There are approximately 400 known neural disorders some of which being due to a disruption or failure of the blood brain barrier (BBB) such as, for example: meningitis (an inflammation of the meninges or membranes surrounding the brain and spinal cord); epilepsy (chronic or acute seizures caused by inflammation); multiple sclerosis (MS - a disease of the immune system or/and the breaking down of the BBB in a section of the brain or spinal cord); Alzheimer disease (AD - a disease in which amyloid beta contained in blood plasma enter the brain and adhere to the surface of astrocytes); possibly prion and prion-like diseases such as Parkinson disease (PD) and AD; HIV encephalitis (a precursor of HIV-associated dementia in which latent HIV can cross the BBB inside circulating monocytes in the blood stream); and systemic inflammation (sterile or infectious) that may lead to effects on the brain, cause sickness behavior and induce or/and accelerate brain diseases such as MS and PD. There are currently active investigations into treatments for a compromised BBB. As a consequence of the growing aging population, many such neurodegenerative diseases, cancer and infections of the brain will become more prevalent.  Of interest here are those disorders requiring treatment by delivery of drugs across the brain protective barriers. 

I will review the difficulties inherent in the delivery of drugs across the BBB in the treatment oif the above neurological disorders, and discuss the mechanisms for drug targeting both “through” and “behind” the BBB. I will also suggest approaches for the enhancement of drug delivery including physiological approaches, chemical and biological delivery, disruption of the BBB system, the use of molecular Trojan horse systems, and the various nanoparticle and nano delivering devices.  

Statement of the Problem: Acanthamoeba is an opportunistic protozoan pathogen and one of the most prevalent organisms in our natural environment (i.e., air, soil and water). It is recognized to cause fatal brain infection (granulomatous encephalitis) and eye infection (blinding keratitis). Treatments for both infections are problematic because of the amoebic cysts resistance to therapeutic agents. That’s why there is no effective anti-amoebic drug available to date. The purpose of the present study was to evaluate in vitro strength of plants extracts on the viability and biological properties of Acanthamoeba castellanii (T4 genotype) and its cytotoxic effects on human corneal epithelial cells (HCEC). Methodology & Theoretical Orientation: Using HCEC, adhesion, cytotoxicity and amoebicidal, amoebisttic and growth assays were performed. Findings: Normally Acanthamoeba exhibited >90 % binding and >80 % cytotoxicity to HCEC cells which was remarkably inhibited by plant extracts to >70 and 60 % respectively. It was also observed that extracts (ranging from 0.1 to 1.5 mg/ml) exhibited amoebicidal effects, i.e., >50 % of trophozoites were killed at 1.5 mg/ml within 1 h. However, the residual amoeba remained static for quite some time. Furthermore, extracts also inhibited >50 % amoeba numbers up to 7 days during growth assay. Furthermore, plant extracts (1 to 30mg/ml) exhibited amoebicidal effects against Acanthamoeba cysts. Furthermore Acanthamoeba encystment was also inhibited in concentration dependent manner with maximum inhibition at 2µg/ml after 48h. Among all Peganum harmala seed extratcs showed optimal activity against amoeba. Our results confirmed that extracts has toxic effects against both cysts and trophozoite. Conclusion & Significance: Overall, we reported for the first time that selested plant extracts exhibited inhibitory effects on biological properties of Acanthamoeba without any toxic effects on HCEC cells in vitro. Recommendations: Further experiments are required with purified fractions of plant extracts to identify the active ingredients and to elucidate the mechanism of action of the effective compounds both in vitro and in vivo which may provide a new series of chemotherapeutic agents.

Given the vast array of applications for protein-based tools and therapeutics inside cells, there is great interest in developing safe and efficient protein delivery platforms that direct biologics into cells. To date, numerous approaches been investigated to facilitate protein entry into the cytoplasm of cells; however, though each capable of delivering protein cargo into cells to varying degrees, general mechanism-based limitations exist for these platforms. In particular, selectivity and/or efficiency remain elusive features for most platforms owing to their shared nonspecific mode of interaction with membranes. Protein toxins, which use host cell-surface receptors to initiate entry into cells, are attractive vectors to consider given their natural tendency to delivery proteins into specific cells with high efficiency. The paucity of development efforts for toxins as protein delivery vectors stem from early studies, which suggested that delivery was restricted to a select few cargo that were largely unfolded prior to translocation; and that the cargo itself greatly diminished the efficiency of translocation of the system. Through careful engineering of the platform, we show that neither of these assertions is true. We show that the diphtheria toxin platform is capable of delivering proteins that are over 100-kDa in size, and of varying structures and stability with exquisite efficiency. In fact, to our surprise, we found that diphtheria toxin could deliver the hyper-stable passenger protein mCherry, which we calculated to have a melting temperature greater than 90 degrees under the translocation conditions, suggesting that even folded proteins could be delivered into cells. Through a rigorous set of experiments we trace the misleading early results to affects of cargo on the readout of translocation, rather than the efficiency of translocation. We also provide functional evidence that the delivered cargo is functional. Using a-amylase as cargo we show that cytosolic glycogen is degraded in a dose- and time-dependent manner. 

Benzopyrano [4,3-d]-pyrimidine Derivatives (3a-c and 4) were prepared via cyclocondensation of 3-ethoxycarbonyl coumarin derivatives (2a-c) with guanidine hydrochloride and thiourea under reflux. Acetylation of (3b, c) with acetic anhydride provided N-acetyl derivatives (5a, b) of benzopyrano-imidazolo-[2,1-a]-benzopyrano [4,3-d]-pyrimidin-1, 6-dione (6) was obtained by treatment of 2-amino-benzopyrano [4, 3-d]-pyrimidine with ethylchloroacetate. Structures of these compounds were established on basis of IR, ¹H-NMR and MS data. Some of the prepared compounds were evaluated for antimicrobial and antitumor activities.

Statement of the Problem: Breast cancer is a one of the main cause of death worldwide. Chemotherapeutic drugs are used as the frontline treatment of breast and other epithelial malignancies. They can be effective solely or in combination with other therapeutic agents [1]. Tamoxifen (TMX) belongs to a class of non-steroidal triphenylethylene derivatives, and is the first selective estrogen receptor modulator[2]. Herceptin has been widely used for treating breast cancer due to overexpression of human epidermal growth factor receptor-2 (HER2) by cells. Combination of TMX with Herceptin can promotes the therapeutic effectiveness of the anti-cancer delivery system. The role of HER2 in the pathogenesis of breast cancer has been well reported [3].

Human serum albumin (HSA) proved to be a promising carrier for targeted drug delivery to tumor cells, as so many therapeutic agents can be encapsulated within HSA[4]. The coupling of the antibody Herceptin to TMX-HSA nanoparticles takes advantage of the capability of HER2-positive cells to incorporate substances binding to HER2. In our present study, we developed TMX-loaded-HSA nanoparticles with desolvation and NAB-technology which were covalently modified on their surface with thiolated Herceptin with special focus on the effectiveness of antibody conjugation. The goal of this study was to show the efficacy of a nanoparticle albumin bound drug delivery system in comparison with an analogue albumin-based nanoparticles’ active targeting with Herceptin conjugation. For this purpose, four different delivery system were investigated based on their biological activity by cell culture. First, tamoxifen-loaded albumin nanoparticles via high pressure homogenizer, as a model containing unchanged albumin secondary structure; second, the tamoxifen-loaded nanoparticles via desolvation method as a model with dramatically changed albumin structure; and finally, the Herceptin-conjugated nanoparticles prepared by these two. 

Ethanol used for enhancing water miscibility of the essential oils for fish anesthesia provides undesirable side effects to the fish. The aim of this study was to develop a water dispersible formulation of Alpinia galanga oil (AGO) self-nanoemulsifying drug delivery systems (SNEDDS) in order to minimize the amount of ethanol in the formulation and to investigate the effects of the AGO and AGO-SNEDDS for fish anesthesia. Response surface methodology was used to investigate how excipients affect the droplet size on AGO-SNEDDS formation. The fish anesthetic activity of AGO-SNEDDS with different droplet sizes was evaluated by the time it took for zebrafish (Danio rerio) to go into surgical anesthesia stage which fish stopped swimming activity, showed loss of equilibrium and responsiveness and subsequent recovery. The predicted contour plots of droplet size indicated that Cremophor RH 40 provided smaller droplet size than Tween 80. The goodness of model fitting (R² > 0.89), prediction power (Q² > 0.72), and the droplet size values between prediction and real measurement showed similar values (%error <10%). Therefore, these models had a good prediction power. Cremophor RH 40, Miglyol 812:Capmul MCM EP=1:1, and AGO concentrations showed the most influential variables affecting the droplet size. The droplet size plays an important role in fish anesthesia. The larger droplet required longer time to take fish to enter surgical anesthesia stage. SNEDDS3 with a droplet size around 200 nm sedated the fish into the anesthetic stage within 270 sec, significantly slower than SNEDDS1 and SNEDDS2 (218 and 212 sec) with droplet sizes around 60 and 110 nm (p < 0.03). All formulations had significantly increased anesthetic activity compared to AGO in an ethanolic solution. In conclusion, the SNEDDS are promising nanodelivery systems of AGO for anesthetic use in zebrafish.

Breast cancer is arguably the most common cancer faced by females today and the second most common cause of death in women in the world.  Indeed, this illness has garnered much attention in the field of pharmacology research.  Modern chemotherapeutic anticancer treatments have come a long way in the fight against breast cancer, thus bringing science closer to a cure.  However, the nature of these drugs is to attack both cancerous and non-cancerous cells at the same time.  With this current approach, a patient’s health, in addition to the cancer, can succumb to chemotherapies. To counter this problem, and increase the efficacy of cancer treatment, methods to customize therapeutic anti-cancer drugs have emerged in the form of targeted drug delivery systems.  In our studies, we present a method of a drug delivery using magnetic polyurethane.   Here, we describe a biocompatible magnetic polymer that can be used to direct chemotherapeutic drugs to cancerous regions in a body using an external magnet.  We show how a co precipitation method with magnetic nano-particles (MNPs) followed by a silica coating process and an in situ polymerization yields the magnetic polyurethanes used in this study. Verification of synthesis for the drug carrier is shown using the characterization techniques of scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), thermal gravimetric analysis (TGA), and a vibrating sample magnetometer (VSM).  The efficiency with drug loading and release of chemotherapeutic medications to the synthesized magnetic polyurethanes is monitored using an HPLC-UV detector.  Our findings present a new biocompatible drug delivery system with a high capacity for loading and directing tow various chemotherapeutic drugs simultaneously to cancer sites with little to no toxicity to the surrounding non-cancerous cells.