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Biomedical subjects

I Fujii

Publications and source records attributed to I Fujii.

At least 73 records · Page 4Linked to original sources
PubMed1995

[Catalytic antibody and its medical applications].

Catalytic antibodies will not only provide new insight into the general potential of natural enzymes, but may also afford novel catalysts to facilitate reactions not catalyzed by natural enzymes. One goal of studying catalytic antibodies is to generate tailor-made catalysts for applications in medicine. An example of a possible use with catalytic antibodies is related to the action of prodrugs. In the rational design of prodrugs, it is necessary to consider (a) what structural modifications of the parent molecule are necessary to reduce or eliminate the particular undesirable effects, and (b) what enzymes are available in vivo to regenerate the parent molecule from the prodrug. However, the design of structurally related analogues of a parent molecule is limited by the enzyme's specificity, the type of reaction catalyzed, and the enzyme distribution and level. Using catalytic antibody technology for the novel design of prodrugs allows for effective structural modifications of the molecule in question. It is also a valuable aid in overcoming the problem of drug delivery by using bi-functional chimeric antibody technology, through which site-specific antibodies are combined with antibodies catalyzing reactions that cannot be accomplished by natural enzymes in vivo. In this review, we describe the first example of prodrug activation via catalytic antibodies as well as future aspects of catalytic antibodies of medical interest.

Anti-Bacterial Agents↗
PubMed1995

Evaluation of binding equation method for prediction of unbound serum valproic acid concentration in pediatric patients with epilepsy.

We retrospectively evaluated the ability of Scatchard binding equation method to predict the unbound serum valproic acid (VPA) concentration in 37 pediatric patients with epilepsy receiving VPA monotherapy. The correlation between predicted and observed unbound serum concentrations was high and significant (r = 0.873, p < 0.001). Mean prediction error (ME), mean absolute prediction error (MAE), and root mean squared error (RMSE) were calculated, and served as a measure of prediction bias and precision. The 95% confidence intervals of ME did not include 0, showing a bias to overpredict unbound concentration. The MAE and RMSE were not small in magnitude (MAE 17.4 mumol/l, RMSE 22.8 mumol/l). The current method using the in vivo population mean binding parameters from healthy young adults may be limited in the predictive performance of unbound serum VPA concentration.

Adolescent↗
PubMed1994

A common ancestry for multiple catalytic antibodies generated against a single transition-state analog.

Immunization with a single haptenic transition-state analog generates a few catalytic antibodies among the dozens of antibodies capable of binding the hapten. The diversity of the immune response has raised some fundamental issues, such as How do catalytic and noncatalytic antibodies differ on a structural basis? To address this issue, the variable region primary sequences of 11 antibodies (including 6 catalytic and 5 noncatalytic antibodies) elicited against a single haptenic transition-state analog were deduced from cDNA sequences. Cluster analyses using phylogenetic trees constructed by the neighbor-joining method have revealed that the amino acid sequences of noncatalytic antibodies bear no relationship to one another, while the catalytic antibodies share significant structural identity. Furthermore, no catalytic antibodies possessing amino acid sequences with high homology to those of noncatalytic antibodies were detected. Five catalytic antibodies examined showed 89-95% and 74-84% sequence homologies in the complete light- and heavy-chain variable regions, respectively. Thus, it seems likely that the catalytic antibodies elicited against a single hapten use the canonical set of variable region genes. Interestingly, one catalytic antibody showed only limited sequence similarity to the other catalytic antibodies and was found to exhibit a distinctly different substrate specificity. From the broad range of their binding constants to the hapten, it is unlikely that highly homologous catalytic antibodies are generated as a result of simple high-affinity choices. These results emphasize the utility of rationally designed transition-state analogs for the induction of antibody molecules with catalytic activity.

Amino Acid Sequence↗
PubMed1994

Development of a novel method for determination of acetyl-CoA:1-alkyl-sn-glycero-3-phosphocholine acetyltransferase activity and its application to screening for acetyltransferase inhibitors. Inhibition by magnolol and honokiol from Magnoliae cortex.

A method was developed for determining the activity of acetyl-CoA:1-alkyl-sn-glycero-3-phosphocholine acetyltransferase (EC 2.3.1.67), a key enzyme in the biosynthesis of platelet-activating factor (PAF, 1-alkyl-2-acetyl-sn-glycero-3-phosphocholine). The assay involves measurement of the radioactivity in the trichloroacetic acid (TCA)-precipitated complex of radioactive product and albumin after incubation of 1-alkyl-sn-glycero-3-phosphocholine and [3H]acetyl-CoA with rat spleen microsomes or membrane fractions of human polymorphonuclear leukocytes (PMNs). The radioactive product associated with the precipitate was identified as PAF using an ultrahigh-sensitivity TV camera system after extraction and separation by TLC. This TCA method was then used to screen the components of crude preparations that inhibited acetyltransferase activity. Major components from the cortex of Magnoliae (magnolol and honokiol), which have anti-inflammatory and anti-bacterial actions, inhibited the acetyltransferase activity in rat spleen microsomes (IC50, 150 and 150 microM, respectively) and membrane fractions of human PMNs (IC50, 70 and 60 microM, respectively). The inhibitory action of magnolol and honokiol was reversible, and similar to or higher than that of nordihydroguaiaretic acid. PAF production in human PMNs stimulated by the ionophore A23187 was also suppressed dose dependently by magnolol and honokiol. These activities may be relevant to the claimed therapeutic effects of the extract from Magnoliae cortex.

Acetyltransferases↗
PubMed1994

Design of the hapten for the induction of antibodies catalyzing aldol reaction.

The transition states of the aldol reaction and their analogues are reported here for the design of the haptens generating catalytic antibodies. The structural and the electrostatic properties were calculated by an ab initio molecular orbital method and were compared using a graphic software. Two transition states that lead to the corresponding stereo isomers of the aldol products were characterized. Also, the suitable transition state analogues were found. It is suggested that the stereo selectivity can be controlled using the catalytic antibodies elicited against the haptens designed here.

Antibodies, Catalytic↗
PubMed1994

Incidence and risk factors in neuroleptic malignant syndrome.

Between October 1981 and September 1989, 564 patients admitted to the Department of Psychiatry of the Hospital of Oita Medical University received antipsychotic treatment. Ten (1.8%) of these patients developed neuroleptic malignant syndrome. This incidence is significantly greater than that reported in a group of other studies (27/17,811, 0.2%). We compared the characteristics of this sample with known risk factors for neuroleptic malignant syndrome. It is possible that Japanese people are at an increased risk for developing neuroleptic malignant syndrome. Psychomotor excitement, refusal of food, a weight loss of greater than 1 kg per week and oral administration of haloperidol at 15 mg/day or above were found to be risk factors for developing the neuroleptic malignant syndrome.

Adolescent↗
PubMed1994

Nucleotide sequence of the aknA region of the aklavinone biosynthetic gene cluster of Streptomyces galilaeus.

A 3.4-kb BamHI fragment that is assumed to be a part of the aklavinone biosynthetic gene cluster of Streptomyces galilaeus 3AR-33 and contains the genes required for the early stage of polyketide biosynthesis was sequenced. The nucleotide sequence of the region that hybridizes to the actIII probe reveals the presence of a gene, aknA, whose deduced protein product is very similar to the ActIII protein and other known oxidoreductases. The predicted AknA protein is believed to be responsible for catalyzing the reduction of the keto group at the ninth carbon from the carboxyl terminus of the assembled polyketide to the corresponding secondary alcohol. The predicted AknA protein has a calculated molecular mass of 27,197 Da (261 amino acids) and the highly conserved sequence Gly-Xaa-Gly-Xaa-Xaa-Ala commonly seen in oxidoreductases. Cloning and sequence analysis of the aknA region of the 2-hydroxyaklavinone-producing strain S. galilaeus ANR-58 identified an alteration in the gene, confirming that the aknA gene is essential for aklavinone biosynthesis.

Amino Acid Sequence↗
PubMed1994

[Drug-induced organic mood disorders].

Firstly, we reviewed the drugs (ex, corticosteroid, interferon) which were thought to be related to organic mood disorders in the literature, and discussed a number of problems with assessment of drug-induced mental disorders. Next, we investigated 1) the antidepressant-induced switch rate from depression to mania and 2) the latency from administration to manic onset in three groups (Major depression: single episode, Major depression: recurrent, Bipolar disorder: depressed). Our data showed, 1) the switch rate was 17.2, 14.3, 32.0% and 2) the latency to onset was 69.9, 65.9, 37.1 days, respectively. The latency to onset showed no significant difference in three groups. (ANOVA, p < 0.05) Clomipramine, amitriptyline, imipramine, and dosulepin had the higher switch rate above 10%.

Antidepressive Agents↗
PubMed1994

In vivo binding characteristics of carbamazepine and carbamazepine 10, 11-epoxide to serum proteins in monotherapy adult patients.

The in vivo serum protein binding characteristics of carbamazepine and carbamazepine 10, 11-epoxide, which was the main metabolite of carbamazepine in plasma, were assessed in sera from 30 adult patients with epilepsy on carbamazepine monotherapy. The binding characteristics of each compound were analyzed according to the two-site binding model. Association constants to the high-affinity binding site on alpha 1-acid glycoprotein (AAG) were 0.053 l/mumol for carbamazepine and 0.013 l/mumol for carbamazepine 10, 11-epoxide. The maximum binding capacities for drug-AAG binding were 49.2 mumol/l for carbamazepine and 48.1 mumol/l for carbamazepine-10, 11-epoxide. The products of the association constant and binding capacity for the lower-affinity site (i.e., the linear component of albumin binding site) were 1.273 for carbamazepine and 0.525 for carbamazepine 10, 11-epoxide. Within the total concentration range of each compound investigated, the contribution of drug-AAG binding to the total serum binding was relatively larger than that of drug-albumin binding.

Adolescent↗
PubMed1994

[Adverse effects of antidepressants and anti-manics/mood stabilizers].

Information on the adverse effects of antidepressants and antimanics/mood stabilizers is presented. The adverse effects of antidepressants include pharmacological side-effects, toxic effects, interactions with other drugs, withdrawal syndrome and "jitteriness" syndrome. The most common adverse effects with lithium treatment are polyuriapoly-dipsia, hypothyroidism, gastrointestinal symptoms, psychological complaints and ECG changes. The adverse effects of carbamazepine and valproate are mild and readily manageable, however problems associated with the hepatic system, the hematopoietic system and the thyroid gland, are important. It seems that no major deficits have been identified in studies of tricyclic antidepressant teratogenicity. Lithium, carbamazepine and valproate carry significant teratogenic effects.

Abnormalities, Drug-Induced↗
PubMed1993

Prodrug activation via catalytic antibodies.

Prodrug activation via antibodies was examined by using the antibiotic chloramphenicol as a model drug. Based on the conformational change between substrate and product, this antibody-catalyzed reaction was designed to prevent product inhibition, thus enhancing turnover. Antibodies elicited against a phosphonate transition-state analogue were found to catalyze hydrolysis of a nonbioactive chloramphenicol monoester as a prodrug at a significantly higher rate above the uncatalyzed background reaction to regenerate chloramphenicol as a parent molecule. The antibody-catalyzed prodrug activation was tested by the paper-disc diffusion method using Bacillus subtilis as an indicator strain. The antibody 6D9 catalyzes the reaction with multiple turnover to generate enough chloramphenicol to inhibit bacterial growth, as indicated by a clear inhibitory zone after incubation with monoester. Using the same method, no inhibition was detected by incubation of either the monoester or the antibody alone. This result reveals that only the antibody hydrolytically activates the monoester, which can be expected to be a suitable prodrug, as it is resistant to the action of bacterial hydrolytic enzymes. The approach in this study demonstrates the use of catalytic antibody technology in medicine and may be applicable to drugs with undesirable effects, particularly in the field of cancer therapy.

Animals↗
PubMed1993

In vivo determinations of carbamazepine and carbamazepine-10, 11-epoxide binding parameters to serum proteins in monotherapy patients.

The in vivo serum protein binding parameters of carbamazepine (CBZ) and carbamazepine-10, 11-epoxide (CBZ-E), which was the main metabolite of CBZ in plasma, were determined in sera from 27 patients on CBZ monotherapy. Based on the results by recent studies, the authors assumed that CBZ and CBZ-E binding to serum proteins were composed of specific binding sites on alpha 1-acid glycoprotein (AAG) and albumin. Therefore, the authors determined the binding parameters of each compound by specific binding equation for two proteins. Association constants for drug-AAG binding were .071 L/mumol for CBZ and .016 L/mumol for CBZ-E. Conversely, those for drug-albumin binding were .00052 L/mumol for CBZ and .00072 L/mumol for CBZ-E. Within the investigated total concentration ranges in each compound, the AAG binding contributes largely to the drug-serum protein interactions. Furthermore, our results indicate that the albumin binding contributes to the nonsaturable serum protein binding of these compounds in the therapeutic range.

Adolescent↗
PubMed1993

Effect of unbound clearance on binding parameters of valproic acid to serum proteins.

Nine healthy subjects received 400 mg sodium valproate orally in the fasting state. Binding parameters of valproic acid to serum proteins were determined by Scatchard analysis for individual series of valproic acid data. Total and unbound (intrinsic) clearances (CLt and CLu) were calculated by dividing the dose by the appropriate area under the serum drug concentration-time curve. Unbound clearance correlated positively with the product of association constant (Ka) and concentration of free protein ((P)) (P < .05). Conversely, no significant correlation was found between CLt and binding parameters. The average unbound concentration correlated negatively with both CLu and ka(P) values. The result indicates an effect of CLu on Ka(P) value of valproic acid.

Administration, Oral↗
PubMed1993

In vivo binding characteristics of carbamazepine and carbamazepine-10,11-epoxide to serum proteins in paediatric patients with epilepsy.

The in vivo serum protein binding characteristics of carbamazepine (CBZ) and carbamazepine-10,11-epoxide (CBZ-E) were assessed in sera from 23 paediatric patients on CBZ monotherapy. We assumed that CBZ and CBZ-E binding to serum proteins comprised specific binding sites on alpha 1-acid glycoprotein (AAG) and non-specific binding sites on serum albumin. Therefore, the binding characteristics of each compound were analysed according to specific and nonspecific binding equations. Association constants for drug-AAG binding were 0.096 l.mumol-1 for CBZ and 0.023 l.mumol-1 for CBZ-E. Within the concentration ranges investigated the specific binding of each compound contributes to the drug-serum protein interactions. Age did not show a significant correlation with the serum unbound fraction of each compound.

Adolescent↗
PubMed1993

X-Neu5Ac: a novel substrate for chromogenic assay of neuraminidase activity in bacterial expression systems.

A chromogenic substrate 1, 5-bromo-4-chloroindol-3-yl 5-acetamido-3,5-dideoxy-alpha-D-glycero-D-galacto-2-nonulopyranosidon ic acid (X-Neu5Ac), has been synthesized to facilitate the screening of bacterial colonies or plaques for the detection of either natural or mutant neuraminidase activity. Substrate 1 was hydrolyzed by neuraminidase isolated from Clostridium perfringens to release a halogenated indol-3-ol 2 that undergoes rapid aerobic oxidation to form the dark blue pigment, 5,5'-dibromo-4,-4'-dichloroindigo 3. Preliminary kinetic studies indicate that this compound is a good substrate (Km 0.89 x 10(-3) M) for neuraminidase and is quite stable under identical conditions in the absence of enzyme. These results suggest that X-Neu5Ac 1 can be useful to screen for bacterially-encoded enzyme production directly on agar plates.

Chromogenic Compounds↗
PubMed1993

Neuroleptics induce penile erection in the rabbit.

1. Intramuscular (i.m.) administration of the neuroleptics chlorpromazine, haloperidol and spiperone at doses ranging from 0.1 to 0.4 mg/kg in male rabbits induced a dose-dependent penile erection. 2. The i.m. administration of the alpha 1-adrenoceptor antagonists prazosin and bunazosin (0.1-0.4 mg/kg), induced a dose-dependent penile erection. However, that of the peripheral dopamine receptor antagonist domperidone (0.4-4.0 mg/kg) and the dopamine receptor agonist apomorphine (0.1-1.0 mg/kg) did not. Penile erection was not induced by i.m. injection of chlorpromazine in combination with intrapenile administration of the alpha 1-adrenoceptor agonist methoxamine. 3. Penile erection was induced by the administration of chlorpromazine (0.25-1.00 mg/body) into the lateral cerebral ventricle. At a low dose, however, the administration of chlorpromazine into the lateral ventricle induced a less notable penile erection than that induced intramuscularly. 4. Penile erection was induced by i.m. injection of the ganglionic blocker hexamethonium (5-20 mg/kg). When chlorpromazine was given after pretreatment with hexamethonium, penile erection was more notable than that induced by either drug given alone. 5. These results suggest that neuroleptics could act locally in the penile structure to cause penile erection by alpha 1-adrenoceptor-blocking actions.

Adrenergic alpha-Agonists↗
PubMed1993

Can the serum protein binding of valproic acid limit the hepatic elimination?

In the previous study, we determined the in vivo binding parameters of valproic acid to serum proteins in seven healthy young adults at steady-state. In this study, we determined the effects of serum protein binding on hepatic elimination with the use of observed data obtained from our previous study of valproic acid. A regression analysis between the binding parameters and the pharmacokinetic parameters was performed. In addition, the relationship between each pharmacokinetic parameter was also analyzed. The order of association constant (K) for valproic acid-serum protein was 10(-2) l/mumol. No significant correlation was found between the binding parameters and the rate of elimination. On the other hand, the average unbound serum concentration was found to be a significantly negative correlation with the unbound (intrinsic) clearance (p = 0.0082). The product of association constant and concentration of free protein (P) correlated positively with the unbound clearance (p = 0.0233) and negatively with the average unbound and total serum concentrations (p = 0.0021 and p = 0.0029, respectively). The results indicate that the membrane permeability of valproic acid is high and that the increase of unbound clearance accompanies directly the decrease of the average unbound and total serum concentrations. Consequently, the KP values are proportional to the unbound clearance due to the rapid changes of the concentration of free protein. Therefore, the dissociation of the valproic acid-serum protein complex is not a rate-limiting factor for hepatic elimination and hence the serum protein binding cannot limit the ability of the liver to extract drug from blood.

Adult↗