Biological and Pharmaceutical Bulletin
Online ISSN : 1347-5215
Print ISSN : 0918-6158
ISSN-L : 0918-6158
19 巻 , 2 号
選択された号の論文の38件中1~38を表示しています
  • Hirofumi INOUE, Yuuko DATE, Kazuya KOHASHI, Hironori YOSHITOMI, Yasuto ...
    1996 年 19 巻 2 号 p. 163-166
    発行日: 1996/02/15
    公開日: 2008/04/10
    ジャーナル フリー
    A method to measure total hydroxyproline (Hyp) and proline (Pro) in human serum and urine by HPLC was developed. Hyp and Pro in acid hydrolysates of serum and urine were derivatized with 4-(5, 6-dimethoxy-2-phthalimidinyl) phenylsulfonyl chloride after treatment with o-phthalaldehyde and cleanup on Bond Elut C18 column. The derivatives of imino acids were separated on a reversed phase column by gradient elution with acetonitrile and phosphate buffer (1 mmol/l, pH 7) and detected by fluorescence measurement at 315 nm (excitation) and 385 nm (emission). Detection limits for both Hyp and Pro were 10 fmol per injection. The within-day and day-to-day relative standard deviations for Hyp and Pro in serum and urine were less than 3.19%. the recoveries of Hyp and Pro added to serum and urine were about 100%. The present method was applied to determine total Hyp and Pro in serum and urine from normal subjects and patients with chronic renal failure.
  • Shigetoshi EDA, Kiyomi KIKUGAWA, Masatoshi BEPPU
    1996 年 19 巻 2 号 p. 167-175
    発行日: 1996/02/15
    公開日: 2008/04/10
    ジャーナル フリー
    The characteristics of the binding of human lactoferrin (LF) to the cells of a human monocytic leukemia cell line, THP-1, were investigated. 125I-Labeled LF (125I-LF) bound to THP-1 cells, and the binding increased markedly as the cells matured into macrophages (THP-1 macrophages) by stimulation with phorbol 12-myristate 13-acetate. Scatchard analysis of the binding of 125I-LF to THP-1 macrophages indicated that high and low affinity receptor sites (Kd=0.57×10-6 and 3.7×10-6M, respectively) are present on the cells. The number of these high and low affinity receptor sites were 2.4×106, and 2.5×106 per cell, respectively. Removal of iron from 125I-LF did not affect its binding to THP-1 macrophages, indicating that the binding is not dependent on Fe (III) ion. The binding of the labeled LF to THP-1 macrophages was markedly decreased following acetylation, suggesting that the amino residues of the polypeptide portion of LF play a major role in the binding. The binding of labeled LF was partially inhibited by the isolated whole oligosaccharides of LF, and by the isolated whole oligosaccharides of band 3 glycoprotein of human erythrocyte membrane which contain poly-N-acetyllactosaminyl saccharide chains, like the LF oligosaccharides. Their inhibitory activity did not depend on the terminal sialyl residues of the saccharide chains. Lacto-N-fucopentaose III and lacto-N-neotetraose, and analogous structure being present in the poly-N-acetyllactosaminyl chains of LF, also partially inhibited the binding of 125I-LF to the THP-1 macrophages. When poly-N-acetyllactosaminyl saccharide chains of 125I-LF were cleaved by endo β-galactosidase, the binding of 125I-LF was partially reduced. These results suggest that binding of LF to THP-1 macrophages is primarily mediated by its protein component, but a short oligosaccharide structure, possibly Galβ1-4GlcNAcβ1-3Gal, which is contained in the nonreducing terminal region of poly-N-acetyllactosaminyl saccharide chains of LF and band 3, and in lacto-N-fucopentaose III and lacto-N-neotetraose is also recognized by THP-1 macrophages, and this recognition partly contributes to the binding of LF to cells.
  • Michinao MIZUGAKI, Akihiko HIROSE, Hideaki SUZUKI, Koichi MIURA, Kiyot ...
    1996 年 19 巻 2 号 p. 176-181
    発行日: 1996/02/15
    公開日: 2008/04/10
    ジャーナル フリー
    The subcellular distribution of 2, 4-dienoyl-CoA reductase (EC. 1.3.1.34, DCR) in rat liver was studied biochemically and immunochemically after the induction of clofibrate. DCR activity was mainly detected in the mitochondrial fraction by sucrose density gradient centrifugation in the livers of both normal and clofibrate-treated rats. It was also shown that the polyclonal antibody against purified DCR detected the enzyme in the mitochondrial fraction. However, the antibodies, which were affinity purified using the purified mitochondrial DCR or were epitope-selected using the fusion-polypeptide expressed from the mitochondrial cDNA clone (λgt11-RDR181), were cross-reacted with the peroxisomal DCR. These results suggest that peroxisomal DCR is immunochemically indistinguishable from mitochondrial DCR.
  • JongBeak PARK, Lisa IMAMURA, Kyoichi KOBASHI
    1996 年 19 巻 2 号 p. 182-187
    発行日: 1996/02/15
    公開日: 2008/04/10
    ジャーナル フリー
    Urease is an important virulence factor of pathogenicity of gastric Helicobacter pylori. The inhibition of H. pylori urease by the novel proton pump inhibitor, rabeprazole, was investigated kinetically. It was found to act as an irreversible noncompetitive inhibitor of the enzyme. The inhibitory potency of rabeprazole was dependent on the pH of reaction mixture and its Ki values were 0.14μM (pH 5.0), 0.34μM (pH 7.0) and 6.10μM (pH 8.5). Progressive inactivation of urease by rabeprazole initially proceeded according to pseudo-first-order kinetics with respect to the remaining enzymatic activity at pH 7.0 and 37°C, with a second-order rate constant of 0.0017μM-1 s-1. When the inactivation half-life was plotted versus the reciprocal of the rabeprazole concentration, a straight line was obtained with a slope of -3.12. From an Arrhenius-plot of the temperature-dependence of the inactivation (over the range of 5-37°C), an activation energy of 13.2 kcal/mol was calculated. Recovery of activity was incomplete for H. pylori urease inhibited by rabeprazole, suggesting that the rabeprazole-urease complex is very stable.
  • Masatoshi BEPPU, Shigetoshi EDA, Mayumi FUJIMAKI, Eisuke HISHIYAMA, Ki ...
    1996 年 19 巻 2 号 p. 188-194
    発行日: 1996/02/15
    公開日: 2008/04/10
    ジャーナル フリー
    The cells of the human monocytic leukemia cell line THP-1 differentiated into macrophages bound to human erythrocytes oxidized with adenosine 5'-diphosphate (ADP)-Fe3+ chelate (ADP/Fe3+) in the absence of serum. The binding was prevented when the cells were treated with ADP/Fe3+ in the presence of antioxidants, indicating that oxidation of the cells is responsible for the increased susceptibility to the THP-1 cell binding. Galactose, fucose, mannose and mannan partially inhibited the binding. Glycoproteins containing poly-N-acetyllactosaminyl saccharide chains such as band 3 glycoprotein isolated from human erythrocyte membrane and lactoferrin, and their oligosaccharides, strongly inhibited the binding. On the other hand, glycoproteins with non-poly-N-acetyl-lactosaminyl saccharide chains such as glycophorin A isolated from the erythrocyte membrane, fetuin and α1-acid glycoprotein, little or partially inhibited the binding. The inhibitory activity of band 3 oligosaccharides and lactoferrin oligosaccharides was little affected by treatment with endo-β-galactosidase, which specifically cleaves poly-N-acetyllactosamine to shorter oligosaccharides. Removal of the nonreducing terminal region of the saccharide chains of band 3 on the eythrocyte surface by treatment of the cells with endo-β-galactosidase resulted in a decrease in the susceptibility of the cells to the THP-1 cell binding. These results suggest that THP-1 cells which have been differentiated into macrophages bind the oxidized erythrocytes primarily through the recognition of poly-N-acetyllactosaminyl saccharide chains of band 3, and the site of the recognition exists in the nonreducing terminal region of the saccharide chains. Clustering of band 3 molecules is proposed as a possible alteration of oxidized erythrocyte membrane which promotes the interaction of the saccharide receptor on THP-1 cells with the saccharide chains of band 3 on erythrocytes.
  • Kentaro SUGIYAMA, YingLi QU, Keiko MARUYAMA, Kaoru HATTORI, Kenichi WA ...
    1996 年 19 巻 2 号 p. 195-198
    発行日: 1996/02/15
    公開日: 2008/04/10
    ジャーナル フリー
    The dissociation rate of amlodipine ((±)-3-ethyl 5-methyl 2-[(2-aminoethoxy) methyl]-4-(o-chlorophenyl)-1, 4-dihydro-6-methyl-3, 5-pyridinedicarboxylate benzenesulfonate) from rat heart and brain membranes preincubated with drugs and washed out with buffer was assessed by radioligand binding assay using 3H-PN200-110 as a radioligand. The remaining KCl-induced contraction in rat aortic strips washed out after treatment with this drug and the pKi (inhibition constant) values of the drug were compared with those of nisoldipine, nifedipine, manidipine and benidipine. The inhibition of 3H-PN200-110 binding induced by nifedipine was reversed by washing, whereas that induced by amlodipine, manidipine, and benidipine was not readily reversed under these conditions. When rat aortic strips were pretreated with Ca2+ antagonists, the rank order of the inhibition of contractions induced by 50mM KCl was manidipine=benidipine>amlodipine>nisoldipine>nifedipine, even though Ca2+ antagonists were not present in the extracellular medium. The pKi values of amlodipine in the heart and brain were 6.86 and 7.41, respectively, and these values were lower than those of the other Ca2+ antagonists. There was a good correlation between the potency of the inhibition of 3H-PN200-110 binding by drugs after the washout of membranes and the inhibition exerted by the drugs in contractions induced by 50mM KCl after the washout of tissues, although this residual inhibition was not correlated with pKi values. Thus, these results suggest that amlodipine has a very slow rate of dissociation from 3H-PN200-110 binding sites, as do manidipine and benidipine, and this property may explain its long-lasting antihypertensive effect.
  • Shinjiro KOBAYASHI, Bao LUO, Motonori OKABE, Ikuko KIMURA, Masayasu KI ...
    1996 年 19 巻 2 号 p. 199-202
    発行日: 1996/02/15
    公開日: 2008/04/10
    ジャーナル フリー
    The influence of the diabetic state on the number and activity of intraperitoneal macrophages was investigated in the GK rat. This rat is a model of non-insulin-dependent diabetes mellitus and was established from the normal Wistar rat. The cell number of a macrophages preparation in diabetic GK rats (10-15 weeks of age) did not differ from that in normal Wistar rats (9-10 weeks of age). The co-cultured macrophages in diabetic GK rats (9-12 weeks of age) significantly increased the tube formation of aortic endothelial cells (EC) compared with that in age-matched Wistar rats. The conditioned medium obtained from a culture of macrophages from GK rats also increased tube formation more than that from a culture of macrophages from Wistar rats. The effect of macrophages in the GK rat was not influenced by interferon-γ (160 and 460pM), but macrophages in normal rats were activated by these concentrations of interferon-γ. In conclusion, the diabetic state increased the activity but not the number of macrophages in the GK rat, to stimulate tube formation of EC. The tube-forming effect of macrophages in the diabetic state may depend on released factors which differ from that of normal macrophages.
  • Abu Bakar Abdul MAJEED
    1996 年 19 巻 2 号 p. 203-208
    発行日: 1996/02/15
    公開日: 2008/04/10
    ジャーナル フリー
    A possible anti-anesthetic effect of idazoxan using the depth versus latency of cortical cellular response and somatosensory evoked potentials as indices of anesthesia was studied. With the administration of 10mg/kg (i.p.) idazoxan, a potent and selective α2-adrenoceptor antagonist, to an anesthetized rat with 1.25-1.5g/kg (i.p.) urethane, the modal latency of somatosensory cortical responses to electrical stimulation of the forepaw (0-90V, 1Hz) was shortened to 87±3.6% (mean±S.D.; n=3) of the baseline value. The number of units firing increased by 259±98.5% (n=3). The combined parameter (1/L×Pi ; L, latency ; Pi, initial positive wave) of the somatosensory evoked potentials was enhanced to 125.0±16.2% (n=19) versus saline (98.9±25.6% ; n=18) during the desynchronized electroencephalogram (EEG). The initial negative component (Ni) of the somatosensory cortical response was increased to 192.0±83.1% (n=19) and 134.8±36.9% (n=19) during the synchronized and desynchronized EEG, respectively. Thus idazoxan appears to produce effects resembling a "lightening of anesthesia."This may provide the impetus for further studies on the possibility of using α2-adrenoceptor antagonists in the recovery from certain types of anesthetic agents.
  • Miyuki SHIMOJI, Yoko ANIYA, M.W. ANDERS
    1996 年 19 巻 2 号 p. 209-213
    発行日: 1996/02/15
    公開日: 2008/04/10
    ジャーナル フリー
    Proteolytic activation of oxidatively modified microsomal GSH S-transferase (GSTm) was investigated. When GSTm was incubated with diamide [diazenedicarboxylic acid bis (N, N-dimethylamide)] or hydrogen peroxide in the presence or absence of glutathione, a protein-glutathione mixed-disulfide and a dimer of the enzyme were formed with a concomitant increase in transferase activity. Although control GSTm was activated 3.4-fold by 3μg/ml of trypsin, the monomeric form of the transferase in which the sulfhydryl group was modified by mixed-disulfide bond formation or by covalent binding with N-ethylmaleimide was futher stimulated by lower concentrations of trypsin than that used in the control. In contrast, no activation of the dimeric transferase was observed with any concentration of trypsin. In immunoblot analysis, a proteolytic product (fragment A) from the dimer transferase was detected after treatment of oxidant-modified microsomes with low concentrations of trypsin, whereas the fragment (fragment B) from the unmodified-monomeric enzyme was observed by high concentrations of trypsin. These results show that oxidatively modified GSTm is sensitive to proteolytic attack by trypsin and that only monomeric transferase is further activated by limited proteolysis.
  • Takaya NITTA, Takashi HOSHINO, Masao KOIDA, Hiromichi NAKAMUTA
    1996 年 19 巻 2 号 p. 214-216
    発行日: 1996/02/15
    公開日: 2008/04/10
    ジャーナル フリー
    It was examined histomorphometrically whether or not and how chronic treatment with nasal salmon calcitonin (SCT) of rats could prevent ovariectomy (OVX)-induced osteoporotic changes in the trabecular bone of tibia. During 7 weeks on a synthetic low Ca diet after OVX, rats developed type 1 osteopenia which was defined by strut analysis as resulting mainly from loss in the connectivity of strut but in the thickness. An intermittent dosing regimen of nasal SCT (10 or 40U/rat/d, 3d/week, for 7 weeks) was able to retard development of osteopenia in a dose-dependent manner. The results indicate that the nasal route would be usable for chronic treatment of experimental osteoporosis with SCT and possibly other peptide anti-osteoporotics.
  • Hiromichi NAKAMUTA, Takaya NITTA, Takashi HOSHINO, Masao KOIDA
    1996 年 19 巻 2 号 p. 217-219
    発行日: 1996/02/15
    公開日: 2008/04/10
    ジャーナル フリー
    A glucocorticoid-induced osteopenia (GC-OP) model was developed in Wistar male rats to use for in vivo screening of anti-osteoporotic candidates. 1) Two week treatment with a wide dose range of methylprednisolone acetate (0.01 to 50mg/kg, s.c., 3d a week) and histomorphometry of the right tibia combined with histological study (n=3) allowed us to select 0.1mg/kg as a proper dose to produce a measurable degree of osteopenia. 2) Eight week treatment with the selected dosing regimen (n=6) of the steroid and histomorphometry including strut analysis measured the development of a characteristic osteopenia which can be described briefly as "uncut but thinning"of trabecular bone structure, and which was prevented by salmon calcitonin (0.33-10U/kg, s.c., 5 times/week) in a dose-dependent manner. The results indicate the usability of this osteopenic model not only for screening of antiosteoporotics but also for study of the mechanism leading to GC-OP.
  • Pascal COUDERT, Fernand LEAL, Eliane DUROUX, Catherine RUBAT, Jacques ...
    1996 年 19 巻 2 号 p. 220-223
    発行日: 1996/02/15
    公開日: 2008/04/10
    ジャーナル フリー
    Ascorbic acid, present in plasma from humans at concentrations of 50 to 200μmol/l, has multiple antioxidant properties. Structural modification of this vitamin by the introduction of lipophilic moieties has allowed to the development of ascorbate esters and ethers active as free radical quenchers. Thus, a new series of ascorbic acid analogues possessing one or two aromatic rings was prepared in an attempt to synthesize potent antioxidants with lipophilic properties. Substituted 3-hydroxy furan-2 (5H)-ones and in some cases, dihydrofuro [3, 4-b] pyrones were prepared. The synthesized compounds were evaluated for their antioxidant activity in vitro. So, 4-(4-methoxybenzoyl)-3-hydroxy-5-phenylfuran-2 (5H)-one 3e (IC50=3.06×10-4M) was found to be the most effective in scavenging the superoxide anion, whereas 4-benzoyl-3-hydroxy-5-(3, 4-dimethoxyphenyl) furan-2 (5H)-one 3d (IC50=1.38×10-4M) was the most active in inhibiting, lipid peroxidation.
  • Hisashi WAKITA, Yoshiki TOKURA, Fukumi FURUKAWA, Masahiro TAKIGAWA
    1996 年 19 巻 2 号 p. 224-227
    発行日: 1996/02/15
    公開日: 2008/04/10
    ジャーナル フリー
    The effects of the macrolide antibiotic, roxithromycin (RXM) on immunological functions of cultured normal human keratinocytes (NHK) were studied. RXM neither modulated the expression of intercellular adhesion molecule-1 (ICAM-1) and human histocompatibility leukocyte antigen (HLA)-DR nor mobilized intracellular free calcium in NHK that were cultured in medium alone. However, the ICAM-1 and HLA-DR expression induced by cultivation with interferon-γ (IFN-γ) was upregulated and suppressed, respectively, by pretreatment of NHK with RXM. Whereas ICAM-1 upregulation was observed with RXM at a concentration as high as 0.1mM, the inhibition of HLA-DR expression by RXM was virtually dose-dependent at doses ranging from 100nM to 0.1mM. Accessory cell function of major histocompatibility complex (MHC) class II-bearing NHK in terms of the T-cell response to staphylococcal enterotoxin B was suppressed by pretreatment of NHK with RXM, indicating the functional consequence of RXM-induced reduction of MHC class II molecules. Furthermore, RXM inhibited IFN-γ-mediated upregulation of IL-1α secretion by NHK. These results show that RXM suppresses immunological functions of keratinocytes triggered by IFN-γ and suggest the therapeutic relevance of RXM to T cell-mediated inflammatory skin diseases and T cell malignancies such as atopic dermatitis, psoriasis and cutaneous T cell lymphoma, which can be exacerbated by keratinocytes immunologically modulated by exposure to IFN-γ.
  • Masahiro NAGAI, Mariko NOGUCHI, Tohru IIZUKA, Kazuyo OTANI, Katsuo KAM ...
    1996 年 19 巻 2 号 p. 228-232
    発行日: 1996/02/15
    公開日: 2008/04/10
    ジャーナル フリー
    A potent vasodilator substance (compound III), [α]D +141°, was isolated from salviae miltiorrhizae radix (dan-shen). This substance was determined to be des (α-carboxy-3, 4-dihydroxyphenethyl) lithospermic acid on the basis of spectrometric and chemical evidence, and was identified with an authentic sample of 8-epiblechnic acid. However, comp. III seemed to be formed from lithospermic acid (LSA) and LSA-B during a chemical procedure to separate active ingredients. It caused a sustained, slowly developing relaxation of rat aortic strips precontracted with norepinephrine (NE) in preparations with or without endothelium. The NE-induced concentration-dependent contraction of aortic strips was significantly attenuated by pretreatment with comp. III. Concentration-response curves for Ca2+-induced contracture of depolarized aortic strips with isotonic high K+ (60mM) were not affected by comp. III. Ca2+-induced contraction of aortic strips, preincubated with 10-6M NE in the presence of 10-6M nicardipine and 0.01mM EGTA in Ca2+-free solution, was slightly inhibited by comp. III. Pretreatment of aortic strips with comp. III slightly inhibited the phorbol ester (PMA)-induced contraction. These results suggest that comp. III inhibits NE-induced contraction of the aortic strips through reduction in Ca2+ mobilization. Since comp. III inhibits NE-induced sustained contraction, this agent may be useful in the treatment of hypertension.
  • Yukihiro OZAKI, Lujian XING, Motoyoshi SATAKE
    1996 年 19 巻 2 号 p. 233-236
    発行日: 1996/02/15
    公開日: 2008/04/10
    ジャーナル フリー
    The present study was carried out to examine the accelerative effect of the ether extract obtained from"Nanshikon"on the proliferation of granulation tissue induced by cotton pellet in rats and to elucidate its active principles. Among naphthoquinone derivatives, the ether extract contained mostly teracryshikonin. At corresponding doses based on the contents of the naphthoquinone derivatives in the ether extract, the accelerative potencies of teracrylshikonin, β, β-dimethylacrylshikonin, and a mixture of α-methyl-n-butylshikonin and isovalerylshikonin were about the same. Also, the accelerative potencies of these compounds were somewhat weaker than that of the ether extract. From these results, it is suggested that the accelerative effect of the ether extract on the proliferation of granulation tissue is mainly due to teracrylshikonin, β, β-dimethylacrylshikonin and a mixture of α-methyl-n-butylshikonin and isovalerylshikonin contained in the ether extract, and the accelerative effect induced by ether extract might be an additive effect of these naphthoquinone derivatives.
  • QiuE ZHAO, Takuma MIHARA, Yukio SUGIMOTO, Chiaki KAMEI
    1996 年 19 巻 2 号 p. 237-240
    発行日: 1996/02/15
    公開日: 2008/04/10
    ジャーナル フリー
    Bradykinin at concentrations higher than 2μM caused a significant histamine release from rat peritoneal mast cells when extracellular Ca2+ was removed from the medium. Under the same experimental conditions, bradykinin increased Ca2+ release from the intracellular Ca store of the rat peritoneal mast cells, and a clear relationship was observed between the magnitude of histamine release and an increase in fluorescence intensity. Addition of Ca2+ to the medium resulted in an inhibition of the response to bradykinin in a concentration-dependent manner. Almost the same results were obtained when Mg2+, Ba2+ and La3+ were added to the medium. Neither B1 nor B2 antagonists caused significant antagonistic effects on histamine release induced by bradykinin. However, B2 antagonists caused a histamine release of the same potency as bradykinin when applied alone. These results indicate that bradykinin-induced histamine release is not attributable to a bradykinin receptor.
  • Mitsutaka SATO, Hiraku ONISHI, Manabu KITANO, Yoshiharu MACHIDA, Tsune ...
    1996 年 19 巻 2 号 p. 241-245
    発行日: 1996/02/15
    公開日: 2008/04/10
    ジャーナル フリー
    Mitomycin C (MMC) was covalently attached to glycol-chitosan (Gly-chitosan) and N-succinyl-chitosan (N-Suc-chitosan) through a spacer of the glutaryl group, and each conjugate was obtained as a water-soluble product. Stability of 1a-(4-carboxybutyryl)-MMC (Glu-MMC) in vitro was investigated by incubation in 1/15M phosphate buffers of pH 5, 6, 7.4, 8 and 9 at 37°C. The release rate of MMC from Glu-MMC was very slow at neutral pH, gradual at basic pH and relatively fast at acidic pH, but the quick decomposition of MMC proceeded simultaneously at acidic pH. Release of MMC from the conjugates in vitro was investigated by incubation in 1/15M phosphate buffers of pH 5, 7.4 and 9 at 37°C. The release rate of MMC from the conjugates was slight at acidic pH, accelerated at basic pH, and was gradual at pH 7.4. In a plasma-buffer (pH 7.4) mixture (1 : 1, v/v), the release rate of MMC from the conjugates was rather similar to that in only the buffer (pH 7.4).
  • Kiyotada YASUI, Hiroshi FUJIOKA, Yasuhiko NAKAMURA
    1996 年 19 巻 2 号 p. 246-251
    発行日: 1996/02/15
    公開日: 2008/04/10
    ジャーナル フリー
    After the administration of the recombinant human tumor necrosis factor (rHuTNF), plasma concentration could be controlled from egg yolk phosphatidylcholine (eggPC)-egg yolk phosphatidic acid (eggPA) liposome (EggPA liposome). The plasma concentration was highest after the rHuTNF solution administration. Although the concentration after the 30% EggPA liposome administration was much lower than that after the solution administration, the antitumor effect against Meth A sarcoma was equal to that of the rHuTNF solution. Non-linear elimination of rHuTNF in rat could explain these phenomena. First, to evaluate the intact absorption profile, we developed a non-linear absorption calculating program (NLAP). The calculation revealed little difference in the total absorption amounts between the solution and liposomes. The absorption rate from the liposomes was lower than that from the solution, and finally, the control of rHuTNF release from liposomes was also confirmed in vivo. These findings suggested that the sustained release of rHuTNF from liposomes maintained high drug concentration in the injected site and decreased the systemic drug concentration.
  • Atsushi KURIHARA, Yoko SHIBAYAMA, Atsuko MIZOTA, Akiko YASUNO, Masaru ...
    1996 年 19 巻 2 号 p. 252-258
    発行日: 1996/02/15
    公開日: 2008/04/10
    ジャーナル フリー
    The effects of i.v. formulations on the pharmacokinetics were examined for two antitumor agents with different lipophilicities : rhizoxin and palmitoyl-rhizoxin (RS-1541). Blood disposition and tissue distributions in rats were evaluated using three formulations : polyethylene glycol 400 (PEG)/dimethylacetamide (DMA) solution, colloidal solution, and lipid emulsions composed of dioctanoyl decanoyl glycerol (ODO) and polyoxyethylene-(60)-hydrogenated castor oil (HCO-60). The effects of emulsion particle size on the pharmacokinetics were also investigated. Rhizoxin rapidly disappeared from the plasma and showed high distribution in the tissues, and in vitro rapidly degraded in the plasma independent of the formulations used. In in vitro plasma, rhizoxin was easily released from the emulsion particles. In contrast to rhizoxin, the pharmacokinetics of RS-1541 with greater lipophilicity changed considerably depending on the formulations. The emulsions showed high and sustained plasma concentrations for RS-1541. RS-1541 was stably incorporated in the emulsion droplets and protected from the degradation when it was applied as an emulsion. Tissue distributions of RS-1541 in rats after an injection as lipid emulsion were strongly affected by the emulsion particle size. Small size emulsions (100-110nm) showed the highest plasma concentrations of RS-1541, though they were unable to suppress distributions of the drug in peripheral tissues. Emulsions larger than 200nm (approx.) in size, on the contrary, effectively inhibited the drug from entering the bone marrow, small intestine and other non-reticuloendothelial system (non-RES) organs, where many cytotoxic compounds showed undesired toxicities. These results indicate that the lipid emulsions composed of ODO and HCO-60 could be a promising and effective DDS carrier for RS-1541, which is highly lipophilic and stabilized in the emulsions. This was not the case for rhizoxin, however, which was less lipophilic than palmitoyl analogue RS-1541. The work described herein has demonstrated that by properly selecting the particle size, these lipid emulsions can control the behavior of a drug in the body.
  • Minoru MACHIDA, Masahiro HAYASHI, Shoji AWAZU
    1996 年 19 巻 2 号 p. 259-262
    発行日: 1996/02/15
    公開日: 2008/04/10
    ジャーナル フリー
    Pulmonary absorption of recombinant human granulocyte colony-stimulating factor (rhG-CSF) was examined in the rat. The relative bioavailabilities of rhG-CSF after intratracheal administration to intravenous and subcutaneous means were 11.6% and 27.4%, respectively, which were evaluated from the ratio of area under the curve (AUC) of the plasma rhG-CSF concentration versus time for 8h. The pharmacological availability after the pulmonary absorption was determined from the AUC for the increase ratio in total blood leukocyte numbers versus time curve. The avaiability was equal to or more than the availabilities after intravenous or subcutaneous administration. Comparison of the two different parenteral administration routes for rhG-CSF, intratracheal and intranasal showed the pharmacological availability after the intratracheal administration to be superior. The pulmonary absorption of rhG-CSF is thus an effective parenteral route of administration.
  • Hirofumi IMOTO, ZiQi ZHOU, Audra L. STINCHCOMB, Gordon L. FLYNN
    1996 年 19 巻 2 号 p. 263-267
    発行日: 1996/02/15
    公開日: 2008/04/10
    ジャーナル フリー
    In vitro skin permeation of buprenorphine (BUP) and three of its alkyl ester prodrugs was evaluated using hairless mouse skin. The three esters selected were the acetyl ester (Ac-BUP), butyl ester (Bu-BUP), and isobutyl ester (Isb-BUP). These drugs were applied on the skin as saturated slurries in three vehicles commonly used to formulate agents for transdermal purposes : propylene glycol, polyethylene glycol 400 (PEG 400), and light mineral oil. Unique solubilities were found for each drug on each vehicle. Fluxes through hairless mouse skin were evaluated for each combination of drug and vehicle using Franz diffusion cells. From PEG 400 formulations, the skin fluxes of BUP, Ac-BUP, Bu-BUP, and Isb-BUP were 0.47±0.08, 1.64±0.31, 0.33±0.05, 0.75±0.20μg/cm2/h, respectively. Thus, among the three potential prodrugs chosen, only Ac-BUP showed significantly higher skin flux than BUP. There were no inter-vehicle differences in the fluxes from saturated slurries between the vehicles. Moreover, all the esters were detected substantially in the form of regenerated parent drug (BUP) in the receptor compartment. Indeed, only Ac-BUP exited the skin in a measurably intact form, but the fraction escaping metabolism in transit was small (approximately 2%). However, based on drug dispositions in the skin, the regeneration of buprenorphine seems to depend on the alkyl chain length of the ester moiety. The molar percentages of regenerated parent drug in whole drug collected from the skin following the permeation experiments were : Ac-BUP, 9.2% ; Bu-BUP, 40.7% ; Isb-BUP, 9.6%, respectively. Thus, only Ac-BUP appears promising as a prodrug of buprenorphine, because it is not overly hydrophilic for skin permeation and is also highly metabolized to the parent compound while in the skin.
  • Shuji KITAGAWA, Yoshio SUGAYA
    1996 年 19 巻 2 号 p. 268-273
    発行日: 1996/02/15
    公開日: 2008/04/10
    ジャーナル フリー
    Characteristics of transport of an oral aminocephalosporin, cefroxadine, in rabbit small intestinal brush border membrane vesicles were examined. Uptake rate of cefroxadine was saturable in the presence of an inward H+ gradient, and kinetic parameters were similar to those of cephradine. However, the uptake rate was almost linear with the concentration in the absence of an inward H+ gradient up to 5mM. Overshoot phenomenon was observed in the presence of an inward H+ gradient at 37°C, but it disappeared with decrease of temperature. The Arrhenius plot of uptake rate constant showed a break point at approximately 30°C. Cefroxadine uptake was optimum in the vicinity of pH 5.5 at 37°C, but the dependence on extravesicular pH disappeared at 15°C. The uptake of cefroxadine in the presence of an inward H+ gradient was markedly inhibited by other aminocephalosporins such as cephalexin, but the inhibition was only slight in the absence of an inward H+ gradient. Alkyl alcohols such as n-hexyl alcohol also inhibited H+-coupled uptake of cefroxadine at the concentration range at which the alcohols increased the membrane fluidity, and overshoot phenomenon diminished, suggesting that H+-coupled transport of cefroxadine is sensitive to the alcohol-induced increase in membrane fluidity. On the other hand, the alcohols rather stimulated its uptake in the absence of an H+ gradient.
  • Fusao KOMADA, Kohshi NISHIGUCHI, Yusuke TANIGAWARA, Taeko AKAMATSU, Xi ...
    1996 年 19 巻 2 号 p. 274-279
    発行日: 1996/02/15
    公開日: 2008/04/10
    ジャーナル フリー
    Human Cu, Zn-superoxide dismutase (hSOD) cDNA was inserted into a eukaryotic expression plasmid (pRc/CMV) under the control of a cytomegalovirus promoter. The hSOD expression plasmid (pRc/CMV-SOD) was transfected into L2 cells by means of lipofection. The integration of the hSOD gene in genomic DNAs in the cells transfected with pRc/CMV-SOD plasmid was examined by Southern blotting using hSOD cDNA as the probe. However, Southern blots of host cells (without transfection) and CMV cells (pRc/CMV plasmid transfection) indicated no hybridization of hSOD cDNA. Western blots indicated that hSOD was expressed in CMV-SOD cells. The SOD activity in CMV-SOD cells was about twice that in host and CMV cells. Furthermore, this SOD activity in CMV-SOD cells was enhanced for 60 d after the selection of cell clones. After exposure to paraquat and catalase, about 90% of the CMV-SOD cells survived compared with the untreated controls, whereas about 60% of the host cells survived. The production of lipid peroxidation in host cells increased significantly after exposure to both paraquat and catalase, whereas that in CMV-SOD cells did not change. The correlation between the surviving cells and lipid peroxidation was inverse. These results indicated that transfection with the SOD gene was effective against superoxide anion induced cytotoxicity.
  • Takashi KONDO, Tetsumi IRIE, Kaneto UEKAMA
    1996 年 19 巻 2 号 p. 280-286
    発行日: 1996/02/15
    公開日: 2008/04/10
    ジャーナル フリー
    Pharmacokinetics of morphine and its glucuronides in plasma were studied after rectal administration of hollow-type oleaginous suppositories containing kneading mixtures of morphine hydrochloride, α-cyclodextrin, and/or xanthan gum in rabbits. In combination with xanthan gum, α-cyclodextrin reduced the first-pass metabolism of morphine in the rectal mucosa and by the liver and improved the apparent rectal bioavailability of the opioid about 4 fold. In vitro permeation studies using an isolated rectal mucosal preparation of rabbits revealed that α-cyclodextrin increased the transepithelial conductance and facilitated the transport of morphine through the rectal mucosa. Furthermore, α-cyclodextrin facilitated its own mucosal permeation and reduced the glucuronidation of morphine during the passage through the rectal mucosa, probably through restricting the formation of a catalytic complex of morphine with glucuronyltransferases, rather than because of the enzyme saturation. The present data suggest that α-cyclodextrin in combination with xanthan gum is particularly effective in improving the rectal bioavailability of morphine from hollow-type suppositories.
  • Jun WATANABE, Kimihiko URANO, Masami HABA, Hiroaki YUASA
    1996 年 19 巻 2 号 p. 287-290
    発行日: 1996/02/15
    公開日: 2008/04/10
    ジャーナル フリー
    Molecular weight dependency in the interaction of fractionated [3H] heparin (FH) with plasma proteins was evaluated by determining the protein binding of low molecular weight fractionated [3H] heparin (LMWFH : 7000 Da) and high molecular weight fractionated [3H] heparin (HMWFH : 16000 Da) by ultrafiltration and the effects of plasma proteins on the uptake in rat hepatocytes in primary culture. The unbound fractions of LMWFH were 0.5 and 0.8 in the presence of α-globulin and albumin, respectively, and were about 10 times larger than those of HMWFH, 0.04 and 0.1, suggesting a reduction in binding with a decrease in molecular weight. However, while the uptake of LMWFH was reduced by these proteins by the extents similar to bound fractions of LMWFH, the uptake of HMWFH was reduced by extents far smaller than bound fractions and comparable with those for LMWFH. Thus, it seemed that, while only unbound LMWFH is available for uptake, HMWFH bound to proteins is to some extent available for uptake (protein-mediated transport). The protein-mediated transport of heparin seemed to reduce with a decrease in molecular weight. It was also shown that the extended uptake of LMWFH was smaller than that of HMWFH not only in the absence of proteins but also in the presence of α-globulin, the major binding protein. The lower uptake of LMWFH is consistent with in vivo suggestion of lower hepatic accumulation.
  • Hideyuki YAMADA, Yasuhiro MINEMATSU, Tetsuya NAKAMURA, Masashi MISE, H ...
    1996 年 19 巻 2 号 p. 291-293
    発行日: 1996/02/15
    公開日: 2008/04/10
    ジャーナル フリー
    We have indicated that strychnine and brucine induce concurrently an unknown P450 (2B-UP) which crossreacts with anti-CYP2B1 antibody (Fujisaki et al., J. Pharmacol. Exp. Ther., 268, 1024-1031, 1994). We purified 2B-UP from brucine-treated rats and characterized it in this study. The purification was achieved by solubilization with sodium cholate followed by successive chromatographic steps with ω-aminooctyl-Sepharose 4B, DEAE-Sephacel and hydroxyapatite. The minimum molecular weight of purified 2B-UP was calculated to be 48000 by sodium dodecyl sulfate-gel electrophoresis. This preparation showed no Soret peak in the ultraviolet absorption spectrum indicating absence of heme. The amino terminal sequence of 2B-UP up to the 10th residue was consistent with the deduced amino acid sequence of CYP2B3 cDNA, but did not agree with the sequences of CYP2B1/2. The result strongly suggests that 2B-UP is CYP2B3. Thus, we indicated here that Strychnos alkaloid, brucine, is a potent inducer of the CYP2B3 or the closely related P450.
  • Tadashi KIHO, Akihiro YAMANE, Ji HUI, Shigeyuki USUI, Shigeo UKAI
    1996 年 19 巻 2 号 p. 294-296
    発行日: 1996/02/15
    公開日: 2008/04/10
    ジャーナル フリー
    A polysaccharide (CS-F30) obtained from the cultural mycelium of Cordyceps sinensis showed potent hypoglycemic activity in genetic diabetic mice after intraperitoneal administration, and the plasma glucose level was quickly reduced in normal and streptozotocin-induced diabetic mice after intravenous administration. Administration of CS-F-30 to normal mice significantly increased the activities of hepatic glucokinase, hexokinase and glucose-6-phosphate dehydrogenase, although the glycogen content in the liver was reduced. Furthermore, CS-F30 lowered the plasma triglyceride level and cholesterol level in mice.
  • JeTae WOO, Yasuo OHBA, Kahori TAGAMI, Koji SUMITANI, Kohji YAMAGUCHI, ...
    1996 年 19 巻 2 号 p. 297-299
    発行日: 1996/02/15
    公開日: 2008/04/10
    ジャーナル フリー
    Effects of concanamycin B, a specific inhibitor of the vacuolar type H+-ATPase (V-ATPase), on the stimulation of bone resorption induced by parathyroid hormone (PTH) were examined in vitro. Concanamycin B was found to inhibit PTH-stimulated osteoclastic pit formation and to suppress the acidification of vacuolar organelles by V-ATPase in the osteoclasts. PTH-stimulated 45Ca release from prelabelled chick embryonic calvariae was also inhibited by concanamycin B in a dose-dependent manner. These results suggest that osteoclastic acidification of lacunae by V-ATPase plays an essential role in mineral dissolution and degradation of the organic matrix during bone resorption.
  • Michihisa TOHDA, Toshihiko MURAYAMA, Shinsuke NOGIRI, Yasuyuki NOMURA
    1996 年 19 巻 2 号 p. 300-302
    発行日: 1996/02/15
    公開日: 2008/04/10
    ジャーナル フリー
    To clarify the quantitative and qualitative changes in type IV phosphodiesterase (PDE IV) with aging, phosphodiesterase (PDE) activity and [3H] rolipram binding in the cytosolic fraction from the brains of young and aged rats were examined. In all areas of the aged (100-week-old) rat brain except for hippocampus, the PDE activity was decreased by about half that in the young (10-week-old) animals. However, inhibition % by 100μM rolipram and by 100μM isobutylmethylxanthine (IBMX) was not significantly different between the aged and young rats. On the other hand, [3H] rolipram binding and rolipram-sensitive PDE activity did not change with aging in the hippocampus, although both were decreased in other brain regions. These results suggest that PDE IV does not decrease with aging and maintains its cAMP degrading activity in the hippocampus. It may be involved in the dysfunction of the hippocampus with aging.
  • Yangzheng FENG, Minoru NARITA, Mizue MAKIMURA, Beth HOSKINS, I.K. HO
    1996 年 19 巻 2 号 p. 303-304
    発行日: 1996/02/15
    公開日: 2008/04/10
    ジャーナル フリー
    The dissociation constant (KD) of [3H] phorbol 12, 13-dibutyrate (PDBu) binding to protein kinase C (PKC) in membranes of rat cortex and midbrain was significantly decreased with no change in the receptor density (Bmax) following 7-d treatment with a κ-opioid agonist, U-50, 488 (20mg/kg/d, i.p.). Neither the Bmax nor KD values in pons/medulla were altered by repreated U-50, 488 treatment. These results suggest that repeated administration of a κ-opioid agonist increases the affinity for PDBu binding to the membrane-bound PKC in rat cortex and midbrain, but not in pons/medulla.
  • Toru MORIGUCHI, Hiroshi SAITO, Nobuyoshi NISHIYAMA
    1996 年 19 巻 2 号 p. 305-307
    発行日: 1996/02/15
    公開日: 2008/04/10
    ジャーナル フリー
    The effects of aged garlic extract (AGE), chronically administered in the diet, on longevity and spatial learning performances were studied using the senescence-accelerated mouse (SAM). A solid diet containing 2% AGE was given to senescence-accelerated-prone mouse 8 (SAMP8) and senescence-accelerated-resistant mouse 1 (SAMR1) from 2 months of age. The survival ratio of SAMP8, a substrain of senescence-accelerated-prone mouse, was significantly lower than that of SAMR1, a substrain of senescence-resistant mouse. In the SAMP8, administration of AGE perfectly prevented the decrease in survival ratio. Moreover, AGE markedly improved the learning deficits of SAMP8 in the Morris water maze test. These results suggest the possibility that AGE prevents physiological aging and age-related memory disorders in human.
  • Shin OHTA, Masahiro MAKINO, Katsuhiro NAGAI, Hiroshi ZENDA
    1996 年 19 巻 2 号 p. 308-310
    発行日: 1996/02/15
    公開日: 2008/04/10
    ジャーナル フリー
    We synthesized a new quaternary ammonium salt, N-alkyl-N-2-hydroxyethyl-N, N-dimethylammonium butyl phosphate (ABP) that does not precipitate in the presence of anionic surfactants by incorporating a paired butyl phosphate anion into cationic surfactants. ABP showed much greater bactericidal activities and antirusting effects than benzalkonium chloride (BAC). In this study, the fungicidal effects of ABP were evaluated in comparison with common disinfectants [BAC, chlorhexidine digluconate (CHX) and alkyldiaminoethylglycine hydrochloride (ADE)]. Fungicidal effects were evaluated in 10 strains of 6 fungal species, namely, 3 Candida albicans, 2 Candida tropicalis, 1 Candida parapsilosis, 1 Aspergillus niger, 2 Aspergillus terreus, and 1 Trichophyton rubrum. ABP and BAC showed the same effects on 2 C. tropicalis and C. parapsilosis, and similar effects on 3 C. albicans, with slight differences among the strains. ABP showed quick fungicidal effects against A. niger and 2 A. terreus of molds in 30 min at 0.1% and in 15 min at 0.2 and 0.4%, but BAC showed slow effects against A. niger at 1h and 2 A. terreus at 30min at 0.2 and 0.4%, respectively. There is one factor which may have caused this difference : the former had a butyl phosphate anion and the latter had a chloride anion as a counter ion. BAC, CHX and ADE needed 24h to cause notable effects on A. niger at 0.1% and 1h at 0.2%. The new synthetic anti-rust ABP is considered to have sufficient fungicidal effects that are comparable to those of BAC, CHX and ADE.
  • Munekazu IINUMA, Hideki TOSA, Toshiyuki TANAKA, Satiyo KANAMARU, Fujio ...
    1996 年 19 巻 2 号 p. 311-314
    発行日: 1996/02/15
    公開日: 2008/04/10
    ジャーナル フリー
    Benzophenone derivatives, garcinol (1) and isogarcinol (2) isolated from the pericarps of Garcinia purpurea (Guttiferae), and xanthochymol (3) and a mixture of isoxanthochymol (4) cycloxanthochymol (5) from the pericarps of G. subelliptica were evaluated for their antibacterial activity against methicillin-resistant Staphylococcus aureus. Among them, 3 showed the lowest minimum inhibitory concentration at 3.1-12.5μg/ml. This concentration is nearly equal to that of the antibiotic, vancomycin.
  • Hiroyuki MASUDA, Ken OHSUMI, Mami KAKO, Toshihiro MIURA, Yumi NISHIYAM ...
    1996 年 19 巻 2 号 p. 315-317
    発行日: 1996/02/15
    公開日: 2008/04/10
    ジャーナル フリー
    The hypolipidemic effect of Senegae Radix, i.e."Senega, "was investigated in normal and hyperlipidemic mice. The n-butanol fraction of the methanol extract of Senegae Radix (SN) (5mg/kg) significantly reduced the blood triglyceride level of normal mice 7h after intraperitoneal administration (p<0.05) and also significantly reduced the blood triglyceride level of cholesterol-fed mice under similar conditions (p<0.05). SN (5mg/kg) also reduced the blood triglyceride and cholesterol levels after repeated administration to cholesterol-fed mice. SN also decreased the blood triglyceride level in Triton-induced hyperlipidemia. It is suggested that this fraction contains one or more hypolipidemic components including the main triterpenoid glycoside, senegin-II, which significantly reduces the levels of blood triglycerides in normal mice.
  • Yukiko NAKAJIMA, Koujirou YAMAMOTO, Yasuhiko YAMADA, Yasufumi SAWADA, ...
    1996 年 19 巻 2 号 p. 318-322
    発行日: 1996/02/15
    公開日: 2008/04/10
    ジャーナル フリー
    Emesis after the administration of cisplatin is a severe complication, and its treatment is an important problem clinically. Cisplatin forces the release of serotonin (5-HT) from enterochromaffin cells in the mucosa, and emesis occurs by the stimulation of 5-HT3 receptors. In this study, we established a simple and simultaneous method of determining 5-HT and its main metabolite, 5-hydroxyindole-3-acetic acid (5-HIAA), in plasma and urine by high performance liquid chromatography with electrochemical detection (HPLC-ECD), and determined the disposition of endogenous 5-HT and 5-HIAA after the administration of cisplatin in rats and dogs. In rats, we found no change in the plasma concentration of 5-HIAA after cisplatin administration, while a distinct increase was shown in the plasma concentration of 5-HT, but it was not significantly different from that of the control rats. The urinary excretion of 5-HIAA was also not different between the two groups. In dogs, we observed intense vomiting in all cisplatin treated dogs. However, we could not detect any change in the 5-HIAA or 5-HT level in the dog plasma. Furthermore, no significant difference in the urinary excretion of 5-HIAA was observed between the cisplatin group and the controls. From these results, the plasma concentration of 5-HT and the urinary excretion of 5-HIAA may not be suitable markers for the evaluation of emesis induced by anticancer drugs in dogs.
  • Yu NAKADA, Koujirou YAMAMOTO, Junichi KAWAKAMI, Yasufumi SAWADA, Tatsu ...
    1996 年 19 巻 2 号 p. 323-325
    発行日: 1996/02/15
    公開日: 2008/04/10
    ジャーナル フリー
    We investigated the effect of acute renal failure on the neurotoxicity of ranitidine in rats. Experimental acute renal failure was produced by bilateral ureteral ligation. Ranitidine was infused into the ureter ligated (UL) and control rats at the rate of 3.25mg/min through the jugular vein until the onset of clonic convulsion. In UL rats, the onset time of convulsion was shorter and ranitidine concentrations in plasma and cerebrospinal fluid (CSF) were lower than those of control rats. However, the ranitidine concentration in the brain at the onset of convulsion was not different between the UL and control rats. From these findings, we concluded that acute renal failure is one of the risk factors for ranitidine neurotoxicity, and the increased sensitivity to the drug on the central nervous system may contribute to the increased toxicity of ranitidine in renal failure.
  • Kazuo OKUSHIO, Natsuki MATSUMOTO, Toshiyuki KOHRI, Masayuki SUZUKI, Fu ...
    1996 年 19 巻 2 号 p. 326-329
    発行日: 1996/02/15
    公開日: 2008/04/10
    ジャーナル フリー
    Following the oral administration of tea catechins, (-)-epicatechin, (-)-epigallocatechin, (-)-epicatechin gallate and (-)-epigallocatechin gallate, respectively, to rats, the presence of these catechins in the portal blood was examined. It was confirmed by HPLC and mass spectrometry analysis that each of the administered catechins was present in the blood. These results clearly indicate that four predominant catechins in fresh tea leaves are absorbed, at least in part, into the rat portal vein.
  • Osamu UEDA, Shigeyuki KITAMURA, Kiyoshi TATSUMI
    1996 年 19 巻 2 号 p. 330-333
    発行日: 1996/02/15
    公開日: 2008/04/10
    ジャーナル フリー
    The further in vivo metabolism of 2-formylaminofluorene (FAF), which is a metabolite of 2-aminofluorene, was examined in rats. The 48 h urine collected following oral administration of FAF to rats was treated with β-glucuronidase/arylsulfatase and extracted with benzene. When the benzene extract was subjected to HPLC, ten peaks due to FAF and its metabolites were detected. Among these metabolites, major metabolites 1, 2, 4 and 5 were isolated and identified unequivocally as 7-hydroxy-2-formylaminofluorene, 7-hydroxy-2-acetylaminofluorene, 5-hydroxy-2-formylaminofluorene and 5-hydroxy-2-acetylaminofluorene, respectively, by comparing their MS and UV spectra, and HPLC and TLC behavior with that of authentic specimens. Minor metabolites, 7-hydroxy-2-aminofluorene, 5-hydroxy-2-aminofluorene, 2-acetylaminofluorene and 2-aminofluorene together with the parent compound were detected by HPLC. These metabolites of FAF were also detected in the feces of rats given FAF.
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