RESUMEN

Autoimmune diseases are characterized by alteration in balance of various cytokines. Rheumatoid arthritis is a well-known inflammatory disease leading to destruction of cartilage at knee and hands. Collagen-induced arthritis (CIA) is a common autoimmune model for rheumatoid arthritis study. Here, we have investigated the therapeutic role of medicarpin, a natural pterocarpan with known anti-osteoclastogenic activities, in postmenopausal polyarthritis model of DBA/1J mice. For this, mice were ovariectomized and CIA was induced in OVx animals with primary immunization. After 21 days, booster dose was injected in Ovariectomy (OVx) mice to develop postmenopausal poly-arthritis mice model. Medicarpin treatment in mice at dose of 10.0 mg/kg/body wt was started after 21 days of primary immunization for one month of time period every day orally. We found that medicarpin prevented alteration of TH-17/Treg ratio in CIA model leading to reduced osteoclastogenesis. Micro Computed Tomography (Micro-CT) analysis demonstrated that medicarpin prevents cartilage erosion in joints and restores loss of trabeculae parameters in distal tibia. Treatment with medicarpin also prevented alteration of various cytokines level by down-regulating various pro-inflammatory cytokines like TNF-α, IL-6 and IL-17A, while up-regulating anti-inflammatory cytokine IL-10 in CIA model of mice. Biological marker of arthritis is cartilage oligomeric matrix protein (COMP). COMP level was up-regulated in CIA induced mice while treatment with medicarpin significantly restored the serum level of COMP compared with untreated groups. Cartilage staining by Safranin-O also indicates that cartilage destruction in joints of CIA mice was prevented by medicarpin treatment. From this study, we can conclude that medicarpin is effective in preventing arthritis in post-menopausal conditions.

RESUMEN

Medicarpin, one of the active constituents isolated from the extract of Butea monosperma, has been shown to have various pharmacological activities including potent anti-osteoporotic properties. The aim of this study was to investigate the oral pharmacokinetics, tissue distribution and excretion of medicarpin following single oral dose administration in female rats. Oral pharmacokinetics was explored at 5 and 20 mg/kg while tissue distribution, urinary and fecal excretion were studied following 20 mg/kg oral dose. Medicarpin was quantified in rat plasma, urine, feces and tissue samples using a validated LC-MS/MS method following reverse-phase HPLC separation on RP18 column (4.6 mm × 50 mm, 5.0 µm) using methanol and 10 mM ammonium acetate (pH 4.0) as mobile phase in the ratio of 80:20 (v/v) at a flow rate of 0.8 mL/min. The oral bioavailability of medicarpin was found to be low with low systemic levels. The concentration in tissues was significantly higher than plasma. Highest tissue concentrations were found in the liver followed by bone marrow. Urinary and fecal excretion of medicarpin was < 1 %. In conclusion, medicarpin was found to be highly distributed in body tissues and minimally excreted via urine or feces.

Asunto(s)

Líquidos Corporales/metabolismo , Osteoporosis/tratamiento farmacológico , Pterocarpanos , Animales , Disponibilidad Biológica , Análisis Químico de la Sangre , Cromatografía Líquida de Alta Presión , Heces , Femenino , Límite de Detección , Extracción Líquido-Líquido , Pterocarpanos/administración & dosificación , Pterocarpanos/síntesis química , Pterocarpanos/farmacocinética , Ratas , Ratas Sprague-Dawley , Espectrometría de Masas en Tándem

RESUMEN

A new dual responsive "turn-on" and "ratiometric" aggregation-induced emission luminogen (AIEgen) 3-formyl-5-(piperidin-1-yl)biphenyl-4-carbonitrile 6 a (FPBC 6 a) for selective detection of hydrazine in solution as well as in vapour phase is described. At a low concentration of 2.5 µm, the probe FPBC 6 a is non-fluorescent (turn-off) but remarkably lights up (turn-on with blue emission) in the presence of hydrazine solution (0.25-25 µm). Interestingly, at higher concentrations, the nanoaggregates of FPBC 6 a (>25 µm, 99 % HEPES in DMSO) displayed ratiometric response in the presence of hydrazine with a remarkable hypsochromic shift from the green (500-550 nm) to blue regions (440-480 nm). Furthermore, a real application of FPBC 6 a was successfully demonstrated through the detection and visualization of hydrazine in live cervical cancer cells as well as using portable test strips.

RESUMEN

A series of new 6H-benzofuro[3, 2-c]chromenes (BFC, pterocarpans) with structure-activity relationships were investigated for their potential use in osteoporosis treatment. One of the BFCs 3-piperidylethoxypterocarpan 20 promotes osteoblast differentiation and mineralization at a dose as low as 1 pM via activation of ER/P38MAPK/BMP-2 pathway. When evaluated for in-vivo osteogenic activity in female Sprague-Dawley rats, BFC 20 increased bone mineral density and new bone formation, compared with control at 1.0 and 10.0 mg/kg/body weight by oral gavage for 30 days. The compound was devoid of any uterotrophic effect and led to the new bone formation in adult ovariectomized osteopenic rats. BFC 20 compound also inhibited bone resorption by reducing Ovx induced increase in urinary CTx, thus exhibiting both bone anabolic and anti-catabolic action. Finally, BFC 20 treatment to Ovx rats led to improved trabecular microarchitectural restoration and exhibited therapeutic potential as a dual acting anti-osteoporotic agent for the management of osteoporosis.

Asunto(s)

Anabolizantes/uso terapéutico , Enfermedades Óseas Metabólicas/tratamiento farmacológico , Hueso Esponjoso/patología , Ovariectomía , Piperidinas/uso terapéutico , Pterocarpanos/uso terapéutico , Fosfatasa Alcalina/metabolismo , Anabolizantes/síntesis química , Anabolizantes/química , Anabolizantes/farmacología , Animales , Biomarcadores/metabolismo , Densidad Ósea/efectos de los fármacos , Enfermedades Óseas Metabólicas/patología , Proteína Morfogenética Ósea 2/metabolismo , Remodelación Ósea/efectos de los fármacos , Calcificación Fisiológica/efectos de los fármacos , Hueso Esponjoso/efectos de los fármacos , Diferenciación Celular/efectos de los fármacos , Femenino , Osteoblastos/efectos de los fármacos , Osteoblastos/metabolismo , Osteoblastos/patología , Fosforilación/efectos de los fármacos , Piperidinas/síntesis química , Piperidinas/química , Piperidinas/farmacología , Pterocarpanos/síntesis química , Pterocarpanos/química , Pterocarpanos/farmacología , ARN Mensajero/genética , ARN Mensajero/metabolismo , Ratas Sprague-Dawley , Reacción en Cadena en Tiempo Real de la Polimerasa , Receptores de Estrógenos/metabolismo , Transducción de Señal/efectos de los fármacos

RESUMEN

Screening of a chemical library of pyranones and their ring transformed fluorophores led to the discovery of a novel 5,6-dihydro-2H-pyrano[3,2-g]indolizine (DPI) class of the luminogen DPI 7, which exhibited unique solution-solid dual emission (SSDE) behavior with an emission color shift from bright-green in solution to a strong red emission in the solid state. The AIE mechanism of these luminogens revealed a well-defined set of noncovalent interactions (CH⋅⋅⋅O and CH⋅⋅⋅N) that block the motion of C2 -flexure leading to restriction of intramolecular vibrations (RIV) in the solid state. DPI-7 is the first example of solution-solid dual emissive RIV-based AIEgen, which has great potential both in biomedical imaging and optoelectronic fields.

RESUMEN

In order to address the existing limitations of the commercially available fluorescent probe CMAC (7-amino-4-chloromethylcoumarin), a new series of highly fluorescent donor-acceptor 7-hydroxy-coumarin derivatives was prepared and these derivatives were used as vacuole specific fluorescent probes for live cell imaging of unicellular parasitic protozoa L. donovani promastigotes and yeast cells S. pombe and S. cerevisiae. The synthesized 7-hydroxy-coumarins exhibited interesting photophysical properties and have advantages such as excitation in the visible region, good water solubility, photo-stability, good quantum yield and low cytotoxicity.

RESUMEN

Osteogenic activity was identified in medicarpin (Med), a natural pterocarpan. Further, it was decided to study the differentially regulated protein expression during osteoblast differentiation in the presence of Med. Using 2D proteomic approach, we found that Med treatment to osteoblasts significantly downregulated GRP78, an ER chaperone with anti-apoptotic properties which also controls the activation of unfolded protein response signaling, a pro-survival strategy for normal ER functioning. However, severe stress leads to triggering of apoptotic responses and signaling switches to pro-apoptotic. In order to elucidate the effect of Med downregulation of GRP78, osteoblasts were transfected with SiGRP78 or SiGRP78+ Med or Med alone. It was seen that mRNA and protein levels of ER stress markers like GRP78, ATF-4, and CHOP were decreased in all the three groups with maximum reduction in SiGRP78+ Med group. Med targets GRP78 by inhibiting mitochondrial-mediated apoptosis which is evident by reduced levels of cytochrome c, caspase-3, Bax/BCL2 ratio, and enhanced expression of survivin. Finally, Annexin-PI staining of apoptotic cells revealed that MED inhibition of GRP78 leads to reduced osteoblast apoptosis and increased osteoblast survival. Altogether, our data show that Med inhibits ER stress-induced apoptosis and promotes osteoblast cell survival by targeting GRP78.

Asunto(s)

Apoptosis/efectos de los fármacos , Estrés del Retículo Endoplásmico/efectos de los fármacos , Proteínas de Choque Térmico/metabolismo , Osteoblastos/metabolismo , Pterocarpanos/farmacología , Animales , Apoptosis/genética , Supervivencia Celular/efectos de los fármacos , Supervivencia Celular/genética , Células Cultivadas , Chaperón BiP del Retículo Endoplásmico , Estrés del Retículo Endoplásmico/genética , Proteínas de Choque Térmico/genética , Ratones , Osteoblastos/citología , Proteómica

RESUMEN

We evaluated the bone regeneration and healing effect of Medicarpin (med) in cortical bone defect model that heals by intramembranous ossification. For the study, female Sprague-Dawley rats were ovariectomized and rendered osteopenic. A drill hole injury was generated in mid femoral bones of all the animals. Med treatment was commenced the day after and continued for 15 days. PTH was taken as a reference standard. Fifteen days post-treatment, animals were sacrificed. Bones were collected for histomorphometry studies at the injury site by micro-computed tomography (µCT) and confocal microscopy. RNA and protein was harvested from newly generated bone. For immunohistochemistry, 5µm sections of decalcified femur bone adjoining the drill hole site were cut. By µCT analysis and calcein labeling of newly generated bone it was found that med promotes bone healing and new bone formation at the injury site and was comparable to PTH in many aspects. Med treatment led to increase in the Runx-2 and osteocalcin signals indicating expansion of osteoprogenitors at the injury site as evaluated by qPCR and immunohistochemical localization. It was observed that med promoted bone regeneration by activating canonical Wnt and notch signaling pathway. This was evident by increased transcript and protein levels of Wnt and notch signaling components in the defect region. Finally, we confirmed that med treatment leads to elevated bone healing in pre-osteoblasts by co localization of beta catenin with osteoblast marker alkaline phosphatase. In conclusion, med treatment promotes new bone regeneration and healing at the injury site by activating Wnt/canonical and notch signaling pathways. This study also forms a strong case for evaluation of med in delayed union and non-union fracture cases.

Asunto(s)

Huesos/patología , Pterocarpanos/farmacología , Receptores Notch/metabolismo , Vía de Señalización Wnt/efectos de los fármacos , Cicatrización de Heridas/efectos de los fármacos , Animales , Biomarcadores/metabolismo , Densidad Ósea/efectos de los fármacos , Enfermedades Óseas Metabólicas/tratamiento farmacológico , Enfermedades Óseas Metabólicas/fisiopatología , Regeneración Ósea/efectos de los fármacos , Regeneración Ósea/genética , Huesos/efectos de los fármacos , Huesos/fisiopatología , Diferenciación Celular/efectos de los fármacos , Femenino , Regulación de la Expresión Génica/efectos de los fármacos , Ovariectomía , Ratas Sprague-Dawley , Coloración y Etiquetado , Células Madre/citología , Células Madre/efectos de los fármacos , Células Madre/metabolismo

RESUMEN

Medicarpin is the active phytoalexin found in the stem bark of Butea monosperma having potent osteogenic activity. An LC-ESI-MS/MS was developed and validated for quantification of medicarpin in rat plasma using liquid-liquid extraction technique and diethyl ether as the extraction solvent. Medicarpin was separated on RP18 column (4.6mm×50mm, 5.0µm) using methanol and 10mM ammonium acetate (pH 4.0) in the ratio of 80:20 (v/v) as mobile phase. The method was linear within the concentration range of 1-500ng/mL and its sensitivity was 1ng/mL. The precision value for intra- and inter-day assays and stability assays was within 0.88-14.22% while the accuracy ranged between 87.46-116.0% at all four QC levels. The validated method was successfully applied to study the preclinical pharmacokinetics of medicarpin in rats. Medicarpin showed multiple peak phenomenon upon oral administration. Its oral bioavailability was 17.43%. It was found to be a rapidly absorbed (Tmax=15min), 81.61% protein bound and pH stable compound. The present study provides important information regarding preliminary pharmacokinetics of medicarpin for its further exploration as a potential therapeutic agent.

Asunto(s)

Cromatografía Liquida/métodos , Pterocarpanos/análisis , Sesquiterpenos/análisis , Espectrometría de Masa por Ionización de Electrospray/métodos , Espectrometría de Masas en Tándem/métodos , Animales , Límite de Detección , Unión Proteica , Pterocarpanos/farmacocinética , Ratas , Reproducibilidad de los Resultados , Sesquiterpenos/farmacocinética , Fitoalexinas

RESUMEN

We report a new bone anabolic and anti-catabolic pterocarpan 9-demethoxy-medicarpin (DMM) for the management of postmenopausal osteoporosis. DMM promoted osteoblast functions via activation of P38MAPK/BMP-2 pathway and suppressed osteoclastogenesis in bone marrow cells (BMCs). In calvarial osteoblasts, DMM blocked nuclear factor kappaB (NFκB) signaling and inhibited the mRNA levels of pro-inflammatory cytokines. DMM treatment led to increased OPG (osteoprotegrin) and decreased transcript levels of TRAP (tartarate resistant acid phosphatase), RANK (receptor activator of NFκB) and RANKL (RANK ligand) in osteoblast-osteoclast co-cultures. Immature female SD rats administered with DMM exhibited increased bone mineral density, bone biomechanical strength, new bone formation and cortical bone parameters. Ovx mice administered with DMM led to significant restoration of trabecular microarchitecture and had reduced formation of osteoclasts and increased formation of osteoprogenitor cells in BMCs. DMM exhibited no uterine estrogenicity. Overall, these results demonstrate the therapeutic potential of DMM for the management of postmenopausal osteoporosis.

Asunto(s)

Conservadores de la Densidad Ósea/farmacología , Densidad Ósea/efectos de los fármacos , Osteoblastos/efectos de los fármacos , Osteoclastos/efectos de los fármacos , Pterocarpanos/farmacología , Animales , Conservadores de la Densidad Ósea/uso terapéutico , Diferenciación Celular/efectos de los fármacos , Femenino , Humanos , Ratones , Osteoblastos/citología , Osteoclastos/citología , Osteoporosis Posmenopáusica/tratamiento farmacológico , Ovariectomía , Pterocarpanos/uso terapéutico , Ratas , Ratas Sprague-Dawley

RESUMEN

A new general methodology for the synthesis of various functionalized privileged oxygen heterocyclic scaffolds, viz. pyrones, coumarins, and benzannulated coumarins, is developed. The synthesis proceeds through carbanion-induced ring transformation of lactones with various methylene carbonyl compounds followed by DDQ-mediated unprecedented oxidative cleavage of oxaylidenes intermediates. Studies of the mechanism of the conversions of oxaylidene intermediates into corresponding carbonyl compounds in the presence of DDQ revealed that the reactions took place via the formation of a Michael adduct instead of an intermolecular charge transfer complex. The methodology offers the fabrication of diverse privileged scaffolds with tolerance for many functional groups onto the oxygen heterocyclic molecular framework.

Asunto(s)

Cumarinas/síntesis química , Oxígeno/química , Pironas/síntesis química , Derivados del Benceno/síntesis química , Ciclización , Lactonas/síntesis química , Lactonas/química , Modelos Moleculares

Asunto(s)

Pterocarpanos/química , Pterocarpanos/clasificación , Técnicas de Química Sintética , Cromatografía Líquida de Alta Presión , Dicroismo Circular , Glicosilación , Espectroscopía de Resonancia Magnética , Fotoquímica/métodos , Pterocarpanos/síntesis química , Estereoisomerismo

RESUMEN

A convenient synthesis of natural and synthetic pterocarpans was achieved in three steps. Optical resolution of the respective enantiomers was accomplished by analytical and semi-preparative HPLC on a chiral stationary phase. For medicarpin and its synthetic derivative 9-demethoxymedicarpin, the absolute configuration was confirmed by a combination of experimental LC-ECD coupling and quantum-chemical ECD calculations. (-)-Medicarpin and (-)-9-demethoxymedicarpin are both 6aR,11aR-configured, and consequently the corresponding enantiomers, (+)-medicarpin and (+)-9-demethoxymedicarpin, possess the 6aS,11aS-configuration. A comparative mechanism study for osteogenic (bone forming) activity of medicarpin (racemic versus enantiomerically pure material) revealed that (+)-(6aS,11aS)-medicarpin (6a) significantly increased the bone morphogenetic protein-2 (BMP2) expression and the level of the bone-specific transcription factor Runx-2 mRNA, while the effect was opposite for the other enantiomer, (-)-(6aR,11aR)-medicarpin (6a), and for the racemate, (±)-medicarpin, the combined effect of both the enantiomers on transcription levels was observed.

Asunto(s)

Conservadores de la Densidad Ósea/síntesis química , Técnicas de Química Sintética/métodos , Pterocarpanos/síntesis química , Animales , Animales Recién Nacidos , Conservadores de la Densidad Ósea/química , Conservadores de la Densidad Ósea/farmacología , Células de la Médula Ósea/efectos de los fármacos , Células de la Médula Ósea/metabolismo , Proteína Morfogenética Ósea 2/biosíntesis , Conformación de Carbohidratos , Diferenciación Celular/efectos de los fármacos , Células Cultivadas , Subunidad alfa 1 del Factor de Unión al Sitio Principal/biosíntesis , Osteoblastos/citología , Osteoblastos/efectos de los fármacos , Osteoblastos/metabolismo , Pterocarpanos/química , Pterocarpanos/farmacología , Ratas , Ratas Sprague-Dawley , Estereoisomerismo

RESUMEN

A rapid, sensitive and selective LC-MS/MS method for the quantitative analysis of 3-hydroxy pterocarpan (S006-1709) in female rat plasma has been developed and validated. A Discovery RP18 column was used for the chromatographic elution using acetonitrile and 0.1% acetic acid in water as mobile phase (80:20 v/v) at the flow rate of 0.5 mL/min. MS/MS analysis was performed using a triple quadrupole mass spectrometer with electrospray ionization in negative ion mode using biochanin as an internal standard (IS). Extraction of S006-1709 and IS from rat plasma was done by liquid-liquid extraction method using diethyl ether. The LC-MS/MS method was sensitive with 1.95 ng/mL as the limit of detection and 3.9 ng/mL as the lower limit of quantification. The method was linear in the concentration range of 3.9-1000 ng/mL. The percentage bias for intraday and interday accuracy was not greater than 4.2 and the %RSD for intraday and interday precision was not greater than 13.2. The recoveries of S006-1709 and IS were 73.9-79.3 and 85.7%, respectively. S006-1709 was found to be stable in various stability studies. The validated LC-MS/MS method was successfully applied for the oral pharmacokinetics study of S006-1709 at 10 mg/kg in female Sprague-Dawley rats.

Asunto(s)

Cromatografía Liquida/métodos , Isoflavonas/análisis , Pterocarpanos/análisis , Espectrometría de Masas en Tándem/métodos , Administración Oral , Animales , Fraccionamiento Químico , Estabilidad de Medicamentos , Estrógenos/análisis , Estrógenos/farmacocinética , Femenino , Isoflavonas/farmacocinética , Modelos Lineales , Pterocarpanos/farmacocinética , Ratas , Ratas Sprague-Dawley , Reproducibilidad de los Resultados , Sensibilidad y Especificidad