34 S: A New Opportunity for the Efficient Synthesis of Stable Isotope Labeled Compounds.

The synthesis of stable isotope labeled (SIL) complex drug molecules with a ≥3 mass unit increase from the parent compound is essential for drug discovery and development. Typical approaches that rely on 2 H, 13 C, and 15 N isotopes can be very challenging or even intractable, and can delay the drug development process. This work introduces a new concept for the synthesis of labeled compounds that relies on the use of 34 S. The synthetic utility of 34 S was demonstrated with the efficient synthesis of [34 S]phosphorothioates [34 S2 ]-PS-ODNs-TTT and [13 C, 15 N, 34 S]-ceftolozane. In addition, a procedure for the direct oxidation of phosphites to [34 S]phosphorothioates using elemental 34 S without isotope dilution was developed.

Moisture determination of tritium tracers utilizing near-infrared spectroscopy.

Tritium tracers are frequently used in biological assays during the drug discovery process because of their high specific activity and relative ease of synthesis. However, this high specific activity, along with other contributing factors, can lead to an increased rate of radiolytic decomposition. As a result, following long-term storage tritium tracers often require purification. Understanding the elements that cause radiolytic decomposition is extremely important to extend the storage life, and consequently reduce unnecessary inventory purifications. One of these elements is the presence of water in tritium tracers. Upon investigation, it was discovered that aside from the relatively common tritium/water exchange that could occur, residual water could also contribute significantly to the decomposition of tritium tracers. A near-infrared method was developed utilizing a portable device to measure the water content in tritium tracers rapidly and without sample destruction. This method proved to be quick, efficient, and achieved an error less than 5% compared to that of traditional Karl Fischer titration. Method validation was performed and good accuracy, linearity, limit of detection and quantitation were all established.

Synthesis of [(14) C]omarigliptin.

An efficient synthesis for [(14) C]Omarigliptin (MK-3102) is described. The initial synthesis of a key (14) C-pyrazole moiety did not work due to the lack of stability of (14) C-DMF-DMA reagent. Thus, a new radiolabeled synthon, (14) C-biphenylmethylformate, was synthesized from (14) C-sodium formate in one step in 92% yield and successfully used in construction of the key (14) C-pyrazole moiety. Regioselective N-sulfonation of the pyrazole moiety was achieved through a dehydration-sulfonation-isomerization sequence. [(14) C]MK 3102 was synthesized in five steps from (14) C-biphenylmethylformate with 25% overall yield.

Enhancing radiolytic stability upon concentration of tritium-labeled pharmaceuticals utilizing centrifugal evaporation.

Tritium radiopharmaceuticals are often used in drug development because of their desirable specific activity. The inherent instability of these radioactive tracers often leads to a requirement to purify prior to use. Purification methodologies such as preparative chromatography and solid/liquid extractions often utilize water as a solvent, which is not suitable for long-term storage and necessitates removal. Rotary evaporation has traditionally been utilized for the removal of this unwanted solvent, however, this method has been shown to lead to decomposition of the tritium species in some cases. Centrifugal evaporation is a milder concentration method which has been demonstrated to effectively remove solvents. In this study, we show that centrifugal evaporation leads to effective concentration of tritium samples without the decomposition typically observed by rotary evaporation.

Development of an automated system for preparation of liquid scintillation counting samples for radiolabeled pharmaceuticals.

Radiolabeled compounds are essential tools in drug development used to obtain critical metabolism and safety information. To support the synthesis and ensure quality of radiolabeled compounds for all programs, bench automation has been implemented in our laboratories. The concept of a platform technology for bench-top automation is not new. A considerable investment in the automation of various critical analytical laboratory workflows to both harmonize the efforts of a large and diverse global organization and minimize capital footprint has been made on our part. Various custom automation techniques and applications have been developed to increase capabilities and productivity of radiochemical analyses at Merck. In this paper, we will present a novel system that is capable of automating the liquid scintillation counting procedure. The system has handled multiple radiolabeled ((3)H, (14)C, and (35)S) pharmaceutical compounds with an accuracy of 5% with a standard deviation of 2% and a cycle time of ~10 min per analysis.

Pharmacology and expression analysis of glycine transporter GlyT1 with [3H]-(N-[3-(4'-fluorophenyl)-3-(4'phenylphenoxy)propyl])sarcosine.

In the central nervous system, re-uptake of the neurotransmitter glycine is mediated by two different glycine transporters, GlyT1 and GlyT2. GlyT2 is found in brainstem and spinal cord, whereas GlyT1 is expressed in rat forebrain regions where it is responsible for most glycine transport activity. Initially, GlyT1 and GlyT2 were pharmacologically differentiated by sarcosine, a weak selective inhibitor of GlyT1. The recently described selective and potent GlyT1 antagonist, NFPS/ALX-5407 provided an important additional tool to further characterize GlyT1 pharmacology. In the present study, we have radiolabeled the racemic form of NFPS (N-[3-(4'-fluorophenyl)-3-(4'-phenylphenoxy)propyl])sarcosine (also known as ALX-5407) to investigate its interaction with GlyT1, as well as define GlyT1 expression in the rat central nervous system. Kinetic studies indicated that [3H]NFPS binds rapidly to rat forebrain membranes and dissociates with a t(1/2) of 28 +/- 5 min. [3H]NFPS labeled a saturable population of sites in rat forebrain with a Kd of 7.1+/-1.3 nM and a B(max) of 3.14 +/- 0.26 pmol/mg protein. Bound [3H]NFPS was fully and potently displaced by unlabeled NFPS, whereas glycine and sarcosine were weak, Na+-dependent inhibitors with IC50 of 1,008 and 190 microM, respectively. Additional saturation experiments indicated that glycine and sarcosine were non-competitive antagonists of [3H]NFPS binding. Functional studies revealed that NFPS was a non-competitive inhibitor of [3H]glycine uptake and does not interact with Na+ and Cl- binding sites of GlyT1. Overall, this work shows that [3H]NFPS is a valuable tool in studying GlyT1 expression and pharmacology and that NFPS interacts with GlyT1 at a site different from the transporter translocation and ion binding sites.

Comunicação entre profissional de saúde e familiares de pacientes em terapia intensiva / Comunicación entre profesional de salud y familiares de pacientes en terapia intensiva / Communication between health workers and relatives of patients in an intensive care

O objetivo do estudo foi avaliar a comunicação entre o profissional de saúde e os familiares dos pacientes internados em uma unidade de terapia intensiva (UTI). Estudo exploratório e de campo realizado na UTI do Hospital Universitário da Universidade de São Paulo entre janeiro e março de 2007. Foram entrevistadas 22 famílias, após a alta do paciente da UTI, com formulário contendo 10 perguntas sobre como foram acolhidos pela equipe de saúde e informados sobre o quadro clínico do paciente durante o horário de visita. Também foi utilizada a técnica da observação participante que permitiu verificar os sinais comunicativos verbais e não verbais dos familiares durante a interação com a equipe. Conclui-se que parte das famílias necessita de mais clareza de informações sobre o ambiente da UTI e de apoio emocional por parte da equipe de saúde.

The purpose of this study was to evaluate communication between health workers and relatives of patients in a intensive care unit (ICU). This exploratory and field study took place at the ICU of São Paulo University Hospital from January to March of 2007. Twenty-two families were interviewed, after patient release from ICU, using a form of ten questions on how they had been received by the health team and informed of the patient’s clinical condition during visiting hours. The technique of participatory observation was also used to discern relatives’ verbal and nonverbal communication signals during interaction with the team. It concludes that some relatives need clearer information on the ICU environment and emotional support from the health team.

Este estudio tuvo el objetivo de evaluar la comunicación entre el profesional de salud y los familiares de los pacientes internados en una unidad de terapia intensiva (UTI). Estudio exploratório y de campo ejecutado en la UTI del hospital escuela de la Universidad de São Paulo-Brasil, entre enero y marzo de 2007. Fueron entrevistadas 22 familias, después de la alta del paciente de la UTI, con 10 preguntas sobre como fueron acogidos por el equipo de salud e informados sobre el cuadro clínico del paciente durante el horario de visita. También fue utilizada la técnica de la observación participante que posibilitó verificar los señales comunicativos verbales y no verbales de los familiares durante la interacción con el equipo. Se concluye que parte de las familias necesita más claridad de informaciones sobre el ambiente de la UTI y de ayuda emocional por parte del equipo de salud.

Comunicación efectiva con familiares de pacientes en cuidados críticos: opinión de los profesionales de salud / Comunicação efetiva com família de pacientes em cuidados críticos: percepção dos profissionais de saúde / Effective communication with family of patients in critical cares: perception of the health professionals

Es importante preparar al equipo de salud para que pueda atender de modo adecuado a los familiares de pacientes en estado crítico. Objetivo: evaluar junto con los profesionales de la salud que trabajan en las Unidades de Cuidados Intensivos(UCI), lo que ellos consideran una comunicación efectiva entre los familiares de pacientes en estado crítico y el equipo de salud, y si ellos la practican. Material y métodos: estudio cualitativo donde se entrevistó con preguntas abiertas a 34 profesionales de una UCI, la mayoría del equipo de enfermería, en la ciudad de Sao Paublo (Brasil). Losdatos fueron categorizados según el análisis de contenido descrito por Bardin. Resultados: se identificó los aspectosverbalesy no verbales de la comunicación, para ejemplificar la postura profesional adecuada. La mayoría consideraba importante recibir información clara y objetiva sobre el paciente y el ambiente de la UCI, además del apoyo emocional,inclusive para el equipo de enfermería; también afirmaron atender sus propias expectativas como profesionales.Quienes declararon no atender a los familiares, se justificaron apelando a la propia dinámica de la unidad. Conclusiones: los profesionales reconocen las necesidades de los familiares, cuando asumen esa posición, no obstante, algunos no logran llenar las expectativas esperadas por la familia.

É importante preparar a equipe de saúde para atender os familiares dos pacientes que estao em estado crítico. Objetivo:o estudo verificou, junto aos profissionais de saúde que trabalham em urna Unidade de Terapia Intensiva (UTI), o que eles consideram urna comunicayao efetiva entre equipe e familiar, e se os mesmos a praticam. Materiais e métodos: estudo de abordagem qualitativa que entrevistou 34 profissionais de urna UTI, na cidade de Sao Paulo (Brasil), sendo a maioria da equipe de Enfermagem, com duas perguntas abertas. Os dados foram categorizados segundo a propostade análise de conteúdo descrita por Bardin. Resultados: foram identificados aspectos verbais e nao-verbais para exemplificar como seria a postura profissional adequada. A maioria considerou importante receber informacoes claras e objetivas sobre o paciente e o ambiente da UTI, e também apoio emocional, inclusive pela equipe de Enfermagem, e afirmam atender as próprias expectativas como profissionais. Os que dizem nao atender se justificam pela dinamica corrida da unidade. Conclusoes: os profissionais reconhecem as necessidades dos familiares, quando se imaginamnessa posicao, porém alguns nao conseguem atingir as expectativas que imaginam ser esperadas pela família.

It is important to prepare the health team to take care of the relatives of the patients who are in critical state.Objective: Evaluate together with health professionals, working in the Intensive Care Units (ICU), what they considereffective communication between the family of patients in critical condition and health team, and if they practice it .Material and methods: Qualitative study of an interview with open questions to 34 professionals of a ICU, in the cityof Sao Paulo (Brazil), being the majority of Nursing staff. The data were categorized according to content analysisdescribed by Bardin. Results: Verbal and nonverbal aspects were identified to exemplify appropriate professionalattitude. Most considered it important to receive clear and objective information about the patient and the environmentof the ICU, in addition to emotional support, including the Nursing staff, also affirms to take care of their ownexpectations as professional. Those who do not take care of relatives justify by appealing the dynamic of the unit.Conclusions: The professionals recognize the needs of the relatives, when they assume that position, however some do not obtain to reach the expectations that imagine to be waited by the family.