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The synthesis of methoxy and hydroxy derivatives of 3,4-dihydro-3-(di-n-propylamino)-2H-1-benzopyran from readily available or commercial o-hydroxybenzaldehydes is described in six steps. The enantiomers of 8-hydroxy-3,4-dihydro-3-(di-n-propylamino)-2H-1-benzopyran 1b have been resolved. It is shown that the compound (−)-1b is a more selective D-2 agonist, compared to apomorphine.

The synthesis of methoxy and hydroxy derivatives of 3,4-dihydro-3-(di-n-propylamino)-2H-1-benzopyran from readily available or commercial o-hydroxybenzaldehydes is described in six steps. The enantiomers of 8-hydroxy-3,4-dihydro-3-(di-n-propylamino)-2H-1-benzopyran 1b have been resolved. It is shown that the compound (−)-1b is a more selective D-2 agonist, compared to apomorphine.

Oxygen Isosteres of hydroxy-di-n-propylaminotetralines. Syntheses of racemic monomethoxy and monohydroxy-3-(di-n-[3H-propyl] amino) chromanes: new Radioligands for 5–HT1A and D2 receptors sites labelling. Condensation of monomethoxy salicylaldehydes: 4a–d with nitroethanol gave methoxy-3-nitro-2H chromenes 5a–d which were reduced with LAH into methoxy-3-amino chromans 6a–d. Two syntheses of 2-hydroxy-6-methoxy-benzal-dehyde: 4a were described. N-alkylations of 6a–d were carried out a/ by Borch procedure b/ by n-propylation with propyl iodide in presence of bases c/ by allylation in presence of bases. Methoxy-3-(diallyl-amino) chromans: 13a–d were catalytically reduced with hydrogen (Pd/C in methanol) into mixtures of methoxy-3 (di-n-propylamino) chromans 2a–d and methoxy-3- (n-propylamino) chromans resulting from the hydrogenolysis of one allyl group. Methoxy-3-dialkylamino chromans 2a–d and 13a were O-demethylated either by boiling 48% HBr or by BBr3 in CH2Cl2. Catalytic hydrogenation with tritium of precursors 13a–d and15a in methanol in presence of 10% Pd/C gave [3H] – 5. OH-DPLC: 3a; [3H] 5. OMe-DPAC: 2a and the isomers: 2b-2d which were purified by HPLC, analyzed by reversed HPLC, 3H-NMR, M.S. (spec. radioactivity: 37.2-80 Ci/μMole. Radiochemical purity: 98.3–99.6%).

La carbonatation avec 14CO2 du Mmethylthiomethyllithium a fourni l'acide methylthio-2 acetique(14C-1) avec un rendement de 83%, dont la reduction par LiA1H4 a conduit au methylthio-2 ethanol-1 (14C-1) qui a ete transforme en chlorure correspondant par action de la tri-n-octylphosphine et CCl4. Le chlorure condense sur le sodio acetamicomalonate d'ethyle a fourni le (methylthio-2 ethyl) acetamidomalonate d'ethyle (14C03) qui a ete saponifie en monoester dont la decarboxylation a produit le DL-methylthio-4 acetamido-2 butyrate d'ethyle (14C-3). L'hydrolyse par l'α-chymotrypsine donne la N-acetyl-L-methionine (14C-3) que l'on separe du methylthio-4 acetamido-2-D-butyrate d'ethyle (14C-3). Par hydrolyse chlorhydrique on aboutit respectivement a la L-methionine (14C-3) et a la D-methionine (14C-3) dactivite specifique: 54 mCi/mM avec des rendements respectifs globaux de 14 et 11% par rapport a 14CO3Ba. Carbonation with 14CO2 of methylthiomethyllithium provides with a 83% yield 2-methylthio-acetic acid 1-14C, the reduction of which with L iAlH4 leads to 2-methylthio-1-ethanol-1-14C which is transformed into the corresponding chloride by action of tri-n-octyl phosphine and CCI4, This chloride condensed with ethyl sodio acetamidomalonate affords ethyl (2-methyl-thioethyl) acetamido malonate which is saponified into to halfester, the decarboxylation of which leads to ethyl DL 4-methylthio-2-acetamidobutyrate-3-14C. The latter is resolved by hydrolysis with a-chymotrypsi which gives rise to N-acetyl-L-methionine-3-14C which is separated from ethyl 4-methylthio-2-acetamido-D-butyrate-3-14C. By hydrochloric hydrolysis are obtained respectively L-methionine-3-14C and D-methionine-3-14C- specific activity: 54 mCi/mMole in respective overall yields 14 and 11% based on bariun carbonate 14C.

Racemic [3-carboxyl 14C] agaric acid triethyl ester was prepared in low yield by treatment of ethyl 2-ethoxalyl octadecanoate 2 with either ethyl lithioacetate or ethyl bromoacetate, zinc and methyl borate. In another approach, lithio ethynyltrimethylsilane 6 was condensed with 2 to give 7 which, with dicyclohexylborane 8, gave the vinyl borane 9; the latter, by hydrogen peroxide oxidation followed by reaction of diazomethane gave the mixed ester 11 in low yield. [3-carboxyl 14C] Agaric acid, specific activity 57 mCi/mmol, was obtained from 5 or 11 by saponification with lithium hydroxide in 1,2-dimethoxyethane. The best overall yield from ethyl [1-14C] stearate was 18%.

For the development of radioimmunoassay procedures for colchicine, three haptens, N-ethylamino-colchiceinamide, 4-formylchochicine - (O-carboxymethyl) oxime and 4-hydroxymethylcolchicine O-hemisuccinic acid were synthetized and characterized by mass and proton magnetic resonance spectrometries. The conjugates obtained by coupling the haptens to bovine serum albumin were employed to immunize rabbits and goats.

Benzoyl chloride-714C is condensed with tris(trimethylsilyl) 1-lithio-1,1,2 propane tricarboxylate, which after hydrolysis gives rise to a 66% yield of 2-methyl-3-(benzoyl-7-14C) propionic acid. After cyclisation with hydrazine, followed by treatment with bromine in acetic acid, 3-hydroxy-4-methyl-6-phenyl pyndazine 6-14C is secured in a 77% yield. This hydroxypyridazine treated with POCl3 gave a 90% yield of 90 % pure 4-methyl-6-phenyl-3-chloropyndazine 6-14C; The latter treated with 2-morpholino-1-aminoethane in presence of sodium iodide has given 3-morpholinoethylamino 4-methyl 6-phenylpyndazine 6-14C. The overall yield based on barium carbonate 14C is 16.7 % (specific activity 18.5 mCi/mMole).

SYNTHESIS OF A LABELLED DERIVATIVE OF METHOMYL WITH POTENTIAL AFFINITY FOR THE BIOLOGICAL ACTIVE SITE OF MITOCHONDRIAS TEXAS The Grignard reagent from 1-chloro-4-methoxy butane was carbonated with 14CO2 giving [carboxyl14C]-5-methoxy pentanoic acid Yield: 80%. Specfic activity : 24,3 mCi/mmole Treated with dry hydrogen bromide in acetic acid, the latter gave [carboxyl 14C]-5-bromopentanoic. acid with was condesed with diphenylphosphoryl azide leading to 5-bromobutyl [carbonyl 14C] isocyanatc. Without isolation, the latter was reacted with methylthioacetoxy giving [carbonyl 14C) S-methyl-N [(4-azidobutyl. carbamoyl) oxy] thivacetimidate with an overall yield of 15,7 %/14CO2 and a radiochemical purity of 99,8 %. Specific activity : 23,7 mCi/mmole.

High specific activity tritiated 3β, 14α -dihydroxy-5β-cholest-7-en-6-one, has been prepared using a precursor which permits rapid and easy labelling. This compound is converted to ecdysone under in vitro conditions by insect prothoracic glands, a well known site of ecdysone biosynthesis.

(2R, 3S); (2S, 3R) [4-14C] Agaric acid and (2R, 3R); (2S, 3S) [4,4,5,5-3H] agaric acid were synthesized from respectively threo methyl 1,2-epoxypropane-1,2,3-tricarboxylate 3 and erythro epoxide 2. Epoxy ring opening of 2 and 3 with [1-14C] hexadecyl copper 10. gave respectively the analog 4 of agaric acid trimethyl ester and methyl [4-14C]agaricate 5, which was hydrolyzed to 1a by lithium hydroxide in 1,2-dimethoxyethane (DME). Epoxy ring trans opening of 2 with diethyl 1-hexadecynylalane 9 gave the alcynyl derivative 7 which was saturated with tritium in presence of Pd/C to lead to 6. After hydrolysis (LiCH + DME), 1b was obtained. The structures of the diastereoisomers (2R, 3R); (2S, 3S) 1b and (2R, 3S); (2S, 3R) 1a rest on 1H-NMR.

A high-affinity (Kd= 5.9 nM) specific binding site for [3H]harmaline was detected in membranes from rat and bovine brains. Studies of the regional and subcellular distributions of this binding indicated its close association with monoamine oxidase type A activity (MAO A) measured with [3H]serotonin ([3H]5-HT) as the substrate. Maximal binding capacity and MAO A activity were found in mitochondrial enriched fractions. Mitochondria of synaptosomal or extra-synaptosomal origin exhibited very similar properties with respect to [3H]harmaline binding characteristics and MAO A activity. Among psychoactive drugs, only monoamine oxidase inhibitors (MAO I) prevented the specific binding of [3H]harmaline. Logit-log inhibition curves of binding by MAO I gave only one slope which was not significantly different from 1.0, suggesting the existence of only 1 category of specific sites for [3H]harmaline in the membrane preparations from rat and bovine brains. Consistent with the preferential inhibition of MAO A by harmaline, other MAO I of this class, i.e. clorgyline and Lilly 51641, were 102-2 × 103 times more efficient than deprenyl and pargyline, two inhibitors of MAO type B, in displacing [3H]harmaline from its specific binding site. Ki and IC50 values for the inhibition of [3H]harmaline binding by MAO I and MAO substrates (tryptamine, 5-HT, norepinephrine) were almost identical with those characterizing their action on MAO A activity with [3H]5-HT as the substrate. In conclusion, the specific binding site for [3H]harmaline exhibited all the expected properties of the active site of MAO A. Like the technique of precipitation with a specific antibody, binding of [3H]harmaline should be of great help for studying the structural characteristics of the active site of MAO A and determining the number of MAO molecules in tissues under various physiological conditions.

The synthesis of an alpha adrenergic antagonist labelled with tritium: [(2,3-3H)-1,4-benzodioxanye] WB 4101 or 2-N[2,6 dimethoxyphenoxyethye] aminomethyl 2,3-3H-1,4 benzodioxan is described. 2-cyano-1,4-benzodioxan: 1 was prepared from catechol and 2-chloroacrylonitrille and then converted to 2-cyano-1,4-benzodioxin: 2 by the successive actions of N-bromosuccinimide in CCl4 and sodium iodide in acetone. 2 was reduced with lithium aluminium hydride into 2-aminomethyl-1,4-benzodioxin: 3 and then converted to 2-[N-(2,6-dimethoxyphenoxyethyl)-aminomethyl]-1,4-benzodioxin: 4a, and its hydrochloride: 4b. The catalytic reduction of 4a, 4b with tritium in various and in presence of various catalysts was studied. The best result was obtained with Wilkinson's catalyst in benzene giving rise to [(2,3-3H)1,4-benzodioxanyl]WB 4101 with a specific activity of 60 CilmMole. 3H.NMR and MS analyses are also described.

14C labelled phospholipid synthesis through the use of “cyclic enediolpyrophosphate”. Ramirez's phrophosphate 1 was used in a one pot convenient synthesis of a few [1-14C]-acyl phospholipids. Reagent 1 was condensed with 1,2-[1-14C] dipalmitoyl Sn-glycerol to the cyclic triester phosphate 4 which without isolation, was reacted with N-tritylethanolamine, 3-0-benzyl Sn-glycerol and glycerol leading to intermediates 5, 6, 7 which were deprotected by triethylamine hydrolysis and other appropriate treatments.

Four methods of syntheses of IDAZOXAN (RX 781094): |2-(2-imidazolinyl)-1,4-benzodioxane| labelled with tritium: 1 at high specific activity, a selective radioligand presynaptic α2- adrenoreceptor antagonist are described. 2-carbethoxy-1,4-benzodioxine 5 was treated with tritium in benzene in presence of Wilkinson's catalyst leading to 2-carbethoxy [2,3-3H]-1,4-benzodioxane: 6 spec. act. 60 Ci/m which was then condensed in toluene with a reagent made from trimethylaluminium and ethylenediamine giving rise to 1 specific activity 44 Ci/mMole. 2-(carbamido N-cyanomethyl)1,4-benzodioxine: 2 was prepared from 2-chlorocarbonyl-1,4-benzodioxine: 10 and 2-aminoacetonitrile. 2 was reduced with tritium in trifluoroacetic acid in presence of Pd/C giving 8 which, without purification was ring closed into 2 by trimethylaminium treatment. Spec. act. 35.5 Ci/mMole. When reduced with tritium in presence of Wilkinson's catalyst, 7 gave 9 spect. act. 55 Ci/mMole. After purification 9 was again reduced with tritium in trifluoroacetic acid in presence of Pd/C, or PtO2, or Rh/Al2O3 giving 8 which was cyclised into 1 by trimethylaluminium treatment in toluene. spec. act. of 1 was 41.5 - 39 Curies/mMole. 1 was purified by successive preparative thin layer chromatography. The radiochemical purity checked by TLC and PLC was better than 99%. Some observations on the rate of self radiolysis of 1 are reported.

Methyl cinnamate -3-14C is reduced smoothly in excellent yield to cinnamyl alcohol -3-14C by aluminum hydride. without attack of the double bond. After purification by column chromatography cinnamyl alcohol -3-14C is obtained with a radioactive overall yield of 65% based on cinnamic acid -3-14C, specific activity = 9 mCi/mM. Le cinnamate de methyle 14C-3 est reduit par l'hydrure d'alumninium en alcool cinnamique 14C-3 avec un excellent rendement. sans attaque de la double liaison. Apres purification par chromatographie sur colonne. on obtient l'alcool cinnamique 14C-3 avec un rendement radioactif global de 65% par rapport a l'acide cinnamique 14C-3. aotivite specifique : 9 mCi/mM.

SYNTHESIS OF 2-METHYL-4,6 DINITRO PHENOL (RING U-14C) (DNOC, DINOC ring U-14C) O-methoxymethylphenol (ring U-14C) 3 Was prepared in 82 % yieltd from phenol (ring U-14C) and methylal in presence of p-toluene sulfonic acid and molecular sieve. Ortholithiation of 3 with n Buli + TMEDA in hexane offorded a lithioderivative 4 which was methylated with methyl iodide in ether - hexane. Deprotection of phenol group lead to o - cresol (ring U-14C) : 6 purified on a silicagal column with a 52% yield based on 3. Nitration of 6 provided DNOC (ring U-14C) with a 28% overall yield from phenol (ring U-14C) - Specific activity was: 70 mCi/mMole.

Methyl iodide-14C was condensed with 2-(1-lithio-hept-1-yn-6-yl) 1,3-dioxolanne 4 in presence of HMPT and benzene to give 2-(oct-2-yn-8-yl-1-14C) 1,3-dioxolanne 5. Catalytic reduction of 5, followed by acid hydrolysis led to pelargonic aldehyde 9-14C 7. Condensation of this aldehyde 7 with an excess of w-carbomethoxy-tetradecylidene), triphenyl phosphorane 11 produced methyl nervonate 24- 14C 12. The latter was isolated from its trans isomer by chromatography on a silver nitrate impregnated silicagel column. Finally, nervonic acid 24-14C 13 was obtained by alkaline hydrolysis (specific activity : 52 mCi/mMole) with an overall yield of 7% based on methyl iodide-14C.

Benzyl chloride-7-14C is condensed with 2-lithio 2-methyl 1,3-dithianes. Deprotection with cupric chloride-copper oxide of the resulting 2-benzyl 2-methyl 1,3-dithianes leads to 1-phenyl-2-propanone-1-14C. The latter with L(-)α-methylbenzylamine in presence of molecuLar sieve gives rise to (-)N-(α-phen-ethyl)phenylisopropylimine which is catalytically reduced into the corresponding amine. Hydrogenolysis of this amine gives D(+) amphetamine-7-14C isolated as the sulfate with an overall yield of 10 % based on benzoic acid-7-14C specific activity: 27 mCi/mM.

La synthese de la dl-norephedrine (methyle 14C) en deux stades, a partir de l'α-N, N-dibenzylaminoacetophenone et de I14CH3 est decrite. Apres purification par chromatographie sur resine echangeuse d'ions AG 50 W-X2 on obtient la dl-norephedrine (methyle 14C) HCl avec un rendement radiochimique de 31% par rapport a 14CO3Ba, activite specifique: 55 mCi/mM. L'analyse par electrophorese sur papier associee a la chromatographie sur papier et en couche mince permet de contrǒler la purete radiochimique de la dl-norephedrine 14C. A two-step synthesis of dl-norephedrine (methyl-14C) from a-N, N-dibenzylaminoacetophenone and 14CH3I is described. After purification by chromatography on an ion exchange resin column AG 50W-X2 dl-nore-phedrine (methyl-14C) HCI is obtained with a radioactive overall yield of 31% based on Ba14CO3, specific activity: 55 mCi/mM. The analysis by paper electrophoresis in conjunction with the paper and thin-layer chromatographies enables control of radiochemical purity of dl-nore-phedrine 14C.

The Grignard reagent made from p-cyclohexyl m-chloroiodobenzene is carbonated with 14CO2 to give 3-chloro 4-cyclohexyl benzoǐc acid (radioactive yield 50 %). This acid is transformed into the corresponding chloride with thionyl chloride. Condensation of the chloride with tris-(trimethylsilyl) 1-lithio-1,1,2 ethane tricarboxylate gives after hydrolysis 4-(3-chloro 4-cyclohexyl phenyl) 4-oxobutyric acid 4-14C with an overall yield of 41 % based on barium carbonate. (Specific activity : 53 mCi/mMole).

1-demethylcolchicine obtained by selective demethylation of colchicine was condensed with (14C) CH3 I to gave (1-methoxy 14C) colchicine 4. After two purifications by silicagel column chromatography 4 was obtained in a 40% yield from (14C) CH3 I and a radiochemical purity of 99% (specific activity : 57 mCi/mmole).

Carbonation of 3,4 methylenedioxy phenylmagnesium bromide with 14CO2 leads to (carboxyl 14C) 3,4 methylenedioxy benzoic acid. Reduction of the latter with LiAlH4 in T H F1 gives rise to 3,4 methylenedioxy benzyl alcohol which reacts with thionylchloride to provide the corresponding chloride. By action of magnesium in T H F 3-4 methylenedioxy benzylmagnesium chloride is obtained; it is then condensed with vinyl bromide in presence of catalytic quantities of FeCl3 to give (1′-14C) safrol, specific activity: 15 mCi/mMole with an overall yield of 40 % based on barium carbonate.

(ring-4,6-14C) Pentamethylmelamine (14C-PMM), a new antitumor agent, was prepared from barium (14C) carbonate by a 6 step synthesis. (14CN) Dimethylcyanamide, prepared from sodium (14C) cyanide with 71% yield and treated with phosgene in the presence of hydrogen chloride, was cyclized to (ring-4,6-14C)-2-chloro-4,6-bis (dimethylamino)-s-triazine. The latter was purified by silicagel low pressure liquid chromatography (15% yield based on nitrile) and condensed with monomethylamine to give 14C-PMM. After purification 14C-PMM was obtained with 91% yield, specific activity = 38 mCi/mMole. The mechanism of the reaction of nitrile with phosgene was discussed.

The distribution of the binding sites of a new, potent agonist of serotonin (5-HT), 8-OH-N,N-dipropoyl-2-aminotetralin (PAT), was studied in the rat brain with the quantitative autoradiographic technique utilizing tritium-sensitive LKB film. The localization of [3H]PAT binding sites was very similar to that of [3H]5-HT binding sites, except in some discrete regions (choroid plexus, striatum, area preoptica lateralis, subiculum, and substantia nigra), which exhibited very low levels of labeling with [3H]PAT and high levels with [3H]5-HT. These results indicate that 5-HT1 receptors are heterogeneous, and that [3H]PAT recognizes only a 5-HT5 subclass (called 5-HT1A).

The carbonation with 14CO2 of the lithium derivative 6 prepared from p-chlorodiphenylmethane 5 and n-butyl lithium in THF in presence of tetramethylenediamine gives (14C-carboxyl) 2-(p-chloro phenyl)-2-phenylacetic acid 7 with 85 % yield based on 14CO2. Action of thionyl chloride gave the corresponding chloride 8 which was condensed with the lithium reagent derived from trimethylsilyl malonate 9 leading after hydrolysis to 1-(p-chlorophenyl)-1-phenyl 2-propanone-2-14C 4 with an overall yield of 72 % based on 14CO2. The ketone 4 reacted with methylphthalate gave rise to 2-((2′-p-chlorophenyl) phenylacetyl-1′-14C)-1, 3-indane dione 1 (chlorophacinone-14C). Overall yield based on 14CO2 was 36 % - specific acitivity : 15 mCi/mMole.

3-([2,6-14C]2-pyridinyldithio) propanoic acid, a disulphide covalent coupling reagent was synthesized in 12 steps from sodium [14C] cyanide. Condensation of sodium [14C] cyanide with 1,3-dibromopropane gave, after hydrolysis [1,5-14C] glutaric acid: 4 which was converted into [1,5-14C] glutarimide : 5 by heating at 280°C with anhydrous ammonia. The imide : 5 was then converted into [2, 6-14C] 2, 6-dichloropyridine: by 6 by treatment with phosphorous penta and trichlorides. Chlorine removal from 6 by hydrogen (Roney Nickel) gave [2, 6-14C] pyridine: 7 which was isolated as the hydrochloride. The latter was tranformed into [2, 6-14C] pyridine-N-oxide :11 by hydrogen peroxide treatment. 11 treated by acetic anhydride (15 hours - 130°C) rearranged into 2-acetyloxy [2, 6-14C] pyridine: 12 which was hydrolysed into [2, 6-14C] 2-pyridone: 13. Thiation of the latter by P2S5 in xylene gave 2-mercapto [2, 6-14C] pyridine: 10 which was then oxidized into [2, 6-14C] 2-dipyridinyldisuplphide: 14 the chlorination of which lead to [2, 6-14C] 2-pyridinylsulfenyl chloride: 15 which was not isolated. It was condensed with 3-mercapto propanoic acid to give 1 with an overall yield of 4% based on sodium cyanide - Specific activity was 55 mCi/mMol. The radiochemical purity checked by TLC and HPLC was 98.7%.

(tetramethyl- 14C)Pentamethylmelamine 6, a new antitumor agent, was prepared by a three-step synthesis starting with (14C)CH3I. Toluene-p-sulphonamide was condensed with (14C)CH3I to give labelled NN-dimethyltoluene-p-sulphonamide. 14C-labelled dimethylamine was released from the dimethylamide by the action of 48% HBr and NaOH, then reacted with 2,4-dichloro-6-methylamino-1,3,5-triazine to afford 6. After purification by silicagel column chromatography 6 was obtained in a 8.5% yield from (14C)CH3I and a radiochemical purity of 99%, specific activity : 78.5 mCi/mmole.

Synthesis of 2-acetoxy 4-trifluoromethyl benzoīc acid [ring U -14C] or “[ring U-14C] Trifusal” U14C)(Ring U-14C) Aniline 1 was converted into (ring U- 14C) idobenzene 2 through the (ring U-14C) phenyldiazonium chloride. Trifluoromethylation of 2 with iodotrifluoromethane in presence of copper gave (ring U-14 C) trifluoromethylbenzene 3 with a 64% yield. Nitration of 3 with sodium nitrate + trifluoromethane sulfonic acid in dichloromethane gave a 86% yield of (ring U-14C) 3-nitro trifluoromethyl benzene 4 which was reduced by iron and HCl into the corresponding amine 5. The latter was transformed into the phenol 6 with a 85% yield by action of Cu (NO3)2 and CuO on (ring U-14C) 3-trifluo romethyl phenyldiazonium sulfate. Kolbe reaction of phenol 6 with carbon dioxide and K2CO3 gave 7 with a 59% yield. Acetylation of 7 gave (ring U-14C) Triflusal 8 with a quantitative yield and a specific activity of 14 mCi/mMole14 radiochemical purity 98.5%. The overall yield from 14C barium carbonate was 10%.

2-Thenoic acid (14C = 0) 1 is prepared in 75 % radioactive yield by carbonation with 14CO2 of 2-thienyL-magnesium bromide. Boiling of 1 with oxalyl chloride gives rise to 2-thenoylchloride (14CO) which is not isolated and is condensed with 2,3 dichLoroanisoLe under FrieLdeL-Grafts conditions. Ketone 3 is demeth Lated with AlCl3 in benzene to give rise to 4- 2-thenoyl (14C = O)-2,3-dichloro phenol 4. The sodium derivative of 4 is condensed in D.M.F. with sodium chloracetate to Lead to the title compound with an overall yield of 16 % based on 14CO2 – TieniLic acid-14c, specific activity: 50 mCi/mMole, has been found to be sensitive to self-irra-diation decomposition in the dry state at - 25%° C. Radiation decomposition is minimized at 24 mCi/mMole in methanol solution.

24,25-3H dihydro lanosterol is obtained by catalytic reduction of lanosterol with tritium at atmospheric pressure. Its specific activity is 23 Ci/mMole. Natural lanosterol impurities do not disturb the synthesis.

dl-Methadone was resolved by crystallization of its ammonium d- α - bromocamphor- π - sulfonate salt to give d-methadone. The latter in ethyl acetate solution was reduced with tritium gas to 1- α -methadol 3H in presence of Adams platinum oxide at normal temperature and pressure. Acetylation of 1- α -carbinol hydrochloride by means of acetyl chloride afforded 1- α -acetylmethadol 3H, specific activity : 20 Ci/mMole. The positions and extent of tritium labelling were determined by 3H NMR spectroscopy.

An efficient procedure is described for alkylation of quercetin with alkylhalides by use of tetraethylammonium fluoride in DMF or HMPT. The method is applied successfully to the preparation of 3,7,3′,4′,tetra-O-ethyl 5 [3H] O-ethylquercetin with a specific radioactivity of 45 Ci/mmol.

Procedures are described for the synthesis of the following metabolites of deltamethrin, the pyrethroid insecticide: 3-phenoxy (carboxyl-14C) benzoic acid, 3-(2′-hydroxyphenoxy) (carboxyl-14C) benzoic acid and the corresponding 3-phenoxybenzyl alcohols, specific activity = 47–57 mCi/mmol.

Cross-coupling of 3,5-dimethoxybenzyl bromide and 4-buten-magnesium bromide in presence of Li2CuCl4 afforded 1-(3,5-dimethoxyphenyl)-4pentene. Demethylation of the latter by methylmagnesium iodide in xylene gave rise to 4′,5′-dehydro-olivetol. The latter, condensed with trans-p-mentha-2,8-dien-1-ol in the presence of either boron trifluoride etherate or p-toluenesulfonic acid, gave 4′,5′-dehydro-Δ9- and Δ8-THC respectively. 4′,5′-dehydro-Δ8-THC was also obtained by isomerization of 4′,5′-dehydro-Δ9-THC. Catalytic selective reduction of these precursors with deuterium or tritium, in the presence of tris(triphenylphosphine)rhodium chloride, afforded Δ9-THC-4′,5′,-2H or Δ9-THC-4′,5′-3H (50 Ci/mMole) and Δ8-THC-4′,5′-2H or Δ8-THC-4′,5′-3H (56 Ci/mMole).

6,7-dimethoxy-4 (parachlorobenzyl) (D-3) isoquinoline : 6 and 6,7-dimethoxy-4 [parachlorobenzyl (methylene 14C)] isoquinoline: 11 were prepared respectively by the condensation of α-deuterio-veratrylaminoacetaldehyde diethylacetal : 4 with parachlorobenzaldehyde and the reaction of parachlorobenzaldehyde (carbonyl 14C) : 9 with veratrylaminoacetaldehyde diethylacetal.

Formaldehyde-14C and 1, 2-dimethyl-4, 5-bis (mercaptomethyl) benzene give a 90% yield of 1, 3 dithiane : 2 b which is metalated by n-butyllithium in ether into the 2, 2-dilithio 1, 3 dithiane : 3 b (yield 70 %). In situ 3 b is treated with ethyl chloroformate to give rise to 4 b in a 70% yield. 4 b is desulfurated by treatment with Raney Nickel in ethanol, giving an 80% yield of ethyl malonate-2-14C. The overall yield from baryum carbonate is about 35%.

Classical phospholipids syntheses were selected and adapted to the preparation of natural glycerophospholipids labelled with carbon 14 in the acyl moiety. 1,2-(14C) diacyl-3-bromo sn-glycerol was the starting material. Saturated 14C-phosphatidyl-ethanolamine, choline and serine were obtained in 30 %, 25 % and 11 % yields relative to the 14C-fatty acid, by condensation of a suitably protected amino-alcohol with a (14C-diacyl) phosphatidic acid in presence of triisopropyl-benzenesulfonyl-chloride followed by deprotection of the amino group. The synthesis of unsaturated (or mixed) 14C phosphatidyl ethanolamine was achieved, with a 28 % yield, by condensation of the 14C-diacyl-bromoglycerol with the silver salt of the N-trityl aminoethyl and benzyl diester of phosphoric acid and subsequent deprotections.

By use of procollagen proline hydroxylas which catalyses the substitution of the hydrogen in the trans position on carbon 4 with an hydroxyl on L-proline residues of procollagen, we have shown that the L-[3H-4] proline product of C.E.N. Saclay contained 29 per cent of the trans and 71 per cent of the cis form. It was also possible to verify the molecular stability of the isomers and that a small amount of tritium was randomly linked to the whole molecule.

Syntnesis of very high specific activity (4,6-14C2) 2-isopropylaminopyrimidine phosphate (I.A.P.P.) (1-14C) Sodium acetate (specific activity : 57 mCi/mMole)- was transformed into ethyl (1- C) acetate with a 76 % yield. The latter was lithiated with lithiodiisopropylamide. The intermediate ethyl (1-14C) lithioacetate was carbonated with 14CO2 (specific activity : 57 mCi/mMole) to give monoethyl (1,3-14C2) malonate which was esterified into diethyl (1,3-14C2) malonate. The latter was condensed with N-isopropylguanidne to give 4,6-dihydroxy 2-iso-propylamino (4,6-14C2) pyrimidine. Treatment with POBr3 gave 4,6-dibromo 2-isopropylamino (4,6-14C2) pyrimidine the hydrogenation of which under controlled conditions gave 2-isopropylamino (4,6-14C2) pyrimidine isolated as the phosphate. Specific activity was 114 mCi/mMole, radiochemical purity (TLC and HPLC) was 99 % - overall yield was 7 % based on (14 C) carbon dioxide.

SYNTHESIS OF N-ACETYLMURAMYL (14C-OXOPROPYL)-L-ALANYL-D- ISOGLUTAMINE (14C - MDP) Condensation of (1-14C) DL-2-bromopropionic acid with benzyl 2-acetamido-4,6-0-benzylidene-2-deoxy-β-D-glucopyranoside 7 in presence of sodium hydride gave 50 % of crystalline compound 3, having the correct stereochemistry of natural muramic acid. The latter, in presence of Woodward's reagent, reacted with L-alanyl-D-isoglutamine benzyl ester hydrochloride, leading to compound 4. Removal of benzylidene and benzyl groups by successive acetic acid exposure and hydrogenolysis gave rise to 2-acetamido-2-deoxy-3-0 (2-D-(14CO) propionyl-L-alanyl-D-isoglutamine)-D-glucopyranose (14C-MDP) Specific activity : 42 mCi/mMole in a 10 % overall yield based on sodium (1-14C) propionate.

A specific binding site for [3H]metergoline characterized by a KD of 0.5–1.0 nM was detected in microsomal and synaptic plasma membranes from various areas of the adult rat brain. Experiments with 5,7-dihydroxy-tryptamine- and kainic acid-induced lesions indicated that this specific binding site was localized post-synaptically with respect to serotoninergic neurons. The pharmacological characteristics of [3H]metergoline binding to microsomal membranes from the whole forebrain strongly suggest that this ligand labels a class of serotonin receptors. This was particularly obvious in the hippocampus in which serotonin was about 400 times more potent than dopamine and norad-renaline for displacing bound [3H]metergoline. In the striatum, serotonin was only 10 times as potent as dopamine in inhibiting [3H]metergoline binding, suggesting that this ligand may also bind to dopamine receptors. Striking similarities between the binding sites for [3H]metergoline and [3H]serotonin were observed in the hippocampus. Thus, not only the total numbers of binding sites for these two ligands in control rats but also their respective increases following intracerebral 5,7-dihydroxytryptamine treatment were very similar. Therefore, at least in the hippocampus, [3H]metergoline might well be the appropriate ligand for studying the characteristics of the ‘antagonist form’ of serotonin receptors postulated by Bennett and Snyder.

Carbonation with 14CO2 of penylethynyl-lithium has given rise to (carboxyl-14C) phenylpropiolic acid with 75 % yield based on 14CO2. The latter condensed with 2-naphthol by treating with polyphosphoric acid afforded (4-14C) β-naphthoflavone, purified by silicagel low pressure liquid chromatography; 22 % overall yield based on 14CO2, SA = 13 mCi/mM. Similar run of 2-(8-14C) naphthol with phenylpropiolic acid gave (5- 14C) β-naphthoflavone; 28 % yield based on radioactive 2-naphthol, SA = 7.2 mCi/mM.

La synthese amelioree de la progesterone 14C-4 est decrite. La purification de la progesterone 14C-4 par chromatographie sur colonne de silicate de Mg et CCM preparative a permis l'isolement de deux sous-produits: isoprogesterone 14C-4 et trioxo-3,5,20-seco A-4,5 pregnane 14C-4, identifies par spectrometrie RMN et de masse. Rendement radioactif global: 40% par rapport a 14CO3Ba, activite specifique: 59 mCi/mM. An improved synthesis of progesterone-4-14C is described. Purification by Mg silicate column chromatography and preparative TLC allowed the isolation of isoprogesterone-4-14C and 3,5,20-trioxo-4,5-seco-A-pregnan-4-14C, as by-products identified by NMR and mass spectrometry. Radioactive overall yield: 40% based on Ba 14CO3, specific activity: 59 mCi/mM.

The two 3H-labeled agonists [3H]8-hydroxy-2-(di-n-propylamino) tetralin ([3H]8-OH-DPAT) and [3H]serotonin ([3H]5-HT) have been used to examine the effects of physico-chemical parameters and modulatory agents on the high affinity 5-HT receptor binding sites in various regions of the rat central nervous system. Sites labeled by [3H]8-OH-DPAT and [3H]5-HT were differentially sensitive to changes in incubation temperature and pH, such that the optimal interaction of [3H]8-OH-DPAT with specific sites in the striatum was at 30 degrees C and pH 7.4, whereas [3H]5-HT sites in the same region were most easily labeled at 2-23 degrees C and pH 8.2. Micromolar concentrations of Mn2+ enhanced [3H]5-HT binding but inhibited markedly [3H]8-OH-DPAT binding to striatal membranes. In contrast, both [3H]5-HT and [3H]8-OH-DPAT binding were increased by the cation in hippocampal membranes. Conversely, GTP reduced the binding of either ligand in the hippocampus but affected only [3H]5-HT binding in the striatum. Furthermore, N-ethylmaleimide inhibited equally [3H]5-HT and [3H]8-OH-DPAT binding to hippocampal membranes, but was markedly less potent against [3H]8-OH-DPAT binding to striatal membranes. These results led to the definition of assay conditions for studying separately [3H]8-OH-DPAT binding to "hippocampal-like" (HL) and "striatal-like" (SL) sites. [3H]8-OH-DPAT HL binding sites were particularly abundant in the hippocampus, septum and cerebral cortex, and exhibited pharmacological properties typical of the postsynaptic 5-HT1A subsites previously characterized with [3H]5-HT as the ligand. The regional distribution of [3H]8-OH-DPAT SL binding sites was strikingly different from that of HL sites, but similar to that of serotoninergic terminals identified by their capacity to take up [3H]5-HT. The selective lesion by 5,7-dihydroxytryptamine of serotoninergic projections induced a marked loss of [3H]8-OH-DPAT SL binding sites in the striatum and the cerebral cortex, indicating that these sites were located presynaptically. In contrast, [3H]5-HT binding sites remained unchanged in lesioned rats, which confirmed further their exclusive postsynaptic location in brain.

{0-methyl 14C} and {0-methyl 3H} bergaptene were prepared by methylation of bergaptol with {14C} methyl iodide and {3H} methyl iodide in radioactive yields respectively 77 % and 59 %. {14C} methyl iodide and 11-hydroxy ibogamine gave {0-methyl C} tabernanthine with a 60 % radioactive yield.

Carbonatation of 3-trifluoromethylthio phenylmagnesium bromide 2 with 14CO2 leads to (carboxyl-14C) 3-trifluoromethylthio benzoic acid 3. Reduction of the latter with borane-methyl sulfide complex (CH3)2 S - BH3 in presence of trimethyl borate gives rise to 3-tri-fluoromethylthio benzyl alcohol 4, which is oxidized by lead tetracetate in pyridine to give 3-trifluoromethylthio benzaldehyde 14CO 5. The latter reacts with nitroethane to give 1-(3′-trifluoro methylthio phenyl)-2-nitropropene-1-14C 6. This nitropropene is reduced by Fe-HCl in presence of FeCl3 into 1-(3′-trifluoromethyl-thio phenyl)-2-propanone-1-14C 7. Condensation of ethylamine with the ketone 7 followed by reduction in situ with sodium borohydride leads to 1-(3′-trifluoromethylthio phenyl)-2-ethylamino propane-1-14C 8 isolated as the hydrochloride. Specific activity: 47 mCi/mMole with an overall yield of 10 % based on barium carbonate-14C.

Three reactions schemes were explored for the preparation of pure isomer free [ring U-14C] parachlorophenylacetonitrile (pCPN). The first route through the cyanomethylation of [U-14C] p-dichlorobenzene with acetonitrile in presence of sodium amide in liquid ammonia gave actually a mixture of meta and para chlorophenylacetonitrile in contradiction with a literature report (5). The second route involved the preparation on a microscale of [U-14C] p-chlorophenylmagnesium chloride in T.H.F, and its hydroxymethylation to [ring U-14C] p-chlorobenzyl alcohol with formaldehyde. This route gave low yields of isomer free pCPN. The third reaction scheme supplied pCPN in 9 steps from [U-14C] aniline. [ring U-14C] acetanilide 11 was brominated and after deacetylation isomer free p-bromoaniline 13 was obtained. 13 was diazotized and transformed into [U-14C] p-chloro bromobenzene: 14 which gave the corresponding Grignard 15 with Mg in ether. Carbonation of 15 gave [ring U-14C] p-chlorobenzoic acid which was reduced (LAH) into the corresponding alcohol which by reaction with bromotrimethylsilane gave [ring U-14C] p-chlorobenzyl bromide 9. Condensation of 9 with KCN in presence of crown ether 18 c 6 gave [ring U-14C] p-chlorophenylacetonitrile which was purified by medium pressure column liquid chromatography. The overall yield from [U-14C] aniline was 26%. The radiochemical purity was better than 99.5% - specific activity : 46 mCi/mmole.