily high CoQ10 loading capacity in the micelles is attributed to good compatibility between CoQ10 and PCL, as indicated by the low Flory−Huggins interaction parameter. Confocal microscopy studies of the fluorescently labeled polymer analog together with the mitochondria-specific vital dye label indicated that the carrier did indeed reach mitochondria. The high CoQ10 loading efficiency allowed testing of micelles within a broad concentration range and provided evidence for CoQ10 effectiveness in two different experimental paradigms: oxidative stress and inflammation. Combined results from chemical, analytical, and biological experiments suggest that the new miktoarm-based carrier provides a suitable means of CoQ10 delivery to mitochondria without loss of drug effectiveness. The versatility of the click chemistry used to prepare this new mitochondria-targeting nanocarrier offers a widely applicable, simple, and easily reproducible procedure to deliver drugs to mitochondria or other intracellular organelles.

Aim: This study investigates the capacity of a potent anti-inflammatory nanomedicine, celastrol, incorporated into poly(amidoamine) dendrimers, to inhibit endotoxin-mediated signaling in microglia. Materials & methods: Celastrol was incorporated into amino (Cel/G4-NH2) and hydroxyl (Cel/G4-OH) terminus poly(amidoamine) (G4) dendrimers. Cell viability, release of nitric oxide, IL-6, TNF- and activation of MAPK (e.g., p38 and JNK) and NF-B were assessed in endotoxin (i.e., lipopolysaccharide) stimulated microglial cells. Results: G4-OH and G4-NH2 increased celastrol aqueous solubility by seven- and 12-fold, respectively. G4-OH and Cel/G4-OH suppressed lipopolysaccharide-mediated release of proinflammatory mediators, such as nitric oxide and IL-6, but not TNF-, without reducing microglial cell viability, while Cel/G4-NH2 potentiated cytotoxicity and cytokine release. Blockade of proinflammatory signaling was accompanied by attenuation of p38 MAPK activation. Conclusion: This study supports the potential use of poly(amidoamine) dendrimers for effective anti-inflammatory therapy in the chronically inflamed CNS.

Near-infrared spectroscopy (NIR) has evolved into an important rapid, direct and non-invasive technique in drugs analysis. In this study, the suitability of NIR spectroscopy to identify two benzodiazepine derivatives, diazepam and flunitrazepam, and a synthetic opiate, methadone hydrochloride, inside USP vials and probe the solid-state form of diazepam presents in tablets has been explored. The results show the potential of NIR spectroscopy for rapid, in situ and non-destructive identification of drugs.

Analysis of fatty acid (FA) is one of the most commonly used tools for investigating microbial populations in ecological studies. Fatty acids can be extracted and esterified to form fatty acid methyl esters (FAME) when analyzed using gas chromatography-mass spectrometry, the resulting profile contains some microbial biomarkers. The aim of the present study to analysis FAME present in two species of earthworms exhibit the same environment. The thin layer chromatography of earthworm Allolobophora caliginosa Savigny showed three major spots corresponding to FAME while that of earthworm Pheretima hawayana Rosa showed two major spots. The GC-MS analysis of Allolobophora caliginosa Savigny extract showed the presence of at least 23 peaks, only two peaks were identified from their Rt and Ms spectrum, hexadecanoic acid methyl ester and octadecenoic acid methyl ester. While the chromatogram of Pheretima hawayana Rosa extract showed at least 20 peaks, five of them were identified; butanedioic acid dimethyl ester, pentadecanoic acid 1 4-methyl ester, 2-hydroxy-hexadecanoic acid methyl ester, 9-octadecenoic acid methyl ester and octadecanoic acid methyl ester. In conclusion, in spite of the two species of earthworms from the same environment, but their content of FAME was different and that may explain why the biological activity of the whole body extract was different.

Analysis of fatty acid (FA) is one of the most commonly used tools for investigating microbial populations in ecological studies. Fatty acids can be extracted and esterified to form fatty acid methyl esters (FAME) when analyzed using gas chromatography-mass spectrometry, the resulting profile contains some microbial biomarkers. The aim of the present study to analysis FAME present in two species of earthworms exhibit the same environment. The thin layer chromatography of earthworm Allolobophora caliginosa Savigny showed three major spots corresponding to FAME while that of earthworm Pheretima hawayana Rosa showed two major spots. The GC-MS analysis of Allolobophora caliginosa Savigny extract showed the presence of at least 23 peaks, only two peaks were identified from their Rt and Ms spectrum, hexadecanoic acid methyl ester and octadecenoic acid methyl ester. While the chromatogram of Pheretima hawayana Rosa extract showed at least 20 peaks, five of them were identified; butanedioic acid dimethyl ester, pentadecanoic acid 1 4-methyl ester, 2-hydroxy-hexadecanoic acid methyl ester, 9-octadecenoic acid methyl ester and octadecanoic acid methyl ester. In conclusion, in spite of the two species of earthworms from the same environment, but their content of FAME was different and that may explain why the biological activity of the whole body extract was different.

Analysis of fatty acid (FA) is one of the most commonly used tools for investigating microbial populations in ecological studies. Fatty acids can be extracted and esterified to form fatty acid methyl esters (FAME) when analyzed using gas chromatography-mass spectrometry, the resulting profile contains some microbial biomarkers. The aim of the present study to analysis FAME present in two species of earthworms exhibit the same environment. The thin layer chromatography of earthworm Allolobophora caliginosa Savigny showed three major spots corresponding to FAME while that of earthworm Pheretima hawayana Rosa showed two major spots. The GC-MS analysis of Allolobophora caliginosa Savigny extract showed the presence of at least 23 peaks, only two peaks were identified from their Rt and Ms spectrum, hexadecanoic acid methyl ester and octadecenoic acid methyl ester. While the chromatogram of Pheretima hawayana Rosa extract showed at least 20 peaks, five of them were identified; butanedioic acid dimethyl ester, pentadecanoic acid 1 4-methyl ester, 2-hydroxy-hexadecanoic acid methyl ester, 9-octadecenoic acid methyl ester and octadecanoic acid methyl ester. In conclusion, in spite of the two species of earthworms from the same environment, but their content of FAME was different and that may explain why the biological activity of the whole body extract was different.

Crinum is a genus of about 130 species belonging to family Amaryllidaceae with wide geographical distribution throughout the tropics, subtropics and warm temperate regions of the world. These plants are not only showy ornamentals but they possess significant folkloric and commercial reputation as well. Long ago, Crinums have been subjected to extensive chemical, cytological and pharmacological investigations. Phytochemical investigations have resulted in isolation of several diverse classes of phytocompounds and have been focused predominantly on alkaloids. The present part of our review work about the phytochemical, biological and toxicological studies on Crinums summarizes crinine-type alkaloids isolated up to now as well as their structural and stereochemical differences, in addition to their distribution in different Crinum species.