Preparation of protamine-hyaluronic acid coated core-shell nanoparticles for enhanced solubility, permeability, and oral bioavailability of decoquinate.

Decoquinate (DQ) has low oral bioavailability owing to its poor water solubility. In this study, a DQ solid dispersion (DQ-SD) was fabricated using mechanochemical technology to encapsulate DQ and improve its oral bioavailability. DQ-SD is easily generated via self-assembly in the aqueous phase to form micelles consisting of disodium glycyrrhizinate (Na2GA) nanoparticles with a protamine (PRM) and anionic hyaluronic acid (HA) layers. The spherical DQ nanoparticles with an average diameter of 114.95 nm were obtained in an aqueous phase with a critical micelle concentration of 0.157 mg/mL, zeta potential of -38.38 mV, polydispersity index of 0.200, and drug loading of 5.66 %. The dissolution rate and cumulative release of DQ-SD were higher than those of pure DQ. Furthermore, the bioavailability of DQ-SD was approximately 6.3 times higher than that of pure DQ. Pharmacokinetic and biodistribution studies indicated that DQ-SD possessed a significantly higher concentration in the blood and preferential liver tissue accumulation, than that of pure DQ. The developed DQ-SD exhibited considerable potential for developing old DQ for a new application as a hematogenous parasite drug and provides a reference for developing more efficient delivery systems for hydrophobic bioactive agents.

Preparation of DNC Solid Dispersion by a Mechanochemical Method with Glycyrrhizic Acid and Polyvinylpyrrolidone to Enhance Bioavailability and Activity.

To exploit aqueous-soluble formulation and improve the anticoccidial activity of 4,4'-dinitrocarbanilide (DNC, active component of nicarbazin), this paper prepared DNC/GA/PVP K30 solid dispersion (SD) with glycyrrhizic acid (GA) and polyvinylpyrrolidone (PVP) K30 by a mechanical ball milling method without using any organic solvent. Fourier transform infrared spectroscopy, X-ray diffraction, differential scanning calorimetry, and scanning electron microscopy were used for the solid state characterization. High performance liquid chromatography, critical micelle concentration, particle characterization, and transmission electron microscopy were used to evaluate the behavior in aqueous solution. In addition, the oral bioavailability, tissue distribution, and anticoccidial activity of DNC/GA/PVP K30 SD were investigated as well. Compared with free drug, the novel formulation not only improved the solubility and dissolution rate of DNC, but also inhibited the fecal output of oocysts and enhanced the therapeutic effect of coccidiosis. According to the experiment results, the DNC/GA/PVP K30 SD increased 4.64-fold in oral bioavailability and dramatically enhanced the concentration in liver which provided a basis for further research in schistosomiasis. In summary, our findings suggested that DNC/GA/PVP K30 SD may have promising applications in the treatment of coccidiosis.

Preparation, physicochemical and pharmacological study of 10-hydroxycamptothecin solid dispersion with complexation agent - xylan-nonanoic acid amphiphilic conjugates.

An amphiphilic conjugate of carboxymethyl xylan-nonanoic acid (CX-NA) was synthesized with molecular weight of 38.35 kDa, HLB value of 13.59, and critical micelle concentration of 23.17 µg/ml. CX-NA could efficiently encapsulate the model drug of 10-hydroxycamptothecin (HCPT). The drug loaded amphiphilic conjugate could self-assembled to micelles with an average diameter of 110 nm, zeta potential of -42.88 mV, and drug encapsulation efficiency of 79.8%. In vitro experiments confirmed that the drug-loaded micelles exhibited excellent stability and permeability in the intestinal environment. Transport pathway demonstrated that HCPT was uptake by cells through clathrin-mediated endocytosis. Intestinal in situ absorption study further confirmed CX-NA vehicle could enhance HPCT to transport across intestinal epithelial cells in colonic tissues. Furthermore, the formulation showed excellent anti-tumor activity in vitro and improved bioavailability of 3.4 times in vivo as comparing with free HCPT. These findings imply that this amphiphilic conjugate is a potential and promising vehicle for delivery anticancer drug.

Preparation of olmesartan medoxomil solid dispersion with sustained release performance by mechanochemical technology.

Hypertension is a common disease for human with high morbidity and mortality, and olmesartan medoxomil (OM) is widely used in the therapy of hypertension. However, poor water solubility and low bioavailability limit its widespread use. To improve the effect of OM, a ternary OM solid dispersion consisting of hydroxypropyl-ß-cyclodextrin (HP-ß-CD) and hydroxypropyl methylcellulose (HPMC) was prepared by mechanochemical method. The best preparation parameters were OM/HP-ß-CD/HPMC-E5 with mass ratio of 1:2.6:1 and milling time of 4 h. Under the optimal preparation conditions, the solubility of the ternary solid dispersion could be increased by 12 times as compared with pure OM. Due to the addition of HPMC-E5, the solid dispersion had sustained release performance with prolonged release time of 12 h. Furthermore, in vivo study demonstrated that the prepared solid dispersion could afford significantly improved bioavailability of ~ 3-fold in comparison with pure drug. Hence, the prepared ternary solid dispersion of OM may be a promise delivery system for clinical application.

Mechanochemical Magnesium-Mediated Minisci C-H Alkylation of Pyrimidines with Alkyl Bromides and Chlorides.

A novel method to synthesize 4-alkylpyrimidines by the mechanochemical magnesium-mediated Minisci reaction of pyrimidine derivatives and alkyl halides has been reported. The reaction process operates with a broad substrate scope and excellent regioselectivity under mild conditions with no requirement of transition-metal catalysts, solvents, and inert gas protection. The practicality of this protocol has been demonstrated by the up-scale synthesis, mechanochemical product derivatization, and antimalarial drug pyrimethamine preparation.

Inositol hexanicotinate self-micelle solid dispersion is an efficient drug delivery system in the mouse model of non-alcoholic fatty liver disease.

Inositol hexanicotinate (IHN) self-micelle solid dispersion (SD) with glycyrrhizic acid (GA) and arabic gum (AG) was prepared by mechanical ball milling process to improve the solubility, stability of amorphous state, and bioavailability of IHN, which enhanced the treatment of IHN on hyperlipidemia and nonalcoholic fatty liver disease (NAFLD). The physicochemical properties of IHN/GA/AG SDs in solid state were characterized by differential scanning calorimetry, X-ray diffraction studies, and scanning electron microscopy. The characteristics of the sample solutions were analyzed by reverse-phase HPLC, particle characterization, critical micelle concentration, and transmission electron microscopy. Further pharmacokinetic study of this SD formulation in rats showed a significant 3.3-fold increase in bioavailability compared to pure IHN. Moreover, biomarkers in serum and liver of NAFLD mice were significantly ameliorated after oral administration of IHN/GA/AG SDs for 15 days. Altogether, these results establish the mechanochemically prepared IHN/GA/AG SDs as an efficacious formulation for the treatment of hyperlipidemia and NAFLD.

Neuroprotective Effects of Oxymatrine on PI3K/Akt/mTOR Pathway After Hypoxic-Ischemic Brain Damage in Neonatal Rats.

Oxymatrine (OMT), a quinolizidine alkaloid extracted from traditional Chinese herb Sophora flavescens Ait, has drawn attention because of its beneficial bioactivities against hypoxic-ischemic brain damage (HIBD). However, the underlying molecular mechanism remains unclear. In this study, we determined the in vivo and in vitro effects of OMT on seven-day old Sprague-Dawley rats with HIBD and in a rat model of primary hippocampal neuron oxygen glucose deprivation reoxygenation (OGD/R). This study was aimed to evaluate whether OMT exerted neuroprotective effects mediated by the (phosphatidylinositol 3-kinase/protein kinase B/mammalian target of rapamycin) PI3K/Akt/mTOR pathway after HIBD. Experimental results showed that the alkaloid significantly improved the early neurofunctional development, brain water content, abnormal pathological changes, and necrosis of neurons after HIBD. Moreover, OMT enhanced the cell viability and stabilized the mitochondrial permeability transition pore in the primary hippocampal neurons after OGD/R. OMT significantly decreased the autophagosome generation, elevated the expression of PI3K, Akt, and mTOR, and simultaneously reversed the mRNA expression of microtubule-associated protein 1-light chain 3 (LC3), Beclin-1, and sequestosomel (P62) induced by hypoxia and ischemia. However, these protective effects against HIBD could be suppressed when rapamycin, a specific inhibitor of mTOR, was included. Hence, the OMT exerted neuroprotective effects against HIBD by attenuating excessive autophagy by mediating the PI3K/Akt/mTOR pathway.

Preparation of camptothecin micelles self-assembled from disodium glycyrrhizin and tannic acid with enhanced antitumor activity.

Natural compounds as carriers for hydrophobic drugs have been increasingly used in drug delivery systems. In this study, disodium glycyrrhizin (Na2GA), tannic acid (TA) and camptothecin (CPT) were firstly used to prepare the camptothecin solid dispersion (CPT SD). When dissolved in a solution medium, Na2GA self-assembled to form micelles and CPT was encapsulated in micelles, meanwhile, TA connected with Na2GA through hydrogen bonds to form a contract shell. The average diameter of the CPT-loaded micelles is 80 nm with the critical micellar concentration of 0.303 mg/mL, the zeta potential of -33 mV, the PDI of 0.25 and drug loading 6.22%. In vitro experiments confirmed that the drug-loaded micelles exhibited excellent stability and permeability in the intestinal environment. Furthermore, the formulation showed excellent anti-tumor activity in vitro and in vivo. These findings imply that this nano-micelles provide a more potential and efficacious oral drug formulation for chemotherapy.

Mechanically induced solvent-free esterification method at room temperature.

Herein, we describe two novel strategies for the synthesis of esters, as achieved under high-speed ball-milling (HSBM) conditions at room temperature. In the presence of I2 and KH2PO2, the reactions afford the desired esterification derivatives in 45% to 91% yields within 20 min of grinding. Meanwhile, using KI and P(OEt)3, esterification products can be obtained in 24% to 85% yields after 60 min of grinding. In addition, the I2/KH2PO2 protocol was successfully extended to the late-stage diversification of natural products showing the robustness of this useful approach. Further application of this method in the synthesis of inositol nicotinate was also discussed.

Preparation, physicochemical and pharmacological study of curcumin solid dispersion with an arabinogalactan complexation agent.

Pharmaceutical solid dispersions (SD) of curcumin (Cur) with macromolecule polysaccharide arabinogalactan (AG) from wood of Larix sibirica were prepared by mechanical ball milling. The physical properties of the dispersed curcumin mixture in solid state were characterized by scanning electron microscope, differential scanning calorimetry and powder X-ray diffraction studies. These methods showed a strong decrease in the degree of crystallinity of Cur and its transformation to amorphization state, accompanied by the formation of the guest-host type complexes. The behavior of the samples in solutions was characterized by reverse phase HPLC, 1H NMR spectroscopy, UV-Visible spectroscopy and gel permeation chromatography (GPC). Mechanochemically prepared complexes demonstrated the increased solubility of Cur up to ~10.5 times in contrast to pure curcumin. The rapid storage test showed high chemical stability of Cur, which depended on mass relations of Cur-AG. Besides, improved membrane permeability of Cur-AG SD was tested by parallel artificial membrane permeability assay. Pharmacokinetic study of Cur-AG SD formulation in rat demonstrated a significant~8-fold enhancement of bioavailability in comparison to pure curcumin. In MTT tests, Cur-AG SD showed moderate cytotoxicity against human glioblastoma cells and immortalized human fibroblasts. Therefore, Cur-AG solid dispersion was a more promising and efficacious formulation for application in oral drug delivery.

Mechanochemical preparation of kaempferol intermolecular complexes for enhancing the solubility and bioavailability.

In this study, complexes of kaempferol (KF) with polysaccharide arabinogalactan (AG) and disodium glycyrrhizinate (Na2GA) were prepared through mechanochemical technique to improve the solubility and bioavailability of KF. The physicochemical properties and the interactions of KF with AG/Na2GA were investigated through dissolution, SEM, XRD, and DSC studies. The reduction of particle sizes and destruction of crystal forms revealed the formation of solid dispersion which may have assisted the dissolution of the drug. The accelerated stability study showed higher stability for KF-Na2GA complex. In vivo pharmacokinetic study was performed to observe the plasma drug concentrations for KF complexes. Mechanochemical complexation of KF with AG/Na2GA improved the pharmacological activity as evident by the inhibitory potential of the complexes towards carbohydrate metabolic enzymes. In vivo studies were performed in STZ-induced diabetic mice, where the group treated with KF-AG complex showed better liver and kidney function and lower blood glucose levels than pure KF. Therefore, mechanochemical complexes of KF with polysaccharide or glycyrrhizate may serve as a promising formulation for the treatment of diabetes.

Preparation of curcumin self-micelle solid dispersion with enhanced bioavailability and cytotoxic activity by mechanochemistry.

An amorphous solid dispersion (SD) of curcumin (Cur) with disodium glycyrrhizin (Na2GA) was prepared by mechanical ball milling. Curcumin loaded micelles were self-formed by Na2GA when SD dissolved in water. The physical properties of Cur SD in solid state were characterized by differential scanning calorimetry, X-ray diffraction studies, and scanning electron microscope. The characteristics of the sample solutions were analyzed by reverse phase HPLC, UV-visible spectroscopy, 1H NMR spectroscopy, gel permeation LC, and transmission electron microscopy. In vitro cytotoxic tests demonstrated that Cur SD induced higher cytotoxicity against glioblastoma U-87 MG cells than free Cur. Besides, an improvement of membrane permeability of Cur SD was confirmed by parallel artificial membrane permeability assay. Further pharmacokinetic study of this SD formulation in rat showed a significant ∼19-fold increase of bioavailability as comparing to free Cur. Thus, Cur SD provide a more potent and efficacious formulation for Cur oral delivery.

The efficacy of the supramolecular complexes of niclosamide obtained by mechanochemical technology and targeted delivery against cestode infection of animals.

Niclosamide is an anthelmintic that is widely used to treat cestode infection of animals. The efficacy of the supramolecular complexes of niclosamide obtained by mechanochemical technology and targeted delivery was studied in hymenolepiosis of mice and monieziosis of sheep. The efficacy of new substances of niclosamide with polyvinylpyrrolidone polymer in different ratios (1:10; 1:5; 1:2) was determined by the results of helminthological necropsy of the small intestine of sheep and mice. Pre-treatment eggs per gram (EPG) were not significantly different (P>0.1) among groups. The controlled test was used to evaluate the efficacy. A high efficacy (>95% efficacy) of the supramolecular complexes of niclosamide with PVP (SCoNwPVP) was shown in different ratios (1:10; 1:5 and 1:2) at a dose of 20mg/kg of body weight at oral administration against Hymenolepis nana in mice and Moniezia expansa in sheep. Whereas the basic drug - substance of niclosamide was effective at a dose of 100mg/kg of b/w. No adverse effects of the drugs on animal health were detected during the study.

Polysaccharide arabinogalactan from larch Larix sibirica as carrier for molecules of salicylic and acetylsalicylic acid: preparation, physicochemical and pharmacological study.

Inclusion complexes of salicylic acid (SA) and acetylsalicylic acid (aspirin, ASA) with polysaccharide arabinogalactan (AG) from larch wood Larix sibirica and Larix gmelinii were synthesized using mechanochemical technology. In the present study, we have investigated physicochemical properties of the synthesized complexes in solid state and in aqueous solutions as well as their anti-aggregation and ulcerogenic activity. The evidence of the complexes formation was obtained by nuclear magnetic resonance (NMR) relaxation technique. It was shown that in aqueous solution the molecules of SA and ASA are in fast exchange between the complex with AG macromolecules and solution. The stability constant of aspirin complex was calculated. It was shown that mechanochemically synthesized complexes are more stable when compared to the complex obtained by mixing solutions of the components. Complexes of ASA show two-fold increase of anti-platelet effect. It allows to reduce the dose of the antithrombotic drug and its ulcerogenic activity. These results substantiate the possibility to design new preparations on the basis of ASA with increased activity and safety.