Analgetikn — Antiphlogistika — Antirheumatika Analgesics — Antiphlogistics — Antirheumatic Drugs Antinociceptive Properties of Thiamine, Pyridoxine and Cyanocobalamin Following Repeated Oral Administration to Mice J. Leuschner Summary Following repeated oral administration for 7 days thia- ‘mine (Uhiamine nirate, CAS 532-43-4), pyridoxine (pyr. doxal hydrochloride, CAS 58.560), and cy lamin (CAS 68-19-5) exhibited at high dose levels alone oF in ‘combination dose related antinociceptive properties in the ‘writhing tes in the mouse. Cyanceabalamin exerted potentiating effect tn the combination ofthe 3 vitamins Zusammenfassung Antinozizepive Eigenschaften von Thiamin, Pyridoxin Koy words: Analgesics » CAS 58-560 - CAS 68.193 Thiamine Vitamin By Vitamin By Vitamin Be 1. Introduction Studies in mouse, rat, and rabbit have established the uncle ve (ena proper of hams py ‘ine, and eyanocobalamin given alone or in combi- nation following intravenous, subcutaneous, and inta- Peritoneal administration. Animal models employed ‘Were the writhing test, heat coil test, hot pate test, and comeal reflex test [, 3, 3,4) Following single oral administration an analgesic effect Of thiamine, pyridoxine, and eyanocobalassin, given lone or in combination has so far only been cemorstra. ted in inflammation-induced pain models employing the ‘arrageenan-induced rat paw or tail ocdema [2], Using EBT Laboratory of Pharmacolagy and Toicolaay, Hamburg (Gnd, Rep. of Germany), and tai fir steicinsche Chore Medizinache Hochschule Hannover, Hannover (ted. Rep: of Gomany 14 ‘und Cyanocobalamin nach wiederholter oraler Verabrei chung bet der Maus ‘Nach wiederholteroraler Vrabreichune iber 7 Tage zeig- ‘en Thiamin (Thiamtin-Nitral, CAS 532-43-4), Pyridosin (Pyrtdoxol-Htydrochlonid, CAS $8.56) und Cyanccoba ‘ain (CAS 6819-9) im hohen Dosishereich bel der Ms im Writhing-Test soweht als Monasubstane als auch in Kombination eine antinozizeptive Dosis Wirkunesbezie. ‘ung. Cyanocobalaran zeigte in der Kombination eine beradditive Wirkung. CAS $32-43-4 - Cyanocobalamin - Pyridoxine the writhing tes, Bartoszyk and Wild [3} were unable to Getect any antinociceptive effect following oral adminis. ‘ration of 2 combination of thiamine, pyridoxing, and eyanocobalamin ata dose level of 100,200, and 0.3 mg! x body weight, respectively to mice, Hence, sim of th study was to determine whether oral administration oF {iamine, pyridoxine, and eyanocabalamin given alone OF in combination for 7 consecutive diys possesses any Antinociveptive effect in the mousecwrithing test. The ‘withing test was chosen as this test system fs the most commonly employed screening test for antinociceptive ‘Properties, favoured because ofits good reproducibility 2, Materials and methods 2.1, Test substances ‘Thamine (Viumin By; thiamine sitate, CAS 532-434), pyc exine (vitamin Bg pyridonlbysrochlonie, CAS $856 020d ‘ganoeabatamin Gitamin By CAS 68-193) wore obtained at BAIVUSP qual The test compounds were suspended in 08 & ‘aqueous aydroxspropy metipellaise el anc ven by gavage 2.2, Animals Female NMRI mice (Lipgische Vevucsterucht, ‘GmbH & Co, Exiertal, FRO) wer weed weighing be nt and24 Mie wore provided with standard mouse dit end water ‘tine onset ofthe main experiment (iting est animals were Tandomiy divided info proupe of, 2.8. Pharmacological methods 2.3.1 Acate toxicity Ing preliminary test the maximum tolerated ote was determi. ted for thiamine, pyridoxine, and eyanceoblamin. The tes fWmpounds were given ones alone or in combination at rato ft 1 Yortiamine and pyridoxine and ota tig ot 10.0005, fortbiamive,pyidorine, and eyanocobaarn, 2.3.2 Writing tast ‘The tet substances were administered so groups ofS mice orally ances day for? consecutive dys, 2h afer the at admis ‘Son the wing rencton sae taaced by tlrapertonelijes- {Hon of 10 mi te aqusous acts aidig body weight The mt ber of withing reactions was monitored fot 20 min after the {njetion.Contol mice received te venile. The doselevels = ployed inthe writhing test are piven In Table Tie Vitamin B ose employe he wigs i po TE Tosa dose a [| rasta ee 10737 49038 2006.5 é | |e 2.3.) Statistical evaluation ‘A Kruskal-Wallis one-way snalyss of varianes in conjunction tha Wilcoxon teosempe test as wed forthe last a> ais 3. Results Ia preliminary acute toxicity test no toxic effets were observed for thiamine, pyridoxine, and cyanocobalamin Ben alone or in combination (ata ratio of 1: | for thia- fine and pyridoxine and at a ratio of ': 1: 0.0025 for thiamine, pyridoxine, and cyanocobalamin) up toa dose level of 5000 ma/kx body weight po. Following repeated oral administration for 7 days ea rnocobalamin alone exerted a slight antinociceptive effect 431 5620 mgkg body weight/day p.o. The number or Wr thing reactions was reduced hy 18 % compared to the control Fig. 1). A combination of thiamine and pyrido- xine ata ratio of 1 1 exerted a more pronounced dose. related antinociceptive effect reducing the number of ‘writhing reactions by 25 and 33% at 1000 and 5620 mg) Kp body weigh/day po. respectively, Addon of Cy Inocobslamia at aralio of 171 0.0025 (thiamine, pyr ‘doxine, and eyanoeobalamin) increased the antinocicep- tive effect even further reducing the number of writhing reactions by 38 and 30 % at 1000 and 5620 me/ke Body weighUday p.o, respectively, 4, Discussion ‘The results of this study revealed a slight dose-related antinociceptive effect for cyanocobalamin at high dose levels in the writhing reaction. A combization of thiae ‘ine and pyridoxine ata ratio of I shoved a higher x24, ‘Re seitig tt ea of eaten haere ls amet m SBT Eisen Bw hema bh ea eB Sarin By (aso 1 00525) "007 cp'e BOS < Oa dogree of dose-related efficacy. Addition of cyanocobae lamin to the thiamine and pyridoxine combination re- sulted in a potentiating effect of cyanocobslamis on the thiamine and pyridoxine combination leading to an in- creased antinociceptive effect. ‘TheB vitamins pay an important rolee.g as co-enzymes inthe synthesis of neurotransmitter, inthe axonal trans- port, or in the excitability of neurons. The underlying ‘europhysiological mechanisms have so far not been elu- gated, Clinical studies have confirmed the therapeutic ticacy of pain relief by using a thiamine/pyridoxine! tyanocobalamin therapy following oral administration in peripheral neuropathy and vertebral syndromes (6, 7) In addtion, B vitamins have been shown to potentiate the antinociceptive effect of non-steroidal ant-inflam- ‘matory drugs sich as diclofense in man following ora administration [2, 8, 9}. 5. References [1 Bantotayk, G, Wile, A, Sor Neurosci. Abst, M4, 324 (1988) ly Herons ©, Wily Ay Newrose Let 101 95 (1939) — ‘Bi Hane A Welacr on 3. Vitam Nu. Re 37,189 (1965) lal erahy HG. Fbig, L. Hotovy, KR, Arenetm. Forsch ‘Drug Res. 11,932 (961) = [5] Wid, A Bartz, Ga it: Ke neck Bedeiang von Visamin By, 84, Bin dar Schinerthe- apie, N- Zoller, H. Fas), 1 Jurna, K'F. Petrik, M. Schatten- [atobner (tsp), 81, Stenkopit- Darmstadt (1988) ~ (6) Le Seman, H, Wiedeley, KD, Therapiewoche 38, 1445 (1988) Sry Sabwger, Git Kliniche Bedeutung von Vitamin B, ‘Be Bye cerSchmeratheraple,N. Zone, Fata, Jura, Fe itis, M. Sehatenkicchner (lag), p. 169, Siciaxepf Bargand(196)—(Shgahlanin Mover.) Keehle, On Klin, Wachensche 6 (1 Briggemann, G, Reet cae EB Wil Wacken 8 TT C986) CCorespondence: Dr. 1. Leuschner, Redderves 8 ‘W208 Hamburg 92 (Fes. Rep. of Germany) ns