Preparation of novel shell-ionotropically crosslinked micelles based on hexadecylamine and tripolyphosphate for cancer drug delivery.

AIMS: Micellar systems have the advantage of being easily prepared, cheap, and readily loadable with bioactive molecular cargo. However, their fundamental pitfall is poor stability, particularly under dilution conditions. We propose to use simple quaternary ammonium surfactants, namely, hexadecylamine (HDA) and hexadecylpyridinium (HDAP), together with tripolyphosphate (TPP) anion, to generate ionotropically stabilized micelles capable of drug delivery into cancer cells. METHODS: optimized mixed HDA/HDAP micelles were prepared and stabilized with TPP. Curcumin was used as a loaded model drug. The prepared nanoparticles were characterized by dynamic light scattering, infrared spectroscopy, transmission electron microscopy, and differential scanning calorimetry. Moreover, their cellular uptake was assessed using flow cytometry and confocal fluorescence microscopy. RESULTS: The prepared nanoparticles were found to be stable under dilution and at high temperatures and to have a size range from 139 nm to 580 nm, depending on pH (4.6-7.4), dilution (up to 100 times), and temperature (25 - 80 °C). They were effective at delivering their load into cancer cells. Additionally, flow cytometry indicated the resulting stabilized micellar nanoparticles to be non-cytotoxic. CONCLUSIONS: The described novel stabilized micelles are simple to prepare and viable for cancer delivery.

The Effect of Weekly 50,000 IU Vitamin D3 Supplements on the Serum Levels of Selected Cytokines Involved in Cytokine Storm: A Randomized Clinical Trial in Adults with Vitamin D Deficiency.

This research aimed to evaluate the effects of high-dose cholecalciferol (VD3) supplements (50,000 IU/week) on selected circulating cytokines associated with cytokine storms in adults with vitamin D deficiency. This clinical trial, based in Jordan, included 50 participants receiving vitamin D3 supplements (50,000 IU/week) for 8 weeks; the exact number was assigned to the control group. Interleukin-6 (IL-6), interleukin-1ß (IL-1ß), interleukin-10 (IL-10), tumor necrotic factor-α (TNF-α), and leptin were measured in serum at baseline and 10 weeks (wash out: 2 weeks). Our results revealed that vitamin D3 supplementation significantly increased the serum levels of 25OHD, IL-6, IL-10, IL-1ß, and leptin compared with baseline. In contrast, the serum level of TNF-α insignificantly increased in the group receiving vitamin D3 supplementation. Although the observations of this trial may refer to a potential negative effect of VD3 supplementation during cytokine storms, further trials are required to clarify the potential benefits of VD3 supplement during cytokine storms.

Co-spray Drying Drugs with Aqueous Polymer Dispersions (APDs)-a Systematic Review.

Aqueous colloidal dispersions of water-insoluble polymers (APDs) avoid hassles associated with the use of organic solvents and offer processing advantages related to their low viscosity and short processing times. Therefore, they became the main vehicle for pharmaceutical coating of tablets and multiparticulates, a process commonly employed using pan and fluidized-bed machinery. Another interesting although less common processing approach is co-spray drying APDs with drugs in aqueous systems. It enables the manufacture of capsule- and matrix-type microspheres with controllable size and improved processing characteristics in a single step. These microspheres can be further formulated into different dosage forms. This systematic review is based on published research articles and aims to highlight the applicability and opportunities of co-spray drying drugs with APDs in drug delivery.

siRNA as a Potential Therapy for COVID-19.

Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV2) is a highly contagious virus causing COVID-19 disease that severely impacted the world health, education, and economy systems in 2020. The numbers of infection cases and reported deaths are still increasing with no specific treatment identified yet to halt this pandemic. Currently, several proposed treatments are under preclinical and clinical investigations now, alongside the race to vaccinate as many individuals as possible. The genome of SARS-CoV2 shares a similar gene organization as other viruses in the Coronaviridae family. It is a positive-sense, single-stranded RNA. This feature suggests that RNA interference (RNAi) is an attractive prophylactic and therapeutic option for the control of this pandemic and other possible future pandemics of the corona viruses. RNAi utilizes the use of siRNA molecules, which are 21-29 nt duplexes RNA molecules that intervene with targeted gene expression in the cytoplasm by a specific mechanism of complementary destruction of mRNA. Previous experience with SARS-CoV and the Middle East respiratory syndrome (MERS) showed that siRNA molecules were effective against these viruses in vitro and in vivo. Moreover, there have been extensive advances in siRNA technology in the past decade from chemistry and target selection considerations; which concluded with the successful approval of two commercial products based on siRNA technology. In addition, the current knowledge of the genome structure and functionality of the corona viruses enables the recognition of conserved sequences to optimize siRNA targeting and avoid viral escape through mutations, either for the current SARS-CoV2 as well as future corona viruses.

The combination of omega-3 fatty acids with high doses of vitamin D3 elevate A1c levels: A randomized Clinical Trial in people with vitamin D deficiency.

PURPOSE: This randomised clinical trial (RCT) was created to assess the influence of 1,25-dihydroxyvitamin D (VD3 ), omega-3 fatty acids (n-3FA) and their combination (D+) on glycated haemoglobin (A1c) levels in Jordanian peoples with vitamin D deficiency (VDD). PARTICIPANTS AND METHODS: This RCT was designed to examine the follow-up (2 months) effect of either 50 000 IU VD3 , 300 mg n-3FA, or the combination of the two supplements on glycated Haemoglobin (A1c) levels in 146 Jordanian women and men with VDD, aged from 25 to 55 years. The eligible participants were randomised into four groups: Control (C); VD3 supplementation (50 000 IU of VD3 was taken weekly) (D3 ); n-3FA supplementation (300 mg of omega-3FA was taken daily) (n-3FA); VD3 and omega-3 supplementation group (D+) with the same protocol as the previous two groups. RESULTS: The combination therapy (n-3FA plus VD3) for 8 weeks significantly increased A1c levels (5.79 ± 0.34 vs 5.41 ± 0.33, P < .001). Tukey test for post hoc comparisons of A1c at follow-up showed that the A1c mean levels were remarkably higher in the D+ study group comparing to the control group (5.78 vs 5.38). CONCLUSION: The intervention of n-3FA alone or in combination with high doses of VD3 may lead to negative effects on glycaemic control or accelerate the insulin resistance's development in susceptible people for diabetes mellitus (type 2).

Hot melt extruded zein for controlled delivery of diclofenac sodium: Effect of drug loading and medium composition.

This study evaluates the potential use of zein as an excipient in hot-melt extrusion for controlled delivery of diclofenac sodium (DS). Mixtures of zein, polyethylene glycol and drug were hot melt extruded and cut into 2 mm extrudates. Extrudates were characterised using differential scanning calorimetry, X-ray powder diffraction and scanning electron microscopy. The drug in the extrudates was found to be in the non-crystalline state, independent of the drug loading. Moreover, the drug release from extrudates was investigated. The release was directly dependent on the drug loading: a controlled and nearly zero-order release was obtained at the lowest drug loading (12.5% w/w), whereas almost immediate release was achieved at higher drug loadings, i.e. 25% and 37.5%. The release was inversely dependent on the ionic strength of the medium. The influence of digestive enzymes on drug release was also studied. Pancreatin, but not pepsin, was found to have a significant influence on the drug release as well as on the microstructure of zein extrudates. These data therefore support the potential use of zein as excipient in hot melt extrusion for controlled release purposes.

The use of design of experiments to develop hot melt extrudates for extended release of diclofenac sodium.

The effect of formulation and processing parameters on processability and release from hot-melt extrusion (HME)-based matrices appears to be API and polymer dependent. Accordingly, the aim of this work was to design an extended-release formulation of diclofenac sodium by using HME technique and design of experiment (DoE). The extrudates were prepared using a vertical lab-scale single screw extruder. A D-optimal design with 16 formulations was employed to evaluate and model the effect of diclofenac sodium, ethyl cellulose and Natrosol L levels on the release profile. The percentage of drug release at 2, 4, 8 and 16 h were the dependent variables. The formulation factors that affect drug release were identified and satisfactorily modeled. The goodness of fit (R2) and goodness of prediction (Q2) parameters obtained for release responses were 0.913 and 0.682 at 2 h, 0.946 and 0.67 at 4 h, 0.942 and 0.658 at 8 h, and 0.892 and 0.673 at 16 h, respectively. The design space of optimal fractions of ethyl cellulose and Natrosol L at various drug levels was successfully constructed by response surface methodology. In conclusion, the DoE approach helped to identify and quantify formulation variables that affect the release of diclofenac sodium from HME-based formulation.

Physicochemical and biological characterization of 1,2-dialkoylamidopropane-based lipoplexes for gene delivery.

Elucidation of the molecular and formulation requirements for efficient lipofection is a prerequisite to enhance the biological activity of cationic lipid-mediated gene delivery systems. To this end, the in vitro lipofection activity of the ionizable asymmetric 1,2-dialkoylamidopropane-based derivatives bearing a single primary amine group as the cationic head group was evaluated. The electrostatic interactions of these cationic lipids with plasmid DNA in serum-free medium were investigated by means of gel electrophoresis retardation and Eth-Br quenching assays. The effect of the inclusion of the helper lipid DOPE in the formulation on these interactions was also considered. The physicochemical properties of these lipids in terms of bilayer fluidity and extent of ionization were investigated using fluorescence anisotropy and surface potential techniques, respectively. The results showed that only the active lipid, 1,2lmp[5], existed in a liquid crystalline state at physiological temperature. Moreover, the extent of ionization of this lipid in assemblies was significantly higher that it's saturated analogues. Inclusion of the helper lipid DOPE improved the encapsulation and association between 1,2lmp[5] and plasmid DNA, which was reflected by the significant boost of lipofection activity of the 1,2lmp[5]/DOPE formulation as compared to the lipid alone. In conclusion, membrane fluidity and sufficient protonation of ionizable cationic lipid are required for efficient association and encapsulation of plasmid DNA and elicit of improved in vitro lipofection activity.

Evaluation of selected biochemical parameters in the saliva of young males using mobile phones.

The biochemical status in the saliva of 12 males before/after using mobile phone has been evaluated. Radio frequency signals of 1800 MHz (continuous wave transmission, 217 Hz modulate and Global System for Mobile Communications [GSM - non-DTX]) with 1.09 w/kg specific absorption rate (SAR) value were used for 15 and 30 min. Cell phone radiation induced a significant increase of superoxide dismutase (SOD); there was a statistically significant effect of talking time on the levels of SOD, F(2, 33) = 8.084, p < 0.05, ω = 0.53. The trend analysis suggests a significant quadratic trend, F(1, 33) = 4.891, p < 0.05; indicating that after 15 min of talking the levels of SOD increased, but as talking time increased the SOD activity started to drop. In contrast to this, there was no statistically significant effect of talking time on the level of salivary albumin, cytochrome c, catalase or uric acid. Results suggest that exposure to electromagnetic radiation may exert an oxidative stress on human cells as evidenced by the increase in the concentration of the superoxide radical anion released in the saliva of cell phone users.

Assessment of oxidant/antioxidant status in saliva of cell phone users.

Hazardous health effects resulting from exposure to radiofrequency electromagnetic radiation (RF-EMR) emitted from cell phones have been reported in the literature. However, the cellular and molecular targets of RF-EMR are still controversial. The aim of this study was to examine the oxidant/antioxidant status in saliva of cell phone users. Saliva samples collected before using a cell phone as well as at the end of 15 and 30 min calls were tested for two commonly used oxidative stress biomarkers: malondialdehyde (MDA) and 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-Oxo-dG). The 8-oxo-dG levels were determined by enzyme-linked immunosorbent (ELISA) competitive assay, while the MDA levels were measured using the OxiSelect MDA adduct ELISA Kit. The antioxidant capacity of the saliva was evaluated using the oxygen radical absorption capacity (ORAC) and the hydroxyl radical averting capacity (HORAC) assays according to the manufacture instructions. The mean 8-oxo-dG and the Bradford protein concentrations (ng/ml and mg/ml, respectively) peaked at 15 min. The levels of HORAC, ORAC and MDA progressively increased with time and reached maximum at 30 min. However, there was no significant effect of talking time on the levels of 8-OxodG and MDA. Similarly, there was no statistically significant effect of talking time on the oxygen and hydroxyl radicals averting capacities, (ORAC) and (HORAC), respectively. These findings suggest that there is no relationship between exposure to radio frequency radiation (RFR) and changes in the salivary oxidant/antioxidant profile.

Understanding and optimizing the dual excipient functionality of sodium lauryl sulfate in tablet formulation of poorly water soluble drug: wetting and lubrication.

PURPOSE: To evaluate and optimize sodium lauryl sulfate (SLS) and magnesium stearate (Mg.St) levels, with respect to dissolution and compaction, in a high dose, poorly soluble drug tablet formulation. METHODS: A model poorly soluble drug was formulated using high shear aqueous granulation. A D-optimal design was used to evaluate and model the effect of granulation conditions, size of milling screen, SLS and Mg.St levels on tablet compaction and ejection. The compaction profiles were generated using a Presster(©) compaction simulator. Dissolution of the kernels was performed using a USP dissolution apparatus II and intrinsic dissolution was determined using a stationary disk system. RESULTS: Unlike kernels dissolution which failed to discriminate between tablets prepared with various SLS contents, the intrinsic dissolution rate showed that a SLS level of 0.57% was sufficient to achieve the required release profile while having minimal effect on compaction. The formulation factors that affect tablet compaction and ejection were identified and satisfactorily modeled. The design space of best factor setting to achieve optimal compaction and ejection properties was successfully constructed by RSM analysis. CONCLUSIONS: A systematic study design helped identify the critical factors and provided means to optimize the functionality of key excipient to design robust drug product.

Functional performance of silicified microcrystalline cellulose versus microcrystalline cellulose: a case study.

BACKGROUND: During the development of a tablet dosage form of an investigational compound, R411, several aspects were identified as critical quality attributes that required optimization. The use of nonsolvent processing prevented the moisture-induced physical changes in the drug product but presented manufacturing challenges related to sticking during compression and slowdown in dissolution after storage at stress conditions. AIM: The aim of this study was to evaluate silicified microcrystalline cellulose (SMCC), microcrystalline cellulose (MCC), and physical mixture of MCC-colloidal silicon dioxide (MCC/CSD at 98:2 ratio) as extragranular compression aids to address the processing and dissolution stability issues of this formulation. METHODS: The compactibility and stickiness upon compression over extended period of time as well as the dissolution of R411 formulations incorporating the aforementioned compression aids were investigated. In addition, the water sorption/desorption properties of these compression aids were determined. RESULTS: All formulations showed comparable compactibility irrespective of the compression aid used. Nevertheless, MCC alone or in a physical mixture with CSD showed sticking of the lower punches, whereas SMCC resulted in clean punch surface during extended compression runs. Furthermore, the three compression aids were compared for their effect on dissolution stability after storage at stress conditions. The formulations containing SMCC provided superior dissolution stability over the other compression aids evaluated in the study. CONCLUSIONS: Novel functionalities of SMCC are presented in terms of sticking prevention while having the most beneficial effect on dissolution stability in R411 formulation.

Physicochemical properties affecting lipofection potency of a new series of 1,2-dialkoylamidopropane-based cationic lipids.

The in vitro transfection activity of a novel series of N,N'-diacyl-1,2-diaminopropyl-3-carbamoyl-(aminoethane) derivatives was evaluated against a mouse melanoma cell line at different +/- charge ratios, in the presence and absence of helper lipids. Only the unsaturated derivative N,N'-dioleoyl-1,2-diaminopropyl-3-carbamoyl-(aminoethane), (1,2lmp[5]) mediated significant increase in the reporter gene level which was significantly boosted in the presence of DOPE peaking at +/- charge ratio of 2. The electrostatic interactions between the cationic liposomes and plasmid DNA were investigated by gel electrophoresis, fluorescence spectroscopy, dynamic light scattering and electrophoretic mobility techniques. In agreement with the transfection results, 1,2lmp[5]/DOPE formulation was most efficient in associating with and retarding DNA migration. The improved association between the dioleoyl derivative and DNA was further confirmed by ethidium bromide displacement assay and particle size distribution analysis of the lipoplexes. Differential scanning calorimetry studies showed that 1,2lmp[5] was the only lipid that exhibited a main phase transition below 37 degrees C. Likewise, 1,2lmp[5] was the only lipid found to form all liquid expanded monolayers at 23 degrees C. In conclusion, the current findings suggest that high in vitro transfection activity is mediated by cationic lipids characterized by increased acyl chain fluidity and high interfacial elasticity.

In vitro lipofection with novel asymmetric series of 1,2-dialkoylamidopropane-based cytofectins containing single symmetric bis-(2-dimethylaminoethane) polar headgroups.

Novel N,N'-diacyl-1,2-diaminopropyl-3-carbamoyl[bis-(2-dimethylaminoethane)] bivalent cationic lipids were synthesized and evaluated for in vitro transfection activity against a murine melanoma cell line. In the absence of the helper lipid DOPE (1,2-dioleoyl-sn-glycero-3-phosphoethanolamine), only the dioleoyl derivative 22 (1,2lb5) elicited transfection activity. The transfection activity of this lipid was reduced when formulated with DOPE. Contrary to that, the dimyristoyl derivative 19 (1,2lb2) mediated no activity when used alone but induced the highest levels of marker gene expression in the presence of DOPE. In an effort to correlate the transfection activity with cationic lipid structures, the physicochemical properties of cationic lipids in isolation and of lipoplexes were studied with surface tensiometry, photon correlation spectroscopy, gel electrophoresis mobility shift assay, and fluorescence techniques. In regard to the lipoplex properties, gel electrophoresis mobility shift assay and EtBr exclusion fluorescence assay revealed that the 1,2lb5 was the only lipid to associate and condense plasmid DNA, respectively. Photon correlation spectroscopy analysis found that 1,2lb5/DNA complexes were of relatively small size compared to all other lipoplexes. With respect to the properties of isolated lipids, Langmuir monolayer studies and fluorescence anisotropy on cationic lipid dispersions verified high two-plane elasticity and increased fluidity of the transfection competent dioleoyl derivative 1,2lb5, respectively. The results indicate that high transfection activity is mediated by cationic lipids characterized by an expanded mean molecular area, high molecular elasticity, and increased fluidity.

Correlation of the physicochemical properties of symmetric 1,3-dialkoylamidopropane-based cationic lipids containing single primary and tertiary amine polar head groups with in vitro transfection activity.

The physicochemical properties of a novel series of symmetric 1,3-dialkylamidopropane-based cationic amphiphiles [M. Sheikh, J. Feig, B. Gee, S. Li, M. Savva, In vitro lipofection with novel series of symmetric 1,3-dialkoylamidopropane-based cationic surfactants containing single primary and tertiary amine polar head groups, Chem. Phys. Lipids 124 (2003) 49-61] were studied by several techniques, in an effort to correlate cationic lipid structure with transfection efficacy. It was found that only the unsubstituted amine and tertiary amine dioleoyl derivatives 1,3lmp5 and 1,3lmt5, respectively, mediated in vitro transfection activity in the absence of helper lipids. This activity pattern was consistent with ethidium bromide fluorescence quenching studies, which indicated that only these two derivatives bound to and efficiently condense plasmid DNA at physiological pH. Dynamic light scattering indicated that lipoplexes made by these two cationic lipids were relatively small particles below 1 microm, in sharp contrast to lipoplexes bigger than 3 microm composed of saturated cationic derivatives. Transmission electron microscopy studies clearly indicated that cationic lipid dispersions made by saturated derivatives form multilamellar tubules at physiological pH. Calorimetric studies showed that cationic amphiphiles with saturated acyl chains longer than 12 carbons exhibit solid-to-liquid crystalline phase transitions above 37 degrees C. In agreement with the microscopy and calorimetry studies, Langmuir film balance experiments indicated that saturated derivatives with hydrophobic chains longer that 12 carbons are not well hydrated and exist at a chain-ordered state at ambient temperature. Calculation of compressibility moduli from monolayer compression isotherms at 23 degrees C suggested that monolayers made by cationic lipids bearing saturated acyl chains are less compressible relative to those of the dioleoyl derivatives 1,3lmp5 and 1,3lmt5. In conclusion, high hydration, increased fluidity and high elasticity of cationic lipid assemblies in isolation, all correlate with high in vitro transfection activity.

Synthesis, in vitro transfection activity and physicochemical characterization of novel N,N'-diacyl-1,2-diaminopropyl-3-carbamoyl-(dimethylaminoethane) amphiphilic derivatives.

A novel series of N,N'-diacyl-1,2-diaminopropyl-3-carbamoyl-(dimethylaminoethane) cationic derivatives was synthesized and screened for in vitro transfection activity at different charge ratios in the presence and absence of the helper lipids DOPE and cholesterol. Physicochemical properties of lipid-DNA complexes were studied by gel electrophoresis, fluorescence spectroscopy and dynamic light scattering. The interfacial properties of the lipids in isolation were studied using the Langmuir film balance technique at 23 degrees C. It was found that only lipoplexes formulated with the dioleoyl derivative, 1,2lmt[5], mediated significant in vitro transfection activity. Optimum activity was obtained with 1,2lmt[5]/DOPE mixture at a +/-charge ratio of 2. In agreement with the transfection results, 1,2lmt[5] was the only lipid found to complex and retard DNA migration as verified by gel electrophoresis. Despite the efficient complexation, no significant condensation of plasmid DNA was observed as indicated by fluorescence spectroscopy measurements. Monolayer studies showed that the dioleoyl derivative 1,2lmt[5] was the only lipid that existed in an all liquid-expanded state with a collapse area and collapse pressure of 59.5 A2 and 38.7 mN/m, respectively. This lipid was also found to have the highest elasticity with a compressibility modulus at monolayer collapse of 80.4 mN/m. In conclusion, increased acyl chain fluidity and high molecular elasticity of cationic lipids were found to correlate with improved transfection activity.