Advanced Pharmaceutical Bulletin
Volume:7 Issue: 1, Apr 2017

Liposomes, due to their various forms, require further exploration. These structures can deliver both hydrophilic and hydrophobic drugs for cancer, antibacterial, antifungal, immunomodulation, diagnostics, ophtalmica, vaccines, enzymes and genetic elements. Preparation of liposomes results in different properties for these systems. In addition, based on preparation methods, liposomes types can be unilamellar, multilamellar and giant unilamellar; however, there are many factors and difficulties that affect the development of liposome drug delivery structure. In the present review, we discuss some problems that impact drug delivery by liposomes. In addition, we discuss a new generation of liposomes, which is utilized for decreasing the limitation of the conventional liposomes.

Many studies have focused on how drugs are formulated in the sol state at room temperature leading to the formation of in situ gel at eye temperature to provide a controlled drug release. Stimuli-responsive block copolymer hydrogels possess several advantages including uncomplicated drug formulation and ease of application, no organic solvent, protective environment for drugs, site-specificity, prolonged and localized drug delivery, lower systemic toxicity, and capability to deliver both hydrophobic and hydrophilic drugs. Self-assembling block copolymers (such as diblock, triblock, and pentablock copolymers) with large solubility variation between hydrophilic and hydrophobic segments are capable of making temperature-dependent micellar assembles, and with further increase in the temperature, of jellifying due to micellar aggregation. In general, molecular weight, hydrophobicity, and block arrangement have a significant effect on polymer crystallinity, micelle size, and in vitro drug release profile. The limitations of creature triblock copolymers as initial burst release can be largely avoided using micelles made of pentablock copolymers. Moreover, formulations based on pentablock copolymers can sustain drug release for a longer time. The present study aims to provide a concise overview of the initial and recent progresses in the design of hydrogel-based ocular drug delivery systems.

Angiogenesis plays an essential role in rapid growing and metastasis of the tumors. Inhibition of angiogenesis is a putative strategy for cancer therapy. Endostatin (Es) is an attractive antiangiogenesis protein with some clinical application challenges including; short half-life, instability in serum and requirement to high dosage. Therefore, production of recombinant endostatin (rEs) is necessary in large scale. The production of rEs is difficult because of its structural properties and is high-cost. Therefore, this review focused on the different expression systems that involved in rEs production including; mammalian, baculovirus, yeast, and Escherichia coli (E. coli) expression systems. The evaluating of the results of different expression systems declared that none of the mentioned systems can be considered to be generally superior to the other. Meanwhile with considering the advantages and disadvantage of E. coli expression system compared with other systems beside the molecular properties of Es, E. coli expression system can be a preferred expression system for expressing of the Es in large scale. Also, the molecular bioengineering and sustained release formulations that lead to improving of its stability and bioactivity will be discussed. Point mutation (P125A) of Es, addition of RGD moiety or an additional zinc biding site to N-terminal of Es , fusing of Es to anti-HER2 IgG or heavy-chain of IgG, and finally loading of the endostar by PLGA and PEGPLGA nanoparticles and gold nano-shell particles are the effective bioengineering methods to overcome to clinical changes of endostatin.

Phytoconstituents have been utilized as medicines for thousands of years, yet their application is limited owing to major hurdles like deficit lipid solubility, large molecular size and degradation in the gastric environment of gut. Recently, phospholipid-complex technique has unveiled in addressing these stumbling blocks either by enhancing the solubilizing capacity or its potentiating ability to pass through the biological membranes and it also protects the active herbal components from degradation. Hence, this phospholipid-complex-technique can enable researchers to deliver the phytoconstituents into systemic circulation by using certain conventional dosage forms like tablets and capsules. This review highlights the unique property of phospholipids in drug delivery, their role as adjuvant in health benefits, and their application in the herbal medicine systems to improve the bioavailability of active herbal components. Also we summarize the prerequisites for phytosomes preparation like the selection of type of phytoconstituents, solvents used, various methods employed in phytosomal preparation and its characterization. Further we discuss the key findings of recent research work conducted on phospholipid-based delivery systems which can enable new directions and advancements to the development of herbal dosage forms.

In the present study the incompatibility of FLM (fluvoxamine) with lactose in solid state mixtures was investigated. The compatibility was evaluated using different physicochemical methods such as differential scanning calorimetry (DSC), Fouriertransform infrared (FTIR) spectroscopy and mass spectrometry.
Non-Isothermally stressed physical mixtures were used to calculate the solid– state kinetic parameters. Different thermal models such as Friedman, Flynn–Wall–Ozawa (FWO) and Kissinger–Akahira–Sunose (KAS) were used for the characterization of the drug-excipient interaction.
Overall, the incompatibility of FLM with lactose as a reducing carbohydrate was successfully evaluated and the activation energy of this interaction was calculated.
In this research the lactose and FLM Maillard interaction was proved using physicochemical techniques including DSC and FTIR. It was shown that DSC- based kinetic analysis provides fast and versatile kinetic comparison of Arrhenius activation energies for different pharmaceutical samples.

Introducing the effect of RNAi in fungi to downregulate essential genes has made it a powerful tool to investigate gene function, with potential strategies for novel disease treatments. Thus, this study is an endeavor to delve into the silencing potentials of siRNA on cyp51A and MDR1 in voriconazole-resistant Aspergillus flavus as the target genes.
In this study, we designed three cyp51A-specific siRNAs and three MDR1- specific siRNAs and after the co-transfection of siRNA into Aspergillus flavus, using lipofectamine, we investigated the effect of different siRNA concentrations (5, 15, 25, 50nM) on cyp51A and MDR1 expressions by qRT-PCR. Finally, the Minimum Inhibitory Concentrations (MICs) of voriconazole for isolates were determined by broth dilution method.
Cyp51A siRNA induced 9, 22, 33, 40-fold reductions in cyp51A mRNA expression in a voriconazole-resistant strain following the treatment of the cells with concentrations of 5, 15, 25, 50nM siRNA, respectively. Identically, the same procedure was applied to MDR1, even though it induced 2, 3, 4, 10-fold reductions. The results demonstrated a MIC for voriconazole in the untreated group (4μg per ml), when compared to the group treated with cyp51A-specific siRNA and MDR1-specific siRNA, both at concentrations of 25 and 50nM, yielding 2μg per ml and 1μg per ml when 25 nM was applied and 2μg per ml and 0.5μg per ml when the concentration doubled to 50 nM.
In this study, we suggested that siRNA-mediated specific inhibition of cyp51A and MDR1 genes play roles in voriconazole-resistant A.flavus strain and these could be apt target genes for inactivation. The current study promises a bright prospect for the treatment of invasive aspergillosis through the effective deployment of RNAi and gene therapy.

Nanostructured lipid carriers (NLCs) composed of solid lipid and oil are a new generation of lipid nanoparticles which have exhibited some merits over traditional used lipid nanoparticles in fortifying food and beverages and nutraceuticals delivery systems such as liposomes and solid lipid nanoparticles.
In this study, Precirol and Compritol as solid lipids, Miglyol and Octyloctanoat as liquid lipids, Tween80, Tween20 and Poloxamer407 as surfactants were used to prepare vitamin D3-loaded NLC dispersion using hot homogenization method. The particle size and size distribution for all formulations were evaluated by immediately after production and during a storage period of 60 days.
The Precirol-based NLC showed superiority over Compritol-based NLC in the point of physical stability. Results clearly suggested that an optimum concentration of 3% of Poloxamer407 or 2% of Tween20 was sufficient to cover the surface of nanoparticles effectively and prevent agglomeration during the homogenization process. Octyloctanoat was introduced for the first time as a good substituent for Miglyol in the preparation of NLC formulations. The vitamin D3 Intestinal absorption enhanced by the incorporating in NLCs.
It was concluded that NLC showed a promising approach for fortifying beverages by lipophilic nutraceuticals such as vitamin D.

The dramatic increase in stress ulcer prophylaxis (SUP) prescribing patterns over the past several years has raised concerns regarding to their appropriate utilization. This prospective study attempted to evaluate the trend of adherence to stress ulcer prophylaxis from admission until discharge in non- Intensive care unit (ICU) setting. Additionally, we attempted to find those variables associated with appropriate SUP administration.
Data collection was performed prospectively to evaluate 195 randomly selected adult patients who received SUP or had indication for that in non-ICU wards of one of the largest referral center in Iran, during 6 months. Adherence was studied according to widely accepted American Society of Health system Pharmacists (ASHP) guideline. Univariate and multivariate logistic regression was also performed to detect associations related to misuse of SUP.
We recognized total inappropriate use of SUP upon admission, during hospitalstay and at discharge were somewhat identical at different time points (61%, 80% and 77.4 respectively). On the other hand, since small number of patients experienced SUP underutilization, unfortunately this was not possible to elucidate factors that may have effect on this flawed behavior. However, increasing age was identified to be significant variable in SUP overutilization.
This prospective study highlighted inappropriate overutilization of SUP within non-critically ill patients and found factors which predicted this behavior. Adherence during hospital stay was also calculated for the first time in this study, which was related to SUP adherence upon hospital admission.

Mammalian target of rapamycin (mTOR)is important in hematopoiesis and affect cell growth,differentiation and survival. Although previous studies were identified the effect of cytokines on the mononuclear cells development however the cytokines effect on mTOR in cord blood mononuclear cells was unclear. The aim of this study was to evaluate mTOR expression in cord blood mononuclear and cord blood stem cells (CD34 cells) in culture conditions for lymphoid cell development.
Isolation of The mononuclear cells (MNCs) from umbilical cord blood were done with use of Ficollpaque density gradient. We evaluated cultured cord blood mononuclear and CD34 cells in presece of IL2, IL7 and IL15 at distinct time points during 21 days by using flow cytometry. In this study, we presented the role of IL2, IL7 and IL15 on the expression of mTOR in cord blood cells.
mTOR expression were increased in peresence of IL2, IL7 and IL15 in day 14 and afterword reduced. However in persence of IL2 and IL15 expression of mTOR significantly reduced. mTOR expression in CD34 cells decreased significantly from day7 to day 21 in culture.
cytokines play important role in mTOR expression during hematopoiesis and development of cord blood mononuclear cells.

Study on gold based therapeutic agents for cancer cells deracination has become under scrutiny in recent years owing to effective treatments are not available for rapidly progressive cancers. The aim of present study was to examine efficiency of radioactive 198Au/PAMAMG4 and non-radioactive 197Au/PAMAMG4 nancomposites against 4T1 and MCF7 breast cancer cell lines.
The PAMAMG4 dendrimer was treated with the gold anions and then, the mixture was chemically reduced by NaBH4. Prepared 197Au/PAMAMG4 was bombarded by thermal neutrons in the Tehran Research Reactor to 198Au/PAMAMG4 be produced. Prepared nanocomposites were characterized by means of FT-IR, 1H NMR, Zeta-potential measurements, TEM and EDX analyses. The radionuclidic purity of the 198Au/PAMAMG4 solution was determined using purity germanium (HPGe) spectroscopy and its stability in the presence of human serum was studied. In vitro studies were carried out to compare toxicity of PAMAMG4, 197Au/PAMAMG4 and 198Au/PAMAMG4 towards 4T1 and MCF7 cancerous cells and C2C12 normal cell lines.
Characterization results exhibited that invitro agents, 197Au/PAMAMG4 and 198Au/PAMAMG4, were synthesized successfully. Cells viability after 24 h, 48 h, and 72 h incubation, using MTT assay showed that the toxicity of 198Au/PAMAMG4 is significantly superior in comparison with 197Au/PAMAMG4 and PAMAMG4. Furthermore, the toxicity of 198Au/PAMAMG4 was higher on cancerous cells especially in higher level of concentrations after 72 hours (P In the current study, the preparation of 197Au/PAMAMG4 and 198Au/PAMAMG4 is described and the cytotoxic properties of them against the MCF7, 4 1 cancerous cells and C2C12 normal cells were evaluated using MTT assay.

Suitable ratio of essential fatty acids has an important role in maintaining good health. There is no pure oil with an ideal fatty acid composition and oxidative stability. The main goal of the present study was to evaluate the physical, chemical and nutritional properties of oil obtained by blending flaxseed oil as a rich source of ω3 fatty acids with sesame and olive oils.
Three different ratios (65:30:5, 60:30:10 and 55:30:15) were prepared using olive, sesame and flaxseed oils. These mixtures were stored at 4°C and 24°C and their quality and physicochemical properties were determined by measuring the fatty acid composition, phenolic compound, peroxide, anisidine values and schaal tests.
Fatty acid composition indicated that adding 10% and 15% flaxseed oil into blends have suitable ratio of essential fatty acids. The sample which contained 5% flaxseed oil had the highest phenolic content among treatments and these compounds showed a significant decrease during storage. A significant increase (p Blending sesame and olive oils with flaxseed oil produced oil blends with a good balance of essential fatty acids. Although peroxide and anisidine values increased during storage of the oil blends; the blends were of a good quality for home and industrial use.

Solubilizers play an important role in dissolution of pharmacological ingredients and should properly dissolve the active principle(s) while preserving its activities. This study investigated the effect of starch, gelatin, methylcellulose and polyvinylpyrrolidone 10000 in the preservation of the androgenic activity of the methanol extract Basella alba (MEBa).
Different groups of male albino rats were orally given the MEBa (1 mg/kg) dissolved into either 1% gelatin (1% gel), %1 methylcellulose (1% MC) and 1% polyvinylpyrrolidone 10000 (1% PVP 10000) or 2% starch solution (2% SS) for 30 days. Thereafter, animals were sacrificed and serum testosterone and creatinine levels as well as alanine aminotransferase (ALT) activity determined. Vital and reproductive organs were dissected out and weighed, while liver thiobarbituric acid reactive substances (TBARS) and glutathione levels were determined.
Different treatments did not affect the animal body and organ weights. The MEBa stimulatory effect on testosterone production was preserved with 2% SS and 1% PVP 10000 as vehicles. Increased liver glutathione and TBARS levels were also observed in the animals fed with the MEBa dissolved in 2% SS and 1% Gel, respectively, while other biochemical parameters remained unchanged.
Starch and polyvinylpyrrolidone 10000 stand as good preservation agents for MEBa androgenic activity, with starch exhibiting additional antioxidant activity through increase of glutathione levels.

Zygophyllum fabago L. (Z. fabago) is a widespread perennial herb which is used as a medicinal plant in traditional medicine of Iran, Turkey and China. The present study was a survey on phytochemical constituents and biological activities of this plant.
Methanolic extract of the roots was fractionated over a C-18 pre-packed cartridge (Sep-pak) and chromatographic separation was performed on a reversed-phase preparative HPLC. Structural elucidation of the isolated compounds was carried out using UV, 1H-NMR and 13C-NMR spectral analyses. Furthermore, the chemical compositions of the essential oil of the aerial parts were identified by GC-MS analysis. Antiproliferative and antioxidant activities of all extracts from aerials were determined by MTT and DPPH assays, respectively.
Phytochemical investigation on the plant roots led to the isolation and identification of two the 60% methanol-water Sep-pak fraction, a prenylated flavone glycoside, 6-C prenyl- 7-O-[ β -D-4'''-O-acetyl-glucopyranosyl-(1'''→2'')-β-D-glucopyranosyl] apigenin, which was named as a Zygocaperoside and also, other flavonoid, was named as the Isorhamnetin -3 O glucoside. None of the extracts showed antiproliferative effect against cancerous cells. However, among the extracts, methanolic extract indicated antioxidant activity. Moreover, essential oils of flowers and leaves of plant have high amounts of sesquiterpene hydrocarbons and diterpenoides.
The results of present study introduce Z. fabago roots as a new source of flavonoid glycosides and suggest it as an appropriate candidate for further pharmacological studies.

This study observed the effect of garlic extracts and amoxicillin against an induced staphylococcal infection model. MIC and MBC were also obtained for aqueous extracts of Allium sativum (Asa) and Allium tuberosum (Atu) against Staphylococcus aureus penicillin-sensitive (PSSA - ATCC 25923) and MRSA (ATCC 33592).
Granulation tissues were induced in the back of 205 rats. After 14 days, 0.5 mL of 108 CFU/mL of PSSA or MRSA were injected inside tissues. After 24h, animals were divided: G1 (Control) – 0.5 mL of NaCl 0.9%; G2 – Asa 100 mg/kg or 400mg/kg; G3 – Atu 100 mg/kg or 400 mg/kg; G4 – amoxicillin suspension 50 mg/kg, considering PSSA infection; and G5 (Control) - 0.5 mL of NaCl 0.9%; G6 – Asa 400mg/kg; G7 – amoxicillin 50 mg/kg; and G8 - Asa 400 mg/kg amoxicillin 50 mg/kg for MRSA. All treatments were administered P.O. every 6h. Animals were killed at 0, 6, 12 and 24h. Samples were spread on salt-mannitol agar. Colonies were counted after 18 h at 37 °C. Atu was not able to inhibit or kill PSSA and MRSA. Considering Asa, MIC and MBC against PSSA were 2 mg/mL and 4 mg/mL, respectively; and 16 mg/mL and 64 mg/mL against MRSA.
No effect was observed in vivo for control, Asa 100 mg/kg and Atu 100 mg/kg, while amoxicillin, Atu 400 mg/kg and Asa 400 mg/kg decreased PSSA counts in all-time points. No effect of any group against MRSA was observed at any time.
Thus, A. sativum and A. tuberosum were able to reduce PSSA infection, but not MRSA infection.

Mesenchymal stem cells (MSCs) have been introduced for cell therapy strategies in osteoarthritis (OA). Despite of their capacity for differentiation into chondrocyte, there are some evidences about their life-threatening problem after transplantation. So, some researchers shifted on the application of stem cells conditioned medium. The goal of this study is to evaluate whether Wharton's jelly derived stem cell conditioned medium (WJSCs- CM) can enhance the gene expression profile by chondrocytes in monolayer and mass culture systems.
Conditioned medium was obtained from WJSCs at fourth passage. Isolated chondrocytes were plated at density of 1×106 for both monolayer and high density culture. Then cells in both groups were divided into control (received medium) and experiment group treated with WJ-CM for 3 and 6 days. Samples were prepared to evaluate gene expression profile of collagen II, aggrecan, cartilage oligomeric matrix protein (COMP) and sox-9 using real-time RT-PCR.
After 3 days, Chondrocytes treated with WJSCs-CM expressed significantly higher level of genes compared to the control group in both culture systems. After 6 days, the expression of genes in monolayer cultivated chondrocytes was decreased but that of the mass culture were up-regulated significantly.
WJ-SCs-CM can increase the expression of cartilage-specific genes and can be introduced as a promoting factor for cartilage regeneration.

PLGA nanoparticles (NPs) have been extensively investigated as carriers of different drug molecules to enhance their therapeutic effects or preserve them from the aqueous environment. Streptokinase (SK) is an important medicine for thrombotic diseases.
In this study, we used electrospray to encapsulate SK in PLGA NPs and evaluate its activity. This is the first paper which investigates activity of an electrosprayed enzyme. Effect of three input parameters, namely, voltage, internal diameter of needle (nozzle) and concentration ratio of polymer to protein on size and size distribution (SD) of NPs was evaluated using artificial neural networks (ANNs). Optimizing the SD has been rarely reported so far in electrospray.
From the results, to obtain lowest size of nanoparticles, ratio of polymer/enzyme and needle internal diameter (ID) should be low. Also, minimum SD was obtainable at high values of voltage. The optimum preparation had mean (SD) size, encapsulation efficiency and loading capacity of 37 (12) nm, 90% and 8.2%, respectively. Nearly, 20% of SK was released in the first 30 minutes, followed by cumulative release of 41% during 72 h. Activity of the enzyme was also checked 30 min after preparation and 19.2% activity was shown.
Our study showed that electrospraying could be an interesting approach to encapsulate proteins/enzymes in polymeric nanoparticles. However, further works are required to assure maintaining the activity of the enzyme/protein after electrospray.

Implication of protein-protein interactions (PPIs) in development of many diseases such as cancer makes them attractive for therapeutic intervention and rational drug design. RON (Recepteur d’Origine Nantais) tyrosine kinase receptor has gained considerable attention as promising target in cancer therapy. The activation of RON via its ligand, macrophage stimulation protein (MSP) is the most common mechanism of activation for this receptor. The aim of the current study was to perform in silico alanine scanning mutagenesis and to calculate binding energy for prediction of hot spots in proteinprotein interface between RON and MSPβ chain (MSPβ).
In this work the residues at the interface of RON-MSPβ complex were mutated to alanine and then molecular dynamics simulation was used to calculate binding free energy.
The results revealed that Gln193, Arg220, Glu287, Pro288, Glu289, and His424 residues from RON and Arg521, His528, Ser565, Glu658, and Arg683 from MSPβ may play important roles in protein-protein interaction between RON and MSP.
Identification of these RON hot spots is important in designing anti-RON drugs when the aim is to disrupt RON-MSP interaction. In the same way, the acquired information regarding the critical amino acids of MSPβ can be used in the process of rational drug design for developing MSP antagonizing agents, the development of novel MSP mimicking peptides where inhibition of RON activation is required, and the design of experimental site directed mutagenesis studies.

A new sensitive sensor was fabricated for the determination of ivabradine hydrochloride (IH) based on modification with multiwalled carbon nanotubes using sodium dodecyl sulfate as micellar medium to increase the sensitivity.
The electrochemical behavior of IH was studied in Britton-Robinson buffer (pH: 2.0-11.0) using cyclic and differential pulse voltammetry.
The voltammetric response was linear over the range of 3.984 x 10-6-3.475 x 10 5 mol L-1. The limits of detection and quantification were found to be 5.160 x 10-7 and 1.720 x 10-6 mol L-1, respectively.
This method is suitable for determination of IH in tablets and plasma.

This study aims to prepare a novel, natural nanoparticle (NP) as a drug carrier, which also has inherent therapeutic effects.
Pistacia khinjuk gum NPs were prepared and Response surface methodology (RSM) was used for statistical analysis of data and optimizing the size of NPs.
NPs were in the range of 75.85–241.3 nm. The optimization study was carried out, and an optimized size (70.86nm) was obtained using DMSO as a solvent. The volume of the organic phase was 111.25μl, and the concentration of gum was 1% w/v. The cell viability assay was performed on the pure gum and NPs toward β-TC3, MCF7, and HT29 cell lines. It was observed that NPs have higher cytotoxic activity in comparison with pure gum, and that the IC50value was achieved at 1% of NPs in β-TC3 cells. The obtained NPs demonstrated antibacterial activity against two bacterial strains (Pseudomonas aeruginosa and Staphylococcus aureus).
Altogether, according to the obtained results, these NPs with inherent cytotoxicity and antibacterial activity are an attractive carrier for drug delivery.