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Resumen El descubrimiento de un nuevo principio activo farmacéutico implica estudios preclínicos, que tienen como objetivo demostrar que es eficaz y seguro para un posterior ensayo en seres humanos. Esto conduce a la necesidad de desarrollar tecnologías que aprovechen las nuevas herramientas analíticas disponi bles dentro de un contexto donde los resultados de las pruebas realizadas, estén plenamente documentados, bajo sistemas de buenas prácticas de laboratorio auditables. En esta revisión se actualizan y describen algunos de los ensayos realizados en la etapa preclínica del desarrollo de un nuevo fármaco y el estado actual de la tecnología analítica empleada para el dosaje de diferentes biomarcadores sanguíneos de interés. Se analizaron los biomarcadores más relevantes, las normativas de validación de las técnicas analíticas empleadas para su determinación y los problemas que se presentan al tratar de aplicarlas.
Abstract New drug discovery involves preclinical studies to demonstrate its effectivity and safety for further tests in humans. This leads to the need to develop technologies that take advantage of the new analytical tools available within a context where the results of the tests carried out are fully documented, under auditable systems of good laboratory practice. This review updates and describes some of the tests carried out in the preclinical stage of the development of a new drug and the current state of the analytical technology used to measure different blood biomarkers of interest. Biomarker parameters were analyzed at the physiological level, considering both the validation regulations of the analytical techniques used for their determination as the problems that arise when trying to apply them, since many of these biomarkers are endogenous compounds in the used matrices.
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Humanos , Biomarcadores , Descubrimiento de DrogasRESUMEN
The early diagnosis of cancer is aimed at intervention, as far as possible in the early stage of the treatment of cancer, thereby reducing the suffering and economic burden of the patient. Circulating tumor cells (CTCs) are tumor cells that fall from the malignant tumor into the circulatory system and form cancer metastasis when they migrate to distant organs. The detection and identification of CTCs is important for the study, monitoring and intervention of cancer metastasis processes. However, the amount of CTCs in the blood is extremely rare, and the environment in the blood is complex, which poses a huge technical challenge for its screening and analysis. Various methods have been developed to enrich CTCs from blood samples of cancer patients. Among them, microfluidic technology has great advantages in the field of CTC detection.
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Miniaturized, integrated and automated microfluidic chips provided a new solution for clinical laboratory technology. However, microfluidic chips still require accessory devices to read, analyze and display results. The portable, widely-used and multi-functional smartphones have provided a simple, portable, cost-effective, and easy-to-use platform for microfluidic chips. The combination of the two technologies is expected to become the next-generation of diagnostic methods, especially the important research direction of point-of-care testing (POCT).
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Malaria remains a major tropical health burden owing to the development of resistance and decreased sensitivity to the frequently used conventional antimalarial drugs. The drug like artemisinin possesses potent antimalarial activities, but has some limitations. Therefore, new strategies are to be implemented for optimal utilization of artemisinin to improve its therapeutic effectiveness and to overcome its limitations. The present review focuses on present scenario of malaria and pharmacological as well as analytical aspects of artemisinin. Data from 2000 to 2018 were collected from NCBI for understanding the various analytical techniques used for estimation of artemisinin. This review will reveal the facts about artemisinin which can be utilized to develop novel drug delivery system either in a combination or as alone for the wellbeing of the patients suffering from malaria.
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@#In recent years, cases of illegal addition of chemical substances into the TCMs and health-care products happened regularly. Therefore, it is particularly important to develop fast, sensitive and accurate analysis methods for detection of the adulterated chemical substances. Through literature survey of relevant papers published in 2016-2017, this article summarizes the application of various analytical techniques for adulterated chemical substances to the TCMs and health-care products with useful information for the further development of new methods and technologies in this field.
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The early diagnosis of cancer is aimed at intervention, as far as possible in the early stage of the treatment of cancer, thereby reducing the suffering and economic burden of the patient. Circulating tumor cells (CTCs) are tumor cells that fall from the malignant tumor into the circulatory system and form cancer metastasis when they migrate to distant organs. The detection and identification of CTCs is important for the study, monitoring and intervention of cancer metastasis processes. However, the amount of CTCs in the blood is extremely rare, and the environment in the blood is complex, which poses a huge technical challenge for its screening and analysis. Various methods have been developed to enrich CTCs from blood samples of cancer patients. Among them, microfluidic technology has great advantages in the field of CTC detection.
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Miniaturized, integrated and automated microfluidic chips provided a new solution for clinical laboratory technology. However, microfluidic chips still require accessory devices to read, analyze and display results. The portable, widely-used and multi-functional smartphones have provided a simple, portable, cost-effective, and easy-to-use platform for microfluidic chips. The combination of the two technologies is expected to become the next-generation of diagnostic methods, especially the important research direction of point-of-care testing (POCT).
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Malaria remains a major tropical health burden owing to the development of resistance and decreased sensitivity to the frequently used conventional antimalarial drugs. The drug like artemisinin possesses potent antimalarial activities, but has some limitations. Therefore, new strategies are to be implemented for optimal utilization of artemisinin to improve its therapeutic effectiveness and to overcome its limitations. The present review focuses on present scenario of malaria and pharmacological as well as analytical aspects of artemisinin. Data from 2000 to 2018 were collected from NCBI for understanding the various analytical techniques used for estimation of artemisinin. This review will reveal the facts about artemisinin which can be utilized to develop novel drug delivery system either in a combination or as alone for the wellbeing of the patients suffering from malaria.
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Flavonoids are a widely distributed group of phytochemicals having benzo-pyrone nucleus, and more than 4,000 different flavonoids have been described and categorized into flavonols, flavones, flavanones, isoflavones, catechins and anthocyanidins. Flavonoids occurs naturally in fruits, vegetables, nuts, and beverages such as coffee, tea, and red wine, as well as in medical herbs. Flavonoids are responsible for the different colors of plant parts and are important constituents of the human diet. Flavanoids have different pharmacological activities, such as antioxidant, anti-allergic, antibacterial, anti-inflammatory, antimutagenic and anticancer activity. Naringenin belongs to the flavanones and is mainly found in fruits (grapefruit and oranges) and vegetables. Pharmacologically, it has anticancer, antimutagenic, anti-inflammatory, antioxidant, antiproliferative and antiatherogenic activities. Naringenin is used for the treatments of osteoporosis, cancer and cardiovascular diseases, and showed lipid-lowering and insulin-like properties. In the present review, detailed pharmacological and analytical aspects of naringenin have been presented, which revealed the impressive pharmacological profile and the possible usefulness in the treatment of different types of diseases in the future. The information provided in this communication will act as an important source for development of effective medicines for the treatment of various disorders.
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Humanos , Antiinflamatorios , Química , Farmacología , Antineoplásicos , Química , Farmacología , Antioxidantes , Química , Farmacología , Flavanonas , Química , Farmacología , Isoflavonas , Química , Farmacología , Neoplasias , QuimioterapiaRESUMEN
Circulating tumor cells ( CTCs ) exists in peripheral blood of patients with a variety of malignant tumors , the accurate detection of CTCs helps early detection of cancer , and can be used to guide individualized treatment of cancer patients , rapid assessment of tumor chemotherapy drugs and detection of tumor recurrence .However , the CTCs in peripheral blood of cancer patients were extremely low , their detection requires efficient and specific enrichment and separation methods .The new microfluidic technology can accurately control the microliquid preciously , combined with the immunomagnetic separation technology , has been applied in detecting CTCs , which can save the blood samples of CTCs , has the advantages of simple operation , fast sorting speed , high specificity and high purity .
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The efficient characterization of genetic and epigenetic alterations in oncology requires highly sensitive and specific high throughput procedures.However, the necessary level of sensitivity and specificity were previously unreachable using conventional testing procedures.By partitioning individual target molecules within separated compartments, digital PCR ( dPCR ) could allow to overcome such limitations and detect with unprecedented accuracy very rare sequences.In such procedure, the sample is diluted such that each individual compartment will contain no more than one target sequences.The assay provides an absolute value and quantitative data.The recent coupling of dPCR procedure with microfluidic systems in commercial platforms should make droplet based digital PCR an essential tool for the management of patients with cancer.Applications range from the analysis of tumor heterogeneity to the analysis of body effluents.Droplet based digital PCR is also particularly suited for the increasing field of liquid biopsy analysis.
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Microfluidics-based digital PCR depends on microfluidic chip to split PCR reaction mixture into many tiny equal-volume units.Quantitative assessment of target DNA template can be obtained by counting the number of fluorescence-positive units after thermocycling.Microfluidics-based digital PCR exhibits many advantages including absolute quantification, high sensitivity and accuracy, and shows great promise in a variety of applications, such as infectious diseases diagnose, early cancer detection and prenatal diagnose. There are already several microfludics-based digital PCR products produced from sereval companies.It is believed that as the technology improves, microfluidics-based digital PCR will find broader applications and become the next-generation tool for genetic tests.
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Microfluidic chip exhibit a great promising development in clinical diagnosis and disease screening due to their advantages of precise controlling of fluid flow,requirement of mini amount sample,rapid reaction speed and convenient integration.A lot of demonstrations on the diagnostic applications related to genes,proteins,and cells have been reported because of their advantages associated with miniaturization and automation.Here,the applications and developments of on-chip nucleic acid amplification and analysis,protein analysis and detection,cell selection and cell drug screening were discussed.Microfluidic chip can provide an easy integration platform for biomarkers in a high throughput and accurate detection.
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@#Pharmaceutical impurities affect the quality of drugs, and their research is essential in drug development, production and sale. Recently, there have been tremendous advancements in instrumental techniques that allow rapid development of technologies and analytical methods in the research of pharmaceutical impurities. This review discusses the origins and analytical methods of pharmaceutical impurities including genotoxic impurities, and mainly describes the analytical techniques and strategies to assess the genotoxicity of impurities, providing technical references for the study of pharmaceutical impurities.
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Objective The micro-vascular element plays a key role in the delivery of nutrients and the regulation of hemodynamic behavior, however, research is often hindered by ethical , economic and technological issues .Therefore, construction of a micro-vascular network in vitro will help to study the related pathological and physiological behavior in microvessels.Methods In this study, a micro-vascular element model with features of a micro-vascular network in vivo was designed based on the network structure of retinal arterioles .A micro-vascular network model in vitro, characterized by network asymmetry and the presence of both bifurcation-and side-branches , was developed by soft lithography technology . The developed microdevice allowed for the quantification of the cell -depletion layer ( CDL) thickness and hematocrit ( Ht) distribution within the microchannel networks .Results and Conclusion The study showed the potential of the developed in vitro model in revealing key hemodynamic features which have been detected for microvascular elements in vivo, including the relationships between CDL thickness , Ht and red blood cell distribution .The present study provides a new strategy and a technology for studying hemodynamics and microvascular system diseases in vitro.
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Major infectious diseases affect human health seriously .In order to prevent , control and treat diseases effectively , the key lies in rapid detection and identification of pathogens .Rapid development and wide application of molecular biological techniques , such as PCR, gene sequencing and bio-chip technology , have greatly improved the prevention , control and treatment of major infectious diseases .The future development trend is to establish an accurate , rapid, high-throughput , portable and intelligent diagnostic technology of molecular biology .
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Antecedentes: Las estatinas presentan principalmente un efecto hipocolesterolémico, pero asimismo acciones biológicas adicionales, como efecto antiinflamatorio e inmunomodulador, actividad antioxidante, reducción de la incidencia de algunos tipos de cáncer, efectos benéficos en el metabolismo óseo y en el tratamiento de enfermedades neurodegenerativas y el SIDA. Se dividen en dos clases: las estatinas naturales y las sintéticas, las cuales surgen como producto de la necesidad de potencializar el efecto de las primeras. Estas propiedades han impulsado las investigaciones encaminadas a la comprensión de su comportamiento químico y sus propiedades fisicoquímicas, así como la comprensión de la relación entre sus estructuras y las ya mencionadas actividades. Su estudio ha permitido el desarrollo de técnicas analíticas eficientes tanto para su determinación en diferentes matrices, como la optimización de los procesos de extracción, separación, cuantificación y elucidación estructural, así como ahondar en el planteamiento de sus rutas biosintéticas, lo que aportará herramientas para poder intervenir, mediante la biotecnología, en los procesos biosintéticos, buscando el aumento de su producción por un organismo específico. Objetivos: El objetivo de este trabajo es presentar una revisión de la química, biosíntesis, farmacocinética y técnicas analíticas para la determinación de las estatinas con el fin de aportar, de manera rápida, conocimientos a quienes realizan investigaciones sobre estos metabolitos. Métodos: La revisión abarcó los últimos 12 años y se efectuó realizando la selección de aquellas investigaciones más relevantes que permiten conocer la química, las variaciones estructurales, las técnicas analíticas empleadas para la determinación de las estaninas y sus rutas biosintéticas. Asimismo, se pretendió abarcar un conocimiento general de sus acciones biológicas, farmacología y farmacocinética, tópicos estrechamente relacionados con sus diferencias estruc...
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Enfermedad Coronaria , Enfermedades Cardiovasculares , Farmacocinética , Inhibidores de Hidroximetilglutaril-CoA ReductasasRESUMEN
Objective A new method for detecting K-ras mutations based liquid chip was used to evaluate K-ras mutations associated with non-small cell lung cancer (NSCLC) patients,to direct the personalized treatment and prognosis evaluation.Methods Take the diagnosis technology research methods,the sensitivity and repeatability of the liquid chip K-ras gene mutation detection method were assessed.A total of 100 NSCLC patients from Nov 2011 to Feb 2012 in Shanghai Chest hospital were included in this study,the fresh tumor tissues were collected for DNA extraction.The 2nd exon 12 and 13 codons,containing 8 K-ras mutations occuring in high frequency were amplified by polymerase chain reaction (PCR),followed by ligation of the PCR products to a series of special probes using ligase detection reaction (LDR),then the PCR-LDR products were analyzed by liquid chip platform.Direct sequencing was applied to compare with the detection results.Results The sensitivity of liquid chip technology detection was 10％-20％,higher than the traditional sequencing method by 1％.Average CV value was 4％-15％ and showed good repeatability.5 K-ras mutations in 100 patients (5％) were detected using multiplex PCR-LDR combined fluid chip methods,including 3 Glyl2Val and 2 Gly12Asp mutations in exon 2.The 5 K-ras mutations were verified accurately by direct sequencing.Conclusions The novel detection method of K-ras mutations based PCRLDR and fluid chip shows high throughput,high sensitivity,good repeatability and the results are reliable and accurate.This method can be used to accurately identified K-ras mutations for NSCLC patients prior to their targeted therapy with TKIs.
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Objective To develop a phenylketonuria (PKU) screening method based on a compact disk (CD) type microfluidic chip capable of generating reciprocating flow within the microchannels that facilitate rapid DNA hybridization. Methods This microfluidic device consists of a two-layer structure: a polydimethylsiloxane (PDMS) top layer containing 12 DNA hybridization microchannels, and a bottom glass layer with immobilized hydrogel conjugated DNA arrays. The DNA arrays included R243Q, V245V and the blank control probes. When the CD device was spun, the PCR products were driven into the hybridization channel by centrifugal force. When the rotation of the CD device was stopped, capillary force pulled the PCR products solution to flow back to the channel. After the on-chip hybridization, the hybridization signals were captured on a fluorescence microscope. The specificity, detection limitation and reproducibility of this device were evaluated. Thirty DNA samples from pregnant women with suspected PKU were detected by this device.Then the results were compared with DNA sequencing results. Results With the compact disk type microfluidic chip, the hybridization time could be reduced to 15 min, sample consume could be as low as 1. 5 μl and the detection limitation was 0. 7 ng/μl. With the chip based method, samples of PKU patients and healthy controls were detected and the results were consistent with DNA sequencing results. Five different batches of chips and five micro-channels of each chip were selected to test one PKU patients with V245V mutation. All the results were positive, indicating good reproducibility. Four cases of V245V mutation and 1 case of R243Q mutation were found in 30 suspected PKU carried pregnant women. Conclusion The compact disk microfluidic device has advantages of simple, rapid and highly sensitive, thus is well suited to PKU screening.
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Objectiye To optimize the depth of the microchannel and the time point for sperm collection,and improve the efficiency of sperm screening on a microfluidic device. Methods Microchannels with four different depths of 25, 50, 100 and 200 μm were tested. Mice sperm were added to the inlet of the microchannel. The relative quantity and motility of sperm in the outlet were recorded at different collection times, i.e. ,5, 15, 30 and 60 min. Statistical method one-way ANOVA and appropriate post-hoc testing were applied to analyze differences between different groups, and further to select the best-fit depth of the microchannel and the time point for collection. Results In microchannels with depths of 25, 50, 100 and 200 μm, the sperm motilities measured in each outlet were (85.4 ± 2.3)%, (85.8 ± 5.8)%,( 87. 2 ± 2. 8 ) %, (76. 5 ± 2. 8 ) % respectively with statistical significance ( F = 5.8, P ＜ 0. 05 ). No obvious differences were found among 25-100 μm channels, however the motility dramatically decreased in the 200 μm group. The relative sperm quantities were (5.2 ±2.0)%, (7.2 ±2.5)%,(12.3 ±2.0)%,(7. 7 ± 1.1 ) % respectively with statistical significance ( F = 6. 9, P ＜ 0. 05), which increased with channel depth from 25 to 100 μm,while it decreased in the 200 μm channel Taking 2 indexes into account, 100 μmwas the most fit channel depth for sperm motility screening. The sperm motility in the outlet gradually decreased with time. At the time points of 5, 15, 30 and 60 min after adding sperm, the sperm motilities were (99. 6 ±0. 7)%, (87.2 ±2. 8)%, (79. 3 ±2. 2)% and (62. 6 ±8.0)% respectively with statistical significance ( F = 37. 3, P ＜ 0. 01 ). Yet the relative quantities of sperm in the outlet increased almost three times in this process. At the time points mentioned above, the relative quantities of sperm were (5.8±1.1)%, (10.6 ± 0.9)%, (12.1 ± 1.7)%, (17.9 ± 3.4)% respectively with statistical significance ( F = 17.8, P ＜ 0. 01 ). Thus 15-30 min was the ideal screening time. Conclusion An effective microdevice for sperm screening with optimized depth and collection time period is developed,which may contribute significantly for the screening of healthy sperm on microfluidic chips.