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Pharmaceuticals

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Open Access journal
ISSN (Print) 1424-8247
Published by MDPI

[90 journals]

Pharmaceuticals, Vol. 6, Pages 579-603: Advances in Mammalian Cell Line Development Technologies for Recombinant Protein Production
- Authors: Tingfeng Lai, Yuansheng Yang, Say Ng
  Pages: 579 - 603
  Abstract: From 2006 to 2011, an average of 15 novel recombinant protein therapeutics have been approved by US Food and Drug Administration (FDA) annually. In addition, the expiration of blockbuster biologics has also spurred the emergence of biosimilars. The increasing numbers of innovator biologic products and biosimilars have thus fuelled the demand of production cell lines with high productivity. Currently, mammalian cell line development technologies used by most biopharmaceutical companies are based on either the methotrexate (MTX) amplification technology or the glutamine synthetase (GS) system. With both systems, the cell clones obtained are highly heterogeneous, as a result of random genome integration by the gene of interest and the gene amplification process. Consequently, large numbers of cell clones have to be screened to identify rare stable high producer cell clones. As such, the cell line development process typically requires 6 to 12 months and is a time, capital and labour intensive process. This article reviews established advances in protein expression and clone screening which are the core technologies in mammalian cell line development. Advancements in these component technologies are vital to improve the speed and efficiency of generating robust and highly productive cell line for large scale production of protein therapeutics.
  PubDate: 2013-04-26
  DOI: 10.3390/ph6050579
  Issue No: Vol. 6, No. 5 (2013)
Pharmaceuticals, Vol. 6, Pages 604-622: Improved Estimation of Protein-Ligand Binding Free Energy by Using the Ligand-Entropy and Mobility of Water Molecules
- Authors: Yoshifumi Fukunishi, Haruki Nakamura
  Pages: 604 - 622
  Abstract: We previously developed the direct interaction approximation (DIA) method to estimate the protein-ligand binding free energy (DG). The DIA method estimates the DG value based on the direct van der Waals and electrostatic interaction energies between the protein and the ligand. In the current study, the effect of the entropy of the ligand was introduced with protein dynamic properties by molecular dynamics simulations, and the interaction between each residue of the protein and the ligand was also weighted considering the hydration of each residue. The molecular dynamics simulation of the apo target protein gave the hydration effect of each residue, under the assumption that the residues, which strongly bind the water molecules, are important in the protein-ligand binding. These two effects improved the reliability of the DIA method. In fact, the parameters used in the DIA became independent of the target protein. The averaged error of DG estimation was 1.3 kcal/mol and the correlation coefficient between the experimental DG value and the calculated DG value was 0.75.
  PubDate: 2013-04-26
  DOI: 10.3390/ph6050604
  Issue No: Vol. 6, No. 5 (2013)
Pharmaceuticals, Vol. 6, Pages 623-633: In Vitro Assessment of Clevidipine Using the Profilin1 Hypertensive Mouse Model
- Authors: Hamdy Hassanain, Mohamed Hassona, Erika Puente, Chengwen Sun, Zeinb Abouelnaga, David Tulman, Sergio Bergese
  Pages: 623 - 633
  Abstract: Hypertension represents a major risk factor for cardiovascular events, associating with vascular hypertrophy and dysfunction in resistance vessels. Clevidipine is a novel antihypertensive drug working as a selective calcium channel antagonist with an ultra-short half-life that lowers arterial blood pressure by reducing systemic arterial resistance. The aim was to assess the effect of clevidipine on the hypertrophic vessels of profilin1 hypertensive transgenic mice compared to sodium nitroprusside (SNP) and labetalol using wire myograph techniques. The effects of clevidipine, SNP and labetalol on the hypertrophic vessels were studied on mesenteric arterial function from 8 profilin1 hypertrophic mice and eight non-transgenic controls. Our results showed a significant difference between the effects of the three drugs on the hypertrophic mesenteric arteries of transgenic profilin1 mice compared to the non-transgenic controls. The half maximal effective concentration (EC50) of clevidipine, SNP and labetalol in profilin1 mice (1.90 ± 0.05, 0.97 ± 0.07, 2.80 ± 0.05 nM, respectively) were significantly higher than the EC50 in non-transgenic controls (0.91 ± 0.06, 0.32 ± 0.06, 0.80 ± 0.09 nM, respectively). Moreover, the increase in the EC50 for clevidipine (2-fold) to produce the same effect on both normal and hypertrophic arteries was less than that of SNP (3-fold) and labetalol (3.5-fold). Therefore, we concluded clevidipine exhibited the lowest dose shift to relax the hypertrophic vessels compared to SNP and labetalol in the profilin1 hypertrophic animal mouse model.
  PubDate: 2013-04-29
  DOI: 10.3390/ph6050623
  Issue No: Vol. 6, No. 5 (2013)
Pharmaceuticals, Vol. 6, Pages 634-649: In Vivo Anti-Leukemia, Quantum Chemical Calculations and ADMET Investigations of Some Quaternary and Isothiouronium Surfactants
- Authors: Ahmed El-Henawy, Manal Khowdiary, Abdelfattah Badawi, Hussein Soliman
  Pages: 634 - 649
  Abstract: Anti-leukemia screening of previously prepared isothiouronium and quaternary salts was performed, and some salts exhibited promising activity as anticancer agents. Quantum chemical calculations were utilized to explore the electronic structure and stability of these compounds. Computational studies have been carried out at the PM3 semiempirical molecular orbitals level, to establish the HOMO-LUMO, IP and ESP mapping of these compounds. The ADMET properties were also studied to gain a clear view of the potential oral bioavailability of these compounds. The surface properties calculated included critical micelle concentration (CMC), maximum surface excess (Γmax), minimum surface area (Amin), free energy of micellization (ΔGomic) and adsorption (ΔGoads).
  PubDate: 2013-04-29
  DOI: 10.3390/ph6050634
  Issue No: Vol. 6, No. 5 (2013)
Pharmaceuticals, Vol. 6, Pages 650-658: Natural Products as a Source for New Anti-Inflammatory and Analgesic Compounds through the Inhibition of Purinergic P2X Receptors
- Authors: Rômulo Soares-Bezerra, Andrea Calheiros, Natiele da Silva Ferreira, Valber da Silva Frutuoso, Luiz Alves
  Pages: 650 - 658
  Abstract: Natural products have reemerged in traditional medicine as a potential source of new molecules or phytomedicines to help with health disorders. It has been established that members of the P2X subfamily, ATP-gated ion channels, are crucial to the inflammatory process and pain signalization. As such, several preclinical studies have demonstrated that P2X2R, P2X3R, P2X4R and P2X7R are promising pharmacological targets to control inflammatory and pain disorders. Several studies have indicated that natural products could be a good source of the new specific molecules needed for the treatment of diseases linked to inflammation and pain disorders through the regulation of these receptors. Herein, we discuss and give an overview of the applicability of natural products as a source to obtain P2X receptors (P2XR) selective antagonists for use in clinical treatment, which require further investigation.
  PubDate: 2013-04-29
  DOI: 10.3390/ph6050650
  Issue No: Vol. 6, No. 5 (2013)
Pharmaceuticals, Vol. 6, Pages 659-680: FedExosomes: Engineering Therapeutic Biological Nanoparticles that Truly Deliver
- Authors: Michelle Marcus, Joshua Leonard
  Pages: 659 - 680
  Abstract: Many aspects of intercellular communication are mediated through “sending” and “receiving” packets of information via the secretion and subsequent receptor-mediated detection of biomolecular species including cytokines, chemokines, and even metabolites. Recent evidence has now established a new modality of intercellular communication through which biomolecular species are exchanged between cells via extracellular lipid vesicles. A particularly important class of extracellular vesicles is exosomes, which is a term generally applied to biological nanovesicles ~30–200 nm in diameter. Exosomes form through invagination of endosomes to encapsulate cytoplasmic contents, and upon fusion of these multivesicular endosomes to the cell surface, exosomes are released to the extracellular space and transport mRNA, microRNA (miRNA) and proteins between cells. Importantly, exosome-mediated delivery of such cargo molecules results in functional modulation of the recipient cell, and such modulation is sufficiently potent to modulate disease processes in vivo. It is possible that such functional delivery of biomolecules indicates that exosomes utilize native mechanisms (e.g., for internalization and trafficking) that may be harnessed by using exosomes to deliver exogenous RNA for therapeutic applications. A complementary perspective is that understanding the mechanisms of exosome-mediated transport may provide opportunities for “reverse engineering” such mechanisms to improve the performance of synthetic delivery vehicles. In this review, we summarize recent progress in harnessing exosomes for therapeutic RNA delivery, discuss the potential for engineering exosomes to overcome delivery challenges and establish robust technology platforms, and describe both potential challenges and advantages of utilizing exosomes as RNA delivery vehicles.
  PubDate: 2013-04-29
  DOI: 10.3390/ph6050659
  Issue No: Vol. 6, No. 5 (2013)
Pharmaceuticals, Vol. 6, Pages 681-688: Evolution of Biologics Screening Technologies
- Authors: Peter Cariuk, Matthew Gardener, Tristan Vaughan
  Pages: 681 - 688
  Abstract: Screening for biologics, in particular antibody drugs, has evolved significantly over the last 20 years. Initially, the screening processes and technologies from many years experience with small molecules were adopted and modified to suit the needs of biologics discovery. Since then, antibody drug discovery has matured significantly and is today investing earlier in new technologies that commercial suppliers are now developing specifically to meet the growing needs of large molecule screening. Here, we review the evolution of screening and automation technologies employed in antibody discovery and highlight the benefits that these changes have brought.
  PubDate: 2013-05-14
  DOI: 10.3390/ph6050681
  Issue No: Vol. 6, No. 5 (2013)
Pharmaceuticals, Vol. 6, Pages 440-468: Design of siRNA Therapeutics from the Molecular Scale
- Authors: Phillip Angart, Daniel Vocelle, Christina Chan, S. Walton
  Pages: 440 - 468
  Abstract: While protein-based therapeutics is well-established in the market, development of nucleic acid therapeutics has lagged. Short interfering RNAs (siRNAs) represent an exciting new direction for the pharmaceutical industry. These small, chemically synthesized RNAs can knock down the expression of target genes through the use of a native eukaryotic pathway called RNA interference (RNAi). Though siRNAs are routinely used in research studies of eukaryotic biological processes, transitioning the technology to the clinic has proven challenging. Early efforts to design an siRNA therapeutic have demonstrated the difficulties in generating a highly-active siRNA with good specificity and a delivery vehicle that can protect the siRNA as it is transported to a specific tissue. In this review article, we discuss design considerations for siRNA therapeutics, identifying criteria for choosing therapeutic targets, producing highly-active siRNA sequences, and designing an optimized delivery vehicle. Taken together, these design considerations provide logical guidelines for generating novel siRNA therapeutics.
  PubDate: 2013-03-25
  DOI: 10.3390/ph6040440
  Issue No: Vol. 6, No. 4 (2013)
Pharmaceuticals, Vol. 6, Pages 469-479: Interactions of a Dopamine D1 Receptor Agonist with Glutamate NMDA Receptor Antagonists on the Volitional Consumption of Ethanol by the mHEP Rat
- Authors: Brian McMillen, Courtney Lommatzsch, Michael Sayonh, Helen Williams
  Pages: 469 - 479
  Abstract: Stimulation of the dopamine D1 receptor is reported to cause the phosphorylation of DARPP-32 at the thre34 position and activates the protein. If intracellular Ca2+ is increased, such as after activation of the glutamate NMDA receptor, calcineurin activity increases and the phosphates will be removed. This balance of phosphorylation control suggests that a D1 receptor agonist and a NMDA glutamate receptor antagonist should have additive or synergistic actions to increase activated DARPP-32 and consequent behavioral effects. This hypothesis was tested in a volitional consumption of ethanol model: the selectively bred Myers’ high ethanol preferring (mHEP) rat. A 3-day baseline period was followed by 3-days of twice daily injections of drug(s) or vehicle(s) and then a 3-day post-treatment period. Vehicle, the D1 agonist SKF 38393, the non-competitive NMDA receptor antagonist memantine, or their combination were injected 2 h before and after lights out. The combination of 5.0 mg/kg SKF 38393 with either 3.0 or 10 mg/kg memantine did not produce an additive or synergistic effect. For example, 5.0 mg/kg SKF reduced consumption of ethanol by 27.3% and 10 mg/kg memantine by 39.8%. When combined, consumption declined by 48.2% and the proportion of ethanol solution to total fluids consumed declined by 17%. However, the consumption of food also declined by 36.6%. The latter result indicates that this dose combination had a non-specific effect. The combination of SKF 38393 with (+)-MK-801, another NMDA receptor antagonist, also failed to show an additive effect. The lack of additivity and specificity suggests that the hypothesis may not be correct for this in vivo model. The interaction of these different receptor systems with intraneuronal signaling and behaviors needs to be studied further.
  PubDate: 2013-03-26
  DOI: 10.3390/ph6040469
  Issue No: Vol. 6, No. 4 (2013)
Pharmaceuticals, Vol. 6, Pages 480-499: NMDA Receptor Antagonists for Treatment of Depression
- Authors: Zeynep Ates-Alagoz, Adeboye Adejare
  Pages: 480 - 499
  Abstract: Depression is a psychiatric disorder that affects millions of people worldwide. Individuals battling this disorder commonly experience high rates of relapse, persistent residual symptoms, functional impairment, and diminished well-being. Medications have important utility in stabilizing moods and daily functions of many individuals. However, only one third of patients had considerable improvement with a standard antidepressant after 2 months and all patients had to deal with numerous side effects. The N-methyl-d-aspartate (NMDA) receptor family has received special attention because of its critical role in psychiatric disorders. Direct targeting of the NMDA receptor could result in more rapid antidepressant effects. Antidepressant-like effects of NMDA receptor antagonists have been demonstrated in different animal models. MK-801 (a use-dependent channel blocker), and CGP 37849 (an NMDA receptor antagonist) have shown antidepressant properties in preclinical studies, either alone or combined with traditional antidepressants. A recent development is use of ketamine clinically for refractory depression. The purpose of this review is to examine and analyze current literature on the role of NMDA receptor antagonists for treatment of depression and whether this is a feasible route in drug discovery.
  PubDate: 2013-04-03
  DOI: 10.3390/ph6040480
  Issue No: Vol. 6, No. 4 (2013)
Pharmaceuticals, Vol. 6, Pages 500-509: The Three Catecholics Benserazide, Catechol and Pyrogallol are GPR35 Agonists
- Authors: Huayun Deng, Ye Fang
  Pages: 500 - 509
  Abstract: Nearly 1% of all clinically used drugs are catecholics, a family of catechol-containing compounds. Using label-free dynamic mass redistribution and Tango β-arrestin translocation assays, we show that several catecholics, including benserazide, catechol, 3-methoxycatechol, pyrogallol, (+)-taxifolin and fenoldopam, display agonistic activity against GPR35.
  PubDate: 2013-04-08
  DOI: 10.3390/ph6040500
  Issue No: Vol. 6, No. 4 (2013)
Pharmaceuticals, Vol. 6, Pages 510-521: Delivery of RNAi-Based Oligonucleotides by Electropermeabilization
- Authors: Sophie Chabot, Sandrine Pelofy, Justin Teissié, Muriel Golzio
  Pages: 510 - 521
  Abstract: For more than a decade, understanding of RNA interference (RNAi) has been a growing field of interest. The potent gene silencing ability that small oligonucleotides have offers new perspectives for cancer therapeutics. One of the present limits is that many biological barriers exist for their efficient delivery into target cells or tissues. Electropermeabilization (EP) is one of the physical methods successfully used to transfer small oligonucleotides into cells or tissues. EP consists in the direct application of calibrated electric pulses to cells or tissues that transiently permeabilize the plasma membranes, allowing efficient in vitro and in vivo. cytoplasmic delivery of exogenous molecules. The present review reports on the type of therapeutic RNAi-based oligonucleotides that can be electrotransferred, the mechanism(s) of their electrotransfer and the technical settings for pre-clinical purposes.
  PubDate: 2013-04-10
  DOI: 10.3390/ph6040510
  Issue No: Vol. 6, No. 4 (2013)
Pharmaceuticals, Vol. 6, Pages 522-535: Disease-Causing Allele-Specific Silencing by RNA Interference
- Authors: Hirohiko Hohjoh
  Pages: 522 - 535
  Abstract: Small double-stranded RNAs (dsRNAs) of approximately 21-nucleotides in size, referred to as small interfering RNA (siRNA) duplexes, can induce sequence-specific posttranscriptional gene silencing, or RNA interference (RNAi). Since chemically synthesized siRNA duplexes were found to induce RNAi in mammalian cells, RNAi has become a powerful reverse genetic tool for suppressing the expression of a gene of interest in mammals, including human, and its application has been expanding to various fields. Recent studies further suggest that synthetic siRNA duplexes have the potential for specifically inhibiting the expression of an allele of interest without suppressing the expression of other alleles, i.e., siRNA duplexes likely confer allele-specific silencing. Such gene silencing by RNAi is an advanced technique with very promising applications. In this review, I would like to discuss the potential utility of allele-specific silencing by RNAi as a therapeutic method for dominantly inherited diseases, and describe possible improvements in siRNA duplexes for enhancing their efficacy.
  PubDate: 2013-04-11
  DOI: 10.3390/ph6040522
  Issue No: Vol. 6, No. 4 (2013)
Pharmaceuticals, Vol. 6, Pages 536-545: Toxicity Studies on Novel N-Substituted Bicyclo-Heptan-2-Amines at NMDA Receptors
- Authors: Natalia Coleman, Zeynep Ates-Alagoz, Boyenoh Gaye, Michelle Farbaniec, Shengguo Sun, Adeboye Adejare
  Pages: 536 - 545
  Abstract: Several novel norcamphor derivatives were designed and synthesized as uncompetitive NMDA receptor antagonists at the phencyclidine (PCP) binding site. Such compounds have potential as ligands for understanding and possibly the treatment of several neurodegenerative disorders and other glutamate-dependent disorders. We examined the toxic effects of the compounds as compared with memantine, an NMDA receptor antagonist that is FDA approved for treatment of Alzheimer’s disease, by testing these compounds on two cell lines: MDCK (to mimic blood brain barrier) and N2a (a neuronal cell line). The compounds showed toxicity profiles similar to those of memantine i.e., dose dependence above 100 μM and IC50 values above 150 μM for each cell line. It is known that the serum level of memantine under therapeutic conditions in patients is about 1 µM, indicting these compounds could have acceptable therapeutic indexes. 2-Phenyl-N-(2-(piperidin-1-yl) ethyl)bicyclo[2.2.1]heptan-2-amine (5a) was found to possess acceptable toxicity profiles in both cell lines. Interestingly, this was the compound identified as a good lead in our previous studies based on binding and anticonvulsant (MES) activity studies. It has thus emerged as an excellent lead compound for further studies.
  PubDate: 2013-04-12
  DOI: 10.3390/ph6040536
  Issue No: Vol. 6, No. 4 (2013)
Pharmaceuticals, Vol. 6, Pages 546-556: An Allosteric Modulator of the Adenosine A1 Receptor Improves Cardiac Function Following Ischaemia in Murine Isolated Hearts
- Authors: Anna Butcher, Peter Scammells, Paul White, Shane Devine, Roselyn Rose'Meyer
  Pages: 546 - 556
  Abstract: The effect of an allosteric modulator of the adenosine A1 receptors was investigated using an ischaemia-reperfusion protocol in murine isolated hearts. Isolated hearts were perfused with Kreb-Henseleit solution gassed with carbogen gas (95% O2 and 5% CO2) in Langendorff mode and electrically paced at 480 bpm. Following 20 min equilibration and 20 min global normothermic ischaemia, the allosteric modulator VCP333 (1 μM) or the adenosine A1 receptor partial agonist VCP102 (10 μM) were infused after 5 min of reperfusion for 15 min. Upon termination of the drug treatment, reperfusion continued for a further 40 min. At the end of 60 min reperfusion, treatment with VCP333 or VCP102 improved the recovery of the left ventricular developed pressure when compared to control group responses (p < 0.05). Neither compound affected end diastolic pressure, coronary flow rates or dP/dtmax values when compared to control tissues during reperfusion (p > 0.05). The infusion of VCP102 or VCP333 during reperfusion reduced cardiac troponin I efflux to 6.7% and 25% respectively of control heart efflux (p < 0.05). This data indicates that the allosteric modulator of the adenosine A1 receptor (VCP333) has similar characteristics to the adenosine receptor partial agonist VCP102 as it improves cardiac function and reduces myocardial cell death following an ischaemic episode.
  PubDate: 2013-04-12
  DOI: 10.3390/ph6040546
  Issue No: Vol. 6, No. 4 (2013)
Pharmaceuticals, Vol. 6, Pages 557-578: Biologics in Dermatology
- Authors: David Chandler, Anthony Bewley
  Pages: 557 - 578
  Abstract: Skin and subcutaneous diseases affect millions of people worldwide, causing significant morbidity. Biologics are becoming increasingly useful for the treatment of many skin diseases, particularly as alternatives for patients who have failed to tolerate or respond to conventional systemic therapies. Biological therapies provide a targeted approach to treatment through interaction with specific components of the underlying immune and inflammatory disease processes. This review article examines the increasing evidence base for biologics in dermatology, including well-established treatments and novel agents.
  PubDate: 2013-04-17
  DOI: 10.3390/ph6040557
  Issue No: Vol. 6, No. 4 (2013)
Pharmaceuticals, Vol. 6, Pages 287-294: RNAi Therapeutics in Autoimmune Disease
- Authors: Kaleb Pauley, Seunghee Cha
  Pages: 287 - 294
  Abstract: Since the discovery of RNA interference (RNAi), excitement has grown over its potential therapeutic uses. Targeting RNAi pathways provides a powerful tool to change biological processes post-transcriptionally in various health conditions such as cancer or autoimmune diseases. Optimum design of shRNA, siRNA, and miRNA enhances stability and specificity of RNAi-based approaches whereas it has to reduce or prevent undesirable immune responses or off-target effects. Recent advances in understanding pathogenesis of autoimmune diseases have allowed application of these tools in vitro as well as in vivo with some degree of success. Further research on the design and delivery of effectors of RNAi pathway and underlying molecular basis of RNAi would warrant practical use of RNAi-based therapeutics in human applications. This review will focus on the approaches used for current therapeutics and their applications in autoimmune diseases, including rheumatoid arthritis and Sjögren’s syndrome.
  PubDate: 2013-03-05
  DOI: 10.3390/ph6030287
  Issue No: Vol. 6, No. 3 (2013)
Pharmaceuticals, Vol. 6, Pages 295-319: Methods for Evaluating Cell-Specific, Cell-Internalizing RNA Aptamers
- Authors: Luiza Hernandez, Katie Flenker, Frank Hernandez, Aloysius Klingelhutz, James McNamara, Paloma Giangrande
  Pages: 295 - 319
  Abstract: Recent clinical trials of small interfering RNAs (siRNAs) highlight the need for robust delivery technologies that will facilitate the successful application of these therapeutics to humans. Arguably, cell targeting by conjugation to cell-specific ligands provides a viable solution to this problem. Synthetic RNA ligands (aptamers) represent an emerging class of pharmaceuticals with great potential for targeted therapeutic applications. For targeted delivery of siRNAs with aptamers, the aptamer-siRNA conjugate must be taken up by cells and reach the cytoplasm. To this end, we have developed cell-based selection approaches to isolate aptamers that internalize upon binding to their cognate receptor on the cell surface. Here we describe methods to monitor for cellular uptake of aptamers. These include: (1) antibody amplification microscopy, (2) microplate-based fluorescence assay, (3) a quantitative and ultrasensitive internalization method (“QUSIM”) and (4) a way to monitor for cytoplasmic delivery using the ribosome inactivating protein-based (RNA-RIP) assay. Collectively, these methods provide a toolset that can expedite the development of aptamer ligands to target and deliver therapeutic siRNAs in vivo.
  PubDate: 2013-03-14
  DOI: 10.3390/ph6030295
  Issue No: Vol. 6, No. 3 (2013)
Pharmaceuticals, Vol. 6, Pages 320-339: Cell Penetration Properties of a Highly Efficient Mini Maurocalcine Peptide
- Authors: Céline Tisseyre, Eloi Bahembera, Lucie Dardevet, Jean-Marc Sabatier, Michel Ronjat, Michel De Waard
  Pages: 320 - 339
  Abstract: Maurocalcine is a highly potent cell-penetrating peptide isolated from the Tunisian scorpion Maurus palmatus. Many cell-penetrating peptide analogues have been derived from the full-length maurocalcine by internal cysteine substitutions and sequence truncation. Herein we have further characterized the cell-penetrating properties of one such peptide, MCaUF1-9, whose sequence matches that of the hydrophobic face of maurocalcine. This peptide shows very favorable cell-penetration efficacy compared to Tat, penetratin or polyarginine. The peptide appears so specialized in cell penetration that it seems hard to improve by site directed mutagenesis. A comparative analysis of the efficacies of similar peptides isolated from other toxin members of the same family leads to the identification of hadrucalcin’s hydrophobic face as an even better CPP. Protonation of the histidine residue at position 6 renders the cell penetration of MCaUF1-9 pH-sensitive. Greater cell penetration at acidic pH suggests that MCaUF1-9 can be used to specifically target cancer cells in vivo where tumor masses grow in more acidic environments.
  PubDate: 2013-03-18
  DOI: 10.3390/ph6030320
  Issue No: Vol. 6, No. 3 (2013)
Pharmaceuticals, Vol. 6, Pages 340-357: A Review of Therapeutic Aptamer Conjugates with Emphasis on New Approaches
- Authors: John Bruno
  Pages: 340 - 357
  Abstract: The potential to emulate or enhance antibodies with nucleic acid aptamers while lowering costs has prompted development of new aptamer-protein, siRNA, drug, and nanoparticle conjugates. Specific focal points of this review discuss DNA aptamers covalently bound at their 3' ends to various proteins for enhanced stability and greater pharmacokinetic lifetimes in vivo. The proteins can include Fc tails of IgG for opsonization, and the first component of complement (C1q) to trigger complement-mediated lysis of antibiotic-resistant Gram negative bacteria, cancer cells and possibly some parasites during vulnerable stages. In addition, the 3' protein adduct may be a biotoxin, enzyme, or may simply be human serum albumin (HSA) or a drug known to bind HSA, thereby retarding kidney and other organ clearance and inhibiting serum exonucleases. In this review, the author summarizes existing therapeutic aptamer conjugate categories and describes his patented concept for PCR-based amplification of double-stranded aptamers followed by covalent attachment of proteins or other agents to the chemically vulnerable overhanging 3' adenine added by Taq polymerase. PCR amplification of aptamers could dramatically lower the current $2,000/gram cost of parallel chemical oligonucleotide synthesis, thereby enabling mass production of aptamer-3'-protein or drug conjugates to better compete against expensive humanized monoclonal antibodies.
  PubDate: 2013-03-19
  DOI: 10.3390/ph6030340
  Issue No: Vol. 6, No. 3 (2013)
Pharmaceuticals, Vol. 6, Pages 358-392: Inactivation of Caliciviruses
- Authors: Raymond Nims, Mark Plavsic
  Pages: 358 - 392
  Abstract: The Caliciviridae family of viruses contains clinically important human and animal pathogens, as well as vesivirus 2117, a known contaminant of biopharmaceutical manufacturing processes employing Chinese hamster cells. An extensive literature exists for inactivation of various animal caliciviruses, especially feline calicivirus and murine norovirus. The caliciviruses are susceptible to wet heat inactivation at temperatures in excess of 60 °C with contact times of 30 min or greater, to UV-C inactivation at fluence ≥30 mJ/cm2, to high pressure processing >200 MPa for >5 min at 4 °C, and to certain photodynamic inactivation approaches. The enteric caliciviruses (e.g.; noroviruses) display resistance to inactivation by low pH, while the non-enteric species (e.g.; feline calicivirus) are much more susceptible. The caliciviruses are inactivated by a variety of chemicals, including alcohols, oxidizing agents, aldehydes, and β-propiolactone. As with inactivation of viruses in general, inactivation of caliciviruses by the various approaches may be matrix-, temperature-, and/or contact time-dependent. The susceptibilities of the caliciviruses to the various physical and chemical inactivation approaches are generally similar to those displayed by other small, non-enveloped viruses, with the exception that the parvoviruses and circoviruses may require higher temperatures for inactivation, while these families appear to be more susceptible to UV-C inactivation than are the caliciviruses.
  PubDate: 2013-03-21
  DOI: 10.3390/ph6030358
  Issue No: Vol. 6, No. 3 (2013)
Pharmaceuticals, Vol. 6, Pages 393-406: Characterization of N-Linked Glycosylation in a Monoclonal Antibody Produced in NS0 Cells Using Capillary Electrophoresis with Laser-Induced Fluorescence Detection
- Authors: Melissa Hamm, Yang Wang, Richard Rustandi
  Pages: 393 - 406
  Abstract: The N-linked glycosylation in recombinant monoclonal antibodies (mAb) occurs at Asn297 on the Fc region in the CH2 domain. Glycosylation heterogeneities have been well documented to affect biological activities such as antibody-dependent cellular cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC) through their interaction with Fc-receptors. Hence, it is critical to monitor and characterize the N-linked glycosylation profile in a therapeutic protein such as a mAb for product consistency. In one approach, the glycans are first released from the mAb using an enzyme specific digestion, such as Protein N-Glycosidase F (PNGase) and subsequently they are labeled using a fluorophore, for example, 8-aminopyrene-1,3,6-trisulfonic acid (APTS) . Here we have applied this approach and used Capillary Electrophoresis with Laser-Induced Fluorescence detection (CE-LIF) to analyze a recombinant mAb produced in murine myeloma (NS0) cells. The technique provides short analysis times, efficient separations, and high sensitivity. CE-LIF peak identification was done by a combination of glycan standards and treatment with various exoglycosidases. Furthermore, the APTS-labeled glycans were also analyzed using hydrophilic interaction chromatography (HILIC) high performance liquid chromatography (HPLC) to aid identification of minor peaks by sample collection and off-line mass spectrometry (MS) analysis.
  PubDate: 2013-03-21
  DOI: 10.3390/ph6030393
  Issue No: Vol. 6, No. 3 (2013)
Pharmaceuticals, Vol. 6, Pages 407-439: Protein Kinase C Inhibitors as Modulators of Vascular Function and Their Application in Vascular Disease
- Authors: Raouf Khalil
  Pages: 407 - 439
  Abstract: Blood pressure (BP) is regulated by multiple neuronal, hormonal, renal and vascular control mechanisms. Changes in signaling mechanisms in the endothelium, vascular smooth muscle (VSM) and extracellular matrix cause alterations in vascular tone and blood vessel remodeling and may lead to persistent increases in vascular resistance and hypertension (HTN). In VSM, activation of surface receptors by vasoconstrictor stimuli causes an increase in intracellular free Ca2+ concentration ([Ca2+]i), which forms a complex with calmodulin, activates myosin light chain (MLC) kinase and leads to MLC phosphorylation, actin-myosin interaction and VSM contraction. Vasoconstrictor agonists could also increase the production of diacylglycerol which activates protein kinase C (PKC). PKC is a family of Ca2+-dependent and Ca2+-independent isozymes that have different distributions in various blood vessels, and undergo translocation from the cytosol to the plasma membrane, cytoskeleton or the nucleus during cell activation. In VSM, PKC translocation to the cell surface may trigger a cascade of biochemical events leading to activation of mitogen-activated protein kinase (MAPK) and MAPK kinase (MEK), a pathway that ultimately increases the myofilament force sensitivity to [Ca2+]i, and enhances actin-myosin interaction and VSM contraction. PKC translocation to the nucleus may induce transactivation of various genes and promote VSM growth and proliferation. PKC could also affect endothelium-derived relaxing and contracting factors as well as matrix metalloproteinases (MMPs) in the extracellular matrix further affecting vascular reactivity and remodeling. In addition to vasoactive factors, reactive oxygen species, inflammatory cytokines and other metabolic factors could affect PKC activity. Increased PKC expression and activity have been observed in vascular disease and in certain forms of experimental and human HTN. Targeting of vascular PKC using PKC inhibitors may function in concert with antioxidants, MMP inhibitors and cytokine antagonists to reduce VSM hyperactivity in certain forms of HTN that do not respond to Ca2+ channel blockers.
  PubDate: 2013-03-21
  DOI: 10.3390/ph6030407
  Issue No: Vol. 6, No. 3 (2013)
Pharmaceuticals, Vol. 6, Pages 124-160: siRNA Genome Screening Approaches to Therapeutic Drug Repositioning
- Authors: Olivia Perwitasari, Abhijeet Bakre, S. Tompkins, Ralph Tripp
  Pages: 124 - 160
  Abstract: Bridging high-throughput screening (HTS) with RNA interference (RNAi) has allowed for rapid discovery of the molecular basis of many diseases, and identification of potential pathways for developing safe and effective treatments. These features have identified new host gene targets for existing drugs paving the pathway for therapeutic drug repositioning. Using RNAi to discover and help validate new drug targets has also provided a means to filter and prioritize promising therapeutics. This review summarizes these approaches across a spectrum of methods and targets in the host response to pathogens. Particular attention is given to the utility of drug repurposing utilizing the promiscuous nature of some drugs that affect multiple molecules or pathways, and how these biological pathways can be targeted to regulate disease outcome.
  PubDate: 2013-01-28
  DOI: 10.3390/ph6020124
  Issue No: Vol. 6, No. 2 (2013)
Pharmaceuticals, Vol. 6, Pages 161-183: Dendrimers for siRNA Delivery
- Authors: Swati Biswas, Vladimir Torchilin
  Pages: 161 - 183
  Abstract: Since the discovery of the “starburst polymer”, later renamed as dendrimer, this class of polymers has gained considerable attention for numerous biomedical applications, due mainly to the unique characteristics of this macromolecule, including its monodispersity, uniformity, and the presence of numerous functionalizable terminal groups. In recent years, dendrimers have been studied extensively for their potential application as carriers for nucleic acid therapeutics, which utilize the cationic charge of the dendrimers for effective dendrimer-nucleic acid condensation. siRNA is considered a promising, versatile tool among various RNAi-based therapeutics, which can effectively regulate gene expression if delivered successfully inside the cells. This review reports on the advancements in the development of dendrimers as siRNA carriers.
  PubDate: 2013-02-04
  DOI: 10.3390/ph6020161
  Issue No: Vol. 6, No. 2 (2013)
Pharmaceuticals, Vol. 6, Pages 184-203: Relationships between Cargo, Cell Penetrating Peptides and Cell Type for Uptake of Non-Covalent Complexes into Live Cells
- Authors: Andrea-Anneliese Keller, Franziska Mussbach, Reinhard Breitling, Peter Hemmerich, Buerk Schaefer, Stefan Lorkowski, Siegmund Reissmann
  Pages: 184 - 203
  Abstract: Modulating signaling pathways for research and therapy requires either suppression or expression of selected genes or internalization of proteins such as enzymes, antibodies, nucleotide binding proteins or substrates including nucleoside phosphates and enzyme inhibitors. Peptides, proteins and nucleotides are transported by fusing or conjugating them to cell penetrating peptides or by formation of non-covalent complexes. The latter is often preferred because of easy handling, uptake efficiency and auto-release of cargo into the live cell. In our studies complexes are formed with labeled or readily detectable cargoes for qualitative and quantitative estimation of their internalization. Properties and behavior of adhesion and suspension vertebrate cells as well as the protozoa Leishmania tarentolae are investigated with respect to proteolytic activity, uptake efficiency, intracellular localization and cytotoxicity. Our results show that peptide stability to membrane-bound, secreted or intracellular proteases varies between different CPPs and that the suitability of individual CPPs for a particular cargo in complex formation by non-covalent interactions requires detailed studies. Cells vary in their sensitivity to increasing concentrations of CPPs. Thus, most cells can be efficiently transduced with peptides, proteins and nucleotides with intracellular concentrations in the low micromole range. For each cargo, cell type and CPP the optimal conditions must be determined separately.
  PubDate: 2013-02-06
  DOI: 10.3390/ph6020184
  Issue No: Vol. 6, No. 2 (2013)
Pharmaceuticals, Vol. 6, Pages 204-222: Elucidating the Function of Penetratin and a Static Magnetic Field in Cellular Uptake of Magnetic Nanoparticles
- Authors: Suman Chaudhary, Carol Smith, Pablo del Pino, Jesus de la Fuente, Margaret Mullin, Andrew Hursthouse, David Stirling, Catherine Berry
  Pages: 204 - 222
  Abstract: Nanotechnology plays an increasingly important role in the biomedical arena. In particular, magnetic nanoparticles (mNPs) have become important tools in molecular diagnostics, in vivo imaging and improved treatment of disease, with the ultimate aim of producing a more theranostic approach. Due to their small sizes, the nanoparticles can cross most of the biological barriers such as the blood vessels and the blood brain barrier, thus providing ubiquitous access to most tissues. In all biomedical applications maximum nanoparticle uptake into cells is required. Two promising methods employed to this end include functionalization of mNPs with cell-penetrating peptides to promote efficient translocation of cargo into the cell and the use of external magnetic fields for enhanced delivery. This study aimed to compare the effect of both penetratin and a static magnetic field with regards to the cellular uptake of 200 nm magnetic NPs and determine the route of uptake by both methods. Results demonstrated that both techniques increased particle uptake, with penetratin proving more cell specific. Clathrin- medicated endocytosis appeared to be responsible for uptake as shown via PCR and western blot, with Pitstop 2 (known to selectively block clathrin formation) blocking particle uptake. Interestingly, it was further shown that a magnetic field was able to reverse or overcome the blocking, suggesting an alternative route of uptake.
  PubDate: 2013-02-06
  DOI: 10.3390/ph6020204
  Issue No: Vol. 6, No. 2 (2013)
Pharmaceuticals, Vol. 6, Pages 223-250: RNAi Therapeutic Platforms for Lung Diseases
- Authors: Yu Fujita, Fumitaka Takeshita, Kazuyoshi Kuwano, Takahiro Ochiya
  Pages: 223 - 250
  Abstract: RNA interference (RNAi) is rapidly becoming an important method for analyzing gene functions in many eukaryotes and holds promise for the development of therapeutic gene silencing. The induction of RNAi relies on small silencing RNAs, which affect specific messenger RNA (mRNA) degradation. Two types of small RNA molecules, i.e. small interfering RNAs (siRNAs) and microRNAs (miRNAs), are central to RNAi. Drug discovery studies and novel treatments of siRNAs are currently targeting a wide range of diseases, including various viral infections and cancers. Lung diseases in general are attractive targets for siRNA therapeutics because of their lethality and prevalence. In addition, the lung is anatomically accessible to therapeutic agents via the intrapulmonary route. Recently, increasing evidence indicates that miRNAs play an important role in lung abnormalities, such as inflammation and oncogenesis. Therefore, miRNAs are being targeted for therapeutic purposes. In this review, we present strategies for RNAi delivery and discuss the current state-of-the-art RNAi-based therapeutics for various lung diseases.
  PubDate: 2013-02-06
  DOI: 10.3390/ph6020223
  Issue No: Vol. 6, No. 2 (2013)
Pharmaceuticals, Vol. 6, Pages 251-268: NMDA Receptor Modulators in the Treatment of Drug Addiction
- Authors: Seven Tomek, Amber Lacrosse, Natali Nemirovsky, M. Olive
  Pages: 251 - 268
  Abstract: Glutamate plays a pivotal role in drug addiction, and the N-methyl-D-aspartate (NMDA) glutamate receptor subtype serves as a molecular target for several drugs of abuse. In this review, we will provide an overview of NMDA receptor structure and function, followed by a review of the mechanism of action, clinical efficacy, and side effect profile of NMDA receptor ligands that are currently in use or being explored for the treatment of drug addiction. These ligands include the NMDA receptor modulators memantine and acamprosate, as well as the partial NMDA agonist D-cycloserine. Data collected to date suggest that direct NMDA receptor modulators have relatively limited efficacy in the treatment of drug addiction, and that partial agonism of NMDA receptors may have some efficacy with regards to extinction learning during cue exposure therapy. However, the lack of consistency in results to date clearly indicates that additional studies are needed, as are studies examining novel ligands with indirect mechanisms for altering NMDA receptor function.
  PubDate: 2013-02-06
  DOI: 10.3390/ph6020251
  Issue No: Vol. 6, No. 2 (2013)
Pharmaceuticals, Vol. 6, Pages 269-286: cGMP-Dependent Protein Kinase Inhibitors in Health and Disease
- Authors: Stefanie Wolfertstetter, Johannes Huettner, Jens Schlossmann
  Pages: 269 - 286
  Abstract: cGMP-dependent protein kinases (PKG) exhibit diverse physiological functions in the mammalian system e.g., in vascular and gastrointestinal smooth muscles, in platelets, in kidney, in bone growth, nociception and in the central nervous system. Furthermore, PKG were found in insects and in the malaria parasite Plasmodium falciparum. Two different genes of PKG exist: a) the PKG-I gene that is expressed as cytosolic PKG-Iα or PKG-Iβ isoform, and b) the PKG-II gene, which expresses the membrane associated PKG-II protein. The enzyme kinetics, the localization and the substrates of these PKG enzymes differ utilizing different physiological functions. Various inhibitors of PKG were developed directed against diverse functional regions of the kinase. These inhibitors of PKG have been used to analyse the specific functions of these enzymes. The review article will summarize these different inhibitors regarding their specificity and their present applications in vitro and in vivo. Furthermore, it will be discussed that the distinct inhibition of the PKG enzymes could be used as a valuable pharmacological target e.g., in the treatment of cardiovascular diseases, diarrhea, cancer or malaria.
  PubDate: 2013-02-07
  DOI: 10.3390/ph6020269
  Issue No: Vol. 6, No. 2 (2013)
Pharmaceuticals, Vol. 6, Pages 25-31: Stability of Recombinant Tissue Plasminogen Activator at &#8722;30 &#176;C over One Year
- Authors: Abdulmalik Alkatheri
  Pages: 25 - 31
  Abstract: Recombinant tissue plasminogen activator (rt-PA) is used to restore patency and avoid inadvertent removal of peripheral and central venous catheters. rt-PA was reconstituted (1 mg/mL) then cryopreserved at −30 °C for 1, 2, 3, 6, 8, and 12 months and, then its stability was determined. After cryopreservation for one and two months, rt-PA kept more than 95% of its activity compared to standard samples, while cryopreservation for three months caused 8% loss of activity. However, after cryopreservation for six months or more, rt-PA retained only 87.5% or less activity compared to standard samples. Therefore, it is recommended that reconstituted rt-PA be cryopreserved at −30 °C for a maximum period of three months.
  PubDate: 2013-01-02
  DOI: 10.3390/ph6010025
  Issue No: Vol. 6, No. 1 (2013)
Pharmaceuticals, Vol. 6, Pages 32-53: The Potential Role of Cell Penetrating Peptides in the Intracellular Delivery of Proteins for Therapy of Erythroid Related Disorders
- Authors: Lefkothea Papadopoulou, Asterios Tsiftsoglou
  Pages: 32 - 53
  Abstract: The erythroid related disorders (ERDs) represent a large group of hematological diseases, which in most cases are attributed either to the deficiency or malfunction of biosynthetic enzymes or oxygen transport proteins. Current treatments for these disorders include histo-compatible erythrocyte transfusions or allogeneic hematopoietic stem cell (HSC) transplantation. Gene therapy delivered via suitable viral vectors or genetically modified HSCs have been under way. Protein Transduction Domain (PTD) technology has allowed the production and intracellular delivery of recombinant therapeutic proteins, bearing Cell Penetrating Peptides (CPPs), into a variety of mammalian cells. Remarkable progress in the field of protein transduction leads to the development of novel protein therapeutics (CPP-mediated PTs) for the treatment of monogenetic and/or metabolic disorders. The “concept” developed in this paper is the intracellular protein delivery made possible via the PTD technology as a novel therapeutic intervention for treatment of ERDs. This can be achieved via four stages including: (i) the production of genetically engineered human CPP-mediated PT of interest, since the corresponding native protein either is missing or is mutated in the erythroid progenitor cell (ErPCs) or mature erythrocytes of patients; (ii) isolation of target cells from the peripheral blood of the selected patients; (iii) ex vivo transduction of cells with the CPP-mediated PT of interest; and (iv) re-administration of the successfully transduced cells back into the same patients.
  PubDate: 2013-01-07
  DOI: 10.3390/ph6010032
  Issue No: Vol. 6, No. 1 (2013)
Pharmaceuticals, Vol. 6, Pages 54-69: A Potent (R)-alpha-bis-lipoyl Derivative Containing 8-Hydroxyquinoline Scaffold: Synthesis and Biological Evaluation of Its Neuroprotective Capabilities in SH-SY5Y Human Neuroblastoma Cells
- Authors: Ivana Cacciatore, Erika Fornasari, Leonardo Baldassarre, Catia Cornacchia, Stefania Fulle, Ester Di Filippo, Tiziana Pietrangelo, Francesco Pinnen
  Pages: 54 - 69
  Abstract: A novel bis-lipoyl derivative containing 8-hydroxyquinoline scaffold (LA-HQ-LA, 5) was synthesized as a new multifunctional drug candidate with antioxidant, chelant, and neuroprotective properties for the treatment of neurodegenerative diseases. We have investigated the potential effectiveness of LA-HQ-LA against the cytotoxicity induced by 6-OHDA and H2O2 on human neuroblastoma SH-SY5Y cell line. Our outcomes showed that LA-HQ-LA resulted in significant neuroprotective and antioxidant effects against H2O2- and 6-OHDA-induced neurotoxicity in human neuroblastoma SH-SY5Y cells, as assessed by MTT assay. In particular, it showed potent neuroprotective effects against 6-OHDA in RA/PMA differentiated cells at all the tested concentrations.
  PubDate: 2013-01-07
  DOI: 10.3390/ph6010054
  Issue No: Vol. 6, No. 1 (2013)
Pharmaceuticals, Vol. 6, Pages 70-84: Clevidipine for Perioperative Blood Pressure Control in Infants and Children
- Authors: Joseph Tobias, David Tulman, Sergio Bergese
  Pages: 70 - 84
  Abstract: Various pharmacologic agents have been used for perioperative BP control in pediatric patients, including sodium nitroprusside, nitroglycerin, β-adrenergic antagonists, fenoldopam, and calcium channel antagonists. Of the calcium antagonists, the majority of the clinical experience remains with the dihydropyridine nicardipine. Clevidipine is a short-acting, intravenous calcium channel antagonist of the dihydropyridine class. It undergoes rapid metabolism by non-specific blood and tissue esterases with a half-life of less than 1 minute. As a dihydropyridine, its cellular and end-organ effects parallel those of nicardipine. The clevidipine trials in the adult population have demonstrated efficacy in rapidly controlling BP in various clinical scenarios with a favorable adverse effect profile similar to nicardipine. Data from large clinical trials regarding the safety and efficacy of clevidipine in children is lacking. This manuscript aims to review the commonly used pharmacologic agents for perioperative BP control in children, discuss the role of calcium channel antagonists such as nicardipine, and outline the preliminary data regarding clevidipine in the pediatric population.
  PubDate: 2013-01-15
  DOI: 10.3390/ph6010070
  Issue No: Vol. 6, No. 1 (2013)
Pharmaceuticals, Vol. 6, Pages 85-107: Nanoparticle-Based Delivery of RNAi Therapeutics: Progress and Challenges
- Authors: Jiehua Zhou, Ka-To Shum, John Burnett, John Rossi
  Pages: 85 - 107
  Abstract: RNA interference (RNAi) is an evolutionarily conserved, endogenous process for post-transcriptional regulation of gene expression. Although RNAi therapeutics have recently progressed through the pipeline toward clinical trials, the application of these as ideal, clinical therapeutics requires the development of safe and effective delivery systems. Inspired by the immense progress with nanotechnology in drug delivery, efforts have been dedicated to the development of nanoparticle-based RNAi delivery systems. For example, a precisely engineered, multifunctional nanocarrier with combined passive and active targeting capabilities may address the delivery challenges for the widespread use of RNAi as a therapy. Therefore, in this review, we introduce the major hurdles in achieving efficient RNAi delivery and discuss the current advances in applying nanotechnology-based delivery systems to overcome the delivery hurdles of RNAi therapeutics. In particular, some representative examples of nanoparticle-based delivery formulations for targeted RNAi therapeutics are highlighted.
  PubDate: 2013-01-16
  DOI: 10.3390/ph6010085
  Issue No: Vol. 6, No. 1 (2013)
Pharmaceuticals, Vol. 6, Pages 108-123: Neurotransmitter CART as a New Therapeutic Candidate for Parkinson&#8217;s Disease
- Authors: Peizhong Mao, Charles Meshul, Philippe Thuillier, P. Reddy
  Pages: 108 - 123
  Abstract: Parkinson’s disease (PD) is one of the most common neurodegenerative diseases. To date, there is no effective treatment that halts its progression. Increasing evidence indicates that mitochondria play an important role in the development of PD. Hence mitochondria-targeted approaches or agents may have therapeutic promise for treatment of the disease. Neuropeptide CART (cocaine-amphetamine-regulated transcript), a hypothalamus and midbrain enriched neurotransmitter with an antioxidant property, can be found in mitochondria, which is the main source of reactive oxygen species. Systemic administration of CART has been found to ameliorate dopaminergic neuronal loss and improve motor functions in a mouse model of PD. In this article, we summarize recent progress in studies investigating the relationship between CART, dopamine, and the pathophysiology of PD, with a focus on mitochondria-related topics.
  PubDate: 2013-01-18
  DOI: 10.3390/ph6010108
  Issue No: Vol. 6, No. 1 (2013)
Pharmaceuticals, Vol. 6, Pages 1-24: Strategies To Modulate Heritable Epigenetic Defects in Cellular Machinery: Lessons from Nature
- Authors: Ganesh Pandian, Hiroshi Sugiyama
  Pages: 1 - 24
  Abstract: Natural epigenetic processes precisely orchestrate the intricate gene network by expressing and suppressing genes at the right place and time, thereby playing an essential role in maintaining the cellular homeostasis. Environment-mediated alteration of this natural epigenomic pattern causes abnormal cell behavior and shifts the cell from the normal to a diseased state, leading to certain cancers and neurodegenerative disorders. Unlike heritable diseases that are caused by the irreversible mutations in DNA, epigenetic errors can be reversed. Inheritance of epigenetic memory is also a major concern in the clinical translation of the Nobel Prize-winning discovery of induced pluripotent stem cell technology. Consequently, there is an increasing interest in the development of novel epigenetic switch-based therapeutic strategies that could potentially restore the heritable changes in epigenetically inherited disorders. Here we give a comprehensive overview of epigenetic inheritance and suggest the prospects of therapeutic gene modulation using epigenetic-based drugs, in particular histone deacetylase inhibitors. This review suggests that there is a need to develop therapeutic strategies that effectively mimic the natural environment and include the ways to modulate the gene expression at both the genetic and epigenetic levels. The development of tailor-made small molecules that could epigenetically alter DNA in a sequence-specific manner is a promising approach for restoring defects in an altered epigenome and may offer a sustainable solution to some unresolved clinical issues.
  PubDate: 2012-12-27
  DOI: 10.3390/ph6010001
  Issue No: Vol. 6, No. 1 (2012)
Pharmaceuticals, Vol. 5, Pages 890-898: Hsp90 Inhibitors and the Reduction of Anti-Cancer Drug Resistance by Non-Genetic and Genetic Mechanisms
- Authors: Xiangyi Lu, Luan Wang, Douglas M. Ruden
  Pages: 890 - 898
  Abstract: In this review, we focus on how inhibitors of Hsp90 can help prevent the resistance to anti-cancer drugs by synergistically increasing their cancer killing abilities and thereby allowing them to function at much lower concentrations than normally used. Hsp90 helps to fold numerous client proteins, such as Akt, Raf, Src, chromatin-modifying proteins, nuclear hormone receptors, and kinetochore assembly proteins. We discuss four mechanisms by which Hsp90 inhibitors can potentially synergize with anti-cancer drugs: by making a drug-resistant protein that is a client for Hsp90 more sensitive to the drug, by increasing chromosomal aneuploidy and the effectiveness of DNA-damaging drugs, by inhibiting Trithorax proteins which trimethylate histone 3 at lysine 4 (H3K4me3) and thereby decreasing expression of tumor promoter genes, and by interacting with the negative elongation factor (NELF) complex in tumors. We also explain how the evolutionary capacitor function of Hsp90 can be exploited with inhibitors of Hsp90 by exposing new protein variants that can be targeted with other drugs, thereby opening new avenues of combination drug therapy to treat cancer. We believe that inhibition of these processes can increase the efficacy of Hsp90 inhibitors with other anti-cancer drugs.
  PubDate: 2012-08-30
  DOI: 10.3390/ph5090890
  Issue No: Vol. 5, No. 9 (2012)
Pharmaceuticals, Vol. 5, Pages 899-924: The Tissue Selective Estrogen Complex: A Promising New Menopausal Therapy
- Authors: Barry S. Komm, Sebastian Mirkin
  Pages: 899 - 924
  Abstract: Menopause is associated with health concerns including vasomotor symptoms, vulvar/vaginal atrophy (VVA), and osteoporosis. Estrogen therapy or combined estrogen-progestin therapy (EPT) are primary treatment options for menopausal symptom relief and osteoporosis prevention. Because EPT has been associated with some safety/tolerability concerns relating to undesirable effects of estrogen and progestin, alternative options are needed. The tissue selective estrogen complex (TSEC) is a novel class of agents pairing a selective estrogen receptor modulator (SERM) with 1 or more estrogens. The TSEC combines the established efficacy of estrogens on menopausal symptoms and bone with the protective effects of a SERM on the reproductive tract. The pairing of bazedoxifene (BZA) with conjugated estrogens (CE) has been evaluated in a series of phase 3 clinical trials. BZA 20 mg/CE 0.45 mg and BZA 20 mg/CE 0.625 mg have shown efficacy in reducing the frequency and severity of hot flushes, relieving VVA symptoms, and maintaining bone mass while protecting the endometrium and breast. These BZA/CE doses have been associated with a favorable safety/tolerability profile, with higher rates of cumulative amenorrhea and lower incidences of breast pain than those reported for EPT. Thus, BZA/CE may be a promising alternative to conventional EPT for treating non-hysterectomized, postmenopausal women.
  PubDate: 2012-09-04
  DOI: 10.3390/ph5090899
  Issue No: Vol. 5, No. 9 (2012)
Pharmaceuticals, Vol. 5, Pages 925-943: Differential Cellular and Molecular Effects of Butyrate and Trichostatin A on Vascular Smooth Muscle Cells
- Authors: Shirlette G. Milton, Omana P. Mathew, Frank M. Yatsu, Kasturi Ranganna
  Pages: 925 - 943
  Abstract: The histone deacetylase (HDAC) inhibitors, butyrate and trichostatin A (TSA), are epigenetic histone modifiers and proliferation inhibitors by downregulating cyclin D1, a positive cell cycle regulator, and upregulating p21Cip1 and INK family of proteins, negative cell cycle regulators. Our recent study indicated cyclin D1 upregulation in vascular smooth muscle cells (VSMC) that are proliferation-arrested by butyrate. Here we investigate whether cyclin D1 upregulation is a unique response of VSMC to butyrate or a general response to HDAC inhibitors (HDACi) by evaluating the effects of butyrate and TSA on VSMC. While butyrate and TSA inhibit VSMC proliferation via cytostatic and cytotoxic effects, respectively, they downregulate cdk4, cdk6, and cdk2, and upregulate cyclin D3, p21Cip1 and p15INK4B, and cause similar effects on key histone H3 posttranslational modifications. Conversely, cyclin D1 is upregulated by butyrate and inhibited by TSA. Assessment of glycogen synthase 3-dependent phosphorylation, subcellular localization and transcription of cyclin D1 indicates that differential effects of butyrate and TSA on cyclin D1 levels are linked to disparity in cyclin D1 gene expression. Disparity in butyrate- and TSA-induced cyclin D1 may influence transcriptional regulation of genes that are associated with changes in cellular morphology/cellular effects that these HDACi confer on VSMC, as a transcriptional modulator.
  PubDate: 2012-09-04
  DOI: 10.3390/ph5090925
  Issue No: Vol. 5, No. 9 (2012)
Pharmaceuticals, Vol. 5, Pages 944-962: Molecular Dynamics Simulations of Hsp90 with an Eye to Inhibitor Design
- Authors: Elisabetta Moroni, Giulia Morra, Giorgio Colombo
  Pages: 944 - 962
  Abstract: Proteins carry out their functions through interactions with different partners. Dynamic conformational switching among different structural sub-states favors the adaptation to the shapes of the different partners. Such conformational changes can be determined by diverse biochemical factors, such as ligand-binding. Atomic level investigations of the mechanisms that underlie functional dynamics may provide new opportunities for the discovery of leads that target disease-related proteins. In this review, we report our views and approaches on the development of novel and accurate physical-chemistry-based models for the characterization of the salient aspects of the ligand-regulated dynamics of Hsp90, and on the exploitation of such new knowledge for the rational discovery of inhibitors of the chaperone.
  PubDate: 2012-09-10
  DOI: 10.3390/ph5090944
  Issue No: Vol. 5, No. 9 (2012)
Pharmaceuticals, Vol. 5, Pages 963-990: Epigenetic Control and Cancer: The Potential of Histone Demethylases as Therapeutic Targets
- Authors: Fernando Lizcano, Jeison Garcia
  Pages: 963 - 990
  Abstract: The development of cancer involves an immense number of factors at the molecular level. These factors are associated principally with alterations in the epigenetic mechanisms that regulate gene expression profiles. Studying the effects of chromatin structure alterations, which are caused by the addition/removal of functional groups to specific histone residues, are of great interest as a promising way to identify markers for cancer diagnosis, classify the disease and determine its prognosis, and these markers could be potential targets for the treatment of this disease in its different forms. This manuscript presents the current point of view regarding members of the recently described family of proteins that exhibit histone demethylase activity; histone demethylases are genetic regulators that play a fundamental role in both the activation and repression of genes and whose expression has been observed to increase in many types of cancer. Some fundamental aspects of their association with the development of cancer and their relevance as potential targets for the development of new therapeutic strategies at the epigenetic level are discussed in the following manuscript.
  PubDate: 2012-09-12
  DOI: 10.3390/ph5090963
  Issue No: Vol. 5, No. 9 (2012)
Pharmaceuticals, Vol. 5, Pages 991-1007: Applications of Cell-Penetrating Peptides for Tumor Targeting and Future Cancer Therapies
- Authors: Jakob Regberg, Artita Srimanee, Ülo Langel
  Pages: 991 - 1007
  Abstract: Cell-penetrating peptides provide a highly promising strategy for intracellular drug delivery. One relevant clinical application of cell-penetrating peptides is cancer therapeutics. Peptide based delivery could increase the uptake of drugs in tumor cells and thereby increase the efficacy of the treatment, either of conventional small molecular drugs or oligonucleotide based therapeutics. This review is focused on the cancer applications of cell penetrating peptides as delivery systems; different aspects of drug loading, cargoes and delivery are discussed together with methods for targeted delivery, activatable cell-penetrating peptides and transducible agents coupled to cell-penetrating peptides.
  PubDate: 2012-09-12
  DOI: 10.3390/ph5090991
  Issue No: Vol. 5, No. 9 (2012)
Pharmaceuticals, Vol. 5, Pages 1008-1020: Heat Shock Protein 90 (Hsp90) Expression and Breast Cancer
- Authors: Flora Zagouri, Evangelos Bournakis, Konstantinos Koutsoukos, Christos A. Papadimitriou
  Pages: 1008 - 1020
  Abstract: Hsp90 is an abundant protein in mammalian cells. It forms several discrete complexes, each containing distinct groups of co-chaperones that assist protein folding and refolding during stress, protein transport and degradation. It interacts with a variety of proteins that play key roles in breast neoplasia including estrogen receptors, tumor suppressor p53 protein, angiogenesis transcription factor HIF-1alpha, antiapoptotic kinase Akt, Raf-1 MAP kinase and a variety of receptor tyrosine kinases of the erbB family. Elevated Hsp90 expression has been documented in breast ductal carcinomas contributing to the proliferative activity of breast cancer cells; whilst a significantly decreased Hsp90 expression has been shown in infiltrative lobular carcinomas and lobular neoplasia. Hsp90 overexpression has been proposed as a component of a mechanism through which breast cancer cells become resistant to various stress stimuli. Therefore, pharmacological inhibition of HSPs can provide therapeutic opportunities in the field of cancer treatment. 17-allylamino,17-demethoxygeldanamycin is the first Hsp90 inhibitor that has clinically been investigated in phase II trial, yielding promising results in patients with HER2-overexpressing metastatic breast cancer, whilst other Hsp90 inhibitors (retaspimycin HCL, NVP-AUY922, NVP-BEP800, CNF2024/BIIB021, SNX-5422, STA-9090, etc.) are currently under evaluation.
  PubDate: 2012-09-12
  DOI: 10.3390/ph5091008
  Issue No: Vol. 5, No. 9 (2012)
Pharmaceuticals, Vol. 5, Pages 1021-1231: Why and How the Old Neuroleptic Thioridazine Cures the XDR-TB Patient
- Authors: Leonard Amaral, Joseph Molnar
  Pages: 1021 - 1231
  Abstract: This mini-review provides the entire experimental history of the development of the old neuroleptic thioridazine (TZ) for therapy of antibiotic resistant pulmonary tuberculosis infections. TZ is effective when used in combination with antibiotics to which the initial Mycobacterium tuberculosis was resistant. Under proper cardiac evaluation procedures, the use of TZ is safe and does not produce known cardiopathy such as prolongation of QT interval. Because TZ is cheap, it should be considered for therapy of XDR and TDR-Mtb patients in economically disadvantaged countries.
  PubDate: 2012-09-17
  DOI: 10.3390/ph5091021
  Issue No: Vol. 5, No. 9 (2012)
Pharmaceuticals, Vol. 5, Pages 1032-1042: Evaluation of the Combined Effects of Stilbenoid from Shorea gibbosa and Vancomycin against Methicillin-Resistant Staphylococcus aureus (MRSA)
- Authors: Dayang Fredalina Basri, Chan Kin Luoi, Abdul Muin Azmi, Jalifah Latip
  Pages: 1032 - 1042
  Abstract: The aim of this study is to determine the combined effects of stilbenoids from Shorea gibbosa and vancomycin against methicillin-resistant Staphylococcus aureus (MRSA). A total of nine pure compounds, five stilbenoid dimers ε-viniferin, ampelopsin A, balanocarpol, laevifonol and diptoindonesin G and four stilbenoid trimers a-viniferin, johorenol A, ampelopsin E and vaticanol G were evaluated for their antibacterial activities against ATCC 33591 and a HUKM clinical isolate. Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) for each active compound were determined using the serial microdilution and plate-streak techniques. The combined effect of stilbenoids with vancomycin against MRSA was evaluated using the checkerboard assay to determine their fractional inhibitory concentration (FIC) index values. The MIC value of a-viniferin on both MRSA strains was 100 μg/mL, whereas those of johorenol A on ATCC 33591 and HUKM strain were 100 μg/mL and 200 μg/mL, respectively. The MIC values of ampelopsin E and vaticanol G were higher than 400 μg/mL. Out of the five stilbenoid dimers, only ε-viniferin was capable of inhibiting the growth of both MRSA strains at MIC 400 μg/mL. The MBC value of ε-viniferin, a-viniferin and johorenol A showed bacteriostatic action against MRSA. The FIC index value of ε-viniferin and a-viniferin in combination with vancomycin showed an additive effect (0.5 < FIC ≤ 2.0) against both MRSA strains. Johorenol A-vancomycin combination was also additive against HUKM strain, but it showed synergistic interaction with vancomycin against ATCC 33591 (FIC < 0.5). Stilbenoid compounds from Shorea gibbosa have anti-MRSA activity and huge potential as an alternative phytotherapy in combating MRSA infections.
  PubDate: 2012-09-20
  DOI: 10.3390/ph5091032
  Issue No: Vol. 5, No. 9 (2012)
Pharmaceuticals, Vol. 5, Pages 1044: Qandil, A.M., Synthesis and Spectroscopic Analysis of Novel 1H-Benzo[d]imidazolylphenylsulfonylpiperazines. Pharmaceuticals 2012, 5, 460-468
- Authors: Amjad M. Qandil
  Pages: 1044 - 1044
  Abstract: We have found the following errors in the title of this article which was recently published in Pharmaceuticals [1].
  PubDate: 2012-09-24
  DOI: 10.3390/ph5091044
  Issue No: Vol. 5, No. 9 (2012)
Pharmaceuticals, Vol. 5, Pages 779-801: Heat Shock Protein 90 and Role of Its Chemical Inhibitors in Treatment of Hematologic Malignancies
- Authors: Ngoc Ho, Adam Li, Shaoguang Li, Haojian Zhang
  Pages: 779 - 801
  Abstract: Heat shock protein 90 (Hsp90) is a conserved and constitutively expressed molecular chaperone and it has been shown to stabilize oncoproteins and facilitate cancer development. Hsp90 has been considered as a therapeutic target for cancers and three classes of Hsp90 inhibitors have been developed: (1) benzoquinone ansamycin and its derivatives, (2) radicicol and its derivates, and (3) small synthetic inhibitors. The roles of these inhibitors in cancer treatment have been studied in laboratories and clinical trials, and some encouraging results have been obtained. Interestingly, targeting of Hsp90 has been shown to be effective in inhibition of cancer stem cells responsible for leukemia initiation and progression, providing a strategy for finding a cure. Because cancer stem cells are well defined in some human leukemias, we will focus on hematologic malignancies in this review.
  PubDate: 2012-07-25
  DOI: 10.3390/ph5080779
  Issue No: Vol. 5, No. 8 (2012)
Pharmaceuticals, Vol. 5, Pages 802-836: Involvement of Multiple Transporters-mediated Transports in Mizoribine and Methotrexate Pharmacokinetics
- Authors: Teruo Murakami, Nobuhiro Mori
  Pages: 802 - 836
  Abstract: Mizoribine is administered orally and excreted into urine without being metabolized. Many research groups have reported a linear relationship between the dose and peak serum concentration, between the dose and AUC, and between AUC and cumulative urinary excretion of mizoribine. In contrast, a significant interindividual variability, with a small intraindividual variability, in oral bioavailability of mizoribine is also reported. The interindividual variability is mostly considered to be due to the polymophisms of transporter genes. Methotrexate (MTX) is administered orally and/or by parenteral routes, depending on the dose. Metabolic enzymes and multiple transporters are involved in the pharmacokinetics of MTX. The oral bioavailability of MTX exhibits a marked interindividual variability and saturation with increase in the dose of MTX, with a small intraindividual variability, where the contribution of gene polymophisms of transporters and enzymes is suggested. Therapeutic drug monitoring of both mizoribine and MTX is expected to improve their clinical efficacy in the treatment of rheumatoid arthritis.
  PubDate: 2012-08-10
  DOI: 10.3390/ph5080802
  Issue No: Vol. 5, No. 8 (2012)
Pharmaceuticals, Vol. 5, Pages 837-852: Role of Transient Receptor Potential Vanilloid 1 in Inflammation and Autoimmune Diseases
- Authors: Fumio Tsuji, Hiroyuki Aono
  Pages: 837 - 852
  Abstract: Transient receptor potential vanilloid 1 (TRPV1), a non-selective cation channel, is a receptor activated by high temperatures and chemical agonists such as the vanilloids and protons. Because of these properties, TRPV1 has emerged as a polymodal nocisensor of nociceptive afferent neurons. TRPV1 is thought to be a central transducer of hyperalgesia and a prime target for controlling pain pharmacologically because it is a point where many proalgesic pathways converge and it is upregulated and sensitized by inflammation and injury. However, whether TRPV1 agonists promote or inhibit inflammation remains unclear. We recently demonstrated that SA13353 (1-[2-(1-adamantyl)ethyl]-1-pentyl-3-[3-(4-pyridyl)propyl]urea), a novel TRPV1 agonist, inhibits tumor necrosis factor-a production by the activation of capsaicin-sensitive afferent neurons and reduces the severity of symptoms in kidney injury, lung inflammation, arthritis, and encephalomyelitis. These results suggest that TRPV1 agonists may act as anti-inflammatories in certain inflammatory and autoimmune conditions in vivo. Given the potential deleterious effects of inhibiting the population of channels with a protective function, caution should be taken in the use of potent TRPV1 antagonists as a general strategy to treat inflammation. Further studies are required to clarify the role of TRPV1 and neuropeptides, which are released because of TRPV1 activation in inflammation and autoimmune diseases.
  PubDate: 2012-08-17
  DOI: 10.3390/ph5080837
  Issue No: Vol. 5, No. 8 (2012)
Pharmaceuticals, Vol. 5, Pages 853-874: Phytomedicine in Otorhinolaryngology and Pulmonology: Clinical Trials with Herbal Remedies
- Authors: Koosha Ghazi Moghadam, Hasan Mete Inançlı, Nazanin Bazazy, Peter K. Plinkert, Thomas Efferth, Serkan Sertel
  Pages: 853 - 874
  Abstract: Phytomedicine has become an important alternative treatment option for patients in the Western world, as they seek to be treated in a holistic and natural way after an unsatisfactory response to conventional drugs. Ever since herbal remedies have been introduced in the Western world, clinicians have raised concerns over their efficacy and possible side-effects. A PubMed (Medline) search was performed covering the last five years (01/07–04/12) and including 55 prospective clinical randomized control trials in the medical specialities Otorhinolaryngology and Pulmonology. In this review, we present evidence-based clinical data with herbal remedies and try to enlighten the question of efficacy and reliability of phytomedicine.
  PubDate: 2012-08-20
  DOI: 10.3390/ph5080853
  Issue No: Vol. 5, No. 8 (2012)
Pharmaceuticals, Vol. 5, Pages 875-881: A Perspective on the Comparative Antileukemic Activity of 5-Aza-2′-deoxycytidine (Decitabine) and 5-Azacytidine (Vidaza)
- Authors: Richard L. Momparler
  Pages: 875 - 881
  Abstract: 5-Aza-2′-deoxycytidine (5-AZA-CdR, decitabine, Dacogen®) and 5-azacytidine (5-AC, Vidaza®) are epigenetic agents that have been approved for the clinical treatment of the hematological malignancy myelodysplastic syndrome (MDS) and are currently under clinical evaluation for the treatment of acute myeloid leukemia (AML). Most investigators currently classify 5-AZA-CdR and 5-AC as inhibitors of DNA methylation, which can reactivate tumor suppressor genes silenced by this epigenetic event. Examination of the pharmacology of these analogues reveals important differences with respect to their molecular mechanism of action. The action of 5-AZA-CdR is due to its incorporation into DNA. 5-AC is a riboside analogue that is incorporated primarily into RNA. A small fraction of 5-AC is converted to its deoxyribose form by ribonucleotide reductase and subsequently incorporated into DNA. The incorporation of 5-AC into RNA can interfere with the biological function of RNA and result in an inhibition protein synthesis. Microarray analysis revealed that both these analogues target the expression of different cohorts of genes. Preclinical studies show that 5-AZA-CdR is a more effective antileukemic agent than 5-AC. One explanation for this observation is that 5-AC blocks the progression of some leukemic cells from G1 into S phase, and this protects these cells from the chemotherapeutic action of this riboside analogue related to its incorporation into DNA. However, differences in chemotherapeutic efficacy of these related analogues have not been clearly demonstrated in clinical trials in patients with hematological malignancies. These observations should be taken into consideration in the design of new clinical trials using 5-AZA-CdR or 5-AC in patients with MDS and AML.
  PubDate: 2012-08-21
  DOI: 10.3390/ph5080875
  Issue No: Vol. 5, No. 8 (2012)
Pharmaceuticals, Vol. 5, Pages 882-889: The Chemical Components of Sesbania grandiflora Root and Their Antituberculosis Activity
- Authors: Noviany Hasan, Hasnah Osman, Suriyati Mohamad, Wong Keng Chong, Khalijah Awang, Anis Safirah Mohd Zahariluddin
  Pages: 882 - 889
  Abstract: Three isoflavanoids, isovestitol (1), medicarpin (2), and sativan (3), along with another known compound, betulinic acid (4), were isolated from the root of Sesbania grandiflora. The structures of the isolated compounds were characterised by means of spectroscopic techniques (UV, IR, MS, 1H- and 13C-NMR, DEPT, COSY, HMQC, HMBC, and MS analysis). All the tested compounds 1–4 exhibited antituberculosis activity against Mycobacterium tuberculosis H37Rv, with MIC values of 50 µg/mL for compounds 1–3, and 100 µg/mL for compound 4, whereas, the methanol extract exhibited antituberculosis activity of 625 µg/mL. This is the first report on the occurrence of isoflavonoids in this plant and their antituberculosis activity.
  PubDate: 2012-08-23
  DOI: 10.3390/ph5080882
  Issue No: Vol. 5, No. 8 (2012)
Pharmaceuticals, Vol. 5, Pages 674-689: Pertuzumab Increases 17-AAG-Induced Degradation of ErbB2, and This Effect Is Further Increased by Combining Pertuzumab with Trastuzumab
- Authors: Juliana Bentes Hughes, Marianne Skeie Rødland, Max Hasmann, Inger Helene Madshus, Espen Stang
  Pages: 674 - 689
  Abstract: ErbB2 is an important oncogenic protein involved in carcinogenesis of, among others, breast, gastric, and ovarian carcinoma. Over-expression of ErbB2 is found in almost 20% of breast cancers, and this results in proliferative and anti-apoptotic signalling. ErbB2 is therefore an important treatment target. Antibodies recognizing full-length ErbB2 are clinically established, and drugs targeting the ErbB2 stabilizing heat shock protein 90 (Hsp90) are under clinical evaluation. We have investigated effects of the ErbB2-binding antibodies trastuzumab and pertuzumab alone and in combination, as well as the effect of the antibodies in combination with the Hsp90 inhibitor 17-AAG. Our results confirm the notion that combination of different ErbB2-binding antibodies more efficiently down-regulates ErbB2 than does one antibody in isolation. Additionally, our data demonstrate that ErbB2 is most efficiently down-regulated upon incubation with anti-ErbB2 antibodies in combination with Hsp90 inhibitors. The combination of anti-ErbB2 antibodies, and especially the combination of antibodies with 17-AAG, did also increase the inhibition of Akt activation of either agent, which could suggest an anti-proliferative effect. In such case, combining these agents could be beneficial in treatment of tumors not responding to trastuzumab only.
  PubDate: 2012-06-28
  DOI: 10.3390/ph5070674
  Issue No: Vol. 5, No. 7 (2012)
Pharmaceuticals, Vol. 5, Pages 690-718: Chemotherapeutic Interventions Against Tuberculosis
- Authors: Neeraj Shakya, Gaurav Garg, Babita Agrawal, Rakesh Kumar
  Pages: 690 - 718
  Abstract: Tuberculosis is the second leading cause of infectious deaths globally. Many effective conventional antimycobacterial drugs have been available, however, emergence of multidrug-resistant tuberculosis (MDR-TB) and extensively drug-resistant tuberculosis (XDR-TB) has overshadowed the effectiveness of the current first and second line drugs. Further, currently available agents are complicated by serious side effects, drug interactions and long-term administration. This has prompted urgent research efforts in the discovery and development of new anti-tuberculosis agent(s). Several families of compounds are currently being explored for the treatment of tuberculosis. This review article presents an account of the existing chemotherapeutics and highlights the therapeutic potential of emerging molecules that are at different stages of development for the management of tuberculosis disease.
  PubDate: 2012-06-28
  DOI: 10.3390/ph5070690
  Issue No: Vol. 5, No. 7 (2012)
Pharmaceuticals, Vol. 5, Pages 719-726: Association of Marijuana Use and Cyclic Vomiting Syndrome
- Authors: Mithun B. Pattathan, Reza A. Hejazi, Richard W. McCallum
  Pages: 719 - 726
  Abstract: Cannabis use has become one of the most commonly abused drugs in the world. It is estimated that each year 2.6 million individuals in the USA become new users and most are younger than 19 years of age. Reports describe marijuana use as high as 40–50% in male Cyclic Vomiting Syndrome patients. It is this interest in cannabis in the World, coupled with recognition of a cyclic vomiting illness associated with its chronic use that beckons a review of the most current articles, as well as a contribution from our own experiences in this area. The similarities we have demonstrated for both cannibinoid hyperemesis syndrome and cyclic vomiting make the case that cannibinoid hyperemesis syndrome is a subset of patients who have the diagnoses of cyclic vomiting syndrome and the role of marijuana should always be considered in the diagnosis of CVS, particularly in males.
  PubDate: 2012-06-29
  DOI: 10.3390/ph5070719
  Issue No: Vol. 5, No. 7 (2012)
Pharmaceuticals, Vol. 5, Pages 727-744: Probiotics and Atopic Dermatitis in Children
- Authors: Fabio Meneghin, Valentina Fabiano, Chiara Mameli, Gian Vincenzo Zuccotti
  Pages: 727 - 744
  Abstract: There is increasing interest in the potential beneficial role of probiotic supplementation in the prevention and treatment of atopic diseases in children. Probiotics are defined as ingested live microorganisms that, when administered in an adequate amount, confer a health benefit to the host. They are mainly represented by Lactobacilli and Bifidobacteria. Several epidemiological data demonstrate that intestinal microflora of atopic children is different from the one of healthy children. Many literature data show that probiotics may modulate the intestinal microflora composition and may have immunomodulatory effect. Based on this hypothesis, probiotics are supposed to confer benefits to allergic diseases. Administration of probiotics when a natural population of indigenous intestinal bacteria is still developing could theoretically influence immune development by favoring the balance between Th1 and Th2 inflammatory responses. For this reason, some studies have evaluated the potential impact of probiotics supplementation in the prevention of atopic dermatitis, with contrasting results. Clinical improvement in immunoglobulin (Ig)E-sensitized (atopic) eczema following probiotic supplementation has been reported in some published studies and the therapeutic effects of probiotics on atopic dermatitis seemed to be encouraging. However, as far as the usefulness of probiotics as a prevention strategy is concerned, results are still inconclusive. In fact, the clinical benefits of probiotic therapy depend upon numerous factors, such as the type of bacteria, dosing regimen, delivery method and other underlying host factors, such as age and diet. More studies are still needed to definitively prove the role of probiotics in the treatment of allergic eczema.
  PubDate: 2012-07-06
  DOI: 10.3390/ph5070727
  Issue No: Vol. 5, No. 7 (2012)
Pharmaceuticals, Vol. 5, Pages 745-757: Synthesis, Reactions and Evaluation of the Antimicrobial Activity of Some 4-(p-Halophenyl)-4H-naphthopyran, Pyranopyrimidine and Pyranotriazolopyrimidine Derivatives
- Authors: Ashraf H. F. Abd El-Wahab
  Pages: 745 - 757
  Abstract: A series of naphthopyran derivatives 3a–f were prepared. Reaction of 2-amino-4-(p-chlorophenyl)-7-methoxy-4H-naphtho[2,1-b]pyran-3-carbonitrile (3b) with Ac2O afforded two products, 2-acetylamino-7-methoxy-4-(p-chlorophenyl)-4H-naphtho-[2,1-b]pyran-3-carbonitrile (4) and 10,11-dihydro-3-methoxy-9-methyl-12-(p-chloro-phenyl)-12H-naphtho[2,1-b]pyran[2,3-d]pyrimidine-11-one (5) and treatment of 3b with benzoyl chloride gave the pyranopyrimidin-11-one derivative 6. While treatment of 3b with formamide afforded 11-amino-3-methoxy-12-(p-chlorophenyl)-12H-naphtho[2,1-b]pyrano[2,3-d]pyrimidine (7). Reaction of 3b with triethyl orthoformate gave the corresponding 2-ethoxymethyleneamino-7-methoxy-4-(p-chlorophenyl)-4H-naphtho-[2,1-b]pyran-3-carbonitrile (8). Hydrazinolysis of 8 in EtOH at room temperature yielded 10-amino-10,11-dihydro-11-imino-3-methoxy-12-(p-chlorophenyl)-12H-naphtho[2,1-b]pyrano-[2,3-d]pyrimidine (9), while aminolysis of 8 with methylamine or dimethylamine gave the corresponding pyranopyrimidine and N,N-dimethylaminomethylene derivatives 10 and 11. Condensation of 9 with some carboxylic acid derivatives afforded triazolopyrimidine derivatives 12–16, while reaction of 9 with benzaldehyde gave 10-benzalamino-10,11-dihydro-11-imino-3-methoxy-12-(p-chlorophenyl)12H-naphtho[2,1-b]pyrano[2,3-d]pyrimidine (17). The structures of the newly synthesized compounds were confirmed by spectral data. The synthesized compounds were also screened for their antimicrobial activity.
  PubDate: 2012-07-12
  DOI: 10.3390/ph5070745
  Issue No: Vol. 5, No. 7 (2012)
Pharmaceuticals, Vol. 5, Pages 758-778: The Influence of Estrogens on the Biological and Therapeutic Actions of Growth Hormone in the Liver
- Authors: Mercedes de Mirecki-Garrido, Borja Guerra, Carlos Mateos-Díaz, Roberto Jiménez-Monzón, Nicolás Díaz-Chico, Juan C. Díaz-Chico, Leandro Fernández-Pérez
  Pages: 758 - 778
  Abstract: GH is main regulator of body growth and composition, somatic development, intermediate metabolism and gender-dependent dimorphism in mammals. The liver is a direct target of estrogens because it expresses estrogen receptors which are connected with development, lipid metabolism and insulin sensitivity, hepatic carcinogenesis, protection from drug-induced toxicity and fertility. In addition, estrogens can modulate GH actions in liver by acting centrally, regulating pituitary GH secretion, and, peripherally, by modulating GHR-JAK2-STAT5 signalling pathway. Therefore, the interactions of estrogens with GH actions in liver are biologically and clinically relevant because disruption of GH signaling may cause alterations of its endocrine, metabolic, and gender differentiated functions and it could be linked to dramatic impact in liver physiology during development as well as in adulthood. Finally, the interplay of estrogens with GH is relevant because physiological roles these hormones have in human, and the widespread exposition of estrogen or estrogen-related compounds in human. This review highlights the importance of these hormones in liver physiology as well as how estrogens modulate GH actions in liver which will help to improve the clinical use of these hormones.
  PubDate: 2012-07-19
  DOI: 10.3390/ph5070758
  Issue No: Vol. 5, No. 7 (2012)
Pharmaceuticals, Vol. 5, Pages 553-590: Gene Therapy for the Treatment of Parkinson’s Disease: The Nature of the Biologics Expands the Future Indications
- Authors: Massimo S. Fiandaca, Krystof S. Bankiewicz, Howard J. Federoff
  Pages: 553 - 590
  Abstract: The pharmaceutical industry’s development of therapeutic medications for the treatment of Parkinson’s disease (PD) endures, as a result of the continuing need for better agents, and the increased clinical demand due to the aging population. Each new drug offers advantages and disadvantages to patients when compared to other medical offerings or surgical options. Deep brain stimulation (DBS) has become a standard surgical remedy for the effective treatment of select patients with PD, for whom most drug regimens have failed or become refractory. Similar to DBS as a surgical option, gene therapy for the treatment of PD is evolving as a future option. In the four different PD gene therapy approaches that have reached clinical trials investigators have documented an excellent safety profile associated with the stereotactic delivery, viral vectors and doses utilized, and transgenes expressed. In this article, we review the clinically relevant gene therapy strategies for the treatment of PD, concentrating on the published preclinical and clinical results, and the likely mechanisms involved. Based on these presentations, we advance an analysis of how the nature of the gene therapy used may eventually expand the scope and utility for the management of PD.
  PubDate: 2012-06-04
  DOI: 10.3390/ph5060553
  Issue No: Vol. 5, No. 6 (2012)
Pharmaceuticals, Vol. 5, Pages 591-612: a-Anilinoketones, Esters and Amides: A Chemical Study
- Authors: Amjad M. Qandil, Lara I. Fakhouri
  Pages: 591 - 612
  Abstract: A group of a-anilinoketones, 2-aminoalcohols, a-anilinoesters and a-anilinoamides were successfully synthesized and characterized by NMR spectroscopy and mass spectrometry. The yields were, in general, moderate to good (up to 75.4%), except for the a-anilinoesters (16.9–35.6%). The a-halocarbonyl starting materials showed different chemical reactivities. a-Haloketones and a-chloroacetates afforded monoalkylation, while small a-chloroamides afforded dialkylation. Finally, NMR spectroscopy revealed interesting structural features about the 2-aminoalcohols and diphenylamides.
  PubDate: 2012-06-05
  DOI: 10.3390/ph5060591
  Issue No: Vol. 5, No. 6 (2012)
Pharmaceuticals, Vol. 5, Pages 613-628: Phytochemical Composition, Antioxidant and Xanthine Oxidase Inhibitory Activities of Amaranthus cruentus L. and Amaranthus hybridus L. Extracts
- Authors: Fernand W. Nana, Adama Hilou, Jeanne F. Millogo, Odile G. Nacoulma
  Pages: 613 - 628
  Abstract: This paper describes a preliminary assessment of the nutraceutical value of Amaranthus cruentus (A. cruentus) and Amaranthus hybridus (A. hybridus), two food plant species found in Burkina Faso. Hydroacetonic (HAE), methanolic (ME), and aqueous extracts (AE) from the aerial parts were screened for in vitro antioxidant and xanthine oxidase inhibitory activities. Phytochemical analyses revealed the presence of polyphenols, tannins, flavonoids, steroids, terpenoids, saponins and betalains. Hydroacetonic extracts have shown the most diversity for secondary metabolites. The TLC analyses of flavonoids from HAE extracts showed the presence of rutin and other unidentified compounds. The phenolic compound contents of the HAE, ME and AE extracts were determined using the Folin–Ciocalteu method and ranged from 7.55 to 10.18 mg Gallic acid equivalent GAE/100 mg. Tannins, flavonoids, and flavonols ranged from 2.83 to 10.17 mg tannic acid equivalent (TAE)/100 mg, 0.37 to 7.06 mg quercetin equivalent (QE) /100 mg, and 0.09 to 1.31 mg QE/100 mg, respectively. The betacyanin contents were 40.42 and 6.35 mg Amaranthin Equivalent/100 g aerial parts (dry weight) in A. cruentus and A. hybridus, respectively. Free-radical scavenging activity expressed as IC50 (DPPH method) and iron reducing power (FRAP method) ranged from 56 to 423 µg/mL and from 2.26 to 2.56 mmol AAE/g, respectively. Xanthine oxidase inhibitory activities of extracts of A. cruentus and A. hybridus were 3.18% and 38.22%, respectively. The A. hybridus extract showed the best antioxidant and xanthine oxidase inhibition activities. The results indicated that the phytochemical contents of the two species justify their traditional uses as nutraceutical food plants.
  PubDate: 2012-06-15
  DOI: 10.3390/ph5060613
  Issue No: Vol. 5, No. 6 (2012)
Pharmaceuticals, Vol. 5, Pages 629-642: Influence of Maternal Bifidobacteria on the Development of Gut Bifidobacteria in Infants
- Authors: Katsunaka Mikami, Moto Kimura, Hedenori Takahashi
  Pages: 629 - 642
  Abstract: Intestinal microbiota plays an important role in human health by influencing metabolic activities that result in the creation of energy and absorbable nutrients, a barrier to the colonization of pathogens, and stimulation of the immune system. The development of fecal microbiota in neonates is crucial because those bacteria are the first to colonize the sterile intestine of the neonates and, thus, have a significant effect on the host. Initial colonization is also relevant to the ﬁnal composition of the permanent microbiota in adults. Bifidobacteria are predominant in the fecal microbiota of infants, and, therefore, they are important to an understanding of how commensal bifidobacteria is established in the intestine of infants. While the mother’s bifidobacteria are considered to significantly influence the infant’s bifidobacteria, it is not clear whether a specific bifidobacterial strain transmits vertically from mother to infant and what factors of the mother before delivery influence the establishment of intestinal bifidobacteria in infants. This review focuses on the impact of maternal bifidobacteria on the development of gut bifidobacteria in the infant and suggests that there is cumulative evidence regarding bifidobacterial transfer from the maternal gut or breast milk to the infant gut.
  PubDate: 2012-06-18
  DOI: 10.3390/ph5060629
  Issue No: Vol. 5, No. 6 (2012)
Pharmaceuticals, Vol. 5, Pages 643-654: Physical Factors Affecting Plasmid DNA Compaction in Stearylamine-Containing Nanoemulsions Intended for Gene Delivery
- Authors: André Leandro Silva, Francisco Alexandrino, Lourena Mafra Verissimo, Lucymara Fassarella Agnez-Lima, Lucila Carmem Monte Egito, Anselmo Gomes de Oliveira, Eryvaldo Socrates Tabosa do Egito
  Pages: 643 - 654
  Abstract: Cationic lipids have been used in the development of non-viral gene delivery systems as lipoplexes. Stearylamine, a cationic lipid that presents a primary amine group when in solution, is able to compact genetic material by electrostatic interactions. In dispersed systems such as nanoemulsions this lipid anchors on the oil/water interface confering a positive charge to them. The aim of this work was to evaluate factors that influence DNA compaction in cationic nanoemulsions containing stearylamine. The influence of the stearylamine incorporation phase (water or oil), time of complexation, and different incubation temperatures were studied. The complexation rate was assessed by electrophoresis migration on agarose gel 0.7%, and nanoemulsion and lipoplex characterization was done by Dynamic Light Scattering (DLS). The results demonstrate that the best DNA compaction process occurs after 120 min of complexation, at low temperature (4 ± 1 °C), and after incorporation of the cationic lipid into the aqueous phase. Although the zeta potential of lipoplexes was lower than the results found for basic nanoemulsions, the granulometry did not change. Moreover, it was demonstrated that lipoplexes are suitable vehicles for gene delivery.
  PubDate: 2012-06-18
  DOI: 10.3390/ph5060643
  Issue No: Vol. 5, No. 6 (2012)
Pharmaceuticals, Vol. 5, Pages 655-664: The Potential Therapeutic Efficacy of Lactobacillus GG in Children with Food Allergies
- Authors: Roberto Berni Canani, Margherita Di Costanzo, Vincenza Pezzella, Linda Cosenza, Viviana Granata, Gianluca Terrin, Rita Nocerino
  Pages: 655 - 664
  Abstract: Food allergy (FA) continues to be a growing health concern for infants living in Western countries. The long-term prognosis for the majority of affected infants is good, with 80–90% naturally acquiring tolerance by the age of five years. However, recent studies suggest that the natural history of FA is changing, with an increasing persistence until later ages. The pathogenesis of FA as well as oral tolerance is complex and not completely known, although numerous studies implicate gut-associated immunity and enteric microflora, and it has been suggested that an altered composition of intestinal microflora results in an unbalanced local and systemic immune response to food allergens. In addition, there are qualitative and quantitative differences in the composition of gut microbiota between patients affected by FA and healthy infants. These findings prompted the concept that specific beneficial bacteria from the human intestinal microflora, designated probiotics, could restore intestinal homeostasis and prevent or alleviate allergy, at least in part by interacting with the intestinal immune cells.
  PubDate: 2012-06-19
  DOI: 10.3390/ph5060655
  Issue No: Vol. 5, No. 6 (2012)
Pharmaceuticals, Vol. 5, Pages 665-673: Removal of Cholera Toxin from Aqueous Solution by Probiotic Bacteria
- Authors: Jari E. Heikkilä, Sonja M. K. Nybom, Seppo J. Salminen, Jussi A. O. Meriluoto
  Pages: 665 - 673
  Abstract: Cholera remains a serious health problem, especially in developing countries where basic hygiene standards are not met. The symptoms of cholera are caused by cholera toxin, an enterotoxin, which is produced by the bacterium Vibrio cholerae. We have recently shown that human probiotic bacteria are capable of removing cyanobacterial toxins from aqueous solutions. In the present study we investigate the ability of the human probiotic bacteria, Lactobacillus rhamnosus strain GG (ATCC 53103) and Bifidobacterium longum 46 (DSM 14583), to remove cholera toxin from solution in vitro. Lactobacillus rhamnosus strain GG and Bifidobacterium longum 46 were able to remove 68% and 59% of cholera toxin from aqueous solutions during 18 h of incubation at 37 °C, respectively. The effect was dependent on bacterial concentration and L. rhamnosus GG was more effective at lower bacterial concentrations. No significant effect on cholera toxin concentration was observed when nonviable bacteria or bacterial supernatant was used.
  PubDate: 2012-06-19
  DOI: 10.3390/ph5060665
  Issue No: Vol. 5, No. 6 (2012)
Pharmaceuticals, Vol. 5, Pages 405-416: Poly(ethylene glycol)-Lipid-Conjugated Antibodies Enhance Dendritic Cell Phagocytosis of Apoptotic Cancer Cells
- Authors: Urara Tomita, Satoshi Yamaguchi, Yoichiro Sugimoto, Satoshi Takamori, Teruyuki Nagamune
  Pages: 405 - 416
  Abstract: A simple method for attaching immunoglobulin G (IgG) on the cell surface was successfully developed for enhancing phagocytosis of apoptotic tumor cells (ATCs) by dendritic cells (DCs) ex vivo. By conjugating with a poly(ethylene glycol) (PEG)-lipid, named the biocompatible anchor for the membrane (BAM), arbitrary IgG could be incorporated into the cell membrane. In particular, when IgG-BAM conjugates were prepared at the optimal molar ratio of IgG to BAM (1 to 20), almost all cells were efficiently modified with IgG by treatment with IgG-BAM. This simple method was successfully applied to four types of mammalian cells. Furthermore, treatment of ATCs with the IgG-BAM conjugate increased the phagocytosis ratio of ATCs by DCs two-fold when compared to no treatment. This phagocytosis-enhancing effect was nearly identical to treatment with a tumor-specific IgG. Thus, without employing the tumor-specific IgG, which is difficult to obtain for any tumor cells and is expensive, the present method could opsonize ATC with the use of arbitrary IgG. The results strongly indicate that IgG-BAM treatment represents a promising method for opsonizing ATC with human serum IgG, and that this approach will lead to objective clinical responses in DC vaccines.
  PubDate: 2012-04-26
  DOI: 10.3390/ph5050405
  Issue No: Vol. 5, No. 5 (2012)
Pharmaceuticals, Vol. 5, Pages 417-446: Pathophysiology of GPCR Homo- and Heterodimerization: Special Emphasis on Somatostatin Receptors
- Authors: Rishi K. Somvanshi, Ujendra Kumar
  Pages: 417 - 446
  Abstract: G-protein coupled receptors (GPCRs) are cell surface proteins responsible for translating >80% of extracellular reception to intracellular signals. The extracellular information in the form of neurotransmitters, peptides, ions, odorants etc is converted to intracellular signals via a wide variety of effector molecules activating distinct downstream signaling pathways. All GPCRs share common structural features including an extracellular N-terminal, seven-transmembrane domains (TMs) linked by extracellular/intracellular loops and the C-terminal tail. Recent studies have shown that most GPCRs function as dimers (homo- and/or heterodimers) or even higher order of oligomers. Protein-protein interaction among GPCRs and other receptor proteins play a critical role in the modulation of receptor pharmacology and functions. Although ~50% of the current drugs available in the market target GPCRs, still many GPCRs remain unexplored as potential therapeutic targets, opening immense possibility to discover the role of GPCRs in pathophysiological conditions. This review explores the existing information and future possibilities of GPCRs as tools in clinical pharmacology and is specifically focused for the role of somatostatin receptors (SSTRs) in pathophysiology of diseases and as the potential candidate for drug discovery.
  PubDate: 2012-04-27
  DOI: 10.3390/ph5050417
  Issue No: Vol. 5, No. 5 (2012)
Pharmaceuticals, Vol. 5, Pages 447-459: The Signalling Role of the avβ5-Integrin Can Impact the Efficacy of AAV in Retinal Gene Therapy
- Authors: Therese Cronin, Daniel C. Chung, Ying Yang, Emeline F. Nandrot, Jean Bennett
  Pages: 447 - 459
  Abstract: Sub-retinal injection of the common AAV2 pseudotypes frequently results in strong transduction of the retinal pigment epithelium (RPE) as well as the retina itself. This has been of benefit to date in human clinical trials using AAV, where the disease target is in the RPE. However, many mutations predisposing to retinal disease are located in the photoreceptor cells, present in the neural retina and not the RPE; in this case the sub-retinal injection route may cause an effective “loss” of therapeutic AAV to the RPE. The avβ5 integrin receptor is highly expressed on the apical surface of the RPE, and is essential to the daily phagocytosis of the outer segment tips of photoreceptor cells. The transduction efficiency of AAV was tested in the retinas of β5−/− mice lacking this receptor and showing defects in photoreceptor outer segment phagocytosis. Following sub-retinal injection of AAV2/5-eGFP, fluorescence was found to be stronger and more widespread in the neural retina of β5−/− mice compared to wild-types with greatly reduced fluorescence in the RPE. Increased levels of the phagocytic signalling protein MFG-E8, the ligand for the avβ5 integrin receptor, is found to have a moderate inhibitory effect on AAV transduction of the retina. However the opposite effect is found when only the integrin-binding domain of MFG-E8, the RGD (Arginine-Glycine-Aspartic acid) domain, was increased. In this case RGD enhanced AAV-mediated retinal transduction relative to RPE transduction. These results are presented for their relevance for the design of AAV-based retinal gene therapy strategies strategies targeting retinal/photoreceptor cells.
  PubDate: 2012-05-02
  DOI: 10.3390/ph5050447
  Issue No: Vol. 5, No. 5 (2012)
Pharmaceuticals, Vol. 5, Pages 460-468: Synthesis and Spectroscopic Analysis of Novel 1H-Benzo[d]imidazoles Phenyl Sulfonylpiperazines
- Authors: Amjad M. Qandil
  Pages: 460 - 468
  Abstract: A group of benzimidazole analogs of sildenafil, 3-benzimidazolyl-4-methoxy-phenylsulfonylpiperazines 2–4 and 3-benzimidazolyl-4-methoxy-N,N-dimethyl- benzenesulfonamide (5), were efficiently synthesized. Compounds 2–5 were characterized by NMR and MS and contrary to the reported mass spectra of sildenafil, the spectra of the piperazine-containing compounds 2–4 showed a novel fragmentation pattern leading to an m/z = 316. A mechanism for the formation of this fragment was proposed.
  PubDate: 2012-05-03
  DOI: 10.3390/ph5050460
  Issue No: Vol. 5, No. 5 (2012)
Pharmaceuticals, Vol. 5, Pages 469-480: Newer Anticoagulants for Non-Valvular Atrial Fibrillation
- Authors: Joseph M. Harburger, Wilbert S. Aronow
  Pages: 469 - 480
  Abstract: Non-valvular atrial fibrillation is a recognized risk factor for stroke and systemic embolism. It has been clearly established that warfarin reduces the risk of stroke and systemic embolism in persons with atrial fibrillation and additional risk factors for stroke. The use of warfarin, however, requires frequent monitoring, and there is great variability in patient response to warfarin. Warfarin interacts with several medications and foods. In addition, warfarin use portends a significant risk of bleeding. For these reasons, warfarin is frequently not prescribed to persons for whom the drug would provide a clear benefit. Over the past decade, attempts have been made to develop drugs that are at least as safe and effective as warfarin for the treatment of atrial fibrillation that do not require monitoring nor have as many interactions. Initial studies of compounds in this regard ultimately failed due to safety concerns, but over the past two years two novel agents have been approved by the United States Food and Drug Association for anticoagulation in non-valvular atrial fibrillation, another drug is under review, and additional compounds are being studied. This article will review the use of warfarin and these new agents in the treatment of non-valvular atrial fibrillation.
  PubDate: 2012-05-04
  DOI: 10.3390/ph5050469
  Issue No: Vol. 5, No. 5 (2012)
Pharmaceuticals, Vol. 5, Pages 481-492: Biochemical Traits, Survival and Biological Properties of the Probiotic Lactobacillus plantarum Grown in the Presence of Prebiotic Inulin and Pectin as Energy Source
- Authors: Filomena Nazzaro, Florinda Fratianni, Pierangelo Orlando, Raffaele Coppola
  Pages: 481 - 492
  Abstract: The viability of the probiotic strain Lactobacillus plantarum subsp. plantarum, after its passage through simulated gastric and pancreatic juices, was evaluated as function of its pre-growth in a medium containing the known prebiotics pectin or inulin, and was compared to glucose used as control. The presence of pectin or inulin did not markedly affect the growth (10.07 log10 colony forming units/mL and 10.28 log10 colony forming units/mL for pectin and inulin respectively versus 10.42 log10 colony forming units/mL obtained for glucose). Pectin and inulin, in contrast to glucose, induced cell stress resistance against gastrointestinal juices (D log101.5 and 2.4 colony forming units/mL respectively, versus D log10 4.0 for glucose). The data were corroborated by the analysis of the protein pattern following stress treatments which, in the case of microbial cells grown with glucose, revealed a more marked protein degradation after the double passage through simulated gastric and intestinal juices. Inulin stimulated the production of the relevant healthy bio-molecule butyrate, which amount was 30% higher respect of growth in the presence of glucose. Inulin and pectin improved cell DPPH scavenging activity, and an impressive hydrophobicity (35.28% and 34.81%, respectively) was observed with respect to the microbial growth in presence of glucose (3.39%).
  PubDate: 2012-05-15
  DOI: 10.3390/ph5050481
  Issue No: Vol. 5, No. 5 (2012)
Pharmaceuticals, Vol. 5, Pages 493-513: Selecting Molecular Recognition. What Can Existing Aptamers Tell Us about Their Inherent Recognition Capabilities and Modes of Interaction'
- Authors: Qian Zhang, Ralf Landgraf
  Pages: 493 - 513
  Abstract: The use of nucleic acid derived aptamers has rapidly expanded since the introduction of SELEX in 1990. Nucleic acid aptamers have demonstrated their ability to target a broad range of molecules in ways that rival antibodies, but advances have been very uneven for different biochemical classes of targets, and clinical applications have been slow to emerge. What sets different aptamers apart from each other and from rivaling molecular recognition platforms, specifically proteins' What advantages do aptamers as a reagent class offer, and how do the chemical properties and selection procedures of aptamers influence their function' Do the building blocks of nucleic acid aptamers dictate inherent limitations in the nature of molecular targets, and do existing aptamers give us insight in how these challenges might be overcome' This review is written as an introduction for potential endusers of aptamer technology who are evaluating the advantages of aptamers as a versatile, affordable, yet highly expandable platform to target a broad range of biological processes or interactions.
  PubDate: 2012-05-18
  DOI: 10.3390/ph5050493
  Issue No: Vol. 5, No. 5 (2012)
Pharmaceuticals, Vol. 5, Pages 514-528: Sphingosine 1-Phosphate Receptor 1 as a Useful Target for Treatment of Multiple Sclerosis
- Authors: Kenji Chiba, Kunitomo Adachi
  Pages: 514 - 528
  Abstract: Sphingosine 1-phosphate (S1P), a lysophospholipid mediator, is generated from sphingosine by sphingosine kinases and binds five known cell surface receptors. S1P receptor 1 (S1P1) plays an essential role in lymphocyte egress from secondary lymphoid organs (SLO), as evinced by the inability of lymphocytes to exit from the SLO in mice lacking lymphocytic S1P1. Fingolimod hydrochloride (FTY720) is a first-in-class, orally active, S1P receptor modulator with a structure closely related to sphingosine. FTY720 was first synthesized by chemical modification of a natural product, myriocin. FTY720 is effectively converted to an active metabolite, FTY720 phosphate (FTY720-P) by sphingosine kinases. FTY720-P shows high affinity to 4 of the S1P receptors (S1P1, S1P3, S1P4, and S1P5). In particular, FTY720-P strongly induces internalization and degradation of S1P1, inhibits S1P responsiveness of lymphocytes in the SLO, and acts as a functional antagonist at lymphocytic S1P1. Consequently, FTY720 inhibits S1P1-dependent lymphocyte egress from the SLO to decrease circulation of lymphocytes including autoreactive Th17 cells and is highly effective in experimental autoimmune encephalomyelitis (EAE), an animal model of multiple sclerosis (MS). Because FTY720 shows a superior efficacy in relapsing remitting MS patients compared to intramuscular interferon-β-1a (Avonex®), S1P1 is presumed to be a useful target for the therapy of MS.
  PubDate: 2012-05-18
  DOI: 10.3390/ph5050514
  Issue No: Vol. 5, No. 5 (2012)
Pharmaceuticals, Vol. 5, Pages 529-552: Cannabidiol in Humans—The Quest for Therapeutic Targets
- Authors: Simon Zhornitsky, Stéphane Potvin
  Pages: 529 - 552
  Abstract: Cannabidiol (CBD), a major phytocannabinoid constituent of cannabis, is attracting growing attention in medicine for its anxiolytic, antipsychotic, antiemetic and anti-inflammatory properties. However, up to this point, a comprehensive literature review of the effects of CBD in humans is lacking. The aim of the present systematic review is to examine the randomized and crossover studies that administered CBD to healthy controls and to clinical patients. A systematic search was performed in the electronic databases PubMed and EMBASE using the key word “cannabidiol”. Both monotherapy and combination studies (e.g., CBD + ∆9-THC) were included. A total of 34 studies were identified: 16 of these were experimental studies, conducted in healthy subjects, and 18 were conducted in clinical populations, including multiple sclerosis (six studies), schizophrenia and bipolar mania (four studies), social anxiety disorder (two studies), neuropathic and cancer pain (two studies), cancer anorexia (one study), Huntington’s disease (one study), insomnia (one study), and epilepsy (one study). Experimental studies indicate that a high-dose of inhaled/intravenous CBD is required to inhibit the effects of a lower dose of ∆9-THC. Moreover, some experimental and clinical studies suggest that oral/oromucosal CBD may prolong and/or intensify ∆9-THC-induced effects, whereas others suggest that it may inhibit ∆9-THC-induced effects. Finally, preliminary clinical trials suggest that high-dose oral CBD (150–600 mg/d) may exert a therapeutic effect for social anxiety disorder, insomnia and epilepsy, but also that it may cause mental sedation. Potential pharmacokinetic and pharmacodynamic explanations for these results are discussed.
  PubDate: 2012-05-21
  DOI: 10.3390/ph5050529
  Issue No: Vol. 5, No. 5 (2012)
Pharmaceuticals, Vol. 5, Pages 339-352: Oral Administration of Shark Type II Collagen Suppresses Complete Freund’s Adjuvant-Induced Rheumatoid Arthritis in Rats
- Authors: Lijuan Chen, Bin Bao, Nanping Wang, Jing Xie, Wenhui Wu
  Pages: 339 - 352
  Abstract: Objective: Shark type II collagen (SCII) is extracted as a glycoprotein from the cartilage of blue shark (Prionace glauca). We aim to confirm the effects of oral tolerance of SCII on inflammatory and immune responses to the ankle joint of rheumatoid-arthritis rats induced by Complete Freund’s Adjuvant (CFA). Materials and Methods: The onset of rheumatoid arthritis (RA) was observed 14 ± x days after injection of CFA. Rats in the control group were treated with acetic acid by oral administration (0.05 mmol kg−1d−1, days 14–28), while rats in experimental groups were treated by oral administration with SCII (1 or 3 mg kg−1d−1, days 14–28), Tripterygium wilfordii polyglycosidium (TWP) (10 mg kg−1d−1, days 14–28), and bovine type II collagen from US (US-CII) (1 mg kg−1d−1, days 14–28), respectively. The severity of arthritis was evaluated by the articular swelling. The immunological indexes observed included delayed type hypersensitivity (DTH) reaction, the level of interleukins 10 (IL-10) in rat blood serum and morphological characterization. Mixed lymphocyte culture (MLC) was performed to investigate the relationship between T cell apoptosis and specific immune tolerance induced by SCII. Results: Treatment with SCII for 2 weeks significantly attenuated the acute inflammation. The rats orally administrated with SCII at the level of 3 mg kg−1d−1 (SCII 3) and US-CII had decreased DTH reaction compared with rats in control group. Rats treated with SCII 3 had the highest level of IL-10 with 102 pg/mL. SCII with concentration of 10 μg/L could help to significantly enhance level of Fas/Apo-1 in T cell in vitro. The result of histological staining indicated that the recovery of the articular membranes of ankle joint in SCII 3 group was greatly enhanced. Conclusions: Our results suggest that appropriate dose of SCII can not only ameliorate symptoms but also modify the disease process of Complete-Freunds-Adjuvant-induced arthritis. Oral administration of SCII might be a potential candidate as a novel drug for further investigation.
  PubDate: 2012-03-28
  DOI: 10.3390/ph5040339
  Issue No: Vol. 5, No. 4 (2012)
Pharmaceuticals, Vol. 5, Pages 353-368: On the Regulatory Approval Pathway of Biosimilar Products
- Authors: Jun Wang, Shein-Chung Chow
  Pages: 353 - 368
  Abstract: Biosimilars (or follow-on biologics) are a new class of medicine which enters the market subsequent to a previously approved version. They have demonstrated similarity to innovator biologic products in terms of quality, safety, and efficacy. The EMA has taken the lead in the regulatory approval framework for biosimilar products, and WHO has published guidelines on the evaluation of biosimilars in order to facilitate the global harmonization. Based on EMA and WHO guidelines, many other countries such as Canada, Japan and Korea have also issued their own guidance for evaluating follow-on biologics. The US FDA was authorized to approve follow-on biologics by the BPCI Act passed by the US Congress on March 23, 2010, and has just issued a draft guidance in early 2012. The basic concepts and main principles of approving biosimilars are similar among various nations, notwithstanding some differences in regard to the scope, the choice of reference product, and the data requirement. This article reviews the regulatory approval pathway of biosimilar products in different regions.
  PubDate: 2012-03-30
  DOI: 10.3390/ph5040353
  Issue No: Vol. 5, No. 4 (2012)
Pharmaceuticals, Vol. 5, Pages 369-383: Epigenetic Mechanisms and Therapeutic Perspectives for Neurodevelopmental Disorders
- Authors: Takeo Kubota, Hirasawa Takae, Kunio Miyake
  Pages: 369 - 383
  Abstract: The number of children with mild neurodevelopmental disorders, such as autism, has been recently increasing in advanced countries. This increase is probably caused by environmental factors rather than genetic factors, because it is unlikely that genetic mutation rates suddenly increased within a short period. Epigenetics is a mechanism that regulates gene expression, depending not on the underlying DNA sequence but on the chemical modifications of DNA and histone proteins. Because mental stress can alter the epigenetic status in neuronal cells, environmental factors may alter brain function through epigenetic changes. However, one advantage of epigenetic changes is their reversibility. Therefore, diseases due to abnormal epigenetic regulation are theoretically treatable. In fact, several drugs for treating mental diseases are known to have restoring effects on aberrant epigenetic statuses, and a novel therapeutic strategy targeting gene has been developed. In this review, we discuss epigenetic mechanisms of congenital and acquired neurodevelopmental disorders, drugs with epigenetic effects, novel therapeutic strategies for epigenetic diseases, and future perspectives in epigenetic medicine.
  PubDate: 2012-04-05
  DOI: 10.3390/ph5040369
  Issue No: Vol. 5, No. 4 (2012)
Pharmaceuticals, Vol. 5, Pages 384-397: Stroke Prevention in Atrial Fibrillation: Latest Clinical Trials and Guidelines
- Authors: Luciana Armaganijan, Dimpi Patel, Cristiano Dietrich, Carlos A. Morillo
  Pages: 384 - 397
  Abstract: Atrial Fibrillation (AF) is the most common sustained arrhythmia and 1/6 strokes is attributed to AF. The cornerstone of treatment remains maintaining sinus rhythm or appropriate ventricular rate control in addition to prevention of stroke. Oral anticoagulation therapy (OAC) with vitamin K antagonists (VKAs) has been the gold standard for almost 50 years and a significant reduction in the risk of stroke in patients with AF has been demonstrated. Nonetheless, only 50% of patients with guideline recommendations for OAC treatment actually receive VKAs and half of these will discontinue therapy within 3 to 5 years with only another half achieving therapeutic ranges more than 50% of the time. The aforementioned limitations in addition with frequent blood monitoring have prompted the development of a series of new OAC therapies. The present review focuses on the current pharmacological management for stroke prevention in patients with AF based on current and emerging evidence.
  PubDate: 2012-04-05
  DOI: 10.3390/ph5040384
  Issue No: Vol. 5, No. 4 (2012)
Pharmaceuticals, Vol. 5, Pages 398-404: Ivabradine Prevents Heart Rate Acceleration in Patients with Chronic Obstructive Pulmonary Disease and Coronary Heart Disease after Salbutamol Inhalation
- Authors: Rustem Zulkarneev, Naufal Zagidullin, Guzel Abdrahmanova, Uta C. Hoppe, Shamil Zagidullin
  Pages: 398 - 404
  Abstract: Accelerated sinus rhythm is an important side effect of inhaled salbutamol which is especially harmful in patients with chronic obstructive pulmonary disease (COPD) and coronary heart disease (CHD). Cross-over, randomized, open label study design. 20 patients (18 males and two females) with COPD stage II–IV and comorbide CHD NYHA class I–III were included. Spirometry with 400 mg salbutamol inhalation was performed at two consecutive days of the study. Patients in group I were prescribed 5 mg ivabradine per os 3 h before salbutamol inhalation solely on the first day of the study and patients of group II received 5 mg ivabradine only on the second day of the study. Salbutamol caused a significant increase of HR by 5.5 bpm (95% CI 0.8; 10.2, p < 0.03). After ivabradine ingestion salbutamol did not change HR significantly by −2.4 bpm (−7.0; 2.3, p = 0.33). The attenuation of HR elevation by ivabradine was significant, p < 0.01. Salbutamol alone increased FEV1 by 6.0% (2.7; 9.3, p < 0.01). This effect was not impaired by ivabradine (FEV1 increase by 7.7% (2.8; 12.6, p < 0.01 versus baseline, p = 0.5 versus no ivabradine). Ivabradine 5 mg per os prevents heart rate acceleration after inhalation of 400 mg salbutamol. Ivabradine has no impact on lung function in patients with moderate-to-very-severe COPD and CHD comorbidity.
  PubDate: 2012-04-16
  DOI: 10.3390/ph5040398
  Issue No: Vol. 5, No. 4 (2012)
Pharmaceuticals, Vol. 5, Pages 1265-1281: &#160; Cell Penetrating Peptoids (CPPos): Synthesis of a Small Combinatorial Library by Using IRORI MiniKans
- Authors: Dominik Kölmel, Daniel Fürniss, Steven Susanto, Andrea Lauer, Clemens Grabher, Stefan Bräse, Ute Schepers
  Pages: 1265 - 1281
  Abstract: Cell penetrating peptoids (CPPos) are potent mimics of the corresponding cell penetrating peptides (CPPs). The synthesis of diverse oligomeric libraries that display a variety of backbone scaffolds and side-chain appendages are a very promising source of novel CPPos, which can be used to either target different cellular organelles or even different tissues and organs. In this study we established the submonomer-based solid phase synthesis of a “proof of principle” peptoid library in IRORI MiniKans to expand the amount for phenotypic high throughput screens of CPPos. The library consisting of tetrameric peptoids [oligo(N-alkylglycines)] was established on Rink amide resin in a split and mix approach with hydrophilic and hydrophobic peptoid side chains. All CPPos of the presented library were labeled with rhodamine B to allow for the monitoring of cellular uptake by fluorescent confocal microscopy. Eventually, all the purified peptoids were subjected to live cell imaging to screen for CPPos with organelle specificity. While highly charged CPPos enter the cells by endocytosis with subsequent endosomal release, critical levels of lipophilicity allow other CPPos to specifically localize to mitochondria once a certain lipophilicity threshold is reached.
  PubDate: 2012-11-23
  DOI: 10.3390/ph5121265
  Issue No: Vol. 5, No. 12 (2012)
Pharmaceuticals, Vol. 5, Pages 1282-1290: Synthesis and COX-2 Inhibitory Activity of 4-[(E)-2-(4-Oxo-3-phenyl-3,4-dihydroquinazolin-2-yl)ethenyl]benzene-1-sulfonamide and Its Analogs
- Authors: Hayun, Sumi Hudiyono, Muhammad Hanafi, Arry Yanuar
  Pages: 1282 - 1290
  Abstract: Some novel 3-phenyl-2-[(E)-2-phenylethenyl]-3,4-dihydroquinazolin-4-one derivatives possessing para-sulfonamides groups on the phenyl ring of the 2-phenylethenyl moiety have been synthesized and their COX-2 inhibitory activity evaluated. The stuctures of the synthesized compounds were confirmed  on the basis of  FT-IR, 1H-NMR, 13C-NMR and mass spectral data. The COX-2 inhibition screening assay revealed that 4-[(E)-2-{3-(4-methoxyphenyl)-4-oxo-3,4-dihydroquinazolin-2-yl}ethenyl]benzene-1-sulfonamide had a  maximum COX-2 inhibition (47.1%), at a concentration of 20 μM.
  PubDate: 2012-11-27
  DOI: 10.3390/ph5121282
  Issue No: Vol. 5, No. 12 (2012)
Pharmaceuticals, Vol. 5, Pages 1291-1331: Multiple Facets of cAMP Signalling and Physiological Impact: cAMP Compartmentalization in the Lung
- Authors: Anouk Oldenburger, Harm Maarsingh, Martina Schmidt
  Pages: 1291 - 1331
  Abstract: Therapies involving elevation of the endogenous suppressor cyclic AMP (cAMP) are currently used in the treatment of several chronic inflammatory disorders, including chronic obstructive pulmonary disease (COPD). Characteristics of COPD are airway obstruction, airway inflammation and airway remodelling, processes encompassed by increased airway smooth muscle mass, epithelial changes, goblet cell and submucosal gland hyperplasia. In addition to inflammatory cells, airway smooth muscle cells and (myo)fibroblasts, epithelial cells underpin a variety of key responses in the airways such as inflammatory cytokine release, airway remodelling, mucus hypersecretion and airway barrier function. Cigarette smoke, being next to environmental pollution the main cause of COPD, is believed to cause epithelial hyperpermeability by disrupting the barrier function. Here we will focus on the most recent progress on compartmentalized signalling by cAMP. In addition to G protein-coupled receptors, adenylyl cyclases, cAMP-specific phospho-diesterases (PDEs) maintain compartmentalized cAMP signalling. Intriguingly, spatially discrete cAMP-sensing signalling complexes seem also to involve distinct members of the A-kinase anchoring (AKAP) superfamily and IQ motif containing GTPase activating protein (IQGAPs). In this review, we will highlight the interaction between cAMP and the epithelial barrier to retain proper lung function and to alleviate COPD symptoms and focus on the possible molecular mechanisms involved in this process. Future studies should include the development of cAMP-sensing multiprotein complex specific disruptors and/or stabilizers to orchestrate cellular functions. Compartmentalized cAMP signalling regulates important cellular processes in the lung and may serve as a therapeutic target.
  PubDate: 2012-11-30
  DOI: 10.3390/ph5121291
  Issue No: Vol. 5, No. 12 (2012)
Pharmaceuticals, Vol. 5, Pages 1332-1345: Integration of Ligand-Based Drug Screening with Structure-Based Drug Screening by Combining Maximum Volume Overlapping Score with Ligand Docking
- Authors: Yoshifumi Fukunishi, Haruki Nakamura
  Pages: 1332 - 1345
  Abstract: Ligand-based and structure-based drug screening methods were integrated for in silico drug development by combining the maximum-volume overlap (MVO) method with a protein-compound docking program. The MVO method is used to select reliable docking poses by calculating volume overlaps between the docking pose in question and the known ligand docking pose, if at least a single protein-ligand complex structure is known. In the present study, the compounds in a database were docked onto a target protein that had a known protein-ligand complex structure. The new score is the summation of the docking score and the MVO score, which is the measure of the volume overlap between the docking poses of the compound in question and the known ligand. The compounds were sorted according to the new score. The in silico screening results were improved by comparing the MVO score to the original docking score only. The present method was also applied to some target proteins with known ligands, and the results demonstrated that it worked well.
  PubDate: 2012-12-04
  DOI: 10.3390/ph5121332
  Issue No: Vol. 5, No. 12 (2012)
Pharmaceuticals, Vol. 5, Pages 1346-1371: Mechanistic and Pharmacological Issues of Aspirin as an Anticancer Agent
- Authors: Melania Dovizio, Stefania Tacconelli, Carlos Sostres, Emanuela Ricciotti, Paola Patrignani
  Pages: 1346 - 1371
  Abstract: Recent findings have shown that aspirin, taken for several years, reduces the long-term risk of some cancers, particularly colorectal cancer. The result that aspirin benefit is detectable at daily low-doses (at least 75mg), the same used for the prevention of cardiovascular disease, positions the antiplatelet action of aspirin at the center of its antitumor efficacy. At low-doses given every 24 h, aspirin is acting by a complete and persistent inhibition of cyclooxygenase (COX)-1 in platelets (in the pre-systemic circulation) while causing a limited and rapidly reversible inhibitory effect on COX-2 and/or COX-1 expressed in nucleated cells. Aspirin has a short half-life in human circulation (approximately 20 min); nucleated cells have the ability to resynthesize the acetylated COX-isozymes within a few hours, while platelets do not. COX-independent mechanisms of aspirin, such as the inhibition of Wnt/ b-catenin and NF-kB signaling and the acetylation of extra-COX proteins, have been suggested to play a role in its chemo-preventive effects, but their relevance remains to be demonstrated in vivo at clinical doses. In conclusion, the results of clinical pharmacology and the analysis of randomized and epidemiological studies suggest that colorectal cancer and atherothrombosis share a common mechanism of disease, i.e. enhanced platelet activation in response to injury at distinct sites.
  PubDate: 2012-12-05
  DOI: 10.3390/ph5121346
  Issue No: Vol. 5, No. 12 (2012)
Pharmaceuticals, Vol. 5, Pages 1372-1392: The Liver as a Target Organ for Gene Therapy: State of the Art, Challenges, and Future Perspectives
- Authors: Frank Jacobs, Stephanie Gordts, Ilayaraja Muthuramu, Bart De Geest
  Pages: 1372 - 1392
  Abstract: The liver is a target for gene therapy of inborn errors of metabolism, of hemophilia, and of acquired diseases such as liver cancer and hepatitis. The ideal gene transfer strategy should deliver the transgene DNA to parenchymal liver cells with accuracy and precision in the absence of side effects. Liver sinusoids are highly specialized capillaries with a particular endothelial lining: the endothelium contains open fenestrae, whereas a basal lamina is lacking. Fenestrae provide a direct access of gene transfer vectors to the space of Disse, in which numerous microvilli from parenchymal liver cells protrude. The small diameter of fenestrae in humans constitutes an anatomical barrier for most gene transfer vectors with the exception of adeno-associated viral (AAV) vectors. Recent studies have demonstrated the superiority of novel AAV serotypes for hepatocyte-directed gene transfer applications based on enhanced transduction, reduced prevalence of neutralizing antibodies, and diminished capsid immune responses. In a landmark clinical trial, hemophilia B was successfully treated with an AAV8 human factor IX expressing vector. Notwithstanding significant progress, clinical experience with these technologies remains very limited and many unanswered questions warrant further study. Therefore, the field should continue to progress as it has over the past decade, cautiously and diligently.
  PubDate: 2012-12-10
  DOI: 10.3390/ph5121372
  Issue No: Vol. 5, No. 12 (2012)
Pharmaceuticals, Vol. 5, Pages 1393-1408: Biosimilars: Company Strategies to Capture Value from the Biologics Market
- Authors: Bruno Calo-Fernández, Juan Martínez-Hurtado
  Pages: 1393 - 1408
  Abstract: Patents for several biologic blockbusters will expire in the next few years. The arrival of biosimilars, the biologic equivalent of chemical generics, will have an impact on the current biopharmaceuticals market. Five core capabilities have been identified as paramount for those companies aiming to enter the biosimilars market: research and development, manufacturing, supporting activities, marketing, and lobbying. Understanding the importance of each of these capabilities will be key to maximising the value generated from the biologics patent cliff.
  PubDate: 2012-12-12
  DOI: 10.3390/ph5121393
  Issue No: Vol. 5, No. 12 (2012)
Pharmaceuticals, Vol. 5, Pages 1147-1159: Metabolism and Pharmacokinetics of the Anti-Tuberculosis Drug Ethionamide in a Flavin-Containing Monooxygenase Null Mouse
- Authors: Amy L. Palmer, Virginia L. Leykam, Andrew Larkin, Sharon K. Krueger, Ian R. Phillips, Elizabeth A. Shephard, David E. Williams
  Pages: 1147 - 1159
  Abstract: Multiple drug resistance (MDR) in Mycobacterium tuberculosis (mTB), the causative agent for tuberculosis (TB), has led to increased use of second-line drugs, including ethionamide (ETA). ETA is a prodrug bioactivated by mycobacterial and mammalian flavin-containing monooxygenases (FMOs). FMO2 is the major isoform in the lungs of most mammals, including primates. In humans a polymorphism exists in the expression of FMO2. FMO2.2 (truncated, inactive) protein is produced by the common allele, while the ancestral allele, encoding active FMO2.1, has been documented only in individuals of African and Hispanic origin, at an incidence of up to 50% and 7%, respectively. We hypothesized that FMO2 variability in TB-infected individuals would yield differences in concentrations and ratios of ETA prodrug and metabolites. In this study we assessed the impact of the FMO2 genetic polymorphism on the pharmacokinetics of ETA after administration of a single oral dose of ETA (125 mg/kg) to wild type and triple Fmo1/2/4-null mice, measuring levels of prodrug vs. metabolites in plasma collected from 0 to 3.5 h post-gavage. All mice metabolized ETA to ETA S-oxide (ETASO) and 2-ethyl-4-amidopyridine (ETAA). Wild type mice had higher plasma concentrations of metabolites than of parent compound (p = 0.001). In contrast, Fmo1/2/4-null mice had higher plasma concentrations of parent compound than of metabolites (p = 0.0001). Thus, the human FMO2 genotype could impact the therapeutic efficacy and/or toxicity of ETA.
  PubDate: 2012-10-25
  DOI: 10.3390/ph5111147
  Issue No: Vol. 5, No. 11 (2012)
Pharmaceuticals, Vol. 5, Pages 1160-1176: Cyclooxygenase (COX) Inhibitors and the Newborn Kidney
- Authors: Francine G. Smith, Andrew W. Wade, Megan L. Lewis, Wei Qi
  Pages: 1160 - 1176
  Abstract: This review summarizes our current understanding of the role of cyclo-oxygenase inhibitors (COXI) in influencing the structural development as well as the function of the developing kidney. COXI administered either during pregnancy or after birth can influence kidney development including nephronogenesis, and can decrease renal perfusion and ultrafiltration potentially leading to acute kidney injury in the newborn period. To date, which COX isoform (COX-1 or COX-2) plays a more important role in during fetal development and influences kidney function early in life is not known, though evidence points to a predominant role for COX-2. Clinical implications of the use of COXI in pregnancy and in the newborn infant are also evaluated herein, with specific reference to the potential effects of COXI on nephronogenesis as well as newborn kidney function.
  PubDate: 2012-10-25
  DOI: 10.3390/ph5111160
  Issue No: Vol. 5, No. 11 (2012)
Pharmaceuticals, Vol. 5, Pages 1177-1209: Improving the Endosomal Escape of Cell-Penetrating Peptides and Their Cargos: Strategies and Challenges
- Authors: Alfredo Erazo-Oliveras, Nandhini Muthukrishnan, Ryan Baker, Ting-Yi Wang, Jean-Philippe Pellois
  Pages: 1177 - 1209
  Abstract: Cell penetrating peptides (CPPs) can deliver cell-impermeable therapeutic cargos into cells. In particular, CPP-cargo conjugates tend to accumulate inside cells by endocytosis. However, they often remain trapped inside endocytic organelles and fail to reach the cytosolic space of cells efficiently. In this review, the evidence for CPP-mediated endosomal escape is discussed. In addition, several strategies that have been utilized to enhance the endosomal escape of CPP-cargos are described. The recent development of branched systems that display multiple copies of a CPP is presented. The use of viral or synthetic peptides that can disrupt the endosomal membrane upon activation by the low pH of endosomes is also discussed. Finally, we survey how CPPs labeled with chromophores can be used in combination with light to stimulate endosomal lysis. The mechanisms and challenges associated with these intracellular delivery methodologies are discussed.
  PubDate: 2012-11-01
  DOI: 10.3390/ph5111177
  Issue No: Vol. 5, No. 11 (2012)
Pharmaceuticals, Vol. 5, Pages 1210-1235: The Role of Transport Mechanisms in Mycobacterium Tuberculosis Drug Resistance and Tolerance
- Authors: Jansy Passiflora Sarathy, Véronique Dartois, Edmund Jon Deoon Lee
  Pages: 1210 - 1235
  Abstract: In the fight against tuberculosis, cell wall permeation of chemotherapeutic agents remains a critical but largely unsolved question. Here we review the major mechanisms of small molecule penetration into and efflux from Mycobacterium tuberculosis and other mycobacteria, and outline how these mechanisms may contribute to the development of phenotypic drug tolerance and induction of drug resistance. M. tuberculosis is intrinsically recalcitrant to small molecule permeation thanks to its thick lipid-rich cell wall. Passive diffusion appears to account for only a fraction of total drug permeation. As in other bacterial species, influx of hydrophilic compounds is facilitated by water-filled open channels, or porins, spanning the cell wall. However, the diversity and density of M. tuberculosis porins appears lower than in enterobacteria. Besides, physiological adaptations brought about by unfavorable conditions are thought to reduce the efficacy of porins. While intracellular accumulation of selected drug classes supports the existence of hypothesized active drug influx transporters, efflux pumps contribute to the drug resistant phenotype through their natural abundance and diversity, as well as their highly inducible expression. Modulation of efflux transporter expression has been observed in phagocytosed, non-replicating persistent and multi-drug resistant bacilli. Altogether, M. tuberculosis has evolved both intrinsic properties and acquired mechanisms to increase its level of tolerance towards xenobiotic substances, by preventing or minimizing their entry. Understanding these adaptation mechanisms is critical to counteract the natural mechanisms of defense against toxic compounds and develop new classes of chemotherapeutic agents that positively exploit the influx and efflux pathways of mycobacteria.
  PubDate: 2012-11-09
  DOI: 10.3390/ph5111210
  Issue No: Vol. 5, No. 11 (2012)
Pharmaceuticals, Vol. 5, Pages 1236-1264: The Biological Role of PI3K Pathway in Lung Cancer
- Authors: Evangelos Sarris, Muhammad Saif, Kostas Syrigos
  Pages: 1236 - 1264
  Abstract: Lung cancer is the primary cause of cancer-related mortality worldwide and although improvements in treatment have been achieved over the last few years, long-term survival rates for lung cancer patients remain poor. Therefore, there is an imperative need for molecularly targeted agents that will achieve long-term disease control. Numerous downstream molecular pathways, such as EGF/RAS/RAF/MEK/ERK and PI3K/AKT/mTOR are identified as having a key role in the pathogenesis of various forms of human cancer, including lung cancer. PI3K/AKT/mTOR signal pathway is an important intracellular signal transduction pathway with a significant role in cell proliferation, growth, survival, vesicle trafficking, glucose transport, and cytoskeletal organization. Aberrations in many primary and secondary messenger molecules of this pathway, including mutations and amplifications, are accounted for tumor cell proliferation, inhibition of apoptosis, angiogenesis, metastasis and resistance to chemotherapy-radiotherapy. In this review article, we investigate thoroughly the biological role of PI3K pathway in lung cancer and its contribution in the development of future therapeutic strategies.
  PubDate: 2012-11-20
  DOI: 10.3390/ph5111236
  Issue No: Vol. 5, No. 11 (2012)
Pharmaceuticals, Vol. 5, Pages 1045-1053: The Use of California Sagebrush (Artemisia californica) Liniment to Control Pain
- Authors: James D. Adams
  Pages: 1045 - 1053
  Abstract: The incidence of arthritis is increasing every year, as does the need for pain medication. The current work reviews an American Indian liniment that is traditionally used for pain therapy. The chemistry, therapeutic use and safety of the liniment are reviewed. The liniment contains monoterpenoids, sesquiterpenes, flavonoids, alkaloids and other compounds.
  PubDate: 2012-09-27
  DOI: 10.3390/ph5101045
  Issue No: Vol. 5, No. 10 (2012)
Pharmaceuticals, Vol. 5, Pages 1054-1063: Cell-Penetrating Recombinant Peptides for Potential Use in Agricultural Pest Control Applications
- Authors: Stephen R. Hughes, Patrick F. Dowd, Eric T. Johnson
  Pages: 1054 - 1063
  Abstract: Several important areas of interest intersect in a class of peptides characterized by their highly cationic and partly hydrophobic structure. These molecules have been called cell-penetrating peptides (CPPs) because they possess the ability to translocate across cell membranes. This ability makes these peptides attractive candidates for delivery of therapeutic compounds, especially to the interior of cells. Compounds with characteristics similar to CPPs and that, in addition, have antimicrobial properties are being investigated as antibiotics with a reduced risk of causing resistance. These CPP-like membrane-acting antimicrobial peptides (MAMPs) are α-helical amphipathic peptides that interact with and perturb cell membranes to produce their antimicrobial effects. One source of MAMPs is spider venom. Because these compounds are toxic to insects, they also show promise for development as biological agents for control of insecticide-resistant agricultural pests. Spider venom is a potential source of novel insect-specific peptide toxins. One example is the small amphipathic α-helical peptide lycotoxin-1 (Lyt-1 or LCTX) from the wolf spider (Lycosa carolinensis). One side of the α-helix has mostly hydrophilic and the other mainly hydrophobic amino acid residues. The positive charge of the hydrophilic side interacts with negatively charged prokaryotic membranes and the hydrophobic side associates with the membrane lipid bilayer to permeabilize it. Because the surface of the exoskeleton, or cuticle, of an insect is highly hydrophobic, to repel water and dirt, it would be expected that amphipathic compounds could permeabilize it. Mutagenized lycotoxin 1 peptides were produced and expressed in yeast cultures that were fed to fall armyworm (Spodoptera frugiperda) larvae to identify the most lethal mutants. Transgenic expression of spider venom toxins such as lycotoxin-1 in plants could provide durable insect resistance.
  PubDate: 2012-09-28
  DOI: 10.3390/ph5101054
  Issue No: Vol. 5, No. 10 (2012)
Pharmaceuticals, Vol. 5, Pages 1064-1079: Statistical Estimation of the Protein-Ligand Binding Free Energy Based On Direct Protein-Ligand Interaction Obtained by Molecular Dynamics Simulation
- Authors: Yoshifumi Fukunishi, Haruki Nakamura
  Pages: 1064 - 1079
  Abstract: We have developed a method for estimating protein-ligand binding free energy (DG) based on the direct protein-ligand interaction obtained by a molecular dynamics simulation. Using this method, we estimated the DG value statistically by the average values of the van der Waals and electrostatic interactions between each amino acid of the target protein and the ligand molecule. In addition, we introduced fluctuations in the accessible surface area (ASA) and dihedral angles of the protein-ligand complex system as the entropy terms of the DG estimation. The present method included the fluctuation term of structural change of the protein and the effective dielectric constant. We applied this method to 34 protein-ligand complex structures. As a result, the correlation coefficient between the experimental and calculated DG values was 0.81, and the average error of DG was 1.2 kcal/mol with the use of the fixed parameters. These results were obtained from a 2 nsec molecular dynamics simulation.
  PubDate: 2012-09-28
  DOI: 10.3390/ph5101064
  Issue No: Vol. 5, No. 10 (2012)
Pharmaceuticals, Vol. 5, Pages 1080-1091: Design, Synthesis and Hydrolytic Behavior of Mutual Prodrugs of NSAIDs with Gabapentin Using Glycol Spacers
- Authors: Monther Mahdi, Hiba Alsaad
  Pages: 1080 - 1091
  Abstract: The free –COOH present in NSAIDs is thought to be responsible for the GI irritation associated with all traditional NSAIDs. Exploitation of mutual prodrugs is an approach wherein the NSAID is covalently bounded to a second pharmacologically active carrier/drug with the ultimate aim of reducing the gastric irritation. In this study some NSAIDs were conjugated with gabapentin via ester bonds using glycol spacers with the expectation of reducing gastric adverse effects and obtaining synergistic analgesic effects. The kinetics of ester hydrolysis were studied in two different non enzymatic buffer solutions at pH 1.2 and 7.4, as well as in 80% human plasma using HPLC with chloroform -methanol as mobile phase. Compounds 9a–c with ethylene glycol spacers showed significant stability at buffer solutions with half lives ranging from about 8–25 h, while the underwent a reasonable plasma hydrolysis (49%–88%) in 2 h. Compound 9d with a propylene glycol spacer shows a higher rate of enzymatic hydrolysis than the corresponding ethylene glycol compound 9c. The result of compounds 9a-c indicate that these compounds may be stable during their passage through the GIT until reaching the blood circulation.
  PubDate: 2012-10-12
  DOI: 10.3390/ph5101080
  Issue No: Vol. 5, No. 10 (2012)
Pharmaceuticals, Vol. 5, Pages 1092-1102: Intranasal Delivery of Camptothecin-Loaded Tat-Modified Nanomicells for Treatment of Intracranial Brain Tumors
- Authors: Hiroyuki Taki, Takanori Kanazawa, Fuminari Akiyama, Yuuki Takashima, Hiroaki Okada
  Pages: 1092 - 1102
  Abstract: The blood-brain barrier is a substantial obstacle for delivering anticancer agents to brain tumors, and new strategies for bypassing it are sorely needed for brain tumor therapy. Intranasal delivery provides a practical, noninvasive method for delivering therapeutic agents to the brain. Intranasal application of nano-sized micelles that have been modified with Tat peptide facilitates brain delivery of fluorescent model materials. In this study, we evaluated a nose-to-brain delivery system for brain tumor therapy. We nasally administered the anti-tumor drug camptothecin (CPT) in solution and in methoxy poly(ethylene glycol) (MPEG)/poly(e-caprolactone) (PCL) amphiphilic block copolymers (MPEG-PCL) and cell penetrating peptide, Tat analog-modified MPEG-PCL (MPEG-PCL-Tat) MPEG-PCL-Tat to rats bearing intracranial glioma tumors and quantified the cytotoxicity against glioma cells, and the therapeutic effects. CPT-loaded MPEG-PCL-Tat micelles showed higher cytotoxicity than CPT-loaded MPEG-PCL. CPT-free MPEG-PCL-Tat didn’t show any cytotoxicity, even at high concentrations (2 mmol/mL). CPT-loaded MPEG-PCL-Tat micelles significantly prolonged the median survival of rats. These results indicate that intranasal delivery of anti-cancer drugs with cell penetrating peptide-modified nanomicelles might be an effective therapy for brain tumors.
  PubDate: 2012-10-15
  DOI: 10.3390/ph5101092
  Issue No: Vol. 5, No. 10 (2012)
Pharmaceuticals, Vol. 5, Pages 1103-1119: Mitochondria-Targeted Antioxidant SS31 Prevents Amyloid Beta-Induced Mitochondrial Abnormalities and Synaptic Degeneration in Alzheimer&rsquo;s Disease
- Authors: Marcus Calkins, Maria Manczak, P. Reddy
  Pages: 1103 - 1119
  Abstract: In neuronal systems, the health and activity of mitochondria and synapses are tightly coupled. For this reason, it has been postulated that mitochondrial abnormalities may, at least in part, drive neurodegeneration in conditions such as Alzheimer’s disease (AD). Mounting evidence from multiple Alzheimer’s disease cell and mouse models and postmortem brains suggest that loss of mitochondrial integrity may be a key factor that mediates synaptic loss. Therefore, the prevention or rescue of mitochondrial dysfunction may help delay or altogether prevent AD-associated neurodegeneration. Since mitochondrial health is heavily dependent on antioxidant defenses, researchers have begun to explore the use of mitochondria-targeted antioxidants as therapeutic tools to prevent neurodegenerative diseases. This review will highlight advances made using a model mitochondria-targeted antioxidant peptide, SS31, as a potential treatment for AD.
  PubDate: 2012-10-16
  DOI: 10.3390/ph5101103
  Issue No: Vol. 5, No. 10 (2012)
Pharmaceuticals, Vol. 5, Pages 1120-1127: Norethindrone Acetate in the Medical Management of Adenomyosis
- Authors: Ozgul Muneyyirci-Delale, Ashadeep Chandrareddy, Siddhi Mankame, Nanna Osei-Tutu, Hans von Gizycki
  Pages: 1120 - 1127
  Abstract: The role of norethindrone acetate (NA) in the management of adenomyosis was evaluated with a retrospective chart review of 28 premenopausal women between 27–49 years of age presenting with moderate to severe pelvic pain and bleeding. Bleeding and dysmenorrhea scores were analyzed using paired T-tests. There was significant improvement of both dysmenorrhea and bleeding after treatment. Age showed no correlation with dysmenorrhea or bleeding. Low dose NA could be considered an effective, well-tolerated and inexpensive medical alternative to surgery for treating symptomatic adenomyosis. Large multicentric studies may help validate our findings.
  PubDate: 2012-10-22
  DOI: 10.3390/ph5101120
  Issue No: Vol. 5, No. 10 (2012)
Pharmaceuticals, Vol. 5, Pages 1128-1146: Advances in Drug Design Based on the Amino Acid Approach: Taurine Analogues for the Treatment of CNS Diseases
- Authors: Man Chin Chung, Pedro Malatesta, Priscila Longhin Bosquesi, Paulo Renato Yamasaki, Jean Leandro dos Santos, Ednir Oliveira Vizioli
  Pages: 1128 - 1146
  Abstract: Amino acids are well known to be an important class of compounds for the maintenance of body homeostasis and their deficit, even for the polar neuroactive aminoacids, can be controlled by supplementation. However, for the amino acid taurine (2-aminoethanesulfonic acid) this is not true. Due its special physicochemical properties, taurine is unable to cross the blood-brain barrier. In addition of injured taurine transport systems under pathological conditions, CNS supplementation of taurine is almost null. Taurine is a potent antioxidant and anti-inflammatory semi-essential amino acid extensively involved in neurological activities, acting as neurotrophic factor, binding to GABA A/glycine receptors and blocking the excitotoxicity glutamate-induced pathway leading to be a neuroprotective effect and neuromodulation. Taurine deficits have been implicated in several CNS diseases, such as Alzheimer’s, Parkinson’s, epilepsy and in the damage of retinal neurons. This review describes the CNS physiological functions of taurine and the development of new derivatives based on its structure useful in CNS disease treatment.
  PubDate: 2012-10-23
  DOI: 10.3390/ph5101128
  Issue No: Vol. 5, No. 10 (2012)