Tumor liberated protein (TLP) has been previously described as a TAA (complex) present in the sera from lung cancer patients with early stage disease.
Since early detection improves overall survival in lung cancer, identification of screening biomarkers for patients at risk for the development of this disease represents an important target. Starting from the peptide epitope RTNKEASI previously isolated from TLP complexes, we generated a rabbit anti-RTNKEASI serum. This antiserum detected and immunoprecipitated a 55kDa protein band in the lysate of the lung cancer cell line A549. This protein band was identified as aldehyde dehydrogenase isoform 1A1 through mass spectrometry, revealing the molecular nature of at least one component of the previously described TLP complex. Next, we screened a cohort of 29 lung cancer patients (all histologies), 17 patients with non-neoplastic lung pathologies and 9 healthy donors for the presence of serum ALDH1A1 and global serum ALDH by enzyme-linked immunosorbent assay. This analysis indicated that the presence of ALDH was highly restricted to patients with lung cancer. Interestingly, the global ALDH test detected more lung cancer patients compared to the ALDH1A1-specific test, suggesting that other ALDH isoforms might add to the sensitivity of the assay. Our data suggest that ALDH levels may therefore be evaluated as part of a marker panel for lung cancer screening.
Finally, the ability of the immune system to recognize a TAA, enables the development of a vaccine approach for preventive and therapeutic application and represents a main target of this field of research.
Tubulin inhibitors (e.g., paclitaxel) are widely used chemotherapeutic drugs in treating various types of cancers, including prostate cancer, breast cancer, lung cancer, ovarian cancer, and pancreatic cancer. However, their clinical efficacy is often limited by the ABC-transporter mediated drug efflux and non-specific tissue distribution, leading to dose limiting toxicity. In addition, existing tubulin inhibitors cannot be taken orally and must be administered by intravenous route in clinical facilities.
We have discovered a new generation of orally bioavailable tubulin inhibitors that: 1) target the colchicine binding site in tubulin and have broad spectrum of potent anticancer activity; 2) effectively circumvent major drug resistance mechanisms that hinder the clinical efficacy with existing tubulin inhibitors; 3) are orally bioavailable and have excellent drug-like properties; and 4) are efficacious against both drug sensitive and drug resistant multiple types of tumors in vivo. We have solved the X-ray crystal structures for many of these compounds to confirm their direct binding to tubulin. We have also developed various nanoparticle formulations for these agents and showed that these targeted drug delivery approaches can improve the anticancer efficacy for these tubulin inhibitors. Currently the leading compound is under Phase I clinical trials for metastatic prostate cancer. This work is mainly supported by NIH grants R01CA148706 and the UTCoP Drug Discovery Center.
To be updated soon...
Drug discovery and development advances in the last decades allowed to find a treatment for many diseases, but children have been poorly benefited from these progresses since most of the new medicines have been developed for adults. Nowadays around 50% of the medicines addressed to children and young patients have been tested specifically for them. Even if paediatricians routinely give drugs to children in a ‘off-label’ prescription, researchers have demonstrated that children often do not respond to medications in the same way as adults.
Until recently, it was assumed that children reacted to medications as ‘small adults’ adjusting dosage to account for a smaller body, with the presuming that clinical effects would be equivalent to those observed in adults. However, the development from preterm to a fully mature individual is a complex process and certain specific disorders are unique to children or occur in children differently.
This “special” population is indeed made of several different age groups with different and variable characteristics. For this reason, it is important to develop better medicines for children and young patients by studying them specifically and/or using extrapolation methods whenever possible.
The EPTRI project arises from the need to find answers to the serious lack of medicines for children in EU and worldwide. It is coordinated by Consorzio per Valutazioni Biologiche e Farmacologiche (CVBF) and is aimed to design the framework for a new Research Infrastructure (RI) to cover the technological gaps in the paediatric research in early stage drug development phases as in the developmental pharmacology and paediatric formulation science to be translated into paediatric use of medicines.
EPTRI activities are focused on the following main scientific domains: paediatric medicines discovery, biomarkers and biosample, developmental pharmacology and paediatric medicine formulations and medical devices.
In order to design the research infrastructure a survey was conducted to map the expertise and competences of the research units in Pan-European countries and to identify high-end apparatus (equipment, resources, research facilities, platforms, methodologies and experimental settings) which can be accessible through the new EPTRI Infrastructure for performing research in a paediatric context.
More than 250 research units answered the survey 63 research units declaring to be able to provide services such as pharmacometrics in the different age and ethnic groups and diseases, paediatric pharmacogenetics, sensitive biofluids assays development of analytical methods adapted to low volumes, markers test and techniques to explore variability in drug disposition and effects, and facilities such as placental platform for drug evaluation.
46 research units answered declared to be able to provide paediatric formulations and drug delivery design for enteral and non-enteral routes of administration, taste masking technologies biopharmaceutics, safety of excipients, palatability assessment in vitro taste tool and in vivo assessment and of these research units, 24 declared to work on medical devices.
Treatment of disorders of the central nervous system such as malignant brain tumours is hindered by the blood-brain barrier (BBB), whose role is to protect the brain from undesirable microorganisms, substances or drugs contained in the blood. Nanotechnology represents a strategy of brain-targeted drug delivery that may overcome this barrier, along with the advantage of carrying and delivering a wide selection of drugs. Chitosan is an appropriate choice of material to formulate nanoparticles because of its advantageous properties. The structure of chitosan has been modified using moieties resembling the structure of alkylglycerols, as these have been previously shown to increase the hydrophobicity profile of nanoparticles, hence improving their crossing through the BBB. We report on the synthesis, characterization, formulation and evaluation of these new materials. Characterization data shows the structure of chitosan was successfully modified. These materials were then further formulated as nanoparticles by means of ionic gelation, producing particles with sizes within suitable ranges for delivery beyond the BBB (i.e. within nano range). Formulations were tested against an in-house in vitro model. Collectively, results indicate that the performed functionalization of chitosan leads to new materials which can be used to formulate better nanoparticulate systems for the delivery of drugs beyond the BBB, when compared to pristine chitosan.
Cancer is a complex disease. Several drug delivery platforms are under development in academia and in industry to deliver therapeutic molecules and new chemical entities to the targeted site in body. Amongst them, nanotechnology both at molecular and supramolecular level is a leading platform and can help to image, detect and treat diseases effectively. Surface modification of nanoparticles by coating or anchoring their surface with special markers, materials, peptide, proteins, antibodies or antigens add extra feature and thereby enhancing their performance. Till date scientists are struggling to understand the complete mechanism and fate of nanoparticles in body and make it safer. The theme of this presentation is to discuss advances in nanoparticles and therapies that are currently used and under development, along with a short discussion on regulatory activities in this direction.
A significant proportion of new drug candidates possesses poor solubility and/or membrane permeability. Several promising techniques have been developed to overcome these disadvantageous properties, including different fiber formation methods. The electrospinning and high-speed rotary spinning are the most commonly used spinning techniques for fiber formation. The fiber properties (high specific surface area, porosity, the possibility of controlling the crystalline-amorphous phase transitions of the loaded drugs) enhance the dissolution rate and apparent solubility of actives and thus their rate and extent of absorption. The hydrophilic polymer-based drug-loaded electrospun nanofibrous orally dissolving webs are promising candidates for rapid drug release, which is due to the advantageous morphological and physical-chemical features of the system. They are also capable of controlled drug delivery over time for local or systemic drug administration. The solubility of the polymer, the fiber diameter and the fiber structure are the primary parameters affecting drug release. In the case of small molecules, developments focus mostly on overcoming the unfavourable physicochemical feature of the active agents. However, the physical and chemical stability of these systems has not yet been thoroughly investigated and thus poses a challenge in their development. Since the stability of these systems is a crucial issue, its sensitive and non-destructive tracking could be of great practical relevance. The presentation focuses on the unique structure-derived advantages of the nanofibrous systems concerning their functionality-related characteristics (drug release, mechanical properties and stability features). The combination of state-of-art morphology (SEM, AFM imaging) and solid-state characterisation methods (ssNMR, PALS, FTIR spectroscopy, XRD, DSC) provide useful information about the state of the drugs embedded to fibers, distinguishing the amorphous solid solutions from the amorphous solid dispersions. The latter is of significant impact on their therapeutic applicability.
Recent developments in nanotechnology has provided new tools for cancer therapy and diagnosis. Among other nanomaterial systems, gold nanoparticles are being used as radiation dose enhancers and anticancer drug carriers. Our studies show that size, shape, and surface properties of NPs can play a major role in their interaction with tumor cells. We have a developed a comprehensive research platform which includes monolayer cell models, multilayer cell models (tissue like models), and in vivo animal models to test the therapeutic efficacy of gold mediated sensitization. It is important to test NP formulations at all three above mentioned levels to optimize their use in future clinical applications. For example, our previous work at monolayer level showed that NPs of diameter 50 nm had the highest cell uptake among the size range 10-100 nm. However, at tissue-level NPs of diameter lower than 50 nm showed the highest tissue penetration. Once these NPs leave the tumor blood vessels, it important that they should be able to penetrate tumor tissue deeper. Hence, we used smaller NPs for our in vivo studies. We were able to achieve more than 12% of the NP formulation within the tumor. We have also shown for the first time that cancer drug loaded gold nanoparticles can reach the nucleus (or the brain) of cancer cells enhancing the therapeutic effect dramatically. Nucleus of the cancer cells are the most desirable target in cancer therapy. In chemotherapy, smart delivery of highly toxic anticancer drugs through packaging using nanoparticles will reduce the side effects and improve the quality and care of cancer patients. In radiation therapy, use of gold nanoparticles as radiation dose enhancer is very promising due to enhanced localized dose within the cancer tissue.
Pharmacy practice includes traditional and extemporaneous products that are well-tolerated by patients, for which shelf-life time cannot be accurately provided. In such cases, the product may not meet the desired requirements even within the expiry date. This problem not only gives rise to uncertainty among patients and pharmacists but is also of quality concern [1, 2]. Moreover, by carrying out certain examinations, even in the small-scale production, an appropriately stable pharmaceutical composition can be prepared.
The aim of this study is focused on the reproduction of a routinely used individual preparation, its physicochemical, accelerated and real-time stability testing to predict the rate of change at a proposed storage temperature.
Five variations of the chosen ointment were freshly prepared and subjected to accelerated stability testing at 40°C; 75±5% relative humidity and 25°C; 60±5% relative humidity. The preparations were monitored, and few units of the reference material were taken at 1, 3 and 6 month intervals. During the stability testing process the following experiments and tests were conducted according to the Hungarian and European Pharmacopoeias: Dropping point and freezing point measurements, extensometric test, microscopic examination, pH measurements of the aqueous phase, rheometric, dissolution and diffusion tests.
The study revealed that the choice of an optimal method of preparation results in a more stable pharmaceutical product than the original preparation. Even similar production methods resulted in ointments with significantly different physicochemical parameters. Based on the study, we can recommend a good manufacturing practice, expiry date, packaging material and storage conditions regarding the chosen formulation. These results confirmed that the physical and chemical stability of the ointments were achieved with the appropriate choice of the preparing conditions.
Since conventional chemotherapy for acute myeloid leukemia (AML) has its limitations, a theranostic platform with targeted and efficient drug transport is in demand. In this study, we developed the first CD123 (AML tumor marker) aptamers and designed a novel CD123-aptamer-mediated targeted drug train (TDT) with effective, economical, biocompatible and high drug-loading capacity. These two CD123 aptamers (termed as ZW25 and CY30, respectively) can bind to a CD123 peptide epitope and CD123 + AML cells with high specificities and KD of 29.41 nM and 15.38 nM, respectively, while has minimal cross reactivities to albumin, IgG and trypsin. Further, TDT is self-assembled from two short primers by ligand-modified ZW25 that acted as initiation position for elongation, while intercalated by doxorubicin (Dox). TDT is capable of transporting high capacity of Dox to CD123 + cells and retains the efficacy of Dox, while significantly reducing drug uptake and eased toxicity to CD123- cells in vitro (p < .01). Moreover, TDT can ease Dox cytoxicity to normal tissues, prolong survivals and inhibit tumor growth of mouse xenograft tumor model in vivo. These suggest that CD123 aptamer and CD123 aptamer-mediated targeted drug delivery system may have potential applications for selective delivery cytotoxic agents to CD123-expressing tumors in AML theranostics.
While vaccines represent the strongest weapons against a contagious disease, several shortcomings still limit their optimal function. This can largely be attributed to the lack of a proper delivery system as well as route of administration for effective activation of the immune system [1-3]. The intradermal route has the potential to be an outstanding route of vaccination due to the localization of potent antigen present cell (A PC ) and other immune cells in the dermis . M N Ps are tiny needles, small enough (100 –1,000 μm) that perforate superficial layers of the skin to deliver a therapeutic agent into the dermis in a relatively painless and invasive manner. M N Ps are generally preferred by patients as compared to traditional hypodermic injections. they take advantage of the potent intradermal immune system, which can create stronger responses than what is typical of the muscle or can generate equivalent responses from lower doses. The majority of works with microneedles has involved bolus delivery of drugs and vaccines using either coated or dissolvable microneedles, but introducing microparticles into microneedles in order to sustained release has received less attention . G iven these objectives and challenges, we designed a multi-compartment dissolvable and degradable polymeric microneedle patches for the sustained release of fd-O VA .
Recently, microRNA-based treatment receives good attention. Decreasing of miR-155-5p expression causes upregulation of HIF1α oncoprotein which worsens the prognosis of the cancer. To delivery miRNA at target cells, the nanoparticle-based packaging method is considered capable of answering the challenges that become one of the problems in the development of this therapy. Cationic chitosan can form nanocomplexes with negatively charged miRNA to carry miRNA to the site of target. The aim of this study was to formulate and evaluate the potential anticancer of chitosan nanoparticles mimic miR-155-5p against SKOV3 ovarian cancer cell line.
Methods: Chitosan nanoparticles (NPs-Cs) were made by ionic gelation method employes sodium tripolyphosphate as a cross-linker. NPs-Cs were characterized using TEM, DLS, electrophoresis and NanoQuant. MTT and qPCR assay were used to determine the effect of CS-NP/miRNA on SKOV3 cells.
Results: From the results of electrophoresis characterization showed that CS-NP can form nanocomplex with anti and mimic miRNA with adsorption efficiency of 57.43%, respectively. The mean diameter of NPs-Cs/mimic miR-155-5p was 261,367 ± 7.16 nm. The nanocomplex particles obtained were positively charged with potential zeta 47,267 ± 0.961. The results of the qPCR analysis showed that NPs-Cs/miRNA was able to send miRNA into SKOV3 cells to balance the levels of previously endregulated endogenous miRNA. Furthermore, the MTT assay results showed that NPs-Cs/miRNA was able to inhibit SKOV3 cell growth by regulating the expression of HIF1α.
Conclusion: Our analysis suggest that chitosan nanoparticle can be mediated delivery of miR-155-5p and to site of target and inhibition ovarian cancer cell.