Biological and metabolic study of naproxen-propyphenazone mutual prodrug.

Naproxen-propyphenazone (NAP-PP) esters were synthesized as prodrugs with the aim of improving the therapeutic index through prevention of gastrointestinal irritation and bleeding. The structures of the synthesized NAP-PP hybrid esters were confirmed by IR and 1H NMR spectroscopy and their purity was established by elemental analysis, HPLC and TLC. The release of NAP as well as PP derivatives, from the ester prodrugs was studied. A validated analytical HPLC method for the estimation of the NAP, and the prodrugs was developed. Also the enzymatic hydrolysis products of the ester were identified by GC-MS and in conjugation with HPLC. The kinetics of ester hydrolysis was studied in two different non-enzymatic buffer solutions, at pH 1.2, and 7.4 as well as in liver homogenates. Study of analgesic and anti-inflammatory properties in comparison with the reference compounds has shown that both analgesic and anti-inflammatory activities were present at the same doses of the investigated compounds. The ester III was found to be less irritating to gastric mucosal membrane than the parent drugs. These results suggest that the synthesized prodrugs are characterized by better therapeutic index than the parent drugs.     

3-Amino-4-arylazopyrazoles: CT Complexation and a Novel Synthesis of Pyrazolo[2,3-a]quinazolines

3,5-Diamino-4-arylazopyrazoles 1a-d form charge-transfer (CT) complexes with chloranil (CHL), fluoranil (TFQ) and react with 2,3-dicyano-5,6-dichlorobenzoquinone (DDQ) to yield pyrazolo[2,3-a]quinazoline derivatives 4 . 5-Amino-3-phenyl-4-arylazopyrazole 2a-d form CT-complexes only with DDQ.     

The human sperm head: a key for successful fertilization

In order to examine the predictive value of determining the sperm head shape, the acrosomal size, the presence of acrosomal vacuoles, and the challenged acrosome reaction (AR) on the outcome of a standard in vitro fertilization (IVF) program, a prospective study was conducted that included 75 couples undergoing IVF treatment. An assessment of sperm morphology was performed using the Hobson Sperm Tracker (Hobson Tracker Limited, Sheffield, United Kingdom). The assessment of the AR was performed before and after adding pooled undiluted human follicular fluid (FF). The outcome measure was an IVF rate of inseminated oocytes. A positive correlation was found between the fertilization rate (FR%) and the proportion of the sperm with a normal (oval) head shape (P <.001), the sperm exhibiting acrosomal vacuoles (P <.003), the sperm with a normal acrosomal size (40%-70% of total head area, P <.025), and the sperm undergoing AR after adding FF (P <.001). Multiple logistic regression analysis revealed that by incorporating the above 4 parameters, the sensitivity of prediction of IVF FR% values was 79%, and the specificity was 93%, with a positive predictive value of 96%. This study shows that the multiparametric assessment of the sperm head is useful in predicting the FR% values of a standard IVF treatment. The automated analysis used in this study is shown to maintain a level of precision and accuracy acceptable for application in a routine semen analysis situation.     

Determination of two ternary mixtures containing phenobarbitone by second derivative of the ratio spectrum-zero-crossing and HPLC methods

A spectrophotometric method is developed for the determination of ternary mixtures with overlapping spectra. The method is based on the use of the second derivative of the ratio spectrum with a zero-crossing technique. The ratio spectrum was obtained by dividing the absorption spectrum of the mixture by that of one of the components. The concentration of the other components are then determined from their respective calibration graphs treated similarly. The method is accurate, non-destructive and do not require resolutions of equations. The method has been applied for the resolution of two ternary mixtures, namely, phenobarbitone, methylphenobarbitone and phenytoin (1), and phenobarbitone, papaverine HCl and piperazine acefyllinate (2). Also, a HPLC method was developed for determination of phenobarbitone, papaverine and HCL and piperazine acefyllinate. The HPLC method depends upon using ODS column with a mobile phase consisting of acetonitrile-5 mM aqueous heptane sulfonic acid sodium salt (50:50, v/v) and adjusted to apparent pH 4 using acetic acid. Quantitation was achieved with UV detection at 220 nm based on peak area. The proposed methods were applied for the determination of the two ternary combinations in synthetic mixtures and in commercial pharmaceutical products. The results obtained were precise and accurate.     

Stargazin differentially controls the trafficking of alpha-amino-3-hydroxyl-5-methyl-4-isoxazolepropionate and kainate receptors

Synaptic plasticity at excitatory synapses in the brain is largely achieved by rapid changes in the number of synaptic alpha-amino-3-hydroxyl-5-methyl-4-isoxazolepropionate (AMPA) receptors. Stargazin, a membrane protein that interacts with AMPA receptors, is believed to play a pivotal role in trafficking AMPA receptors to the plasma membrane and targeting them to the synapse. However, it is unclear whether the trafficking of kainate receptors, which are structurally very similar to AMPA receptors, is also dependent on stargazin. Here we show that in both cerebellar granule cells and in Xenopus laevis oocytes expression system, surface delivery of kainate receptor is independent of stargazin. These results suggest that stargazin action is highly selective for AMPA receptors.     

Photodynamic selectivity of 5-aminolevulinic acid to prostate cancer cells

OBJECTIVE: To determine the selectivity of 5-aminolevulinic acid (5-ALA) as a photosensitizer to malignant prostatic cells in men undergoing radical retropubic prostatectomy. PATIENTS AND METHODS: Nineteen patients with localized prostate cancer were included in the study. Eighteen patients received 5-ALA and one patient did not receive it and was used as a control. The dose was 20mg /kg body weight, 15 patients received 5-ALA 4 hours before radical prostatectomy, two patients received it 2 hours before prostatectomy through a Ryle tube, and one patient received 5-ALA 12 hours before the operation. The removed prostates were examined for protoporphyrin IX (PpIX) fluorescence macroscopically, by fluorescence microscopy and by light microscopy. RESULTS: All carcinomas showed a clear evidence of PpIX-enrichment except in the control case. The enrichments were strong (++) in 15 cases and weak (+) in 3 cases. Two of those three cases were given 5-ALA two hours through a Ryle tube before excision of the prostate as well as the patient who was given 5-ALA 12 hours preoperatively. No PpIX enrichment was observed in the stroma of the prostate gland or in the benign tissue sections in any case (0/19). CONCLUSION: Oral 5-ALA is selectively concentrated in malignant cells of the prostate. This may lead to the clinical application of photodynamic therapy for localized prostate cancer.     

The pharmacokinetics of the prostaglandin E1 analogue misoprostol in plasma and colostrum after postpartum oral administration

OBJECTIVE: To study pharmacokinetics of prostaglandin E1 analogue, misoprostol in plasma and colostrum after postpartum oral administration. STUDY DESIGN: Twenty women received 600 microg doses of misoprostol orally after delivery. Plasma levels of the principal metabolite, misoprostol acid, were measured at 2, 10, 20, 30, 40, 50, 60, 90, 120, 180, 240 and 300 min (48 samples). Colostrum was expressed from the breasts to measure misoprostol acid at 60, 120, 180, 240, and 300 min (24 samples). Assay was done using isotope dilution gas chromatography (GC)/negative ion chemical ionisation mass spectrometry (MS). RESULTS: The plasma concentration of misoprostol acid rose quickly. Two minutes after oral administration its mean level was 91.5 pg/ml, peaked at 20 min (344 pg/ml), then fell steeply by 120 min (27.8 pg/ml) and remained low for the duration of the study. Misoprostol acid in colostrum reached maximum concentration of 20.9 pg/m within 1h after oral administration. It then declined gradually to 17.8 pg/ml at 2h, 2.8 pg/ml at 4h and to <1 pg/ml at 5h. Areas under misoprostol concentration versus time curves up to 5h were 290.1 pgh/ml in the plasma and 51.4 pgh/ml in colostrum, respectively. CONCLUSION: Misoprostol acid is secreted in colostrum within 1h of oral administration of 600 microg of misoprostol; the pharmacokinetics of misoprostol after oral administration during postpartum is similar to that of other pregnancy periods.     

Prospective study of the ability of serial measurements of serum chromogranin A and gastrin to detect changes in tumor burden in patients with gastrinomas

BACKGROUND: Assessment of tumor burden changes is essential for the management of patients with neuroendocrine gastrointestinal (GI) tumors. Chromogranin A (CgA) is a tumor marker for such tumors; however, to the authors' knowledge, there is little information on whether serial assessments can assess changes in tumor burden. In this prospective study of patients with gastrinomas, serial changes in serum CgA levels were compared with changes in levels of the specific tumor marker gastrin to determine whether they reflected changes in tumor burden. METHODS: In 72 consecutive patients, the mean CgA and gastrin levels from three determinations were measured on each visit. Changes in markers were correlated with changes in tumor burden determined by imaging. By assessing daily changes, significance changes in CgA and gastrin levels were determined. RESULTS: During 103 follow-up visits (mean, 9.6 months), an increased tumor size occurred in 25% of patients, no change occurred in 62% of patients, and a decrease occurred in 13% of patients. In patients who had increasing tumor size, CgA levels increased numerically in 77% of patients, gastrin levels increased in 54% of patients, and the increases were significant in 60-80% of patients. In patients who had tumor stabilization, CgA levels in 63% of patients and gastrin levels in 73% of patients did not show a significant change. Decreased tumor size postresection showed a significant decrease in CgA and gastrin levels in all patients. The sensitivity of CgA and gastrin was as follows: sensitivity for detecting an increase, 62% for CgA and 31% for gastrin; sensitivity for detecting no change, 42% for CgA and 75% for gastrin; and sensitivity for detecting a decrease in tumor size, 85% for CgA and 85% for gastrin. The specificity varied from 53% to 99% for CgA and from 49% to 93% for gastrin. CONCLUSIONS: In patients with gastrinomas, serum CgA and gastrin levels varied considerably from day to day, and this must be taken into consideration. Both markers had low sensitivity and specificity for detecting tumor increases and stabilization. For large tumor decreases postresection, both markers had high sensitivity and specificity. The current results suggest that these markers do not have sufficient sensitivity to replace serial imaging studies for detecting important smaller changes in tumor burden in patients with gastrinomas.     

High complete pathological response in locally advanced breast cancer using paclitaxel and cisplatin

Background.In an earlier study, we have demonstrated a high response rate in metastatic breast cancer using paclitaxel (P) and cisplatin (C). A phase II study using the same regimen (PC) has been conducted in locally advanced breast cancer (LABC). Methods.A total of 72 consecutive patients with non-inflammatory LABC (T24cm, T3 or T4, N0–N2, M0). Patients were scheduled to receive 3–4 cycles of the neoadjuvant PC (paclitaxel 135mg/m² and cisplatin 75mg/m² on day 1) every 21 days. Patients were then subjected to surgery and subsequently received 6 cycles of FAC (5-fluorouracil 500mg/m², doxorubicin 50mg/m², and cyclophosphamide 500mg/m²) or 4 cycles of AC (doxorubicin 60mg/m², and cyclophosphamide 600mg/m²). Patients then received radiation therapy, and those with hormone receptor positive tumors were given adjuvant tamoxifen intended for 5 years. Results.The median age was 39 years (range, 24–78). Clinically, 7%, 58%, and 35% of patients had T24cm, T3, and T4, respectively. Disease stage at diagnosis was IIB (33%), IIIA (27%), and IIIB (40%). Complete and partial clinical response to PC was demonstrated in 13 (18%), and 52 (72%) patients, respectively. Of those patients with evaluable pathologic response (68 patients), complete pathologic response (pCR) was achieved in 15 (22%) patients. At a median follow-up of 22 (±3.5) months, 58 (81%) were alive with no recurrence, nine (12%) were alive with evidence of disease, and five (7%) were dead. None of the patients achieving pCR has developed any relapse. The median overall survival has not been reached for all 72 patients with a projected 3-year survival (±SE) of 90% (±4%). The median progression-free survival (PFS) was 42.1 (±4.8) months with a projected PFS of 74%±7% at 3-years (for 68 patients). Conclusions.PC regimen in LABC produced a high pCR. The contribution of the other added modalities to survival could not be assessed.