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1.
J Biol Chem ; 295(50): 17169-17186, 2020 12 11.
Artigo em Inglês | MEDLINE | ID: mdl-33028635

RESUMO

We have observed overexpression of PACS-1, a cytosolic sorting protein in primary cervical tumors. Absence of exonic mutations and overexpression at the RNA level suggested a transcriptional and/or posttranscriptional regulation. University of California Santa Cruz genome browser analysis of PACS-1 micro RNAs (miR), revealed two 8-base target sequences at the 3' terminus for hsa-miR-34a and hsa-miR-449a. Quantitative RT-PCR and Northern blotting studies showed reduced or loss of expression of the two microRNAs in cervical cancer cell lines and primary tumors, indicating dysregulation of these two microRNAs in cervical cancer. Loss of PACS-1 with siRNA or exogenous expression of hsa-miR-34a or hsa-miR-449a in HeLa and SiHa cervical cancer cell lines resulted in DNA damage response, S-phase cell cycle arrest, and reduction in cell growth. Furthermore, the siRNA studies showed that loss of PACS-1 expression was accompanied by increased nuclear γH2AX expression, Lys382-p53 acetylation, and genomic instability. PACS-1 re-expression through LNA-hsa-anti-miR-34a or -449a or through PACS-1 cDNA transfection led to the reversal of DNA damage response and restoration of cell growth. Release of cells post 24-h serum starvation showed PACS-1 nuclear localization at G1-S phase of the cell cycle. Our results therefore indicate that the loss of hsa-miR-34a and hsa-miR-449a expression in cervical cancer leads to overexpression of PACS-1 and suppression of DNA damage response, resulting in the development of chemo-resistant tumors.


Assuntos
Dano ao DNA , Resistencia a Medicamentos Antineoplásicos , MicroRNAs/metabolismo , RNA Neoplásico/metabolismo , Neoplasias do Colo do Útero/metabolismo , Proteínas de Transporte Vesicular/metabolismo , Feminino , Fase G1 , Células HeLa , Humanos , MicroRNAs/genética , RNA Neoplásico/genética , Pontos de Checagem da Fase S do Ciclo Celular , Proteína Supressora de Tumor p53/genética , Proteína Supressora de Tumor p53/metabolismo , Neoplasias do Colo do Útero/genética , Neoplasias do Colo do Útero/patologia , Proteínas de Transporte Vesicular/genética
2.
AAPS PharmSciTech ; 20(1): 16, 2018 Dec 19.
Artigo em Inglês | MEDLINE | ID: mdl-30569266

RESUMO

Lipids have been extensively used in formulations to enhance dissolution and bioavailability of poorly water-soluble as well as water-soluble drug molecules. The digestion of lipid-based formulations, in the presence of bile salts, phospholipids, and cholesterol, changes the lipid composition in vivo, resulting in the formation of different colloidal phases in the intestine. Therefore, in vitro characterization and evaluation of such formulations are critical in developing a successful formulation. This review covers comprehensive discussion on in vitro characterization techniques such as solubility, drug entrapment, thermal characterization, dissolution, and digestion of lipid-based formulations.


Assuntos
Sistemas de Liberação de Medicamentos , Lipídeos/química , Administração Oral , Animais , Química Farmacêutica , Composição de Medicamentos , Tamanho da Partícula , Fosfolipídeos , Solubilidade
3.
Artigo em Inglês | MEDLINE | ID: mdl-28807910

RESUMO

Vancomycin has been associated with acute kidney injury in preclinical and clinical settings; however, the precise exposure profiles associated with vancomycin-induced acute kidney injury have not been defined. We sought to determine pharmacokinetic/pharmacodynamics indices associated with the development of acute kidney injury using sensitive urinary biomarkers. Male Sprague-Dawley rats received clinical-grade vancomycin or normal saline as an intraperitoneal injection. Total daily doses between 0 and 400 mg/kg of body weight were administered as a single dose or 2 divided doses over a 24-h period. At least five rats were utilized for each dosing protocol. A maximum of 8 plasma samples per rat were obtained, and urine was collected over the 24-h period. Kidney injury molecule-1 (KIM-1), clusterin, osteopontin, cystatin C, and neutrophil gelatinase-associated lipocalin levels were determined using Milliplex multianalyte profiling rat kidney panels. Vancomycin plasma concentrations were determined via a validated high-performance liquid chromatography methodology. Pharmacokinetic analyses were conducted using the Pmetrics package for R. Bayesian maximal a posteriori concentrations were generated and utilized to calculate the 24-h area under the concentration-time curve (AUC), the maximum concentration (Cmax), and the minimum concentration. Spearman's rank correlation coefficient (rs ) was used to assess the correlations between exposure parameters, biomarkers, and histopathological damage. Forty-seven rats contributed pharmacokinetic and toxicodynamic data. KIM-1 was the only urinary biomarker that correlated with both composite histopathological damage (rs = 0.348, P = 0.017) and proximal tubule damage (rs = 0.342, P = 0.019). The vancomycin AUC and Cmax were most predictive of increases in KIM-1 levels (rs = 0.438 and P = 0.002 for AUC and rs = 0.451 and P = 0.002 for Cmax). Novel urinary biomarkers demonstrate that kidney injury can occur within 24 h of vancomycin exposure as a function of either AUC or Cmax.


Assuntos
Injúria Renal Aguda/induzido quimicamente , Antibacterianos/efeitos adversos , Antibacterianos/farmacocinética , Vancomicina , Animais , Área Sob a Curva , Biomarcadores/sangue , Biomarcadores/urina , Moléculas de Adesão Celular/sangue , Clusterina/sangue , Cistatina C/sangue , Lipocalina-2/sangue , Masculino , Osteopontina/sangue , Ratos , Ratos Sprague-Dawley , Vancomicina/efeitos adversos , Vancomicina/sangue , Vancomicina/farmacocinética
4.
Antimicrob Agents Chemother ; 60(10): 5742-51, 2016 10.
Artigo em Inglês | MEDLINE | ID: mdl-27431226

RESUMO

Vancomycin has been associated with acute kidney injury (AKI). However, the pharmacokinetic/toxicodynamic relationship for AKI is not well defined. Allometrically scaled vancomycin exposures were used to assess the relationship between vancomycin exposure and AKI. Male Sprague-Dawley rats received clinical-grade vancomycin in normal saline (NS) as intraperitoneal (i.p.) injections for 24- to 72-h durations with doses ranging 0 to 200 mg/kg of body weight divided once or twice daily. Urine was collected over the protocol's final 24 h. Renal histopathology was qualitatively scored. Urinary biomarkers (e.g., cystatin C, clusterin, kidney injury molecule 1 [KIM-1], osteopontin, lipocalin 2/neutrophil gelatinase-associated lipocalin 2) were assayed using a Luminex xMAP system. Plasma vancomycin concentrations were assayed by high-performance liquid chromatography with UV detection. A three-compartment vancomycin pharmacokinetic model was fit to the data with the Pmetrics package for R. The exposure-response in the first 24 h was evaluated using Spearman's nonparametric correlation coefficient (rs) values for the area under the concentration-time curve during the first 24 h (AUC0-24), the maximum concentration in plasma during the first 24 h (Cmax0-24 ), and the lowest (minimum) concentration in plasma after the dose closest to 24 h (Cmin0-24 ). A total of 52 rats received vancomycin (n = 42) or NS (n = 10). The strongest exposure-response correlations were observed between AUC0-24 and Cmax0-24 and urinary AKI biomarkers. Exposure-response correlations (rs values) for AUC0-24, Cmax0-24 , and Cmin0-24 were 0.37, 0.39, and 0.22, respectively, for clusterin; 0.42, 0.45, and 0.26, respectively, for KIM-1; and 0.52, 0.55, and 0.42, respectively, for osteopontin. However, no differences in histopathological scores were observed. Optimal sampling times after administration of the i.p. dose were 0.25, 0.75, 2.75, and 8 h for the once-daily dosing schemes and 0.25, 1.25, 14.5, and 17.25 h for the twice-daily dosing schemes. Our observations suggest that AUC0-24 or Cmax0-24 correlates with increases in urinary AKI biomarkers.


Assuntos
Injúria Renal Aguda/induzido quimicamente , Injúria Renal Aguda/urina , Biomarcadores/urina , Vancomicina/efeitos adversos , Vancomicina/farmacocinética , Animais , Rim/efeitos dos fármacos , Rim/patologia , Masculino , Ratos Sprague-Dawley
5.
J Biol Chem ; 289(50): 34921-37, 2014 Dec 12.
Artigo em Inglês | MEDLINE | ID: mdl-25331947

RESUMO

The molecular mechanism of p16-mediated senescence in cisplatin-treated cancer cells is not fully understood. Here we show that cisplatin treatment of head and neck cancer cells results in nuclear transport of p16 leading to a molecular modification of NFκB. Chromatin immunoprecipitation assays show that this modification is associated with the inhibition of NFκB interacting with its DNA binding sequences, leading to decreased expression of NFκB-transcribed proteins. LCMS proteomic analysis of LAP-TAP-purified proteins from HeLa cells containing a tetracycline-inducible GFP-S peptide-NFκB expression system identified gigaxonin, an ubiquitin E3 ligase adaptor, as an NFκB-interacting protein. Immunoblotting and siRNA studies confirmed the NFκB-gigaxonin interaction and the dependence of this binding on p16-NFκB binding. Using gel shift assays, we have confirmed p16-NFκB and gigaxonin-NFκB interactions. Furthermore, we have observed increased NFκB ubiquitination with cisplatin treatment that is abolished in the absence of p16 and gigaxonin expression. Analysis of 103 primary tumors has shown that increased nuclear p16 expression correlates with enhanced survival of head and neck cancer patients (p < 0.0000542), indicating the importance of nuclear p16 expression in prognosis. Finally, p16 expression is associated with reduced cytokine expression and the presence of human papilloma virus in chemoradiation-sensitive basaloid tumors. However, the absence of p16 expression is associated with enhanced cytokine expression and the absence of human papilloma virus in aggressive tumors. These results clearly demonstrate that nuclear p16 and gigaxonin play an important role in chemosensitivity of head and neck cancers through ubiquitination of NFκB.


Assuntos
Antineoplásicos/farmacologia , Senescência Celular/efeitos dos fármacos , Cisplatino/farmacologia , Inibidor p16 de Quinase Dependente de Ciclina/metabolismo , Proteínas do Citoesqueleto/metabolismo , NF-kappa B/metabolismo , Ubiquitinação/efeitos dos fármacos , Transporte Ativo do Núcleo Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Núcleo Celular/efeitos dos fármacos , Núcleo Celular/metabolismo , Ciclina D1/metabolismo , Inibidor p16 de Quinase Dependente de Ciclina/genética , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Neoplasias de Cabeça e Pescoço/diagnóstico , Neoplasias de Cabeça e Pescoço/metabolismo , Neoplasias de Cabeça e Pescoço/patologia , Neoplasias de Cabeça e Pescoço/virologia , Papillomavirus Humano 16/fisiologia , Humanos , Prognóstico
6.
Drug Dev Ind Pharm ; 41(12): 2077-88, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26289004

RESUMO

The objective of our investigational work was to develop a proliposomal formulation to improve the oral bioavailability of valsartan. Proliposomes were formulated by thin film hydration technique using different ratios of phospholipids:drug:cholesterol. The prepared proliposomes were evaluated for vesicle size, encapsulation efficiency, morphological properties, in vitro drug release, in vitro permeability and in vivo pharmacokinetics. In vitro drug-release studies were performed in simulated gastric fluid (pH 1.2) and purified water using dialysis bag method. In vitro drug permeation was studied using parallel artificial membrane permeation assay (PAMPA), Caco-2 monolayer and everted rat intestinal perfusion techniques. In vivo pharmacokinetic studies were conducted in male Sprague Dawley (SD) rats. Among the proliposomal formulations, F-V was found to have the highest encapsulation efficiency of 95.6 ± 2.9% with a vesicle size of 364.1 ± 14.9 nm. The in vitro dissolution studies indicated an improved drug release from proliposomal formulation, F-V in comparison to pure drug suspension in both, purified water and pH 1.2 dissolution media after 12 h. Permeability across PAMPA, Caco-2 cell and everted rat intestinal perfusion studies were higher with F-V formulation as compared to pure drug. Following single oral administration of F-V formulation, a relative bioavailability of 202.36% was achieved as compared to pure valsartan.


Assuntos
Desenho de Fármacos , Pró-Fármacos/administração & dosagem , Pró-Fármacos/farmacocinética , Valsartana/administração & dosagem , Valsartana/farmacocinética , Administração Oral , Animais , Disponibilidade Biológica , Células CACO-2 , Humanos , Lipossomos , Masculino , Ratos , Ratos Sprague-Dawley
7.
Cancer Res Commun ; 4(3): 706-722, 2024 03 08.
Artigo em Inglês | MEDLINE | ID: mdl-38421310

RESUMO

Gigaxonin is an E3 ubiquitin ligase that plays a role in cytoskeletal stability. Its role in cancer is not yet clearly understood. Our previous studies of head and neck cancer had identified gigaxonin interacting with p16 for NFκB ubiquitination. To explore its role in cancer cell growth suppression, we analyzed normal and tumor DNA from cervical and head and neck cancers. There was a higher frequency of exon 8 SNP (c.1293 C>T, rs2608555) in the tumor (46% vs. 25% normal, P = 0.011) pointing to a relationship to cancer. Comparison of primary tumor with recurrence and metastasis did not reveal a statistical significance. Two cervical cancer cell lines, ME180 and HT3 harboring exon 8 SNP and showing T allele expression correlated with higher gigaxonin expression, reduced in vitro cell growth and enhanced cisplatin sensitivity in comparison with C allele expressing cancer cell lines. Loss of gigaxonin expression in ME180 cells through CRISPR-Cas9 or siRNA led to aggressive cancer cell growth including increased migration and Matrigel invasion. The in vitro cell growth phenotypes were reversed with re-expression of gigaxonin. Suppression of cell growth correlated with reduced Snail and increased e-cadherin expression. Mouse tail vein injection studies showed increased lung metastasis of cells with low gigaxonin expression and reduced metastasis with reexpression of gigaxonin. We have found an association between C allele expression and RNA instability and absence of multimeric protein formation. From our results, we conclude that gigaxonin expression is associated with suppression of epithelial-mesenchymal transition through inhibition of Snail. SIGNIFICANCE: Our results suggest that GAN gene exon 8 SNP T allele expression correlates with higher gigaxonin expression and suppression of aggressive cancer cell growth. There is downregulation of Snail and upregulation of e-cadherin through NFκB ubiquitination. We hypothesize that exon 8 T allele and gigaxonin expression could serve as diagnostic markers of suppression of aggressive growth of head and neck cancer.


Assuntos
Neoplasias de Cabeça e Pescoço , Humanos , Animais , Camundongos , Regulação para Baixo/genética , Linhagem Celular Tumoral , Neoplasias de Cabeça e Pescoço/tratamento farmacológico , Transição Epitelial-Mesenquimal/genética , Caderinas/genética
9.
Genes Chromosomes Cancer ; 50(6): 434-41, 2011 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-21432940

RESUMO

Approximately 25,000 ovarian cancers are diagnosed in the United States annually, and 75% of cases are in the advanced stage when they are largely incurable. There is a critical need for improved early detection tools and development of novel treatments. Recently, we showed that among 20q13-amplified genes in ovarian cancer, ADRM1 overexpression was the most highly correlated with amplification and was significantly upregulated with respect to stage, recurrence, and metastasis. In addition, overexpression of ADRM1 correlated significantly with shorter time to recurrence and overall survival. Herein, array-CGH and microarray expression of ovarian cancer cell lines provides evidence consistent with the primary tumor data that ADRM1 is a 20q13 amplification target. Knockdown of ADRM1 in amplified ovarian cell-line OAW42 results in downregulation of growth factor GIPC1 and upregulation of tumor-suppressor RECK RNA and protein. In our dataset of 141 ovarian primary tumors, ADRM1 overexpression significantly correlates with GIPC1 overexpression. In addition, there is a significant anticorrelation between ADRM1 overexpression and RECK expression. Further research is necessary to determine whether targeting knockdown of ADRM1 in 20q13-amplified ovarian cancers results in growth inhibition and tumor suppression via downstream targets GIPC1 and RECK.


Assuntos
Proteínas Adaptadoras de Transdução de Sinal/genética , Proteínas Ligadas por GPI/genética , Glicoproteínas de Membrana/genética , Neoplasias Ovarianas/genética , Neoplasias Ovarianas/patologia , Linhagem Celular Tumoral , Feminino , Amplificação de Genes , Regulação Neoplásica da Expressão Gênica , Técnicas de Silenciamento de Genes , Humanos , Peptídeos e Proteínas de Sinalização Intracelular , Estadiamento de Neoplasias
10.
Curr Drug Deliv ; 5(4): 306-11, 2008 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-18855601

RESUMO

Elementary osmotic pumps (EOP) are well known for delivering moderately soluble drugs at a zero order rate. A push-pull osmotic system was developed and commercialized for poorly water-soluble drugs [Procardia XL (Nifedipine), Glucotrl XL (Glipizide)]. However, the technology is complex comprising of bilayer compression and the suspension of drug formed in the core has more viscosity and has to withstand the osmotic pressure within the tablet, for which the membrane must be thicker than that of EOP. The aim of the present study was to develop a solid dispersion based EOP system for a poorly water-soluble drug, nifedipine and deliver it in a zero order fashion over an extended period of time. Solid dispersions were prepared by hot melt technique using Poloxamer-188 at various ratios of drug and polymer (1:1, 1:5 and 1:10, on weight basis) and investigated for solubility study. Formation of complex and decrease in crystallinity was confirmed from differential scanning calorimetry (DSC) and X-ray crystallography (XRD) study. Core tablets using solid dispersions were prepared and coated with cellulose acetate and PEG-400. An orifice was drilled manually to create passage for drug release. The system was optimized for amount of osmogent, membrane weight gain, amount of plasticiser and diameter of the orifice, to achieve desired release profile. The osmotic system was found to deliver nifedipine at a zero order rate for 20 h. The drug release from the developed formulation was independent of pH and agitational intensity.


Assuntos
Sistemas de Liberação de Medicamentos , Nifedipino/administração & dosagem , Química Farmacêutica , Concentração de Íons de Hidrogênio , Nifedipino/química , Osmose , Plastificantes/química , Poloxâmero/administração & dosagem , Poloxâmero/química , Solubilidade , Difração de Raios X
11.
Cancer Res ; 66(3): 1630-9, 2006 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-16452222

RESUMO

Lapatinib (GW572016) is a selective inhibitor of both epidermal growth factor receptor (EGFR) and HER-2 tyrosine kinases. Here, we explore the therapeutic potential of lapatinib by testing its effect on tumor cell growth in a panel of 31 characterized human breast cancer cell lines, including trastuzumab-conditioned HER-2-positive cell lines. We further characterize its activity in combination with trastuzumab and analyze whether EGFR and HER-2 expression or changes induced in the activation of EGFR, HER-2, Raf, AKT, or extracellular signal-regulated kinase (ERK) are markers of drug activity. We report that concentration-dependent antiproliferative effects of lapatinib were seen in all breast cancer cell lines tested but varied significantly between individual cell lines with up to 1,000-fold difference in the IC(50)s (range, 0.010-18.6 micromol/L). Response to lapatinib was significantly correlated with HER-2 expression and its ability to inhibit HER-2, Raf, AKT, and ERK phosphorylation. Long-term in vivo lapatinib studies were conducted with human breast cancer xenografts in athymic mice. Treatment over 77 days resulted in a sustained and significant reduction in xenograft volume compared with untreated controls. For the combination of lapatinib plus trastuzumab, synergistic drug interactions were observed in four different HER-2-overexpressing cell lines. Moreover, lapatinib retained significant in vitro activity against cell lines selected for long-term outgrowth (>9 months) in trastuzumab-containing (100 microg/mL) culture medium. These observations provide a clear biological rationale to test lapatinib as a single agent or in combination with trastuzumab in HER-2-overexpressing breast cancer and in patients with clinical resistance to trastuzumab.


Assuntos
Anticorpos Monoclonais/farmacologia , Protocolos de Quimioterapia Combinada Antineoplásica/farmacologia , Neoplasias da Mama/tratamento farmacológico , Quinazolinas/farmacologia , Receptor ErbB-2/antagonistas & inibidores , Animais , Anticorpos Monoclonais/administração & dosagem , Anticorpos Monoclonais Humanizados , Neoplasias da Mama/enzimologia , Neoplasias da Mama/patologia , Ciclo Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Interações Medicamentosas , Ensaios de Seleção de Medicamentos Antitumorais , Receptores ErbB/biossíntese , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Feminino , Humanos , Lapatinib , Camundongos , Camundongos SCID , Proteína Oncogênica v-akt/metabolismo , Fosforilação , Quinazolinas/administração & dosagem , Receptor ErbB-2/biossíntese , Trastuzumab , Ensaios Antitumorais Modelo de Xenoenxerto , Quinases raf/metabolismo
12.
Int J Pharm ; 337(1-2): 357-60, 2007 Jun 07.
Artigo em Inglês | MEDLINE | ID: mdl-17270373

RESUMO

Erythropoietin (EPO) loaded carbon nanotubes (CNTs) with surfactant as an absorption enhancer were prepared for the oral delivery of EPO using two types of CNTs, long and short fiber length CNTs, and the effect of CNT fiber length on the absorption efficiency of EPO was studied. After Labrasol, PEG-8 caprylic/capric glycerides, as absorption enhancer was adsorbed into long fiber CNTs of which mean fiber length was 20-80 microm, as a carrier, EPO and casein as protease inhibitor and Explotab (sodium starch glycolate) as a disintegrating agent, were mixed. The resulting solid preparation was administered into the rat jejunum and serum EPO levels were measured by ELISA. The dose of EPO, CNTs, casein and Explotab were 100 IU/kg, 5mg/kg, 25mg/kg and 2.5mg/kg, respectively. Serum EPO level reached to C(max), 69.0+/-3.9 mIU/ml, at 3.5+/-0.1h and AUC was 175.7+/-13.8 mIU h/ml. These values were approximately half of that obtained with short fiber length CNTs of which C(max) was 143.1+/-15.2 mIU/ml and AUC was 256.3+/-9.7 mIU h/ml. When amphoteric surfactant, Lipomin LA, sodium beta-alkylaminopropionic acid, was used to accelerate the disaggregation of long fiber length CNTs, C(max) was 36.0+/-4.9 and AUC was 96.9+/-11.9, which showed less bioavailability (BA) of EPO. These results suggest that the short fiber length CNTs deliver more both EPO and absorption enhancer to the absorptive cells of the rat small intestine and the aggregation of CNTs is not the critical factor for the oral delivery of EPO.


Assuntos
Portadores de Fármacos , Eritropoetina/farmacocinética , Hematínicos/farmacocinética , Absorção Intestinal , Intestino Delgado/metabolismo , Nanotubos de Carbono/química , Administração Oral , Animais , Área Sob a Curva , Disponibilidade Biológica , Química Farmacêutica , Composição de Medicamentos , Ensaio de Imunoadsorção Enzimática , Eritropoetina/administração & dosagem , Eritropoetina/sangue , Eritropoetina/química , Excipientes/química , Hematínicos/administração & dosagem , Hematínicos/sangue , Hematínicos/química , Masculino , Taxa de Depuração Metabólica , Tamanho da Partícula , Ratos , Ratos Wistar , Tensoativos/química
13.
Am J Health Syst Pharm ; 73(4): 241-6, 2016 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-26843502

RESUMO

PURPOSE: The compatibility of vancomycin and piperacillin-tazobactam in concentrations typically used in extended-infusion dosing schemes was evaluated. METHODS: Piperacillin-tazobactam was reconstituted and diluted to concentrations of 33.75, 45, 50, 60, 67.5, 80, and 90 mg/mL. Vancomycin was diluted to concentrations of 4-8, 10, and 12 mg/mL. The resultant admixtures were visually observed after preparation against black and white backgrounds each hour between hours 1 through 4 and after 24 hours. Frozen products of each medication and brand-name Zosyn powder for reconstitution also were studied. Each combination of products and concentrations was tested for precipitation using simulated Y-site administration. Absorbance and microscopic analyses were performed to discern less perceptible incompatibilities in combinations that did not result in visual precipitation. Changes in absorbance were evaluated using two-way repeated-measures analysis of variance with post hoc Bonferroni corrections. RESULTS: No tested concentrations of piperacillin-tazobactam showed precipitations with vancomycin up to concentrations of 7 mg/mL. Piperacillin-tazobactam 80-90 mg/mL formed reversible precipitation with vancomycin 8 mg/mL. All tested concentrations of piperacillin-tazobactam formed a reversible precipitate with vancomycin 10 mg/mL. Irreversible precipitation was noted with all combinations of piperacillin-tazobactam and vancomycin 12 mg/mL. No significant changes in absorbance analyses were identified for all tested piperacillin-tazobactam concentrations and vancomycin 4-10 mg/mL compared with 0.9% sodium chloride injection (p > 0.05). Similar results were observed using frozen preparations and brand-name Zosyn. CONCLUSION: Visual, microscopic, and absorbance analyses showed no evidence of incompatibility when piperacillin-tazobactam 33.75-90 mg/mL was combined with vancomycin ≤7 mg/mL. Reversible and irreversible precipitates formed when piperacillin-tazobactam was combined with vancomycin ≥8 mg/mL.


Assuntos
Antibacterianos/química , Ácido Penicilânico/análogos & derivados , Vancomicina/química , Antibacterianos/administração & dosagem , Precipitação Química , Composição de Medicamentos/métodos , Incompatibilidade de Medicamentos , Humanos , Ácido Penicilânico/administração & dosagem , Ácido Penicilânico/química , Piperacilina/administração & dosagem , Piperacilina/química , Combinação Piperacilina e Tazobactam , Cloreto de Sódio/química , Fatores de Tempo , Vancomicina/administração & dosagem
14.
Mol Cell Biol ; 36(12): 1776-92, 2016 06 15.
Artigo em Inglês | MEDLINE | ID: mdl-27090639

RESUMO

We and others have shown that the cystatin E/M gene is inactivated in primary human tumors, pointing to its role as a tumor suppressor gene. However, the molecular mechanism of tumor suppression is not yet understood. Using plasmid-directed cystatin E/M gene overexpression, a lentivirus-mediated tetracycline-inducible vector system, and human papillomavirus 16 (HPV 16) E6 and E7 gene-immortalized normal human epidermal keratinocytes, we demonstrated intracellular and non-cell-autonomous apoptotic growth inhibition of tumor cell lines and that growth inhibition is associated with cytoplasmic retention of NF-κB. We further demonstrated decreased phosphorylation of IκB kinase (IKKß) and IκBα in the presence of tumor necrosis factor alpha (TNF-α), confirming the role of cystatin E/M in the regulation of the NF-κB signaling pathway. Growth suppression of nude mouse xenograft tumors carrying a tetracycline-inducible vector system was observed with the addition of doxycycline in drinking water, confirming that the cystatin E/M gene is a tumor suppressor gene. Finally, immunohistochemical analyses of cervical carcinoma in situ and primary tumors have shown a statistically significant inverse relationship between the expression of cystatin E/M and cathepsin L and a direct relationship between the loss of cystatin E/M expression and nuclear expression of NF-κB. We therefore propose that the cystatin E/M suppressor gene plays an important role in the regulation of NF-κB.


Assuntos
Cistatina M/metabolismo , Citoplasma/metabolismo , Proteínas I-kappa B/metabolismo , NF-kappa B/metabolismo , Fator de Necrose Tumoral alfa/metabolismo , Neoplasias do Colo do Útero/patologia , Animais , Catepsina L/metabolismo , Linhagem Celular Tumoral , Proliferação de Células , Cistatina M/genética , Doxiciclina/administração & dosagem , Feminino , Regulação Neoplásica da Expressão Gênica , Vetores Genéticos/farmacologia , Células HeLa , Humanos , Lentivirus/genética , Camundongos , Camundongos Nus , Transplante de Neoplasias , Fosforilação , Transdução de Sinais , Neoplasias do Colo do Útero/genética , Neoplasias do Colo do Útero/metabolismo
15.
Oncogene ; 23(7): 1354-63, 2004 Feb 19.
Artigo em Inglês | MEDLINE | ID: mdl-14691457

RESUMO

Gene amplification is common in solid tumors and is associated with adverse prognosis, disease progression, and development of drug resistance. A small segment from chromosome 17q11-12 containing the HER-2/Neu gene is amplified in about 25% of breast cancer. HER-2/Neu amplification is associated with adverse prognosis and may predict response to chemotherapy and hormonal manipulation. Moreover, HER-2/Neu amplification may select patients for anti-HER-2/Neu-based therapy with Herceptin. We and others recently described a common sequence element from the HER-2/Neu region that was amplified in breast cancer cells. In addition, most, if not all, of the amplified genes from this region display overexpression. This raises the intriguing possibility that genes immediately adjacent to HER-2/Neu may influence the biological behavior of breast cancer carrying HER-2/Neu amplification and serve as rational targets for therapy. By extracting sequence information from public databases, we have constructed a contig in bacterial artificial chromosomes (BACs) that extends from HER-2/Neu to a phosphotidylinositol phosphate kinase (PIPK), Pip4k2beta from 17q11-12. Although a role of PI-3-kinase and AKT in cancer biology has been previously described, PIPK has not been previously implicated. We show that Pip4k2beta, initially known as Pip5k2beta, is amplified in a subset of breast cancer cell lines and primary breast cancer samples that carry HER-2/Neu amplification. Out of eight breast cancer cell lines with HER-2/Neu amplification, three have concomitant amplification of the Pip4k2beta gene--UACC-812, BT-474 and ZR-75-30. Similarly, two out of four primary breast tumors with HER-2/Neu amplification carry Pip4k2beta gene amplification. Intriguingly, one tumor displays an increase in the gene copy number of Pip4k2beta that is significantly more than that of HER-2/Neu. Moreover, dual color FISH reveals that amplified Pip4k2beta gene may exist in a distinct structure from that of HER-2/Neu in ZR-75-30 cell line. These studies suggest that Pip4k2beta may reside on an amplification maximum distinct from that of HER-2/Neu and serve as an independent target for amplification and selective retention. Pip4k2beta amplification is associated with overexpression at the RNA and protein level in breast cancer cell lines. Stable expression of Pip4k2beta in breast cancer cell lines with and without HER-2/Neu amplification increases cell proliferation and anchorage-independent growth. The above observations implicate Pip4k2beta in the development and/or progression of breast cancer. Our study suggests that Pip4k2beta may be a distinct target for gene amplification and selective retention from 17q11-12.


Assuntos
Neoplasias da Mama/metabolismo , Divisão Celular/fisiologia , Fosfotransferases (Aceptor do Grupo Álcool)/genética , Neoplasias da Mama/genética , Cromossomos Humanos Par 17 , Feminino , Genes erbB-2 , Humanos , Hibridização in Situ Fluorescente , Antígenos de Histocompatibilidade Menor , Fosfotransferases (Aceptor do Grupo Álcool)/biossíntese , Mapeamento Físico do Cromossomo
16.
Biomaterials ; 26(34): 7154-63, 2005 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-15967493

RESUMO

In the present study, an attempt was made to study the feasibility of nanoparticulate adsorbents in the presence of an absorption enhancer, as a drug delivery tool for the administration of erythropoietin (EPO) to the small intestine. Liquid filled nano- and micro-particles (LFNPS/LFMPS) were prepared using solid adsorbents such as porous silicon dioxide (Sylysia 550), carbon nanotubes (CNTs), carbon nanohorns, fullerene, charcoal and bamboo charcoal. Surfactants such as a saturated polyglycolysed C8-C18 glyceride (Gelucire 44/14), PEG-8 capryl/caprylic acid glycerides (Labrasol) and polyoxyethylene hydrogenated castor oil derivative (HCO-60) were used as an absorption enhancer at 50mg/kg along with casein/lactoferrin as enzyme inhibitors. The absorption of EPO was studied by measuring serum EPO levels by an ELISA method after small intestinal administration of EPO-LFNPS preparation to rats at the EPO dose level of 100 IU/kg. Among the adsorbents studied, CNTs showed the highest serum EPO level of 62.7 +/- 11.6 mIU/ml. In addition, with the use of casein, EPO absorption was improved, C(max) 143.1 +/- 15.2 mIU/ml. Labrasol showed the highest absorption enhancing effect after intra-jejunum administration than Gelucire 44/14 and HCO-60, 25.6 +/- 3.2 and 22.2 +/- 3.6 mIU/ml, respectively. Jejunum was found to be the best absorption site for the absorption of EPO from LFNPS. The use of CNTs as LFNPS, improved the bioavailability of EPO to 11.5% following intra-small intestinal administration.


Assuntos
Materiais Revestidos Biocompatíveis/química , Portadores de Fármacos/química , Eritropoetina/administração & dosagem , Eritropoetina/farmacocinética , Intestino Delgado/efeitos dos fármacos , Intestino Delgado/metabolismo , Nanotubos/química , Absorção , Animais , Eritropoetina/sangue , Eritropoetina/química , Estudos de Viabilidade , Masculino , Teste de Materiais , Nanotubos/ultraestrutura , Ratos , Ratos Wistar , Soluções
17.
Curr Pharm Biotechnol ; 16(4): 303-12, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-25601600

RESUMO

Proliposomes are phospholipid based drug delivery systems that are finding important applications in the field of pharmaceutics. Proliposomes have been extensively studied as a potential carrier for oral delivery of drugs with poor bioavailability, but the mechanism of absorption and cellular uptake pathways has not yet been clearly understood. An in-depth insight into the physical and biological behavior of proliposomes is necessary for designing an effective delivery system for enhancing the availability of drug at the intended site. Reformulation of sub optimal drugs using proliposomes has given an opportunity to improve the therapeutic indices of various drugs predominantly by altering their uptake mechanism. This work reviews the proliposomal drug delivery field, summarizes the success of proliposomes for the oral delivery of drugs with poor bioavailability; indicating the key issues to be addressed to affirm that proliposomes can effectively work as a drug carrier in clinical settings with a clear understanding of its behavior in biological environment, as they are now an established platform technology with considerable clinical acceptance.


Assuntos
Lipossomos , Administração Oral , Animais , Humanos , Lipídeos/química , Preparações Farmacêuticas/administração & dosagem , Preparações Farmacêuticas/química
18.
Oncotarget ; 6(21): 18504-17, 2015 Jul 30.
Artigo em Inglês | MEDLINE | ID: mdl-26098778

RESUMO

Head and neck squamous cell carcinoma (HNSCC) is the sixth most common cancer, with 600,000 new cases every year worldwide. Although chemotherapeutics exist, five-year survival is only 50%. New strategies to overcome drug resistance are required to improve HNSCC treatment. Curcumin-difluorinated (CDF), a synthetic analog of curcumin, was packaged in liposomes and used to evaluate growth inhibition of cisplatin resistant HNSCC cell lines CCL-23R and UM-SCC-1R generated from the parental cell lines CCL-23 and UM-SCC-1 respectively. Growth inhibition in vitro and expression levels of the CD44 (cancer stem cell marker), cytokines, and growth factors were investigated after liposomal CDF treatment. The in vivo growth inhibitory effect of liposomal CDF was evaluated in the nude mice xenograft tumor model of UM-SCC-1R and the inhibition of CD44 was measured. Treatment of the resistant cell lines in vitro with liposomal CDF resulted in a statistically significant growth inhibition (p < 0.05). The nude mice xenograft study showed a statistically significant tumor growth inhibition of UM-SCC-1R cells and a reduction in the expression of CD44 (p < 0.05), indicating an inhibitory effect of liposomal CDF on CSCs. Our results demonstrate that delivery of CDF through liposomes may be an effective method for the treatment of cisplatin resistant HNSCC.


Assuntos
Carcinoma de Células Escamosas/tratamento farmacológico , Proliferação de Células/efeitos dos fármacos , Curcumina/farmacologia , Neoplasias de Cabeça e Pescoço/tratamento farmacológico , Hidrocarbonetos Fluorados/farmacologia , Células-Tronco Neoplásicas/efeitos dos fármacos , Animais , Antineoplásicos/farmacologia , Carcinoma de Células Escamosas/genética , Carcinoma de Células Escamosas/metabolismo , Linhagem Celular Tumoral , Cisplatino/farmacologia , Curcumina/análogos & derivados , Citocinas/metabolismo , Resistencia a Medicamentos Antineoplásicos/efeitos dos fármacos , Feminino , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Neoplasias de Cabeça e Pescoço/genética , Neoplasias de Cabeça e Pescoço/metabolismo , Humanos , Receptores de Hialuronatos/genética , Lipossomos , Camundongos Nus , Células-Tronco Neoplásicas/metabolismo , Células-Tronco Neoplásicas/patologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Carga Tumoral/efeitos dos fármacos , Carga Tumoral/genética , Ensaios Antitumorais Modelo de Xenoenxerto
19.
Curr Med Chem ; 9(24): 2243-70, 2002 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-12470245

RESUMO

Design and synthesis of metabolically stable peptide analogs that can either mimic or block the bioactivity of natural peptides or enzymes is an important constituent of bioorganic and medicinal chemistry research. Isosteric replacement of a scissile peptide bond represents a viable and popular approach in the rational design of peptidomimetics. Peptidomimetics find applications as drugs, in protein engineering and so on. This is evident from the wealth of therapeutically useful peptidomimetic leads incorporating any of the peptide isosteres that are currently available. In this review, we have given a brief account of the types of peptide isosteres widely known till date. With this background, we have described some of the recent developments in synthetic approaches. This includes methods involving a common intermediate to synthesize different possible isosteres and their peptide analogs, solid phase synthesis and combinatorial approach. One such method involving stereoselective nitrile oxide cycloaddition as the key step has been studied extensively in our research laboratory. Finally, we have also discussed about some of the recent reports on the design and inhibitory activities of peptidic or non-peptidic analogs against aspartic proteases (HIV-1, renin, ACE and pepsin) and peptide analogs of an immunomodulating hexapeptide.


Assuntos
Inibidores Enzimáticos/síntese química , Peptídeos/síntese química , Inibidores da Enzima Conversora de Angiotensina/síntese química , Inibidores da Enzima Conversora de Angiotensina/química , Inibidores da Enzima Conversora de Angiotensina/farmacologia , Ácido Aspártico Endopeptidases/antagonistas & inibidores , Técnicas de Química Combinatória , Inibidores Enzimáticos/química , Inibidores Enzimáticos/farmacologia , Protease de HIV/metabolismo , Inibidores da Protease de HIV/síntese química , Inibidores da Protease de HIV/química , Inibidores da Protease de HIV/farmacologia , Pepsina A/antagonistas & inibidores , Peptídeos/química , Peptídeos/farmacologia , Relação Estrutura-Atividade , Especificidade por Substrato
20.
Curr Med Chem ; 10(19): 1973-91, 2003 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-12871099

RESUMO

In recent years modified oligonucleotides (oligos) have become an area of increased research activity. These synthetic biopolymers have long been used as molecular probes to understand better the interaction between proteins and nucleic acids at molecular level, which in fact is responsible for all aspects of cellular or viral gene expression. Modified oligos have also been used as therapeutic and diagnostic agents. For example, the antisense oligos are used to block gene expression by binding especially with complimentary sequences of target mRNA. Also, over the past decade efforts are made to modify the native phosphodiester linkages to improve nuclease resistance and membrane permeation of naturally occurring oligonucleotide for therapeutic applications. Therefore, design and synthesis of modified oligonucleotides for a wide range of applications are of interest for bioorganic chemists and biologists. The advent of the phosphoramidite chemistry has accelerated the development of oligos with modified nucleobases, phosphate-protecting groups and modified sugars. In this review, we have presented the recently reported nucleoside based phosphoramidites with modified base, sugar or backbone units and their subsequent conversion into modified DNA and RNA analogs. The effect of such modification on structural, thermodynamic, and hybridization properties of the modified oligos and their duplex with natural complementary oligos have also been discussed for some cases. Similarly, we have highlighted the role of modified nucleoside phosphoramidite building blocks in synthesis of oligos modified with optically or electrochemically active molecules. Nucleoside phosphoramidites or small oligos immobilized over solid supports through different linkages or groups and their biological applications has also been mentioned.


Assuntos
Amidas/química , Oligonucleotídeos/síntese química , Ácidos Fosfóricos/química , Estrutura Molecular , Conformação de Ácido Nucleico , Ácidos Nucleicos/química , Oligonucleotídeos/química , Oligonucleotídeos/metabolismo
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