In order to know the organ distribution of Chinoform, I4C.Chinoform was injected into the tail vein of the mice, and radioactivity was measured in the chloroform soluble fractions in some organs and tissues containing non.conjugated Chinoform. The results obtained are as follows. 1. Uptake of Chinoform by the visceral organs was found to be in the following ascending order: fat tissue, kidney, spleen, liver, small intestine, (blood), muscle and eye, and marked uptake by the fat tissue and kidney was observed. 2. The presence of radioactive Chinoform in the chloroform soluble fraction of the central nervous system was recognized and it was almost in the same degree of specific radioactivity as that of blood. 3. A higher uptake in the chloroform soluble fraction of the sciatic nerve than that of central nervous system was recognized, and the value of the former was about 3 to 8 times as high as that of the latter. 4. The presence of Chinoform in the chroloform soluble fraction of the bile, although it increased after incu bation of the bile with j9-glucuro.
nidase was observed. High radiogctivity of chinoform in the total fraction of the bile suggests a possible presence of &#34;liver-intestine-circulation&#34; of
the drug.</p

Kuroda, Shigetoshi

Ogata, Masana

Otsuki, Saburo

Tateishi, Jun

Tomokuni, Katsumaro

Watanabe, Shosuke

Okayama University Digital Information Repository

Metabolism of 14 C-iodochlorohydroxyquinoline (chinoform) in mice

Okayama University Scientific Achievement Repository

Acta Medica OkayamaVolume 25, Issue 3 1971 Article 5JUNE 1971Metabolism of 14C-iodochlorohydroxyquinoline (chinoform) inmiceMasana Ogata∗ Shosuke Watanabe† Jun Tateishi‡Shigetoshi Kuroda∗∗ Katsumaro Tomokuni†† Saburo Otsuki‡‡∗Okayama University,†Okayama University,‡Okayama University,∗∗Okayama University,††Okayama University,‡‡Okayama University,Copyright c©1999 OKAYAMA UNIVERSITY MEDICAL SCHOOL. All rights reserved.Metabolism of 14C-iodochlorohydroxyquinoline (chinoform) inmice∗Masana Ogata, Shosuke Watanabe, Jun Tateishi, Shigetoshi Kuroda, KatsumaroTomokuni, and Saburo OtsukiAbstractIn order to know the organ distribution of Chinoform, I4C.Chinoform was injected into thetail vein of the mice, and radioactivity was measured in the chloroform soluble fractions in someorgans and tissues containing non.conjugated Chinoform. The results obtained are as follows. 1.Uptake of Chinoform by the visceral organs was found to be in the following ascending order: fattissue, kidney, spleen, liver, small intestine, (blood), muscle and eye, and marked uptake by thefat tissue and kidney was observed. 2. The presence of radioactive Chinoform in the chloroformsoluble fraction of the central nervous system was recognized and it was almost in the same degreeof specific radioactivity as that of blood. 3. A higher uptake in the chloroform soluble fraction ofthe sciatic nerve than that of central nervous system was recognized, and the value of the formerwas about 3 to 8 times as high as that of the latter. 4. The presence of Chinoform in the chroloformsoluble fraction of the bile, although it increased after incu bation of the bile with j9-glucuro.nidase was observed. High radiogctivity of chinoform in the total fraction of the bile suggests apossible presence of “liver-intestine-circulation” of the drug.∗PMID: 4263521 [PubMed - indexed for MEDLINE] Copyright c©OKAYAMA UNIVERSITYMEDICAL SCHOOLActa Med. Okayama 25, 199-204 (971)METABOLISM OF 14C.IODOCHLOROHYDROXYQUINOLINE(CHINOFORM) IN MICEMasana OGATA*, Shosuke WATANABE**, Jun TATEISHI**,Shigetoshi KURODA **, Katsumaro TOMOKUNI*,and Saburo OTSUKI**Department of Public Health*, and N euro-Psychiatry**,Okayama University Medical School, Okayama, jabanReceived for publication, May 2, 19715-Chloro.7.iodo-8-hydroxyquinoline (Chinoform) is a well.known in-testinal antiseptic in the world.wide use. BERGGREN, L. et ai. (l) drewattention by reporting that optic atrophy has followed long. term medica.tion with Chinoform. It is often stated that Chinoform is not absorbedin the gastrointestinal tract. But urinary excretion of the glucuronic andsulfuric acid conjugates of Chinoform given orally to rabbit (2) and man(3~5) has been reported.In Japan, so.called subacute myelo.optico.neuropathy with abdominalsymptoms (SMON) was first reported by T AKASAKI (6) in 1961 in MiePrefecture, central part of Japan. Since then, the number of patientsof SMON has been found increasing in many places throughout Japan,and the number of patients reported during 1967 to 1968 reached 4, 280(7). The clinical symptoms of SMON show signs of lateral corticospinaltracts and of posterior column disturbances (8) ensuing from abdominalsymptoms, and 89.5 per cent of patients show sensory disturbances and60.8 per cent motor disturbances and 23.5 per cent are accompanied byvisual disturbances (7).Recently, it is suspected that long term administration of Chinoformmay be one of the causal factors of this syndrome (9). In the present studythe distribution in the visceral and nervous tissues and bile-excretion ofChinoform in mice were investigated. The results are briefly described here.MATERIALS AND METHODSMaterials:ddN-Strain female mice weighing about 20 grams were used. Animals werekept in a stock on ad libitum diet. The day before the experiment, animals wereadministered ad libitum 1 per cent iodine solution, orally as thyroid blockingagent.1991Ogata et al.: Metabolism of 14 C-iodochlorohydroxyquinoline (chinoform) in miceProduced by The Berkeley Electronic Press, 1971200 M.OCATA, S. WATANABE, J. TATEISHI, ~. KURODA, K. TOMOKUNI and S.OTSUKISubstance: Chinoform -2, 3, 4, .14C (specific activity: 1.63 mCi/mmole) waspurchased from Daiichi Pure Chemicals Co., Ltd. Purity analyses by radio.thinlayer chromatography on silica gel in methanol: benzene: acetone: aceticacid [4: 14 : 1 : IJ and benzene: methanol: acetic acid [45: 8: 9J showed thatradioactive, 14C.Chinoform has 99 per cent purity. The radioactive Chinoformwas emulsified in carboxylmethyl cellulose suspension and served as materialsfor injection. Each mouse was injected 0.3 mCi Chinoform into tail vein.Methods:Mice were sacrificed one hour after the injection by means of blood drawingfrom orbital vein. Organs and tissues were separated respectively, and theirwet weight was measured. Each sample was homogenized with 3 volumes ofwater in a teflon homogenizer for 5 minutes, thereafter, 3 ml chloroform wasadded, rehomogenized and centrifuged. Then chloroform soluble layer contain.ing non.conjugated Chinoform (3) was separated by centrifugation, transferredto a counting vial and dried by blowing air. The dried material thus obtainedwas dissolved in 0.5 ml of methyl alcohol.hyamine solution, added 15 ml oftoluene containing PPO and POPOP, and radioactivity was measured in aPackard liquid scintillation spectrometer.The radioactivity was expressed as dpm by external standardization method.RESULTSTable 1 shows the distribution of radioactivity (dpm/gr, wet weight)in the chloroform soluble fractions of various organs. Between two mice,there is no significant difference in the distribution in visceral and nervousorgans.In the kidney and fat tissue of mice, uptake is most marked, 14,326dpm/gr., w. w. and 14,692 dpm/gr., w. w. respectively. These valuesare about 6 to 7 times as much as much as radioactivity of blood (2, 384dpm/gr., w. w,). The order of radioactivity in other visceral organs is asfollows: spleen, liver, small intestine, muscle and eye. The level ofradioactivity in these organs is similar to that of blood. The presence ofradioactivity in various parts of the central nervous system was recognized,and the values of brain stem (2,397 dpm/gr., w. w,), spinal cord (cervical,2,877 dpm/gr., w. W.; thoracic and lumbar, 3,159 dpm/gr., w. w,) werehigher than those of cerebrum (1,469 dpm/gr., w. w,) and cerebellum(2,103 dpm/gr., w. w,). A great deal of radioactivity was found in thesciatic nerve (10, 936 dpm/gr., w. w,). The value was about 3 to 5 timesas high as that of central nervous system or 5 times as high as that ofblood. This fact was also found in our previous experiment of rat ad.ministered 131I-Chinoform (9, 10), and agrees with the experiments withrabbits (2) or rats (3), in which experimental neuritis of the sciatic nerve2Acta Medica Okayama, Vol. 25 [1971], Iss. 3, Art. 5http://escholarship.lib.okayama-u.ac.jp/amo/vol25/iss3/5Metabolism of Chinoform in MiceTABLE 1. DISTRIBUTION OF RADIOACTIVITY IN THE CHLOROFORM SOLUBLE FRACTIONOF VARIOUS ORGANS IN THE MICE AFTER INJECTION OF 14C.CHINOFORM[dpm/gr. wet weight]organ mouse I mouse 2 meancerebrum 1,370 1,568 1,469cerebellum 1,917 2,288 2,103brain stem 2,285 2,508 2,397cervical cord 3,001 2, 752 2,877thoracic & lumbar cord 2,929 3,388 3,159sciatic nerve 11, :179 10,493 10,936eye 1,673 1,001 1,337liver 2,983 2,565 2,774kidney 11,817 16,835 14,326spleen 4,713 4,591 4,652small intestine 2, 785 2,862 2,734fat tissue 13,588 15,796 14,692muscle 1,435 2,506 1,971blood 2,875 1,892 2,384bile 509,934* 17,543* 263,739** Radioactivity in the total fractions (chloroform soluble and insoluble fractions)201was induced by the administration of Chinoform, The data indicate thatmuch higher specific radioactivity in the total fraction (Chloroform solubleand insoluble fraction) was found in the bile (263, 739 dpm/gr., w. w,).In the experiment, the total radioactivity of bile was about 110 times as highas that of blood. In the case of mouse 2, the ratio of radioactivity in thechloroform soluble fraction to insoluble fraction in the bile was 1 : 6 andthat in the bile after incubation with i1-g1ucuronidase (Fishman unit, 1000,for 45 min. at 37°C in phosphate buffer 0.15 M, pH 6,0) was 2: 1. There-fore, it is considered that large portion of Chinoform in bile conjugatedwith glucuronic acid.DISCUSSIONIn the present experiment, radioactivity was measured with thechloroform soluble fractions obtained from various organs. The values showthe distribution of most of free Chinoform, which can be combined withmetals and exhibit high toxicity to the various organs. It is by now wellknown that administration of Chinoform substantially elevates the iodinecontent in the blood. LIEWENDAHL et at. (4) showed in their experimentadministred mI.Chinoform orally, that iodine corresponding to 0.4 per3Ogata et al.: Metabolism of 14 C-iodochlorohydroxyquinoline (chinoform) in miceProduced by The Berkeley Electronic Press, 1971202 M. OGATA, S. WATANABE, ]. TATEISHI, S. KURODA, K. TOMOKUNI and S.OTSUKIcent of Chinoform could be liberated from this agent within 24 hours inman. In our experiment, we used 14C.Chinoform instead of 125I.Chino·form and also administered iodine to mice before and during the experi.ment to prevent Chinoform binding to thyroxine and liberation of iodinefrom the Chinoform. Therefore, the effect of iodine liberation will besmallest. Our results suggest that some part of Chinoform is excreted intobile in the form of glucuronic acid conjugates, as the free Chinoformcontent in the bile increases after incubation with ,9.g1ucuronidase. Thisresult agrees with the finding in our experiment described previously (10),in which 131I-Chinoform was used as a tracer, and the presence of somefree chinoform in the bile to be excreted into intestine suggests some poss.ibilities of chinoform reabsorption. Therefore, a presence of a liver.intestine circulation of this drug is conceivably possible. A high level ofspecific radioactivity in the fat tissue indicates a possible accumulation ofChinoform in this tissue.The organs measured in this experiment contain a small amount ofblood. LAJTHA et ai. (14) described that the maximum value of bloodcontained in the central nervous system, liver and muscle is, 0.9 per cent,3.2 per cent and 0.9 per cent respectively by measurement of CO·Hb inthese organs. And BOSSE et ai. (15) reported that the quantity of bloodcontained in the organs was 1.98±0.1l (m+O') per cent in cerebrum,2.7 ±O .18 per cent in cerebellum, 3. 02± 0.22 per cent in medulla obIon.gata, 2.45 ± 0.22 per cent in spinal cord, and 2.17 ± 0.24 per cent insciatic nerve.In the visceral organs and also in the central and peripheral nervoussystems, uptake of 14C.Chinoform was demonstrated by the presence oftrue radioactivity after subtracting blood radioactivity from measuredradioactivity in studying the chronic intoxication of Chinoform. Theproblem, whether the uC.Chinoform can penetrate blood brain barrier ornot, is yet to be reported. Chloroform soluble extract of contents of asmall intestine containing 14.4% of total activity with a new mouse willbe expressed on our next paper.CO~CLUSIONIn order to know the organ distribution of Chinoform, I4C.Chinoformwas injected into the tail vein of the mice, and radioactivity was measuredin the chloroform soluble fractions in some organs and tissues containingnon.conjugated Chinoform.The results obtained are as follows.4Acta Medica Okayama, Vol. 25 [1971], Iss. 3, Art. 5http://escholarship.lib.okayama-u.ac.jp/amo/vol25/iss3/5Metabolism of Chinoform in Mice 2031. Uptake of Chinoform by the visceral organs was found to be inthe following ascending order: fat tissue, kidney, spleen, liver, smallintestine, (blood), muscle and eye, and marked uptake by the fat tissueand kidney was observed.2. The presence of radioactive Chinoform in the chloroform solublefraction of the central nervous system was recognized and it was almostin the same degree of specific radioactivity as that of blood.3. A higher uptake in the chloroform soluble fraction of the sciaticnerve than that of central nervous system was recognized, and the valueof the former was about 3 to 8 times as high as that of the latter.4. The presence of Chinoform in the chroloform soluble fraction ofthe bile, although it increased after incu bation of the bile with j9-glucuro.nidase was observed. High radiogctivity of chinoform in the total fractionof the bile suggests a possible presence of "liver-intestine-circulation" ofthe drug.REFERENCE1. BERGGREN, L. and HANSSON, 0.: Treating acrodermatitis enteropathica. Lancet 1, 52,19662. HASKINS, W. T. and LUTTERMOSER, G. W.: Urinary excretion of Vioform and Diodoguinin rabbits. j. Pharmacol & Exper. Therap. 109, 201, 19533. BERGGREN, L. and HANSSON, 0.: Absorption of intestinal antiseptics derived from 8-hy-droxyquinolines. Clin. Pharmacol. Therap. 9, 67, 19684. LIEWENDAHL, K. and LAMBERG, B. A.: Metabolism of I25Iodochloroxyquinoline in man.I. Absorption, binding and excretion. Nucl. Med. 6, 20, 19675. LIEWENDAHL, K., KIVIKANGAS, V. and LAMBTRG, B. A.: Metabolism of I25Iodochloroxy-quinoline in man. II. Absorption, binding and excretion. Nucl. Med. 6, 32, 19676. TAKASAKI, H.: A case report of lower extremity paralysis appeared in the course ofintestinal disease. j. jap. Soc. Intern. Med. 50, 171, 1961 (in Japanese)7. SMON Research Association (edited ty Itsuzo SHIGEMATSU). The integrated results ofSMON patients in Japan, 1970 (in Japanese)8. TAKASAKI, H.: Nonspecific cerebra-spinal infection. Handbook of Modern InternalMedicine Supplement, (a). Nakayama Shoten, Tokyo, 19679. TSUBAKI, T., HONMA, Y. and HOSHI, T.: Ehtiology of "SMON". Read at Kanto Pro-vincial Meetins of Japan Academy of Neurolo,?,y. 5. Sept. 197010. OGATA, M., TOMOKUNI, K., HAYASHI, S., WATANABE, S., TATEISHI,J., KURODA, S., OTSUKI,S. and ANDO, A.: Accumulation and excretion of radioactive Quinoform in mice andrats. Igaku to Seibutsugaku 83, 1971 in press (in Japanese)11. WATANABE, S., TATEISHI, J., KURODA, S.,OTSUKI, S., OGATA, M., TOMOKUNI, K. andANDO, A.: IncoTJ:oration of radioactive Quinofrom into the nerve tissues of rats. Igakuto Seibutsugaku 83, 1971 in press (in Japanese)12. IGATA, T. and TOYOKURA, Y.: Experiment on disturbance of nervous system inducedby Chinoform. Igaku no Ayumi 75, 309, 1970 (in Japanese)13. OUCHI, K., SATO, T. and TSUBAKI, T.: Electron Microscope of peripheral nerve of rats5Ogata et al.: Metabolism of 14 C-iodochlorohydroxyquinoline (chinoform) in miceProduced by The Berkeley Electronic Press, 1971204 M. OGATA, S. WATANABE, J. TAKEISHI, S. KURODA, K. TOMOKUNI and 5.0TSUKI~oisoned with Chinoform. Record of SMO~ Research Association. p 55, 197114. LAJTHA,A., FURST, S., GERSTEIN,A. and WAELSCH,H.: Amino acid and protein meta-bolism of the Brain: Turnover of free, and protein bound lysine in brain and otherorgans: j. Neurochem. 1, 289, 195715. BOSSE, J. A. and WASSERMAN, 0.: On the blood content of guinea-pig tissues: Pharma-cology 4, 273, 19706Acta Medica Okayama, Vol. 25 [1971], Iss. 3, Art. 5http://escholarship.lib.okayama-u.ac.jp/amo/vol25/iss3/5

Metabolism of 14 C-iodochlorohydroxyquinoline (chinoform) in mice

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