Biomedical subjects

W A Lee

Publications and source records attributed to W A Lee.

At least 19 recordsLinked to original sources

PubMed1999

Enhanced chemical stability of the intracellular prodrug, 1-[((S)-2-hydroxy-2-oxo-1,4,2-dioxaphosphorinan-5-yl)methyl]cytosine, relative to its parent compound, cidofovir.

Degradation kinetics of cyclic HPMPC (cHPMPC), 1-[((S)-2-hydroxy-2-oxo-1,4,2-dioxaphosphorinan-5-yl)methyl]cytosi ne, and its parent compound cidofovir (also known as HPMPC) were conducted in the pH range of 2-11 at 70 degrees C. cHPMPC manifested greater chemical stability than cidofovir, except under alkaline conditions (pH?9). Three degradation products-cidofovir, cyclic HPMPU and HPMPU-were identified for cHPMPC, and the product distribution was characterized via a stability-indicating HPLC assay. Cyclic HPMPU and HPMPU are the uracil analogs of cHPMPC and cidofovir, respectively, formed through a hydrolytic deamination pathway. The deamination and hydrolysis rate constants for cHPMPC under acidic conditions were derived from the degradation product curves. The deamination rate constants for cHPMPC were about 8-fold slower compared to that for cidofovir. The enhanced chemical stability for cHPMPC relative to cidofovir is attributed to the absence of intramolecular catalysis with cHPMPC.

Antiviral Agents↗

PubMed1998

Effect of oral probenecid coadministration on the chronic toxicity and pharmacokinetics of intravenous cidofovir in cynomolgus monkeys.

In animals and humans, intravenous administration of the antiviral nucleotide analogue cidofovir results in a dose-limiting nephrotoxicity characterized by damage to the proximal tubular epithelial cells. Probenecid, a competitive inhibitor of organic anion transport in the proximal tubular epithelial cells, was evaluated for its effect on the chronic toxicity and pharmacokinetics of cidofovir. Cynomolgus monkeys (5/sex/group) received cidofovir for 52 consecutive weeks as a once weekly intravenous bolus injection at 0 (saline), 0.1, 0.5, or 2.5 mg/kg/dose alone or at 2.5 mg/kg/dose in combination with probenecid (30 mg/kg/dose via oral gavage 1 h prior to cidofovir administration). Cidofovir-associated histopathological changes were seen only in the kidneys, testes, and epididymides. Nephrotoxicity (mild to moderate cortical tubular epithelial cell karyomegaly, tubular dilation, basement membrane thickening) was present only in monkeys receiving 2.5 mg/kg/dose cidofovir without probenecid. The incidence and severity of testicular (hypo- and aspermatogenesis) and epididymal (severe oligo- and aspermia) changes were increased in monkeys administered cidofovir at 2.5 mg/kg/dose, either alone or in combination with oral probenecid. Renal drug clearance was decreased between Weeks 1 and 52 in the 2.5 mg/kg/dose groups and resulted in an increased systemic exposure to cidofovir (as measured by AUC) that was significantly greater in monkeys administered cidofovir alone (312% increase in males, 98% in females) than in those coadministered probenecid (32% increase in males, 3% in females). These results demonstrate that oral probenecid coadministration protects against the morphological evidence of nephrotoxicity and the accompanying decrease in renal clearance in monkeys receiving chronic intravenous cidofovir treatment.

Administration, Oral↗

PubMed1998

Biexponential decomposition of a neuraminidase inhibitor prodrug (GS-4104) in aqueous solution.

PURPOSE: To examine the degradation kinetics and identify the degradation products of a neuraminidase inhibitor prodrug. GS-4104. METHODS: Degradation was studied as a function of pH and temperature using a stability-indicating RP-HPLC assay. Degradation products were isolated by RP-HPLC and identified by NMR. Specific rate constants were calculated based on a scheme defined by products(s) analysis. RESULTS: Three distinct degradation products were observed in the pH region studied (pH 2-8): isomer I, GS-4071, and isomer II. Isomer I resulted from the N, N-migration of the acetyl group. Gs-4071 was formed by the hydrolysis of the ethyl ester. Both GS-4071 and isomer I degraded further to isomer II by N, N-acyl migration and ester hydrolysis, respectively. The N, N-acyl migration reaction was characterized using two dimensional heteronuclear multiple bond correlation (HMBC) NMR. The decomposition kinetics of GS-4104 follow a biexponential decay at pH 2-7. The degradation kinetics of Gs-4104 at pH 4.0, 70 degree C were independent of the initial GS-4104 concentration. CONCLUSIONS: The degradation profile indicates that development of solution or solid dosage from of GS-4104 with adequate shelf-life stability at room temperature is feasible.

Amines↗

PubMed1998

Pharmacokinetics and bioavailability of the anti-human immunodeficiency virus nucleotide analog 9-[(R)-2-(phosphonomethoxy)propyl]adenine (PMPA) in dogs.

The pharmacokinetics, bioavailability, and metabolism of the anti-human immunodeficiency virus nucleotide analog 9[(R)-2-(phosphonomethoxy)propyl]adenine (PMPA) were determined in beagle dogs following intravenous, intraperitoneal, and oral administration. Fasted male beagle dogs (n = 5) were pretreated with pentagastrin and received PMPA (10 mg/kg of body weight) by the intravenous and oral routes with a washout period of 1 week between doses. A further group of male dogs received PMPA as a single dose via the intravenous (1 mg/kg; n = 5) and the intraperitoneal (10 mg/kg; n = 3) routes, with 1-week washout period between doses. The concentrations of PMPA in plasma and urine were determined over 48 h postdosing by fluorescence derivatization and high-performance liquid chromatography (HPLC). The potential for metabolism or biliary excretion of PMPA was evaluated in a dog with a chronic indwelling bile cannula. Urine, feces, and bile were collected at intervals over 48 h following the intravenous administration of [14C]PMPA (10 mg/kg; 55 microCi/kg). The concentrations of PMPA in plasma after intravenous injection were best described by an open two-compartment model with a terminal half-life of approximately 10 h. PMPA was excreted unchanged in urine (70%); recovery in feces (0.42%) or bile (0.26%) was negligible. The plasma clearance of PMPA (0.28+/-0.05 liter/h/kg) was substantially greater than the glomerular filtration rate in this species, suggesting active tubular secretion of PMPA. No metabolites of [14C]PMPA were observed in urine, feces, or bile on the basis of HPLC with radioactive flow detection. The remainder of the dose was probably excreted unchanged in urine beyond 48 h postdosing. The mean+/-standard deviation observed bioavailabilities of PMPA following oral and intraperitoneal administration at 10 mg/kg were 17.1%+/-1.88% and 73.5%+/-10.5%, respectively.

Adenine↗

PubMed1997

Learned changes in the complexity of movement organization during multijoint, standing pulls.

This paper tests the hypothesis that the central nervous system (CNS) learns to organize multijoint movements during a multijoint 'bouncing pull' task such that, after practice, motion of the anterior-posterior center of mass (CMAP) more closely resembles that of a conservative, one degree of freedom (DF), inverted pendulum model. The task requires standing human subjects to produce precise peak pulling forces on a handle while maintaining balance-goals that can be easily accomplished if movement is organized as in the model. Ten freely standing subjects practiced making brief, bouncing pulls in the horizontal direction to target forces (20-80% of maximum) for 5 days. Pulling force, body kinematic and force plate data were recorded. An eight-segment analysis determined sagittal-plane CM motion. We compared the effects of practice on the regression-based fit between actual and model-simulated CMAP trajectories, and on measures of CMAP phase plane symmetry and parameter constancy that the model predicts. If the CNS learns to organize movements like the inverted pendulum model, then model fit should improve and all other measures should approach zero after practice. The fit between modeled and actual CMAP motion did not improve significantly with practice, except for moderate force pulls. Nor did practice increase phase plane symmetry or parameter constancy. Specifically, practice did not decrease the differences between the pre-impact and rebound positions or speeds of the CMAP, although speed difference increased with pulling force. CMAP at the end of the movement was anterior to its initial position; the anterior shift increased after practice. Differences between the pre-pull and balance-recovery ankle torque (TA) impulses were greater on day 5 and correlated with the anterior shift in CMAP. These results suggest that practice separately influenced the force production and balance recovery phases. A modified model with damping could not explain the observed behaviors. A modified model using the actual time-varying TA profiles improved fit at lower force levels, but did not explain the increased postural shift after practice. We conclude that the CNS does not learn to organize movements like the conservative, inverted pendulum model, but rather learned a more complex form of organization that capitalized on more time-varying controls and more intersegmental dynamics. We hypothesize that at least one additional DF and at least one time-varying parameter will be needed to explain fully how the CNS learns to organize multijoint, bouncing pulls made while standing.

Adult↗

PubMed1997

Bioavailability and metabolism of cidofovir following topical administration to rabbits.

The bioavailability and metabolism of the antiviral nucleotide analog cidofovir (HPMPC) were examined in New Zealand white rabbits following topical administration to normal and abraded skin. Male rabbits (four per group) received 14C-cidofovir (100 microCi/kg) intravenously (1 mg/kg) as a solution or topically (2 mg/animal) as a 1% w/w gel containing hydroxyethylcellulose (HEC) with or without propylene glycol (PG). The same PG/HEC formulation was applied topically to an abraded skin site in a fourth group of animals. All radioactivity detected in plasma and skin was accounted for by cidofovir. Plasma concentrations of radioactivity declined multiexponentially following intravenous administration, with a terminal half-life of 5.4 h. For intact skin, the absolute bioavailabilities of the HEC and PG/HEC formulations were 0.2 and 2.1%, respectively. For abraded skin, the bioavailability for the PG/HEC gel was 41%. Radioactivity in kidneys was attributed to cidofovir ( > 95%) and cyclic HPMPC. Concentrations in kidney following topical administration of cidofovir to normal skin were < 4% of those following intravenous dosing. Topical application of cidofovir to intact skin led to negligible systemic exposure to the drug. The topical bioavailability and hence the flux of cidofovir through intact skin was enhanced by the presence of PG in the formulation. Abrasion of the skin removed the principal barrier to absorption and led to significant systemic exposure to cidofovir.

Administration, Topical↗

PubMed1997

Oral formulations of adefovir dipivoxil: in vitro dissolution and in vivo bioavailability in dogs.

The effect of formulation on oral bioavailability of the antiviral nucleotide analogue adefovir from the prodrug adefovir dipivoxil was examined in beagle dogs. A suspension formulation of adefovir dipivoxil granules was administered to five fasted male beagle dogs (250 mg prodrug per dog; 135.7 mg-equiv of adefovir per dog). Tablets prepared from the same granulation (batch B94) were administered at 2 x 125 mg prodrug per dog. In addition, the same tablets were administered to dogs in the fed state or following pentagastrin pretreatment. Two further tablet batches (H94 and D501) with slight formulation changes were also evaluated in pentagastrin pretreated dogs (n = 5). Concentrations of adefovir in plasma were determined by HPLC following fluorescence derivatization. Tablet dissolution was examined at pH 2.0. One batch of adefovir dipivoxil tablets showed a 5-fold slower dissolution rate in vitro (B94 = H94 >> D501). Adefovir dipivoxil was completely converted to adefovir following oral absorption in dogs. The oral bioavailability of adefovir from the suspension was 35.0 +/- 8.9%. The oral bioavailability of adefovir from the tablet formulation was 34.7 +/- 10.3%, 37.2 +/- 4.5%, and 44.9 +/- 5.9% in fasted dogs, fed dogs and fasted dogs pretreated with pentagastrin, respectively. All three tablet batches had equivalent bioavailability in dogs. Oral bioavailability of adefovir from the prodrug in dogs (35-46%) was unaffected by formulation, food, or the acidic pH of the gastrointestinal tract. In vitro dissolution of adefovir dipivoxil tablets did not correlate with oral bioavailability. Oral bioavailability of adefovir dipivoxil appears to be limited by low permeability and biological conversion of the prodrug to adefovir.

Adenine↗

PubMed1997

Metabolism and pharmacokinetics of novel oral prodrugs of 9-[(R)-2-(phosphonomethoxy)propyl]adenine (PMPA) in dogs.

PURPOSE: A series of prodrugs designed to enhance the oral bioavailability of the antiretroviral agent 9-[(R)-2-(phosphonomethoxy)propyl]adenine (PMPA; 1) have been synthesized, including a bis-(acyloxymethyl) ester 2 and a series of bis-(alkoxycarbonyloxymethyl) esters 3-9. The in vitro biological stability and in vivo pharmacokinetics of these prodrugs were evaluated to support selection of a prodrug candidate for clinical evaluation. METHODS: The in vitro biological stability of the prodrugs was examined in dog tissues (intestinal homogenate, plasma and liver homogenate). The apparent half-lives were determined based on the disappearance of prodrug using reverse-phase HPLC with UV detection. Oral bioavailability of PMPA from each prodrug was determined in fasted beagle dogs. Concentrations of PMPA in plasma were determined by HPLC following fluorescence derivatization. Data for prodrugs were compared to historical data for intravenous PMPA. RESULTS: All prodrug were rapidly hydrolyzed in dog plasma and tissues (t1/2 < 60 min). In fasted beagle dogs, bis-[(pivaloyloxy)methyl] PMPA (bis-POM PMPA) 2 had the highest oral bioavailability as PMPA (37.8 +/- 5.1%). The oral bioavailabilities of PMPA from bis-(alkoxycarbonyloxymethyl) esters ranged from 16.0% to 30.7% and PMPA was the major metabolite formed. CONCLUSIONS: There was a correlation between oral bioavailability and intestinal stability of bis-(alkoxycarbonyloxymethyl) ester prodrugs (r2 = 0.96). Lipophilicity (log P) was not a good predictor of oral bioavailability. The most labile prodrugs in dog intestinal homogenates, bis-(n-butyloxycarbonyloxymethyl) PMPA 5 and bis-(neo-pentyloxy-carbonyloxymethyl) PMPA 8 (t1/2 < 5 min) had the lowest oral bioavailabilities. Based on good oral bioavailability (30.1%), chemical and intestinal stability bis-(isopropyloxycarbonyloxymethyl) PMPA (bis-POC PMPA) 4 was selected as a candidate for clinical evaluation.

Adenine↗

PubMed1997

Prolonged extracorporeal life support (ECLS) for varicella pneumonia.

OBJECTIVE: To review the institutional experience of a national tertiary referral center for extracorporeal life support (ECLS) in severe varicella pneumonia. DATA SOURCES: Hospital records and ECLS flow sheets. STUDY SELECTION: All pediatric (nonneonatal) and adult patients who were treated for varicella pneumonia with ECLS at the University of Michigan Medical Center between 1986 and 1995. DATA EXTRACTION: Diagnosis of varicella pneumonia was made by history of recent exposure to chickenpox, progressive dyspnea, fever, a characteristic diffuse, vesicular rash, and a supporting chest roentgenogram. Indications for ECLS included a shunt fraction of > 30% or PaO2/FlO2 ratio of < 80 despite maximal conventional therapy, which included aggressive diuresis, blood transfusions to optimize oxygen-carrying capacity, pressure-controlled/inverse-ratio ventilation, and intermittent prone positioning. DATA SYNTHESIS: Between 1986 and 1995, 191 patients were referred for ECLS. Among these patients, there were 51 (27%) cases of viral pneumonia, of which nine cases were due to acute varicella-zoster infection. Intravenous acyclovir was administered to eight of the nine patients. Of the nine patients, two patients improved using conventional ventilator management, and seven patients underwent ECLS. Overall survival on ECLS was 71% (5/7). The mean (+/-SD) alveolar-arterial oxygen gradient and PaO2/FlO2 ratio were 533 +/- 101 torr (71.3 +/- 13.5 kPa) and 67 +/- 24, respectively. The median duration of mechanical ventilation before ECLS and the subsequent duration of ECLS were 4 and 12.8 days, respectively. One of the deaths was from progressive right heart failure secondary to pulmonary hypertension and the other death was from overwhelming Pseudomonas sepsis. CONCLUSIONS: Early recognition of imminent pulmonary failure and rapid institution of ECLS are critical in the successful management of severe, life-threatening varicella pneumonia.

Acyclovir↗

PubMed1997

Pharmacokinetics and metabolism of selected prodrugs of PMEA in rats.

The oral bioavailability of PMEA [9-[2-(phosphonomethoxy)ethyl]adenine; adefovir) has been determined in rats from three bisester prodrugs of PMEA: bis-(pivaloyloxymethyl) PMEA (bis-POM PMEA), bis-(phenyl) PMEA, and bis-(o-ethoxyphenyl) PMEA. The prodrugs were each administered to 9 male rats as solutions in PEG 400 at a dose of 10 mg-equivalent of PMEA per kg. Plasma samples were obtained over the course of 12 hr and concentrations of PMEA were determined by fluorescence derivatization and analysis by HPLC. Concentrations of PMEA observed in plasma following oral administration of PMEA prodrugs were compared with levels observed for intravenous PMEA. The observed oral bioavailabilities of PMEA from bis-POM PMEA, bis-(phenyl) PMEA, and bis-(o-ethoxyphenyl) PMEA were 38.2%, 2.46%, and 40.1%, respectively. PMEA was the only metabolite formed after oral administration of bis-POM PMEA. Three metabolites were detected after oral administration of either bis-(phenyl) PMEA or bis-(o-ethoxyphenyl) PMEA to rats: PMEA, the corresponding monoester, and 2-adenylacetic acid. The major metabolite of bis-(phenyl) PMEA was 2-adenylacetic acid following oral administration. 2-Adenylacetic acid appears to have been formed from the intact prodrugs by a P450 mediated oxidation of the ethyl side chain.

Adenine↗

PubMed1996

Overexpression of the 67 kD laminin receptor correlates with the progression of gastric carcinoma.

This retrospective study was designed to investigate the relationship between overexpression of the 67 kD laminin receptor (67LR) using immunohistochemistry, and several clinicopathological parameters including overall survival in human gastric adenocarcinoma. We stained paraffin-embedded sections of 93 resected primary gastric adenocarcinomas using a polyclonal antibody specific for the 67LR as well as monoclonal antibodies for p53 protein, epidermal growth factor receptor, proliferating cell nuclear antigen, carcinoembryonic antigen and chromagranin. The results showed statistically significant correlations between overexpression of the 67LR and types of early or advanced gastric carcinoma (p < 0.001), depth of invasion (p < 0.001), WHO histopathologic classification (p < 0.001), stage (p = 0.001), expression of p53 protein (p = 0.019), expression of epidermal growth factor receptor (p < 0.001) and proliferating cell nuclear antigen labeling index (p = 0.002). A lower proportion of signet ring cells revealed a higher percentage of overexpression of 67LR in both early (p < 0.002) and advanced (p < 0.001) gastric carcinomas. Intestinal type adenocarcinoma (according to Lauren's classification) revealed a higher percentage of overexpression of the 67LR than the diffuse type in both early (p = 0.057) and advanced (p < 0.001) gastric carcinomas. The correlations between overexpression of the 67LR and lymph node metastasis were statistically significant (p < 0.07). Although the overexpression of the 67LR tended to correlate with lower survival rates, the correlation was not statistically significant due to the limited sample size. Our data revealed that overexpression of the 67LR is correlated with the progression of gastric carcinoma. The expression of the 67LR may be important as one of the steps which determines invasiveness during the progression of cancer.

Adenocarcinoma↗

PubMed1996

Distribution and metabolism of intravitreal cidofovir and cyclic HPMPC in rabbits.

PURPOSE: This study was designed to evaluate the intraocular distribution and metabolism of the antiviral nucleotide analogs cidofovir and cyclic 1-[(S)-3-hydroxy-2-(phosphonomethoxy) propyl]cytosine (HPMPC) in New Zealand white rabbits following intravitreal administration. METHODS: Male rabbits received either 14C-cidofovir or 14C-cyclic HPMPC by intravitreal injection into both eyes (50 micrograms/eye, 11 microCi/eye). Two animals per group were sacrificed at 24, 48, 72 or 240 h post-dose. Ocular tissues, kidney and liver were oxidized to determine total radioactivity and metabolites were determined by HPLC. RESULTS: At 24 h post-dose, total radioactivity was 9.96 and 5.18 micrograms-equiv/g for cidofovir and cyclic HPMPC, respectively, in vitreous and 20.9 and 3.54 micrograms-equiv/g, respectively, in retina. Although the initial vitreal clearance was 2-fold faster for the cyclic analog, the estimated terminal elimination half-lives in vitreous (42 hr) and in retina (66-77 hr) were similar for both drugs. By 240 h post-dose, radioactivity in all ocular tissues was approximately ten-fold higher for cidofovir. Radioactivity in vitreous at 240 h after intravitreal dosing with either drug contained cidofovir, cyclic HPMPC and cidofovir-phosphocholine. CONCLUSIONS: The long retinal half-life observed presumably reflects formation of phosphorylated cidofovir within retinal cells. Cidofovir achieved a ten-fold higher level of phosphorylated drug in retina than cyclic HPMPC: Therefore, intravitreal cidofovir may be expected to suppress progression of retinitis for a longer period than an equivalent intravitreal dose of cyclic HPMPC: The intravitreal half-life of cidofovir was 20-fold longer than that of ganciclovir in the same animal model.

AIDS-Related Opportunistic Infections↗

PubMed1996

Pharmacokinetics of cidofovir in monkeys. Evidence for a prolonged elimination phase representing phosphorylated drug.

After intravenous administration of [14C]cidofovir to African green monkeys (43 mg/kg, 29.5 microCi/kg), the drug distributed rapidly into extracellular fluid. Concentrations of radioactivity in plasma were described by a three-compartment model with alpha, beta, and gamma half-lives of 0.67, 3.02, and 36.0 hr, respectively (N = 3). These phases are believed to represent renal elimination, efflux of free cidofovir from cells, and efflux from cells of cidofovir produced from dephosphorylation of metabolites, respectively. Less than 5% of the dose was phosphorylated, based on the proportion of total AUC in the gamma-phase. The clearance of cidofovir (211 +/- 16.6 ml/hr/kg) was dependent on dose and exceeded the theoretical glomerular filtration rate. Concentrations of cidofovir in kidney declined with a half-life of 23 hr and were > 1,000-fold higher than plasma levels by 120 hr. Clearance of cidofovir after multiple intravenous doses of 4.9 mg/kg/day (18.5 microCi/kg/day) decreased significantly by day 10, consistent with the observed nephrotoxicity. Oral and subcutaneous bioavailabilities of cidofovir were 21.8 +/- 9.44 and 98.5 +/- 15.8%, respectively. After intravenous administration of [14C]cidofovir to cynomolgus monkeys (10 mg/kg, 100 microCi/kg) alone or 1 hr after oral probenecid (30 mg/kg), mean +/- SD (N = 3) urinary recoveries of total radioactive dose were 91.4 +/- 11.3% and 94.4 +/- 29.8%, respectively, at 7 days postdose. The mean +/- SD half-lives of the terminal elimination phases were 33.3 +/- 10.6 and 24.4 +/- 5.0 hr, respectively. Cidofovir accounted for 98% of the radioactivity recovered in urine; the remainder was attributed to cidofovir phosphocholine. The prolonged elimination phase observed in monkeys is consistent with the long intracellular half-life of phosphorylated cidofovir in vitro and supports infrequent dosing of the drug for antiviral therapy.

Administration, Cutaneous↗

PubMed1996

Pharmacokinetics, bioavailability, metabolism, and tissue distribution of cidofovir (HPMPC) and cyclic HPMPC in rats.

The pharmacokinetics, distribution, and metabolism of [14C]cidofovir and its cyclic analog {[2-14C]-1-[((S)-2-hydroxy-2-oxo-1,4, 2-dioxaphosphorinan-5-yl)methyl]cytosine; [14C]cyclic HPMPC} were compared after administration to Sprague-Dawley rats. After intravenous (iv) administration of [14C]cidofovir (3 or 5 mg/kg) or [14C]cyclic HPMPC (5 mg/kg), plasma concentrations of total radioactivity declined in a multiexponential manner for both drugs with terminal half-lives of 7-11 hr. No metabolites of cidofovir were observed in plasma. However, iv [14C]cyclic HPMPC was converted in vivo to cidofovir (3.9% of the AUC of total radioactivity). The corrected half-life of cyclic HPMPC was 0.53 +/- 0.05 hr. The clearance of iv cidofovir (0.60-0.79 liter/hr/kg) was significantly less than that of cyclic HPMPC (1.10 +/- 0.12 liter/hr/kg). Cyclic HPMPC was more efficiently secreted by the kidney than cidofovir. The steady-state volume of distribution of cyclic HPMPC (0.32 +/- 0.5 liter/kg) was substantially less than that of cidofovir (0.9-3.0 liter/ kg). Concentrations in kidney at 24 hr after iv administration of 5 mg/kg were approximately 20-fold higher for cidofovir (6.6 micrograms-eq/g) than for cyclic HPMPC, whereas levels of radioactivity in the remaining tissues were similar for both drugs. This is consistent with the improved therapeutic index observed for cyclic HPMPC in animals. After iv administration of [14C]cidofovir (3 mg/kg), concentrations in most tissues declined with half-lives > 12 hr, presumably reflecting the long intracellular half-life of phosphorylated drug. Kidneys and liver of animals given iv [14C]cidofovir (5 mg/kg) contained unchanged cidofovir (35-65%), metabolite I (attributed to cidovofir phosphocholine) (29-60%), and a peak coeluting with cyclic HPMPC on three different HPLC systems (metabolite II) (5-7%). Kidneys, liver, and lung of animals given iv [14C]cyclic HPMPC contained cyclic HPMPC (1-6%), cidofovir (44-50%), and metabolite I (44-54%). The subcutaneous bioavailability of cidofovir was 91.5%. The subcutaneous and oral bioavailabilities of cyclic HPMPC were 82.7% and 3.50 +/- 1.07%, respectively, based on total radioactivity.

Administration, Cutaneous↗

PubMed1996

Effect of probenecid on the distribution, metabolism, and excretion of cidofovir in rabbits.

The effect of concomitant probenecid on the tissue distribution, metabolism, and urinary excretion of cidofovir was examined in New Zealand white rabbits. Two groups of six male rabbits received intravenous [3H]cidofovir (5 mg/kg, 20 mu Ci/kg) alone or with concomitant intravenous probenecid (90 mg/kg). Radioactivity in kidney at 15 min postdose was decreased 70% by probenecid; plasma levels at 15 min postdose were increased 65% by probenecid. These effects were diminished at later time points. The estimated elimination half-life of cidofovir from the kidney was 16 hr in the presence of probenecid and 11 hr without probenecid. Two additional groups of six rabbits received intravenous [14C]cidofovir (15 mg/kg, 100 mu Ci/kg) alone or 1 hr after oral administration of probenecid (90 mg/kg). Radioactivity was highest in the kidney (approximately 700 mu g-eq/g at 30 min postdose). Probenecid did not affect the gross distribution of radioactivity. However, autoradiography of left kidneys revealed localization of the drug in the renal cortex; radioactivity in the cortex at 30 min postdose was decreased 50% by probenecid. These data are consistent with inhibition of tubular secretion of cidofovir by probenecid. More than 73% of the cidofovir dose was recovered in the urine in 24 hr. Urine contained unchanged cidofovir (>97%) and a metabolite coeluting with authentic cidofovir phosphocholine (2%). This metabolite also accounted for approximately 1-4% of the radioactivity in rabbit kidney.

Administration, Oral↗

PubMed1995

Optimization of in vitro flux through hairless mouse skin of cidofovir, a potent nucleotide analog.

The in vitro flux (4-8 h) of cidofovir (1-[(S)-3-hydroxy-2-(phosphonomethoxy)propyl]cytosine) was measured across full-thickness hairless mouse skin to evaluate potential formulations for local treatment of herpes virus infections. The effects of propylene glycol, isopropyl alcohol, oleic acid, pH, and cidofovir concentration were examined. In addition, several prototype aqueous gel formulations were studied. Flux values (4-8 h) increased linearly with cidofovir concentration in both solution and gel formulations. Removal of the stratum comeum by tape stripping increased the flux by approximately 400-fold, whereas pH (4.5 versus 7) had little effect on flux. The presence of propylene glycol, isopropyl alcohol, or their combination did not significantly increase mean flux (p > or = 0.05). Pretreatment of the skin with oleic acid resulted in a significant enhancement of cidofovir flux (p < or = 0.01). From the measured flux values, the calculated concentration of cidofovir achievable in the viable epidemis from a 1% cidofovir gel formulation was approximately 14 micrograms/mL, which is comparable to the in vitro 50% inhibitory dose (ID50) values for herpes simplex viruses HSV-1 and HSV-2.

Animals↗

PubMed1995

A novel oligodeoxynucleotide inhibitor of thrombin. I. In vitro metabolic stability in plasma and serum.

PURPOSE: To determine the degradation rates and pathways of GS-522, a potent oligodeoxynucleotide (GGTTGGTGTGGTTGG) inhibitor of thrombin, in serum and plasma. METHODS: A stability-indicating, anion-exchange HPLC method was developed and used to determine concentrations of GS-522 and metabolites. RESULTS: In monkey plasma at 2 microM or below, the degradation of GS-522 can be fit to a first-order exponential with a kpobs approximately 0.01 min-1. At 3 microM and above the degradation process deviates from a monoexponential decay profile. An initial fast degradation process is followed by a slower phase with an observed rate constant equal to that observed at 2 microM and below. In monkey serum, the KM and Vmax are 8.4 microM and 0.87 microM min-1, respectively. CONCLUSIONS: The kinetics are consistent with an equilibrium binding of GS-522 to prothrombin in plasma (Kd = 50 nM) which saturates at GS-522 concentrations > 2 microM. Compared to a scrambled sequence (GGTGGTGGTTGTGGT), with no defined tertiary structure, GS-522 is 4-fold more stable in serum. The metabolic profile in plasma is consistent with a 3'-exonuclease catalyzed hydrolysis of GS-522.

Aptamers, Nucleotide↗

PubMed1995

A novel oligodeoxynucleotide inhibitor of thrombin. II. Pharmacokinetics in the cynomolgus monkey.

PURPOSE: To determine the pharmacokinetics of GS-522, an oligodeoxynucleotide (GGTTGGTGTGGTTGG) inhibitor of thrombin, after constant infusion and bolus administration in the cynomolgus monkey. METHODS: Using a stability indicating HPLC method, the GS-522 plasma concentration versus time data were obtained after constant infusion (0.1, 0.3, 0.5 mg/kg/min) and bolus administration (11.25 and 22.5 mg/kg). Plasma data after bolus administration was fit to a three-compartment model. RESULTS: The half-lives for the alpha and beta phases were 1.4 and 5.4 min, respectively. Steady state GS-522 concentrations were reached within 10 minutes after initiation of constant infusions. Termination of infusions resulted in a rapid elimination of GS-522 with an average elimination half-life equal to 1.5 min. The Vss calculated from both the constant infusion and bolus data approximated the blood volume of the monkey. Substitution of the phosphodiester backbone at the 3' end of GS-522 with two phosphorothioate linkages did not substantially effect the elimination half-life upon termination of infusion. CONCLUSIONS: These data in conjunction with published biodistribution data suggest that oligodeoxynucleotides are rapidly cleared from plasma by tissue uptake and that little efflux back into blood takes place. Additionally, strategies designed to increase oligodeoxynucleotide resistance to exonucleases will not dramatically increase plasma half-lives.

Animals↗