Selective synthesis of N,N,N-trimethylated chitosan derivatives at different degree of substitution and investigation of structure-activity relationship for activity against P. aeruginosa and MRSA.

Two new cationic chitosan derivatives were synthesized using a combination of Boc and TBDMS protection strategies. This included a series of six samples of the TMCNH2/TM derivative, where some of the amino groups were N,N,N-trimethylated and the remaining was in the primary state. A series of six samples of the TACin derivative, where some of the amino groups were N-acetylated with quaternary 2-(N,N,N-trimethylammoniumyl) acetyl group and the remaining fully N-acetylated, were also synthesized. The degree of substitution (DS) for quaternary amino groups in these series ranged from 0.06-0.89. TMCDM/TM derivatives with a mix of N,N,N-trimethylated and N,N-dimethylated groups were also synthesized according to a published procedure but in this case, it was more difficult to control the DS than with the TBDMS protection strategy. Broth microdilution assay revealed a markedly different structure-activity relationship (SAR) for the two derivatives. The activity for the TMC derivatives reached a plateau above 0.2-0.3 DS whereas the activity increased continuously with DS for TACin. The highest DS TMCNH2/TM was more active than the highest DS, TACin, against Gram-positive MRSA but less active against the Gram-negative P. aeruginosa.

[Adverse drug reaction reports in Iceland from 2013 to 2016. A comparison with other Nordic countries].

INTRODUCTION: Information regarding adverse drug reactions (ADRs) of new medications is based on clinical studies of selected populations. The reporting of ADRs from real-life use following the marketing of new active substances is instrumental for the continuous evaluation of their benefit-risk balance. The aim of this study was to determine the number and nature of ADR reports in Iceland and compare with other Nordic countries. MATERIALS AND METHODS: Reports of ADRs from 2013 to 2016 were examined using the Icelandic Medicines Agency´s database. The total number and seriousness of ADRs by ATC-classification of drugs were compared with data published in the 2013 to 2015 annual reports from the Swedish, Danish and Norwegian Medicines Agencies. Comparison of sales between countries was examined. RESULTS: The number of ADR reports in Iceland was between 36 to 104 per 100 thousand inhabitants/year, with less than 10% defined as serious. This compares to 58 to 133 ADR reports per 100 thousand inhabitants in the other Nordic countries, with 38% to 64% of ADRs classified as serious. In Iceland, ADR reports were more common for medications in ATC-class A and less common for classes B, J and L compared to the other Nordic countries. Sales of medications were comparable between these nations. CONCLUSION: There is great variability in the number of ADRs reported annually in Iceland. The drugs reported are within different ATC-classes and the proportion of serious ADRs is low compared to the other Nordic countries. This is not explained by different sales volumes. Key words: adverse drug reaction, pharmacovigilance, ADR reporting, side effects. Correspondence: Hrefna Guðmundsdóttir, Hrefna.Gudmundsdottir@lyfjastofnun.is.

Alternatives to antibiotics-a pipeline portfolio review.

Antibiotics have saved countless lives and enabled the development of modern medicine over the past 70 years. However, it is clear that the success of antibiotics might only have been temporary and we now expect a long-term and perhaps never-ending challenge to find new therapies to combat antibiotic-resistant bacteria. A broader approach to address bacterial infection is needed. In this Review, we discuss alternatives to antibiotics, which we defined as non-compound approaches (products other than classic antibacterial agents) that target bacteria or any approaches that target the host. The most advanced approaches are antibodies, probiotics, and vaccines in phase 2 and phase 3 trials. This first wave of alternatives to antibiotics will probably best serve as adjunctive or preventive therapies, which suggests that conventional antibiotics are still needed. Funding of more than £1·5 billion is needed over 10 years to test and develop these alternatives to antibiotics. Investment needs to be partnered with translational expertise and targeted to support the validation of these approaches in phase 2 trials, which would be a catalyst for active engagement and investment by the pharmaceutical and biotechnology industry. Only a sustained, concerted, and coordinated international effort will provide the solutions needed for the future.

Characterization of cold-adapted Atlantic cod (Gadus morhua) trypsin I--kinetic parameters, autolysis and thermal stability.

Atlantic cod trypsin I is a highly active cold-adapted protease. This study aimed at further characterization of this enzyme with respect to kinetic parameters, sites of autolysis and stability. For that purpose, trypsin I was purified by anion exchange chromatography. Its purity and identity was verified by SDS-PAGE analysis and mass spectrometry. Concomitantly, another cod trypsin isozyme, trypsin X, previously only described from its cDNA sequence was detected in a separate peak from the ion exchange chromatogram. There was a stepwise increase in the catalytic efficiency (k(cat)/K(m)) of cod trypsin I obtained with substrates containing one to three amino acid residues. As expected, the activity of trypsin I was maintained for longer periods of time at 15 degrees C than at higher temperatures. The residues of the trypsin I molecule most sensitive to autolysis were identified using Edman degradation. Eleven autolytic cleavage sites were detected within the trypsin I molecule. Unfolding experiments demonstrated that autolysis is a contributing factor in the stability of trypsin I. In addition, the data shows that cod trypsin I is less stable towards thermal unfolding than its mesophilic bovine analogue.

Immunization prevents DDT buildup in mouse tissues.

DDT is used for pest control, causing health and environmental hazards in some parts of the world. The goal of this study was to assess whether immunization against a toxic compound could reduce the toxicant uptake of an organism, specifically to develop a DDT immunization that promotes the production of specific antibodies and assess whether it reduces DDT levels in the bodies of mice that are exposed to DDT by intake. BALB/c mice were immunized with DDT-keyhole limpet hemocyanine (DDT-KLH) conjugate (n=10) or unconjugated KLH (n=10), which was used as a control. After the immunization specific DDT antibodies in the mouse serum were determined by ELISA and then the mice were fed chow containing 40 mg/kg of DDT for 45 days. Finally, the concentration of DDT and its metabolites, DDE and DDD, in various tissues was measured by gas chromatography. Specific DDT antibody levels were significantly higher in the DDT immunized group than in the control group. DDT, DDE and DDD levels in adipose tissue, blood, brain and spleen were significantly reduced in the DDT immunized animals relative to control animals. However, DDT and DDD levels were higher in the liver compared to the control group. The findings indicate that the DDT immunization reduces the total uptake of DDT in animal tissues, which is reflected by the lower levels in adipose tissue, blood, brain and spleen. The elevated levels in liver suggest that DDT-antibody complexes in mouse serum are delivered to the liver.