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M Chiba

Publications and source records attributed to M Chiba.

At least 163 records · Page 9Linked to original sources
PubMed1997

In vitro metabolism of indinavir in the human fetal liver microsomes.

In vitro microsomal formation of primary metabolites of indinavir (CRIXIVAN, MK-0639, L-735,524), an HIV protease inhibitor, were qualitatively similar among the different developmental stages in humans, although the fetal liver had a lower capability to form the metabolites than the pediatric and adult liver. The lower activity of fetal liver was mainly owing to a decrease in the Vmax values. The Vmax value in the fetus was about one-third of that in the adult human, while no significant difference was found in Km values between groups. The liver microsomes were also characterized using P450 markers to examine the development-associated alteration in P450 functional activities. Debrisoquine 4-hydroxylase activity was comparable among the three age groups. In contrast, tolbutamide methyl hydroxylase activity, as well as the CYP3A marker, testosterone 6beta-hydroxylase activity, in the fetal liver microsomes was much lower than in the pediatric and adult by more than 40-fold. However, the difference in testosterone 2beta-hydroxylase and nifedipine N-oxidase activities between fetus and adult was markedly smaller. The ratio of indinavir metabolism in pediatric or adult liver to fetus was 1.7 for pediatric and 3.6 for adult liver microsomes. Similarly, testosterone 2beta-hydroxylase and nifedipine N-oxidase activities showed smaller differences between adult (or pediatric) and fetal liver microsomes than testosterone 6beta-hydroxylase activity. The reason for the observed marked differences in the development-associated alteration may lie in the differences of substrate specificities between CYP3A isoforms.

Adult↗
PubMed1997

Metabolic profiles of montelukast sodium (Singulair), a potent cysteinyl leukotriene1 receptor antagonist, in human plasma and bile.

Montelukast sodium [1-([(1(R)-(3-(2-(7-chloro-2-quinolinyl)-(E)- ethenyl)phenyl)-3-(2-(1-hydroxy-1-methylethyl)phenyl)propyl)thio]methyl)cyclopropylacetic acid sodium salt] (MK-476, Singulair) is a potent and selective antagonist of the cysteinyl leukotriene (Cys-LT1) receptor and is under investigation for the treatment of bronchial asthma. To assess the metabolism and excretion of montelukast, six healthy subjects received single oral doses of 102 mg of [14C]montelukast, and the urine and feces were collected. Most of the radioactivity was recovered in feces, with </=0.2% appearing in urine. Based on these results and the reported modestly high oral bioavailability of montelukast, it could be concluded that a major part of the radioactivity was excreted via bile. A second clinical study was conducted to identify biliary metabolites of montelukast. The bile was aspirated using a modified procedure involving a nasogastric tube placed fluoroscopically near the ampulla of Vater, after an oral dose of 54.8 mg of [14C]montelukast. This technique appears to be a new application for drug metabolism studies. The study was conducted with fasted and nonfasted subjects, with the bile being aspirated continuously under suction over periods of 2-8 hr and 8-12 hr after the dose, respectively. Two hours before the end of the collection procedure, cholecystokinin carboxyl-terminal octapeptide was administered iv to stimulate gallbladder contraction. Plasma samples also were collected periodically over 10 hr. Due to the nature of the collection procedure and the limited sampling time, recovery of radioactivity in bile was incomplete and varied from 3 to 20% of the dose. Radiochromatographic and LC-MS/MS analyses of bile showed the presence of one major and several minor metabolites, along with small amounts of unchanged parent drug. The minor metabolites were identified, by LC-MS/MS comparison with synthetic standards or by NMR, as acyl glucuronide (M1), sulfoxide (M2), 25-hydroxy (a phenol, M3), 21-hydroxy (diastereomers of a benzylic alcohol, M5a and M5b), and 36-hydroxy (diastereomers of a methyl alcohol, M6a and M6b) analogs of montelukast. The major metabolite was characterized as a dicarboxylic acid (M4), a product of further oxidation of the hydroxymethyl metabolite M6. Chiral LC-MS/MS analyses of M4 revealed that this diacid, like M5 and M6, was formed in both diastereomeric forms. The levels of metabolites in the systemic circulation were low in the fed as well as fasted subjects, with <2% of the circulating radioactivity being due to metabolites M5a, M5b, M6a, and M6b. Overall, this bile aspiration technique, which is less invasive than either T-tube drainage or fine-needle percutaneous puncture, provided a convenient and expedient means of identifying the biliary metabolites of montelukast, relatively free of contributions from colonic microflora.

Acetates↗
PubMed1996

Retinoic acid stimulates expression of the functional osteoclast integrin alpha v beta 3: transcriptional activation of the beta 3 but not the alpha v gene.

The capacity of osteoclasts to resorb bone depends, in part, on the surface expression of the integrin alpha v beta 3. We have investigated whether the steroid hormone retinoic acid, known to stimulate bone resorption, regulates the appearance of the alpha v beta 3 complex in avian osteoclast presursors. Using surface labeling, followed by immunoprecipitation with a alpha v beta 3-specific antibody, we show that retinoic acid increases surface expression of the heterodimer in a dose- and time-dependent manner. Northern analysis reveals that the high basal steady-state levels of alpha v mRNA do not change, while those for beta 3 rise significantly from their initially low levels. Nuclear run-on studies confirm that steroid treatment stimulates transcription of the beta 3, but not the alpha v gene. Osteoclast precursors treated with retinoic acid exhibit increased multinucleation and expression of the osteoclast marker enzyme tartrate-resistant acid phosphatase. However, the fused cells do not have an increased capacity to resorb bone. In summary, multinucleated cells generated in this study do not represent fully differentiated bone-resorbing polykaryons. These results suggest that treatment of osteoclast precursors with retinoic acid is necessary, but insufficient, for expression of the mature osteoclast phenotype.

Animals↗
PubMed1996

The importance of preserving disc structure in surgical approaches to lumbar disc herniation.

STUDY DESIGN: This study examined the clinical and radiographic results in patients with lumbar disc herniation treated surgically with one of four procedures: two different methods of herniotomy using a posterior approach and two different methods of percutaneous nucleotomy. OBJECTIVES: The authors goal was to identify the structural changes in the disc after each of the procedures and to correlate them with the clinical outcome. SUMMARY OF BACKGROUND DATA: The details of herniotomy procedure via a posterior approach or by percutaneous nucleotomy as described by many authors have not been consistent. Furthermore, previous reviews of these studies have been less than satisfactory. METHODS: One hundred fifty-seven herniotomies using a posterior approach and 94 percutaneous nucleotomies were studied with a follow-up of at least 2 years. Each of these groups was divided into two subgroups to evaluate the role of the remaining nucleus pulposus in the central area of the disc. RESULTS: Changes seen in imaging studies, such as a decrease in disc height and an increase of intervertebral instability in plain radiographs and a decrease in signal intensity of the disc in magnetic resonance images, were significantly less common in the subgroups of herniotomy and percutaneous nucleotomy in which the nucleus pulposus in the central area of the disc was preserved when compared with the subgroups in which a complete removal of the disc was attempted. The changes seen in the imaging studies were significant in patients younger than 40 years of age and correlated closely with the clinical results. CONCLUSIONS: Preserving the nucleus pulposus during the surgical treatment of lumbar disc herniation in patients younger than 40 years of age resulted in better radiographic and clinical results.

Adolescent↗
PubMed1996

Microtiter plate immunoassay for the evaluation of platelet adhesion to fibronectin.

Investigations of platelet adhesion to adhesive proteins have been pursued to understand the basic mechanisms of hemostasis and thrombosis. Most assays used to determine platelet adhesion under stasis conditions rely on radiolabeled platelets. We describe a new microtiter immunoassay to study platelet adhesion to adhesive proteins under stasis conditions. Direct comparison of platelet adhesion to fibronectin using a standard platelet adhesion assay based on 51Cr-labeled platelets and the new immunoassay showed that the optical density values obtained with the immunoassay are directly proportional to the number of platelets bound. The choice of platelet suspension buffer crucial for the design of such experiments, because the adhesion of resting platelets to fibronectin is increased in response to thrombin stimulation. This increase buffer rather than Tris buffer. Platelet adhesion to fibronectin is increased in response to thrombin stimulation. This increase can be inhibited by synthetic RGD peptides. The thrombin-induced increase of platelet adhesion to fibronectin could be detected with antibodies against actin and glycoprotein IIb-IIIa, but not against the alpha-granule constituent platelet factor 4 (PF4). This assay is very versatile, because it avoids the use of radioactivity, and allows the parallel processing of a large number of samples. In addition, the parallel use of antibodies against different platelet antigens allows the screening for platelet activation events associated with the measured platelet adhesion.

Amino Acid Sequence↗
PubMed1996

Outcome of anterior decompression and stabilization for thoracolumbar unstable burst fractures in the absence of neurologic deficits.

STUDY DESIGN: This study investigated the outcome of anterior decompression and stabilization for "unstable" burst fractures without initial neurologic deficits in the thoracolumbar spine. OBJECTIVE: To determine the pain and work status of the patients, who underwent the anterior decompression and stabilization, to evaluate the relationship between residual back pain and the degree of kyphosis, and to analyze the advantages and disadvantages of surgery for burst fractures without initial neurologic deficits in the thoracolumbar spine. SUMMARY OF BACKGROUND DATA: A retrospective review was conducted on 45 patients with unstable burst fractures in the thoracolumbar spine treated by anterior decompression and stabilization in our institutions from 1982 to 1993. Nineteen patients were available for this study. Ten patients were men and 9 were women. Their age at surgery ranged from 15 to 76 years (average, 46 years). The duration of follow-up ranged from 24 to 84 months (average follow-up, 54 months). METHODS: All clinical charts and radiologic data of these patients were reviewed by the first author. Their pain, work, and employment statuses were assessed by the scales proposed by Denis in 1984. RESULTS: Nine patients were rated P1, 7 were rated P2, 2 were rated P3, 1 was rated P5, 11 were rated W1, 3 were W2, 1 was W3, and 4 were rated W5. Even the physical laborers' return to work averaged 5 months after surgery (range, 3-12 months). The postoperative kyphotic angle ranged from -6 degrees to 45 degrees (average, 11 degrees), and it remained -5 degrees to 45 degrees (average, 12 degrees) at final follow-up. There was no statistical correlation between the kyphotic angle and the pain scale. CONCLUSIONS: Anterior decompression and stabilization for unstable burst fracture without initial neurologic deficits in the thoracolumbar spine has some advantages in the view of anatomical reduction and rigid stability that allows patients an early rehabilitation, return to work, and gainful employment.

Adolescent↗
PubMed1996

Short-segment pedicle instrumentation. Biomechanical analysis of supplemental hook fixation.

STUDY DESIGN: This biomechanical study of fractures in cadaver vertebrae used specially designed pedicle screws to determine screw strains during loading of two different fixation constructs. OBJECTIVES: The authors determined the relative benefit of adding offset sublaminar hooks to standard pedicle screw constructs to reduce screw bending moments and prevent fixation failure and sagittal collapse. SUMMARY OF BACKGROUND DATA: Clinical studies have demonstrated a high incidence of early screw failure in short-segment pedicle instrumentation constructs used to treat unstable burst fractures. Strategies to prevent early construct failure include longer constructs, anterior strut graft reconstruction, and use of offset sublaminar hooks at the ends of standard short-segment pedicle instrumentation constructs. METHODS: Human cadaver spines with an L1 burst fracture were instrumented with a standard short-segment pedicle instrumentation construct using specially instrumented pedicle screws. Mechanical testing was carried out in flexion, extension, side bending, and torsion, and stiffness and screw bending moments were recorded. Offset hooks were applied initially, then removed and testing repeated. Stiffness data were compared to intact and postfracture results, and between augmented and standard constructs. RESULTS: Addition of offset laminar hooks, supralaminar at T11 and infralaminar at L2, to standard short-segment pedicle instrumentation constructs increased stiffness in flexion by 268%, in extension by 223%, in side bending by 161%, and in torsion by 155% (all were significant except torsion). Sublaminar hooks also reduced pedicle screw bending moments to roughly 50% of standard in both flexion and extension (P < 0.05). CONCLUSIONS: Supplemental offset hooks significantly increase construct stiffness without sacrificing principles of short-segment pedicle instrumentation, and absorb some part of the construct strain, thereby reducing pedicle screw bending moments and the likelihood of postyield deformation and clinical failure.

Aged↗
PubMed1996

Measurement of the force needed to restrain eruptive movement of the rat mandibular incisor.

This study describes the precise determination of the extrusive force in the rat mandibular incisor measured for a relatively longer period of time under artificial respiration with halothane anaesthesia. Following restraint of the eruptive movement of the incisor for a period of 20 h, the pushing force increased gradually and was maintained as an extremely gentle slope toward the end of the experimental period. The maximum pushing forces were estimated to be 9.1 +/- 2.5 (SD) mN in impeded incisors and 9.8 +/- 2.3 mN in unimpeded incisors. When force was converted to pressure, values were 30 +/- 8 (SD)mmHg and 32 +/- 8 mmHg, respectively; the difference was not significant. The maximum pushing force decreased rapidly--almost in parallel--with the fall of systemic arterial blood pressure after death of the animals; the amounts of reduction were 7.2 +/- 1.8 mN (24 +/- 6 mmHg) in impeded incisors and 7.0 +/- 2.0 mN (23 +/- 7 mmHg) in unimpeded incisors. Residual pushing forces were 1.7 +/- 1.0 mN (5.6 +/- 3.3 mmHg) in impeded incisors and 2.5 +/- 0.6 mN (8.2 +/- 2.0 mmHg) in unimpeded incisors; the difference was significant (P < 0.05). These results suggest that, under these experimental conditions, the pushing force of the rat incisor originates primarily from localized blood pressure within the incisor socket and, in part, from tissue growth.

Alveolar Process↗
PubMed1996

Molecular nature of colon tumors in hereditary nonpolyposis colon cancer, familial polyposis, and sporadic colon cancer.

BACKGROUND & AIMS: Microsatellite instability (replication error [RER]) is a characteristic of tumors in hereditary nonpolyposis colon cancer (HNPCC), but the mechanism of HNPCC carcinogenesis is not yet understood. To clarify the nature of HNPCC tumors, RER and genetic changes were compared between HNPCC and non-HNPCC tumors. METHODS: RER and genetic changes were analyzed in 21 HNPCC, 389 familial adenomatous polyposis, and 206 sporadic tumors using polymerase chain reaction, single-strand conformation polymorphism, sequencing, and Southern hybridization. RESULTS. in HNPCC, 95% tumors at all stages showed RER positivity (altered loci, 4.3 of 5). In familial adenomatous polyposis and sporadic tumors, RER positivity (1.7 of 5) was 3% in adenoma and intramucosal carcinoma, 13%-24% in invasive carcinoma, and 35% in carcinoma metastasized to liver. Fifty percent of RER-positive HNPCC tumors had both germline and somatic mutations of hMSH2 or hMLH1 gene, whereas 6% of RER-positive non-HNPCC had somatic mutation. APC, p53, and K-ras-2 mutations and loss of heterozygosity of tumor-suppressor genes were significantly less frequent (P = 0.03 to 0.0006) but transforming growth factor beta type II receptor mutation was significantly more frequent (P = 0.000001) in HNPCC than in non-HNPCC. CONCLUSIONS: RER positivity occurs from an early stage of carcinogenesis in HNPCC but in later stages in non-HNPCC. Most HNPCC tumors may develop through different genetic changes from those in the adenoma-carcinoma sequence, although a certain percentage develops through APC mutation.

Adaptor Proteins, Signal Transducing↗
PubMed1996

Indices of lead-exposure in blood and urine of lead-exposed workers and concentrations of major and trace elements and activities of SOD, GSH-Px and catalase in their blood.

Seventy male factory workers were studied. The lead concentrations in their blood (Pb-B) were 16.55 +/- 11.53 micrograms/100 ml (range 1.5 to 50.2 micrograms/100 ml). The subjects were divided into three groups according to Pb-B (in microgram/100 ml): group A, Pb-B < or = 10 (n = 22); group B, 10 < Pb-B < or = 20 (n = 30); group C, Pb-B > 20 (n = 18). The mean +/- S.D. in each group was 5.57 +/- 2.53, 15.02 +/- 2.75, and 32.52 +/- 9.49 micrograms/100 ml, respectively. Pb in plasma was 0.011 +/- 0.010, 0.017 +/- 0.033, and 0.021 +/- 0.021 microgram/liter, and Pb in the RBC was 0.281 +/- 0.246, 0.701 +/- 0.325, and 1.626 +/- 0.861 micrograms/g Hb, respectively. In addition to Pb concentration, the concentrations of 34 elements in the plasma or in the RBC were determined. Se concentrations in RBC in each group were 0.618 +/- 0.139, 0.670 +/- 0.207, and 0.728 +/- 0.200 microgram/g Hb, and the mean values were significantly different between groups A and C (p < 0.05). For Se concentration in plasma, the mean +/- S.D. in each group was 0.132 +/- 0.035, 0.130 +/- 0.031, and 0.126 +/- 0.021 microgram/ml, respectively, and there was no significant difference between groups. On the other hand, when the activities of total SOD, Mn-SOD, Cu, Zn-SOD, and catalase in the plasma and the activities of GSH-Px both in the plasma and in the RBC were assayed, some differences were found. The activities in GSH-Px in RBC were 17.19 +/- 5.03, 17.59 +/- 3.95, and 15.25 +/- 3.18 mumol/g Hb/min, and those in plasma were 0.069 +/- 0.032, 0.081 +/- 0.023, and 0.080 +/- 0.028 mumol/ml/min. In group C, GSH-Px activity was lower in the RBC and higher in the plasma than those in group A, and it was observed that the Se concentration was higher in RBC, and that there was no remarkable change in the plasma. Catalase activity in group C was 3.58 +/- 0.81 mgH2O2/ml/30 min, which was significantly higher than that in group A (2.81 +/- 0.90 mgH2O2/ml/30 min). Further investigation is necessary in order to explain the above results. The regular indices used for evaluating lead exposure, showed significant correlations with Pb-B: r = -0.786 vs delta-Aminolevulinic acid (ALA) dehydratase activity in blood, r = 0.927 vs. inhibition rate, and r = 0.339 vs. ALA in urine.

Aminolevulinic Acid↗
PubMed1996

[Concentrations of essential trace elements in blood and introduction of analytical techniques].

Research on trace elements has been paid attention recently. In order to determine concentrations of various kinds of trace elements in biological materials, atomic absorption spectrophotometer is widely used. This instrument allows single element analysis at ppb levels. Microwave induced plasma-mass spectrophotometer (MIP-MS) or inductively coupled plasma-mass spectrophotometer (ICP-MS) permits multielement analysis at sub-ppb or ppt levels. These analytical instruments accept liquid samples only. Biological materials such as organs, tissue and blood must be digested in prior to apply into the instrument. Digesting methods, simplified wet ashing and microwave ashing are introduced.

Humans↗
PubMed1996

Uptake of sulfate conjugates by isolated rat hepatocytes.

The uptake of estrone sulfate (E1S; 1 to 400 microM), harmol sulfate (HS; 5 to 900 microM), and 4-methylumbelliferyl sulfate (4MUS; 5 to 1000 microM) was investigated in isolated rat hepatocytes in the presence or absence of inhibitors. Uptake of all of the sulfate conjugates was rapid and exhibited saturation kinetics, best characterized by saturable and nonsaturable (linear transmembrane clearance) transport systems. The KM's were: 16 +/- 6, 123 +/- 28, and 64 +/- 6 microM for E1S, HS, and 4MUS, respectively, with corresponding Vmax's of 0.85 +/- 0.56, 0.48 +/- 14, and 0.42 +/- 0.07 nmol/min/10(6) cells. The nonsaturable uptake clearances, which displayed concentration-independent uptake, were 3 +/- 2, 1 +/- 0.1, 0.5 +/- 0.1 microliter/min/10(6) cells, respectively. Uptake of E1S was inhibited by ouabain (1 mM) and replacement of sodium by choline, whereas HS was insensitive to the addition or substitution. Uptake of both E1S and HS was significantly reduced by metabolic inhibitors (antimycin A, 2.7 microM, rotenone, 30 microM, and KCN, 2 mM) and temperature reduction (from 37 to 27 degrees C). 4,4'-Diisothiocyanostilbene-2-2-'disulfonic acid (2 mM), an inhibitor of anion transport, reduced E1S and HS uptake; E1S uptake was also reduced by HS. HS uptake by both saturable and nonsaturable transport components was depressed by 4MUS (300 microM); the apparent KM was increased by 83% while the Vmax remained unaltered, and the nonsaturable component was decreased by 48%. The data strongly suggest that multiple pathways exist for the uptake of E1S, HS, and 4MUS. E1S uptake is sodium-dependent, requires energy, and is inhibited by anions such as 4,4'diisothiocyanostilbene-2-2'-disulfonic acid and other sulfate conjugates. HS uptake, while being energy dependent, is not sodium dependent, and is inhibited by 4MUS in a competitive fashion. At least one of these pathways is shared.

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗
PubMed1996

Role of cytochrome P450 3A4 in human metabolism of MK-639, a potent human immunodeficiency virus protease inhibitor.

MK-639 (L-735,524) is a potent human immunodeficiency virus protease inhibitor under investigation in the treatment of acquired immunodeficiency syndrome. Five in vitro approaches have been used to identify the cytochrome P450 isoform(s) responsible for the human microsomal oxidative metabolism of MK-639. These approaches are: 1) chemical inhibition; 2) immunochemical inhibition; 3) metabolism by cDNA-expressed human cytochrome P450 enzymes; 4) a correlation analysis; and 5) competitive inhibition of marker activities. Ketoconazole and troleandomycin, both selective inhibitors for cytochrome P450 3A4 (CYP3A4), markedly inhibited the formation of all oxidative metabolites of MK-639; whereas other inhibitors (furafylline, sulfaphenazole, quinidine, S-mephenytoin, and diethyldithiocarbamate) had little effect on MK-639 metabolism. This suggested the involvement of CYP3A4 in MK-639 metabolism. Consistent with this, an anti-rat CYP3A1 rabbit polyclonal antibody, which shows a cross-reactive inhibition of CYP3A4-dependent testosterone 6beta-hydroxylation in human liver microsomes, completely inhibited MK-639 metabolism. Human recombinant CYP3A4 showed a high metabolic activity to form all MK-639 metabolites found in native human liver microsomes. In addition, the formation of individual MK-639 metabolites correlated well with each other and with testosterone 6beta-hydroxylation in 12 different human liver microsomes, whereas no correlation was observed between MK-639 metabolite formation and bufuralol 1'-hydroxylation (or tolbutamide methyl hydroxylation). Furthermore, MK-639 strongly inhibited testosterone 6beta-hydroxylation in a concentration-dependent manner. Kinetic analysis showed that MK-639 is a very potent competitive inhibitor for testosterone 6beta-hydroxylation, with a Ki value of approximately 0.5 mu M. Collectively, these results consistently indicate that CYP3A4 is the isoform responsible for the oxidative metabolism of MK-639 in human liver microsomes.

Anti-Bacterial Agents↗
PubMed1996

[Present status of endoscopic therapy for esophageal varices--endoscopic variceal ligation (EVL)].

Recently, EVL was developed to provide the endoscopist a safer method for the treatment of esophageal varices. The purpose of this report is to reveal our data and the present status. From 1989 to 1995, we performed EVL with EIS using Aethoxysklerol in a total of 214 patients who had bleeding and/or risky esophageal varices. Our technique of EVL is as follows; For prophylactic and elective patients, a spiral ligation method was employed and followed by EIS to obtain the complete eradication. When active bleeding was observed, direct ligation was performed onto the bleeding point and if the bleeding point was not found, spiral ligation method was done. Eradicating effect was observed in 97% of the entire patients and 29 active/spurting bleedings were well controlled by this technique. Complications were experienced in 12 cases (2 stenosis and 10 post EVL ulcedr bleeding), however, there were no patients who could not be managed by endoscopic treatment. Variceal recurrence after EVL/EIS were occurred in 36 of 214 patients (16.8%). Recurrent period was ranged from 1 month to 24 months and mean period was 10 months. We conclude that EVL/EIS is safe, easy and effective mean for the treatment of esophageal varices.

Combined Modality Therapy↗
PubMed1996

Species differences in the pharmacokinetics and metabolism of indinavir, a potent human immunodeficiency virus protease inhibitor.

Indinavir, a potent and specific inhibitor of human immunodeficiency virus protease, is undergoing clinical investigation for the treatment of acquired immunodeficiency syndrome. The studies described herein were designed to characterize the absorption, distribution, metabolism, and excretion of the drug in rats, dogs, and monkeys. Indinavir exhibited marked species differences in elimination kinetics. The plasma clearance was in the rank order: rat (107 ml/min/kg) > monkey (36 ml/min/kg) > dog (16 ml/min/kg). Significant differences in the bioavailability of indinavir also were observed. When given orally as a solution in 0.05 M citric acid, the bioavailability varied significantly from 72% in the dog to 19% in the monkey, and 24% in the rat. These differences in bioavailability were attributed mainly to species differences in the magnitude of hepatic first-pass metabolism. The distribution of indinavir was studied only in rats, both intravenously and orally. Intravenously, indinavir was distributed widely throughout the body. Brain uptake studies showed that indinavir penetrated the blood-brain barrier, but that the penetration was limited. After oral administration, indinavir was distributed rapidly into and out of the lymphatic system. The rapid lymph transfer is of clinical relevance, because a primary clinical hallmark of acquired immunodeficiency syndrome is the depletion of CD4 lymphocytes. Biliary and urinary recovery studies revealed that metabolism was the major route of indinavir elimination in all species, and N-dealkylation, N-oxidation, and hydroxylation seemed to be the major pathways. Although limited to qualitative aspects, the metabolite profile obtained from in vitro microsomal studies generally reflected the in vivo oxidative metabolism of indinavir in all species studies. Results from the chemical and immunochemical inhibition studies indicated the possible involvement of isoforms of the CYP3A subfamily in the oxidative metabolism of indinavir in rats, dogs, and monkeys. This is consistent with our previous studies, which have shown that CYP3A4 is the isoform responsible for the oxidative metabolism of indinavir in human liver microsomes. Furthermore, the in vivo oxidative metabolism of indinavir in rats, dogs, and monkeys was qualitatively similar to that in humans. The high degree of similarity in the metabolite profiles of drug metabolism between animals and humans validates the use of these animal models for toxicity studies of indinavir. Attempts were made to quantitatively extrapolate in vitro metabolic data to in vivo metabolism. With the application of the well-stirred and parallel-tube models, the hepatic clearance and hepatic extraction ratio were calculated using the in vitro Vmax/Km values. In rats, the predicted hepatic clearance (31 ml/ min/kg) and hepatic extraction ratio (0.47) agreed well with the observed in vivo hepatic clearance (43 ml/min/kg) and hepatic extraction ratio (0.68). In addition, the hepatic clearance of indinavir was predicted reasonably well in dogs and monkeys. Based on the in vitro intrinsic clearance of human liver microsomes, a small but significant hepatic first-pass metabolism (ca. 25%) is expected in humans.

Animals↗