AU639669B2 - Process for the specific synthesis of new dithiocarbamic acid esters by substituting hydroxylated sites on mono- or polyhydroxylated molecules; products obtained by this process and their application - Google Patents

Description

639669 COMMONWEALTH OF AUSTRALIA PATENTS ACT 1952 COMPLETE SPECIFICATION NAME ADDRESS OF APPLICANT: Institut Merieux International 17, rue Bourgelat Lyon F-69002 France NAME(S) OF INVENTOR(S): Denis Ghislain POSTEL Gino Lino RONCO Pierre Joseph VILLA Guy Andre VILLE Robert PLAN ADDRESS FOR SERVICE: DAVIES COLLISON Patent Attorneys 1 Little Collins Street, Melbourne, 3000.

COMPLETE SPECIFICATION FOR THE INVENTION ENTITLED: Process for the specific synthesis of new dithiocarbamic acid esters by substituting hydroxylated sites on mono- or polyhydroxylated molecules; prod ucts obtained by this process and their application The following statement is a full description of this invention, including the best method of performing it known to me/us:- (C~r.

1A This invention relates to a process for specific preparation of new dithiocarbamic acid esters from mono- and polyhydroxylated molecules.

It also relates to products obtained by this process as well as their applications, notably as drugs.

It is known that N,N-dialkyl-dithiocarbamic acids and their salts are used as chelating and complexing agents, notably for transition metals. It is known that these compounds also have interesting biological properties and that they are used as radioprotectors, as antiviral and antitumorai agents.

It is known that the synthesis of dithiocarbamic acid esters can only be made by direct action of dithiocarbamic acids on the OH group(s) of alcohols or o2 polyols.

2 Une of the aims of this invention is to suggest a process for specific preparation of dithiocarbamic acid esters from mono- or polyhydroxylated molecules with high yields even on the secondary hydroxylated, other than anomeric, sites, and which may easily be put into practice.

Another aim of this invention is to provide new dithiocarbamic esters whicn, through their biological properties, find a particularly interesting application I as immunomodulating and antiviral drugs.

The inventive process is characterized in that in a preliminary step at least one hydroxyl group of mono- or polyhydroxylated molecules is transformed into a leaving group, and in that this leaving ,5 group is displaced by the anion N C S wherein R 1 and R 2 represent saturated or unsaturated substituted or unsubstituted alkyl radicals, or else forming with the nitrogen atom a saturated or unsaturated ring which may or not bear heteroatoms, and which is 'S'0 U introduced as a salt in the appropriate medium.

The leaving group is preferably made up by an halogenide such as an iodide or else by a sulfonate such as mesylate, tosylate, triflate, imidazole sulfonate.

In order to prepare intermediate products denoted MY resulting from the transformation of hydroxyl groups into leaving groups one can make use of known processes.

One can use for instance for iodized derivatives, the process as described in French Patent Application No 88 05230 file4 A- rei stred- April 20, 1988 in the name of the University of Picardy .0 for chlorinated and brominated derivatives the process as described by HANESSIAN in Carbohydrate Research 24, p. 45 (1972), for suifonic esters, the process as described in French Patent Application NO 88 05231 rogic~tered April 20, 1988 in the name of the University of Picardy In the case of polyhydroxylated molecules denoted ISu(OH)nI, one may obtain a dithiocarbamatation on one or several hydroxylated sites. If dithiocarbamatation is desired on one OH site only, one must protect the other OH groups.

As applied to the prearation of dithLocarbamic acid monoesters from polyols ISu(OH)nj, the process comprises sequentially the selective protection of OH groups with the help of a heton or an aldehyde to yield ISu(O-A)n-I-OH], the tranformation of the OH group into a leaving group to yield ISu(O-A)n-1-Y], denoted 3u ISu P-Y], the esterification by reacting with [Su P-YJ a salt of dithiocarbamic acid having formula 4

R

I

S

II

N C SH, to yield the ester 2 S R 11

I

Su S C N n-i and liberation of previously protected hydroxylated sites in the form of dioxolane.

The regeneration of hydroxylated sites may be a partial or total one so as to obtain dithiocarbamic monesters having different physicalchemical properties, notably as to their solubility in water and oil.

Among polyhydroxylated molecules which are particularly suitable for carrying out the invention, are glycerol, galactose, glucose, mannose, fructose, itols and, more generally, mono-, di- or polysaccharides.

The preparation of dithiocarbamic esters from the halogenide or sulfonate ISu(O-A)n- 1 YI denoted ISu comprises the following steps contacting the halogenide or sulfonate [Su P-YJ, and a salt of dithiocarbamic acid in the presence of a polar aprotic solvent, the acid salt being used in excess to the stochiometric amount, eliminating the solvent and absorbing the residue with an organic solvent, eliminating the residual dithiocarbamichsalt, evaporating the organic phase containing the dithiocarbamic ester, purifying said ester, and, perhaps partially or totally deprotecting OH -Citcr which ara protected4in the form dioxolane.

The dithiocarbamic acid salt which is reacted with intermediary (Su P-Y| is advantageously made up of an alKaline metal salt such as a lithium, sodium, i0 or potassium salt.

This dithiocarbamic acid salt is added to the polar aprotic solvent in a molar ratio generally between 1.1 and 2, and preferably about 1.5 in relation to the intermediary ISu(O-A)n-l-Yj.

As a polar aprotic solvent, one can use acetone, hexamethyl-phosphorotriamide (HMPA), 'imethylformamide (DMF), dimethoxYethane (DME), dimethylsulfoxide (DMSO) or a mixture of these solvents. One can also associate one of these solvents or the mixture of these solvents with another organic solvent which may be an aromatic solvent such as toluene, xylene, benzene or a saturated hydrocarbon, or a mixture of saturated hydrocarbons.

After reacting the dithiocarbamic acid salt with intermediary (Su P-YI, and eliminating the aprotic solvent, the residue is absorbed by an organic solvent such as ethyl ether which solubilizes the ester, the unreacted dithiocarbamic acid being eliminated by filtration.

After evaporating the organic phase containing the ester, the later is purified by recrystallization or by silica gel liquid phase chromatography.

In the case where dithiocarbamic esters are obtained by reacting the dithiocarbamic acid salt with a sulfonic ester (the leaving group is a sulfonate), one carries out a filtration step S when the reaction is ended so as to eliminate the saline sulfonate which is formed.

In the case when one starts from polyhydroxylated molecules whose OH sites,which must not be esterified,have been protected, the partial or total deprotection of these protected OH sites is carried out either in heterogeneous acid-catalysis by passing a solution of the dithiocarbamic ester in a mixture of alKanol/water solvants, in pure alKanol or a mixture of alkanols through a column packed with an acid resin which is maintained at a controlled temperature, or in an homogeneous acidcatalysis in the same solvents in the presence of a strong mineral acid at a concentration between 0.05 and IN, while stirring at the same controlled temperature and during the appropriate length of time. One obtains according to operating conditions, a partial or total deprotection of hydroxyl functions.

The dithiocarbamic esters obtained by the inventive process and which are new esters, preser, similar properties toSthe- of sodium diethyldithiocarbamate (DEDTC-Na), which is commercially available, but with a delayed therapeutic action, a lesser toxicity and an administration which is eased by the modulation of chemical and physical properties due to the choice of the starting mono- or polyhydroxylated molecule.

7 The inventive dithiocarbamic esters find an interesting application in the preparation of immunomodulator products. Such drugs may advantageously be used for the treatment and prevention of diseases linked with immune disorders of the child or infant, and notably mucoviscidosis, recurring respiratory infections, juvenile polyarthritis. These drugs may also be useful in the case of auto-immune adult diseases and notably ±0 rhumatoid polyarthritis, systemic lupus erythemato sus, long term or recurring bacterial infections, septic complications of chronic bronchitis, bird short retinopathy and lymphocyte deficiencies.

These dithiocarbamic esters may also be interestingly applied in the preparation of a drug which is active in the prevention and 'he treatment of viral diseases, notably those due to HIVi or 2 viruses.

The inventive dithiocarbamic acid esters 2U are admniizttr at doses which are generally between 1 and 500 mg/kg body weight.

The administration of these drugs may be made for example orally, either in the form of a solution or in solid foi'm, for example in the form of gelatin capsules with gastric protection.

A few examples for carrying out the invention are given below as illustrations but not as limitations.

For these examples, the type ISu(OA)n-l-OHI intermediary is either a commercial product or a product prepared by protecting hydroxylated sites in the form of dioxolane from polyhydroxylated ISu(OH)nj molecules according to conventional liferature methods.

-8- The type [Su P-y] intermediate is either a tosylate, or a mesylate, a triflate, or an iodide.

The tosylate is prepared according to French Patent 88 05231 One adds into a flask containing 200 ml of anhydrous acetone maintained at 0°C, 0.1 mol hydroxylated [Su(OA)n- 1 -OH] molecule, 1.11 mol triethylamine and 0.11 mol paratoluene sulfonyl chloride, while stirring.

One leaves the mixture to return to room temperature while stirring during 10 hours. The triethylamine hydrochloride crystals are eliminated by filtration. The filtrate is evaporated under reduced pressure, the residue is thrice recrystallised in an hexane/ethyl ether mixture, 50/50 The pure tosylate yields are between 60 and The mesylate is prepared from methane sulfonyl chloride by replacing in the preceding protocol acetone with ethyl ether/hexane, 50/50 and bringing back the reaction time to one hour instead of 10 hours. When the melting point of the mesylate is low, purification is carried out on silica gel. The pure mesylate yields are between 75 and The triflate is prepared according to the method given by FLECHNER (Carbohydrate Research 77, (1979), 262- 266) by adding [Su(OA)n-.-OH] to a triflic anhydride i ft cooled'solution in dichloromethane in the presence of pyridin. After 90 minutes reaction at -10 0 C, one neutralizes with NaHCQ 3 the organic phase is washed with an aqueous 3% HC1 solution, and it is purified by silica 30 gel chromatography or by recrystallisation.

The iodide is prepared according to French Patent 88 05230 0.1 mol [Su(OA)n-i-OH] hydroxylated molecule, 0.105 mol triphenylphosphine and 0.2 mol imidazol are 35 added in a refrigerated flask containing 500 ml xylene.

To this mixture, which is brought to 80 0 C with stirring, are added 0.1 mol iodine, and the whole is brought to 92091EEDAT.038,a\4S528pas.page,8 -9reflux during 3 hours. The reaction mixture is then transferred to a 2 liter erlenmeyer, wherein 500 ml of saturated HNaC0 3 aqueous solution are added. After stirring during 10 minutes, iodine is added till the organic phase is coloured, then the necessary amount of sodium thiosulfate in aqueous solution is added to observe a decolorizatioin The organic phase is decanted, dried on Na 2

SO

4 filtrated and evaporated under reduced pressure. The residue is purified either by two successive recrystallizations in a hexane/ethyl ether mixture, 90/10 or by silica gel chromatography.

The yields are between 60 and Example 1 Synthesis of deoxy-1 N,N-diethyl dithiocarbamate O-isopropylidene-2,3 glycerol from 0-isopropylidene-2,3 glycerol sulfonate and deoxy-1 iodo-l-0-isopropylidene- 2,3-glycerol.

g (8.2x10 2 mol) of O-isopropylidene-2,3 glycerol tosylate (solketal tosylate) and 27.75 g (12.3x10 2 mol) of frihydrated sodium N,N-diethyl dithiocarbamate (DEDTC- Na) are placed in a 1 liter three neck flask containing 500 ml acetone. While stirring, the mixture is brought to reflux the progress of the reaction is monitored by

HPLC.

25 8 hours after the beginning of reflux, one observes Sa solketal-tosylate conversion ratio of 96%.

After return to room temperature, the filtrate containing the diethyldithiocarbamic derivative is filtrated on sintered MN4, the precipitate being sodium S 30 tosylate.

The acetone is eliminated under reduced pressure.

The residue is absorbed with ethyl ether which selectively solubilizes the ester, the DEDTC-Na being eliminated by filtration.

The ethyl ether is eliminated under reduced pressure, the raw product (24.09 g) which is observed in the form of an oil, is purified by HPLC.

92091 1 EEDAT.038a:\45228pas.page,9 The raw product is introduced into the eluent which is a technical hexane/acetone 93/7 mixture on a silica gel column. One obtains 17.85 g of 0isopropylidene-2, 3 glycerol N, N-diethyl. dithiocarbamate (solketal DEDTC) in pure form according to HPLC, 1 H and 1CMNR analyses.

The synthesis was also m.acde according to the above described p -otocol, from O-isopropylidene-2,3 glycerol rnesylate (solketal mesylate). After 24 hours reaction, W0 the yield in solketal diethyldithiocarbamate is By replacing the pure acetone solvent with an ace~one/HMPA 80/20 mixture one obtains, after 24 hours, a solketal mesylate conversion ratio of The synthesis was also made according to the above described protocol from deoxy-l iodo-1 O-isoprppylidene- 2,3 glycerol (iodo-solketal). After 12 hours ref luxing in acetone the conversion of iodo solketal 1.9 total. One otitains after purifying 5~6% solketal N,N-diethy'Ldithioce~rbamate.

The 1H and 1CMNR spectral characteristics of the obtained solketal DEDTC are respectively given in tables I and II.

From tosylate, mesylate and iodo solketal, are prepared, according -Lo this same protocol, deoxy-. N,N dimethyl dithiocarbamate 0-isopropylidenei 2,3 glycerol (solketal DMDTC) with yields above 90% (purity controlled by HPLC and MNR).

Example 2 OP 30Syn~thesis of deoxy-6 X, N-di ethyl di thiocarbamate di- -isopropylidene'-12 3,4-a-D galactop-ranos6 from deoxy-6 ipdo-6 di-O-isopropylidene 1,2:3, 4-a-D galactopyranose.

g (6.7ul10 2 mol) deoxy-6 iodo-6 di-Oisopropylidene 4-a-D-galactopyranose (iodo diacetone galactose) and 22.78 g (10,2x10 2 nol) DEDTC-Na are placed in a 1. liter three neck flask containing 500 ml acetone. While stirring the mixture is brought to U41 g9'l91,E-EDAT,038,a\452ia.Fage10 00 O 11 ref lux co~ntrol. of the reaction in progress is made ,y HPLC. 24 hours af ter the beginning of ref lux, a total disappearance oZ iodo diacetone galactose is observed.

Acetone is evaporated under reduced prcessure. The residue is absorbed with ethyl ether unreacted DEIYX-Na is eliminated by filtration. The filtrate is evaporated under reduced pressure. 26 g raw; product are obtained.

The purification is carried out through a dotible recrystallization in technical hexane.

One obtains 24.1 g pure diacetone galactose N,N-diethyldithiocarbamate acci~rding to HPLC, 1 H and 13

C

MNR analyses.

F E4 0 C [a [20 13.60 (CHdC1 3 Trhe 1 H and 1CMNR spectral characteristi.cs of tk~e obtained DEDTC diacetone galactose are given in tables III aknd IV, respectively.

'I Example 3: Synthesis of deoxy-6 N,X-dimethyldithiocarbamate di- 20 O-isopropylidene-l,2:3,4-a-D galactopyranose from deoxy-6 iodo-6 di-0-isopropylidene 1, 2:3,4-a-D galactopyranose.

2.9 g (7.84x10- 3 mk deoxy-6 iodo-6 di-O- :;.oprc~pylidene l,2:3,4-a-D galactopyranose and 2.1 g (11 ,7x1O- 3 mol) dihydrated sodium dimethyldithiocarbamate are placed In a 100 ml flask containing 58 ml acetone.

While stirring, the mixture is brought to reflux.

After 24 hours, the complete disappearance of the substrate is observed, the reaction is stopped, the mixture is extracted according to the above deccribed protocol (example The purification, is obtained by a 9 double recrystallization in technical hexane.

g (87.8 pure diacetone galactose N,Ndimethyldithiocarbanate are obtained according to 1 H and 1CMNR and HPLC analyses.

920911,EEAT.03,a452pas.page,lI 12 F 128-129'C 20 =10.30 c~ 3 Example 4 Synithesis of deoxy-6 N,N dimethyl dlthiocarbamate and deoxy N,N diethyl dithiccarbamate di-O- A A.

A

A AC 7 s

A)

92091 1,EEDA AO038,a N45228pn5page, 12 isopropylidene 1,2 3,4-a-D galactopyranose. from tosylate-6 di-0-isopropylidene 1,2:3,4-a-Dgalactopyranose (diacetonegalactose tosylate) and corresponding lithium dithiocarbamates.

The lithium N,N dimethyl dithiocarbamate and N,N diethyl dithiocarbamate (DMDTC-Li and DEDTC-Li) are first prepared as follows to a solution containing 11. g Li 2

SO

4 and 45 g DEDTC-Na, 3 in 110 ml water, stirred for 30 minutes, are added 500 ml ethanol/water 95/5 The Na 2

SO

4 precipitate is discarded by filtration. The filtrate which is evaporated under reduced pressure and dried up during 12 hours in a dessicator in the presence of P 2 0 5 yields 34 g DEDTC-Li of a purity above 90% after control by flame photometry. A purity above 99% was obtained by passing the above filtrate on an ion exchangecolu.n (saturated in lithium ions).

A solution containing 11 g Li 2

SO

4 and 35.8 g DMDTC-Na, 2 H20 treated in the above conditions yields 29,8 g DMDTC-Li of a purity above 90%. The purity above 99% was obtained after passing the filtrate on an ion exchange column.

Preparation of deoxy-6 N,N diethyl dithiocarbamate-6 di-0-isopropylidene 1,2:3,4-a-D galactopyranose.

0.10 md diacetone galactose tosylate and 0.13 mol DEDTC-Li are placed in a flask containing 450 ml toluene/HMPA 66/34 solvent, after 2 hours reaction with stirring at 110 0 C, extraction and purification, the product is obtained with a 94% yield.

-14- Preparation of deoxy-6 N,N dimethyl dithiocarbamate- 6 di-0-isopropylidene 1,2:3,4-a-D galactopyranose.

0.10 mol diacetone galactose tosylate and 0.13 mol DMDTC-Li are treated as above. After purification, the product is obtained with a 93.5% yield.

Example 5 Synthesis of deoxy-3 N,N diethyldithiocarbamate di- 0-isopropylidene-1,2:5,6-a-D glucofuranose from deoxy-3 iodo-3 di-O-isopropylidene-1,2:5,6-a-D allofuranose.

10 g (2.7x10-2 mol),deoxy-3 iodo-3 di-Oisopropylidene-1,2:5,6-a-D allofuranose and 1;.18 g (5.4x10 2 mol) trihydrated sodium N,N-diethyldithiocarbamate (DEDTC-Na) are placed in a 250 ml three neck flask containing 150 ml hexamethyl phosphorotriamide (HMPA). While stirring, the mixture is brought to The reaction is stopped after 24 hours reaction, the conversion being 95% according to HPLC monitoring.

An hexane/ethyl ether 90/10 mixture is added to the reaction mediu, then the resulting mixture is washed with iced water. The aqueous phase is dried up on anhydrous sodium sulfate and evaporated under reduced pressure. 18 g raw product are obtained.

Purification is obtained by passing on a silica SK column; after elution with an hexane/ethyl ether 85/15 mixture, 8.6 g pure diacetone glucose N,Ndiethyldithiocarbamate are isolated according to HPLC, 1

H

and 13C MNR analyses.

F 83-84 0 C [a D 44.3" (CHC1 3 D Tables V and VI respectively give 1 H and 13 C MNR 'spectral characteristics of the obtained diacetone glucose DEDTC.

By replacing pure HMPA solvent with-an acetone/HMPA 50/50 mixture one obtains after 48 hours, a conversion ratio of iodo diacetone allose of 920911,EEDAT.O38,a: 45228paspage,14 By replacing the HMPA solvent with toluene/HMPA (50/50) mixture, one obtains after 18 hours at 110*C an iodo-diacetone allose conversion ratio of 96%.

The dimethyls are obtained with the DMDTC salts.

By operating in the toluene/HMPA (50/50)(v/v) mixture and by replacing DEDTC-Na with the corresponding lithium salt one obtains after 2 hours at 110°C an iododiacetone allose conversion ratio of By taking trifluoromethyl sulfonate-3 di-Oisopropylidene 1,2:5,6 a-D glucofuranose in the place of iodo-3 diacetone allose and by operating in the •HMPA/toluene (50/50) mixture with DEDTC-Li in excess, one obtains deoxy-3 diethyl dithiocarbamate-3 di- O-isopropylidene 1,2:5,6 a-D allofuranose.

Example 6 Preparation of deoxy-l N,N diethyl dithiocarbamate di-O-isopropylidene 2,3 a-D fructopyranose (diacetone fructose DEDTC) 0.1 mol deoxy-1 iodo-l di-0-isopropylidene 2,3 a-D-fructopyranose (iodo diacetone fructose) and 0.13 mol DEDTC-Na are placed in a flask containing 250 ml HMPA.

After 8 hours reaction at 70°C the conversion ratio of iodo diacetone fructose is 88%, after extraction and purification, the product is obtained with a 74% yield.

S; 20 D -5.02° (CHC13) F 100 0

C

i <i 9291,EEDAT.038,aA\45228pas.page,15 Th e 1 H and 13C MNR spectra are given in tables VII and VIII.

By operating with the HMPA/toluene (50/50) (v/v) mixture at 110'C, the conversion ratio of iodo diacetore fructose is 82% after 18 hours reaction.

Example 7 Products obtained by deprotection of hydroxylated sites.

The deprotection of products of the formula .L0 S R Su-(OA) n--S-C-N

R

2 was carried out in ethanol/water 95/5 by acidcatalysis either in heterogeneous phase on an AMBERLIST 11+ Wet (ROHM HAAS)colurnn or in homogeneous phase in the presence of H 2 S0 4 at a concentratioai of 0.2N.

Preparation in heterogeneous phase of deoxy- 1 N,N-diethyl dithiocarbamate-l glycerol (DEDTCglycerol) from deoxy-l N,N(diethyl dithiocarbarnate-l 0-isopropylidene-2,3 glycerol (DEDTC-solketal) and deoxy- 1 N,N dimethyl dithiocarbamate-1 glycerol (DMDTCglycerol) from deoxy-l N,N dimethyl dithiocarbamate- 1 0-isopropylidene 2,3 glycerol (DMDTC solketal).

A solution of 10 g DEDTC-solketal in 100 ml 950 alcoho., is passed with a flow rate of 2 mi/minute on the thermostated 65%C column containing 1',00 g ANIBERLIST H* resin. The columti is rinsed with 100 ml 950 alcohol.

After evaporating the solvent and purifying on silica gel, 50/50 8.4 g DEDTC glycerol are obtained accords A -to (92% yield), a liquid at room temperature, pure afaet HPLC,1H and 1 3 C MNR analyses.

InI D 1.5803.

20 The H and 1C MNR spectral characteristics of obtained glycerol DEDTC are given in tables IX and X, respectively.

Under the same conditions, DMDTC glycerol is obtained from DMDTC solketal with an 88% yield.

Preparation of di-O-butanoyl-2,3-deoxy-l N,N diethyl dithiocarbamate-l glycerol (DEDTC glycerol dibutyrate).

x 10-2 mol DEDTC glycerol are placed in 15 ml pyridine/toluene (.50/50) mixture and x 10- 2 mol butyryl chloride dissolved in 10 ml toluene are added dropwise while stirring after 5 hours reaction at room temperature, the reaction mixture is washed once with a IN HC1 aqueous solution and twice with water, the organic phase is dried on Na 2

SO

4 then evaporated, the residue is purified on silica gel with the technical acetone/hexane mixture as an eluent. After purification, an 80% yield is obtained.

(nj 2 0 1.5142 1H and 13C MNR spectra are given in tables XI and XII.

-18- Preparation of di-O-palmitoyl-2,3 deoxy-l N,N diethyl dithiocarbamate-1 glycerol (DEDTC glycerol dipalmitat:e).

One proceeds under the same conditions as for DEDTC glycerol dibutyrate. The residue, after extraction and evaporation of the solvent, is purified by recrystallization in technical hexane.

An 88% yield is obtained.

The 1 H and 13 C MNR spectra are given in tables XV and XVI.

-Preparation in heterogeneous phase of deoxy-3 N,N diethyl dithiocarbamate-3 O-isopropylidene-1,2 a-Dglucofuranose (DEDTC-MaGlu) and deoxy-3 N,N diethyl dithiocarbamate -3 D-glucopyranose (DEDTC glucose) from deoxy-3 N,N diethyl dithiocarbamate-3 di O-isopropylidene 1,2:5,6 a-glucofuranose (DEDTC-DAGlu).

The deprotection of hydroxylated sites by acid catalysis on a H+ resin column was carried out under the same solvent concentration, flow rate, and temperature conditions as above. After rinsing the column with alcohol and evaporating the solvent one obtains from 10 g starting product 8.5 g raw product. Silica gel chromatography (60 g) yields 3 fractions 0.58 g starting product, eluated with the hexane/acetone, 85/15 mixture.

-6.8 g DEDTC-MAGlu, eluated with the hexane/acetone, 70/30 mixture, pure after HPLC, 1

H

and 13 C MNR analyses.

4 F 78-80 0 C, +90.8" (CHC1 3 4 0.6 g DEDTC glucose, eluated with acetone alone, pure according to HPLC analysis.

For a given starting substrate concentration and a given amount of resin, the proportion of totally unprotected derivative, (DEDTC glucose) may be increased 92O911 4 EBDAT.038,ak45228pas.page,18 -19at the expense of the monodeprotected derivative (DEDTC- MAGlu) by lowering the flow rate and/or increasing the temperature for instance at 70 0 C with a flow rate of ml/minute, one obtains the complete disappearing of the starting product and the DEDTC-MAGlu and DEDTCglucose products in the proportion of 30/70.

The same operating conditions allowed one to prepare deoxy-3 N,N dimethyl dithiocarbamate-3 0-isopropylidene 1,2 a-D glucofuranose (DMDTC-MAGlu) and deoxy-3 N,N dimethyl dithiocarbamate-3 D glucopyranose (DMDTCglucose) from DMDTC diacetone glucose with similar yields.

Preparation in homogeneous phase, in the presence of H 2 S0 4 of DEDTC-MAGlu and DEDTC-glucose from DEDTC- DAGlu.

Deprotection of hydroxylated sites by acid catalysis in the presence of 0.2N H 2

SO

4 was carried out in ethanol at the controlled temperature of 50 0 C, from 10 g DEDTC-DAGlu (25.5 mmol) in 100 ml solvent, while stirring. After one hour, 96% of the substrate having reacted, one proceeds to a neutralization at pH 7, with 8N caustic soda lye. The formed sodium sulfate is filtrated on sintered felt number 4 and the evaporated filtrate gives 8.3 g of a raw product which is subjected to a chromatography as above, to yield 0.35 g starting product 6.3 g pure DEDTC-MAGlu S* 0.5 g pure DEDTC-Glu.

SBy carrying out for 20 hours the acid catalysis 30 under the same temperature, solvent and acidity conditions, the deprotaction of hydroxylated sites of S: DEDTC-DAGlu is obtained in a proportion above 90%, but the percentage of DEDTC-Glu does not exceed 60%, while such a reaction time triggers a degradation of the product by the solvent.

The 1 H and 13 C MNR spectral characteristics of the obtained monoacetone glucose DEDTC (DEDTC-MAGlu) are V'21,EPDAT.038,a: \4S228paspagce,19 20 respectively given in tables XV and XVI.

The 1CMNR spectral characteristic of the obtained glucose DEDTC are given in table XVIII.

Example 8 Preparation in pseudohomogeneous phase of deoxy-6 N, N-diethyl dithiocarbamate D-galactopyranose (Dh'DTCgalactose) and deoxy-5 N,N diinethyl dithiocarbamate Dgalactopyranose (DMDTC-galactose).

2 M DEDTC-diacetone galactose are added into a flask containing 40 nml dioxane and 10 ml 12N aqueous HCl.

Aifter stirring during 1 hour at 50 0 C, the solution becomes limpid and the DEDTC diacetone galactose 'totally disappe ared according to thin layer chromatography controls. One then proceeds to neutralization by slowly adding triethylamine (TEA) and the TEA hydrochloride prec:Lpitate is separated by filtration. The filtrate is then evaporated and absor~bed with water to precipitate the expected product. The DEDTC-galactose is obtained w2.th a 60% yiel~d. The 1 3 C MNR c,,ectrum is given in table 920911,E5DAT.O38,a:\45228pa&page,20 DMDTC-galactose is obtained under the same operating conditions with a 58% yield.

The same process may be applied to the preparation of deoxy-1 NN diethyl (dimethyl) dithiocarbamate of glucose, fructose, mannose or other monosaccharides from di-O-isopropylidene precursors.

Example 9 The biological properties, (mice LD50) of dithiocarbamic esters described in examples 2,3,4, 5 and 8, are compared with those of saline diethyldithiocarbamate DEDTC-Na, commercially available.

The obtained results are given in table XIX.

One may conclude from these results that the inventive dithiocarbamic acid esters have a notably lower toxicity than the commercially available DEDTC-Na.

Furthrmorit-turns -4 that-- the-intermedIte lithium dithiocarbamates products, notably lithium dime dithiocarbamate and lithium diethyldithiocarbamate are active, have a very low toxicity and may be used pur associated, as drugs for the same applicatignw, for mple with similaZ doses to those which are used in the e of sodi-um The use of lithium dithiocarbamates in the process for the preparation of inventive esters is preferred to that of other saline dithiocarbamates because the reactions are quicker at lower temperature, and one can use a wider range of solvents.

TABLE I H MNR SPECTRUM OF SOLKETAL DEOTC FORMULA PROTONS ti- H-1 H-C-SR H-2 li O,.CH: H-3 12

C

i HC~i H 3 i :H S CH.-C8 2 It CH j CH.-CH3 CH 2 SCH2 iS CH n CH

I

3 :of the signal 6 0n porn/IMS in COcI c.oupl~ing constant$ d m dd ad S 3.58 4 41 4,10 3, 74 3 1-.2 =5.9 3 2-3 E6,1 3 3

I

1, 4 1, 34 S m m t t 3,96 3.68 1 30 1, 27 3 =7.3 3 7.2 3 7.1 23 TABLE I I 13MNR SPECTRUM OF SOLKETAL

DEDTC

C FORMULA CARBON5 6 inpprn/TMS (in. CDC1 3 H C-1 39.99 H-CT-SR C -2 '04.44 C H ;COc C-3 6 6. 42 H-CT0

'CH,

H H S C P.-CH: R-N

"CH.-CHI

2CH 3 (isoprop.) 26.61-25,LL0 CH Is L6 4 2 1 CH( 12,42 (ICH 114 I i94~,51 cs 2 TABLE III H MNR SPECTRUM OF DFDTC DIAC ETONEGALACTOSE (DEDTC-DAGal) i~i-r r-r :Nature F 'ORMULA PROTONS in ppff/XmS Coupling of the in CD1 constants(I) C :signal H-1 d 5.51 3 1-2 z H-2 q A RS- ,6 6' H-6 j

'I

N,,

1 2CH

CH

r (isoprp.): 3 (CH nq1): cm t 3 CH~ 3 4,29 4,62 3.92 3.@31 3 -63 3 2-3 a 2.4 3 3-4 z 7.95 3 i1-5 r 3 6-V 3 A-6 6 .9 1.131 1, 36 1.-47 1- 48 3 4 2 4.08 1.24 1)29 0 a- 11i 17 r ,I i-- ~I TABLE IV 13MNR SPECTRUM OF DIACETONEGALACTOSE

DEDTC

CFORMULA CARBONS 6 in ppm/TMS c in CDCI 3 C..1 96.48) ,6'C- 2 70,90) 0 70.56 C Hj- C-4 0,76

CH

3 ~1C-5 72 .37 3 C C636.88 ol

CH

3 (C-7 109 .27 C-8 108,78 S CH.-CH3 4CH (isoprop.) 24,37-25 05-25.90 RC-N 25 .00 'CI (s C 2 195.27 (CH 2 SI-1) 400#38 c F, Z Wfl-1) 40o.70 cCH 3 sl 12.45 CH C 3 ni 1b,43 TABLE V ~fl MNR SPECTRUM OF DEDTC DIACETONEGLUCOSE (DEDTC-DAGlu) Nature :Q1ifl ppffdTSS Coupling FORMULA :PROTONS :of the in COC13 costnt signalCosat

I

H.'1 d 5.75 :312=3.~2 me 0 ,a)

C

C H-3 dd 4.70 :3 -4 =3.7 c~ H 2 A dd 4 63 3 1 o 7: H-5 m 4,19:J H-6 q 4.01 366 H-6 9 3.95' 5-61 'CH-PH3 4CH 3 3 1.26 1.26 1.49

C

C

C

C

C

C

C

I

C

C

4- (iioprop.): 3 :CH( ni me 3.6 1 2 CH z(nl' m nl) t *c W 3.s98 1. 20 1,26 fl -SV-1:6.4 TABLE VI CMNR SPECTRUM OF DIACETONSGLUC0SE DEDTC (DEDTC-DAGlu) FORMULA *CARBONS 6i p/M in CDC1 3 104.47 Me 0 *86.69) Me 0- ~c-3 57.12 C- 78.96 73- 87 I- 0 67.44 mo c -7 112.#12 S *CH.-CH3 R-CNC-8 109.43 R C CH CH22i'soprop.) 20".29-26.52-25.20- CH nl-1) 49.29) (CH 2(f-1 P. 46463) CH'( f)*12.44 CH 3 (Sl) 11.43 *s 2 TABLE VII HMNR SPECTRUM OF DEDTC-l of DIACETONE FRUCTOSE Nature 6 in PPMf'17S Coupling FORMUJLA PROTONS of the Constants signal i n CDC1 3

MI

H'd 3,81. J1. 2-1,2HZ H d 4,29 J3-4-2.5 H4 *dd 4,52 J4-3x2.4 J4-5-7.9 66HS dd 4416 JB-6-1.9 ,2 H6 dd 3.81 J5-6-1.9 0 2CH 3 S 1.46 s

CH

3 s 1.39 1 1 CHi 1 CH 3

QCH

3 C8 2 in1.23 Jfl-cn-1) zIot 1.21 computable in 3,92 Jj7-Cjj-j) not cornputttbln TABLE VIII 13, ,MNR SPECTRUM OF DEDTC-1 OF DIACETONE FRUCTOSE a in ppmThiS FORMULA CARBON*.

i~n CMci 46. 49 102.68 73,16 70,43 70.43 61 ,39 616, 109.01 SR 108.38 0.

S

it CHz-CH 3 R C WCHf CH3 J *L 26.29 2S,.88- 25.08 24.04 12.S0 11.47 49,67 46.69 lY53. 29 CH3-CH2 TABLE IX HMNR SPECTRUM OF THE DEDTC GLYCEROL Nature 6in pm~r, TMS Coupling C FORMULA PROTONS of the in C~DC 3 ConstantsC) signal: H 3 H..1 3.4 2

P)

12 3 dd3.49. 0 2-.3 13 R CHi- waJ I. R (I1 q 3.98 J CH q 3.7

C

3 f CH S1 ).2 (3 TABLE X 13MNR SPECTRUM OF.DEDTC GLYCEROL ppm/TMS FORMULA CARBONS ~n CDCI 3

H

I'

H-C-OWR

H-C -0H 13

H

c-1 C-2 C -3 *39.06 71.36 64.51 S CH.-CH: R- C.-N 'cHrc CH- 3 '-1 CH M-1) CH .1) CH nl CH fSV Cs 2 50.16 47,07 12.38 11,47 195,75 TABLE XI OF DIBUTYRATE 1MNR

SPECTRUM

DEDTC GLYCEROL FOMUAPRTOS Natur e of 4 i n pMrDS Coupling FOR~l POTNSthe Constants signal in C=c. I H-1 H-1' 3- 6S J1-1 '-14.2 J1-2 -4.9 XJ1-14.1 J1 '-2*7.4 3.35 H-3 m S 7 1 HC cLSCACHI CH3 a ICH 2

CH

3 I U2 I2 11 1 0 01 12 13 Hc-OC, CH 2

CHICH

3 a2 i 0 H-3 I H-7,71 H-8,8'1 H- 10010' H-1i 111,12 12! 5.17 4.23 4.02 3.59 2.15 2.14 1150 1 .13 1.1 0,79S J3-3'-11 .9 J3-2u3.5 J3-2-3, J-6. 8 J-.*39 J- 7, 31 J-7.38 J-6 .81 J-7.31 7.35 TABLE XII 13,MNR SPECTRUM OF DEDTC GLYCEROL DIBUTYRATE 6 'in ppm/ThM FORMzULjA CARBON n1 HC! C NICHj'CH 3 2 ICHf CH 3 2 1 HC-OC CH:CHzCH 3 3 1010' 12 13 HC~0CCHICHZCH 3 0 c6 =0 c s=0

C

4

CH

2 C(7) CR (1) 2 CH2 (13

CR

3 (14)

CH

3 (1 5)

CR

3 (16) 37.23 69.83 63.68 172.85 172.34 193.67 49-76 46.64 35.95 35.78 18,22 12,36 11,34 13~.46 V 34 TABLE XIII lH MNR SPECTRUM OF DEDTC GLYCEROL DIPALMITATE Nature of FORMULA PROTONS the 6 in pMrM Coupling signal Constai-s in D1 H-1 H-1 3,74 H-2 H-3 3.46 5.28 4 M3 4 .13 I CHrCH3 Ic oc UH~c l ciImc 31 i 0 J1 -1 0 J1-2-5. 0 J1'-1 -14.2 31 -2-7.4 J3-3'-12,1 J3-2-3.6 33'-3-1.2.0 33 '2-5 .7 J-7. 0 J-6 .8 Jw7, 3w.-4 H-3' ard H-~71 H-8 and H-8' H1-9 and H-9' arnd H-10' 3.97 3.69 2.27 2.26 1rn TABLE XIV 13 C MNR SPECTRUM OF DEDTC GLYCEROL DIPALMITATE

FORMULA-

CARBON

6 in ppm/TMS i n CDC1 3 37.39 69.95 63.84 c 6 m0 C 5 0

C

4 5 S, 7 to H~~t CLSCN' CH3 13 15424 ;IC-CC. H4H)HCH IIIu3 1 3 0 0 1j3 6 s13 2143 HCO-C CH 3 cH CH:)CH1,CHtCHt 11 37 31 40 0

CH

2 cM 2 CH 2

CH

2

CH

2 CH 2

CH

3

CH

3 (9) (10) Cl3and 14) (15 to 24 anid 2 7and3i) (2 5and 3 7) (26 and 38) (3 9and 4 0) (7) 172.74 197.90 34,25 34 .87 31 29.61 to 2 8 9 24.87 22,62 14.04 49.87 46. 69 12.45 11.45 36 TABLE XV H MNR SPECTRUM OF DEDTC-3 MONOACETONE GLUCOSE (DEDTC-MAGlu) i FORMULA PROTONS Ct H-1 c H-2 1 i0o 0,6 H-3 110. 9 C NI -4 SR H-3 2 (Me: H-6 C S RS ,CH.-CH c Nl CM :n 2CM 3 s C (isoprop.J; Nature 6 in PRPRUHS Coupling of the in COCI signal Congtants d 5.73 :3 1-2 x 3.6 d 473 3 06 dd 4.83 3-4 x 3. 4 dd 4.35 8. 8 3 K-'i3..55-3,87 6V 56'

S

3 1 .26 1,49

(S)

C CH l n-i) qd 3.93 CH (no-1) qd 4,00 CH t 1.23 CH (n t 1. r26 on 41 7. 1 .*3 OH5* 5 3.46

OH

6 3 2.51 TABLE XVI 3C MNR SPECTRUM OF DEDTC-3 MONOACETONE GLUCOSE (DEDTC-MAGlu) FORMUJLAE CARBONS 6 inppm/TMS e CDC1 ~P I C C-1 104.47 C HO 6,6' SHc -2 85.86 5 0 58.69 4 C-4 78 .98 C 70.30 3 7 C-6 64,13 0 m Mc C -7 112.26 s Cc C73 CH.-NH3ZCH 3 (isoprop.) 26.24-Z6.42 C H 2-1 47,41 CH 50.34 21 CH(1% $1 11.46 8fCH 3 (n 12,49 Cs 2 192.73 38 TAf3LE XVII 13 C MNR SPECTRUM OF GLUCOSE DEDTC FORMULA CARBONS 6 in ppm/TMS in

CDCI

3 91.77

C

1 :'98.11 HO 4 H 70)72 c 2 70.14 4 S-R 57.33 H3 59.S 3 C 69.93

OH

C 73.15 72,44 S Ci-HIU 62.18 R- C-N e62 c C CH 3 61,5' t wrw CS 196.36 2 195.99 CH(1-1) 50. 37 2 CH W 47 .84 CH 3 n 11.62 CH 3 fnl 12. 57 TABLE XVIII 1CMNR SPECTRUM OF GALACTOSE DEDTC-6 FORM4ULA CARBON 6 in ppmffl4 i n MMc 3 RS 6,6' 0 0

OH

S

R I IJCHeCH3 R- CCHf~CH 3 8Form C 4 C 6 a Fojrn C 2 c3 C 4 c. S 0. and a Forms 012 01 3 CH

I

74.52 75.57 72.S53 76.48 39.39 197.33 95..24 73.04 Ili .01 72.53 71,96 197.33 53,16 50-S5 14,25 13 ,C4 TABLE XIX MICE P.O0. I.P.

SOLVENT

(g/kg) (g/kg) DEDTC-Na com~ercial water DM50 4.00 ±0.32 1.90 ±0.34 1 1.5± 0.01

DE.DTC

diacetone glucose DR540 8 4 20 460.48

DEDTC

diacetone

DMDTC

diacetone

DMDTC

glycerol galactose galactose DM0 2 65 t 0 .52 2.15 t: 0.3S DM50 DMSO 4.00 :t 0.30 1,4 0.28 Dipa.'.mitate DEDTC glycerol Dibuty-rate DEDIC glycerol DMS0 4 2 i 0 .46 DM50 8 35±4 0,42

Claims (31)

1. A process for preparation of dithiocarbamic acid esters having the formula R, S II N C SH R in which R 1 and R 2 are saturated or unsaturated substituted or unsubstituted alkyl radicals or else forming with the nitrogen atom a saturated or unsaturated ring which may contain heteroatoms starting from alcohols or polyols, characterized in that at least one *4 4 hydroxylated group of mono- or polyhydroxylated molecule S1 is transformed at first into a leaving group and in that the leaving group is displaced by the anion R, S 11 N C- S Ii I wherein R 1 and R 2 have the above quoted meaning, as introduced in the form of a salt in a solvent or mixture of solvents, and in that the leaving group may be an iodide or a tosylate or a mesylate or a triflate or an Simidazole sulfonate which, notably in the case of mono- or di-saccharides, may be grafted unto a secondary site other than anomeric 930419,EEDAT.038,a:45228pas.page,41 as well as unto a primary or anomeric site.

2. A process according to claim 1, characterized in that, when it is applied to the preparation of a dithiocarbamic acid monoester from polyols, denoted ISu(OH)nl, the process comprises the sequence of following steps: selective protection of OH groupsto give ISu(O-A)n--OHJ, preferably in the form of dioxolane, transformation of the OH group into a leaving group to yield ISu(O-A)n-l-Yj, denoted ISu P-Y) esterification through the action of a salt preferably a lithium salt, of dithiocarbamic acid R.I S II N C SH R 2 on ISu(O-A)n-1-Yl denoted ISu P-YJ which yields S R, I /1 Su (0-A)n S C N R 2 and partial or total regeneration of protected selected sites.

3. A process according to any of claims 1 and 2, characterized in that the leaving group is made up by an halogenide.

4. A process according to claim 3, characterized in that the halogenide is iodide. A process according to any of claims 1 and 2, characterized in that the leaving group is made up by a sulfonate. -43-

6. A process according to claim 5, characterized in that the sulfonate is chosen among the group made up by mesylates, tosylates, triflates, imidazole sulfonate.

7. A process according to any of claims 1 to 6, characterized in that the preparation of the dithiocarbamic ester includes the following steps contacting the intermediate [Su P-Y] and a dithiocarbamic acid salt in the presence of a polar aprotic solvent, the dithiocarbamic acid salt being used in excess to the stoichiometric amount, reacting the mixture while stirring during the necessary time to yield a conversion ratio of [Su(O-A)n-1-Y] denoted [Su P-Y] above eliminating the polar aprotic solvent, absorbing the residue by an organic solvent, eliminating the residual dithiocarbamic acid salt, evaporating the organic phase containing the dithiocarbamic ester, purifying this ester.

8. A process according to claim 7, characterized in that the dithiocarbamic acid salt is an alkaline metal salt. 44 4* 444*

9. in that 25 acid is A process according to claim 8, characterized the alkaline metal salt of the dithiocarbamic the lithium, sodium or potassium salt. 92D91 EEDATO38,aA45228paspage,43 B1w00 lilqlll A process according to any of claims 7-9, characterized in that the dithiocarbamic acid salt is added to the polar aprotic solvent in a molar ratio generally between 1.1 and 2, and preferably about to the intermediate (Su(O-A) n-l Y denoted ISu P-Y).

11. A process according to any of claims 7-10, characterized in that the polar aprotic solvent is chosen among the group made up of acetone, hexamethyl phosphorotriamide, dimethylformamide, dimethoxyethane, dimethylsulfoxide or else a mixture of these solvents.

12. A process according to any of claims 7-11, characterized in that the polar aprotic solvent or mixture of polar aprotic solvents is associated with another organic solvent.

13. A process according to claim 12, characte- rized in that the organic solvent associated with the polar aprotic solvent is an aromatic solvent such as toluene, xylene, benzene.

14. A process according to claim 12, characteri- zed in that the organic solvent associated with the polar aprotic solvent is a saturated hydrocarbon or else a mixture of saturated hydrocarbons. A process according to claim 7, characte- rized in that the organic solvent into which the residue is absorbed, after eliminating the polar aprotic solvent is ethyl ether.

16. A process according to claim 7, characterized in that the residual dithiocarbamic acid salt is eliminated by filtration after absorbing the residue in the organic solvent;

17. A process according to claim 7, characterized in that the dithiocarbamic ester is purified by recrYstallization or by silica gel liquid phase chromatography.

18. A process according to claim 7, characterized, in the case of sulfonic esters, in that one carries out a filtration so as to eliminate the sulfonate which is formed.

19. A process according to any of claims 1 and 2, characterized in that the polyol Su(OH) is chosen among glycerol, glucose, galactose, mannose, fructose, itols or mono-, di- or polysaccharides.

20. A process according to any of preceding claims, characterized in that it comprises the selective or total depr tection of protected OH sites of the dithiocarbarmic ester by homogeneous acid catalysis in a water/alkanol mixture, in pure alkanol or in a mixture of alkanol or by a I S 15 acid catalysis in pseudohomogeneous phase or in 0" heterogeneous phase on a resin. S21. A process according to claim 20, characterized in that the selective or total deprotection of protected OH sites of the dithiocarbamic ester is carried out by passing a solution of the dithiocarbamic ester in a water/alkanol solvent mixture, in pure alkanol or in a mixture of alkanols on a column packed with an acid resin S' maintained at a controlled temperature. A dithiocarbamic acid ester having the formula R1 S N C SH in which R 1 and R 2 are saturated or unsaturated substituted or unsubstituted alkyl radicals or forming with the nitrogen atom a saturated or unsaturated ring which may contain heteroatoms and glycerol, in the acetal or deprotected form, excluding glycerol DEDTC. 930419,EEDAT.038,a:\45228pas.page,45 -46-

23. A dithiocarbamic acid ester having the formula RI S N C SH in which R, and R 2 have the meaning given in claim 22 and galactose, in the form of acetal or deprotected, excluding diacetone galactose DMDTC.

24. A dithiocarbamic acid ester having the formula R 1 S N C SH in which R, and R 2 have the meaning given in claim 22, and glucose in the form of acetal, or deprotected. A dithiocarbamic acid ester having the formula N C SH in which R, and R 2 have the meaning given in claim 22 and itols, in the acetal or deprotected form.

26. Deoxy-l N,N-cliethyl dithiocarbamate-l O-isopropylidene-2,3 glycerol.

27. Deoxy-l N,N-dimethyl dithiocarbamate-l glycerol.

28. Di-O-butanoyl (or palmitoyl)-2,3 deoxy-l N,N diethyl (or dimethyl)dithiocarbamate-l-glycerol.

29. Deoxy-3 diethyl (or dimethyl) dJithiocarbamate-3 di-O-isopropylidente 1,2:5, 6a-D allofuranoses. ncy-l N,N diethyl. dithiocarbamate-1 di-O- isopropylidene 2,3 4,5a-D fructopyranose.

31. Deoxy-6 N,N-diethyl dithiocarbamate-6 di-O- isopropylidene-1,2 3,4 -a-D galactopyranose. 92091 ,EEDAT.038, 4 45M2panpage,46 -47-

32. Deoxy-3 N,N-diethyl (or dimethyl) dithiocarbamate-3 di-O-isopropylidene-l,2 glucofuranoses.

33. Deoxy-3 N,N-diethyl (or dimethyl) dithiocarbamate-3 0-isopropylidene 1,2-a-D glucofuran- oses.

34. Deoxy-3 N,N-diethyl (or dimethyl) dithiocarbamate-3 D-glucopyranoses. Immunomodulating drug characterized in that it contains as an active substance one of the dithiocarbamic esters as defined in claims 22 to 34 associated with a pharmaceutically acceptable carrier.

36. A drug for the treatment of viral diseases, notably those that are due to HIV 1 or 2 viruses, characterized in that it contains as an active substance one of the dithiocarbamic esters as defined in claims 22 to 34 associated with a pharmaceutically acceptable carrier.

37. A process according to claim 1 substantially as hereinbefore described with reference to any one of the "examples.

38. An ester according to any one of claims 22 to DATED this llth day of September, 1992. PASTEUR MERRIEUX SERUMS ET VACCINS By Its Patent Attorneys 30 DAVIES COLLISON CAVE t t S 92911EEDAT.038,ak45228pasipage,47 INTERNATIONAL SEARCH REPORT International Application No PCT/FR 89/00569 I. CLASSIFICATION OF 5U1IJKCT MATTER (it several cialfication symbol& apply, Indicate all) According to internationht Patent £Aasalficatlon (IPC) or to both National Classification and IPC Int.Cl. 5 .C 07 H 13/12, A 61 K 31/70, C 07 D 317/24, A 61 K 31/335, *C 07 C 333/16, 333/18, 333/20, A 61 K 31/27 Ii. FIELDS SEARCHED Minimum Documentation SearchedI Documentation Searched other than Minimum Documentation to the Extent that such Documents ere Inctuded in the Fields Searched 6 Ill, DOCUMENTS CONSIDERED TO SE RELIVANTO Category Clitation Of Doc-umntn, 1% with indication, wheat& of'vta the relevant psmape It Relevant to Claim No. 13 X "The Chemistry of the Thiol Group", part 1, 1974, 1-21 editor Saul Patai, published by John Wiley Sons, (London, GB), pages 210-211 see pages 21(-211 X Recueil, vol. 8B, 1969, C.W. Pluijgers et al.: 1-21,24 "Synthesis and antifungal activity of some carbohydrate and amino acid derivatives of dimethyldithiocarbanic acid and of pyridine-2- thiol 1-oxide"l, pages 241-253, see page 242, table 1, compounds 1,2; page 246, table 3, compounds 1,2 X Chemical Pharmaceutical Bulletin, vol. 15. No. 3, 1-21,23,24 March 1967, S. Teji'ma et al.: "Thiosugars. X. Studies on glycosyl N,N-dialkyldithio- carbamates", pages 255-263 see the whole document X Journal of Chromatography, vol. 143, 1977, Elsevier Scientific Publishing Company, (Amsterdam, NL), D. Munger et al.: "High- performance liquid chromatographic analysis CSpecial categories at cited documentsl It later document published atr the interatiowal filing date ocu~nIdefnin thegenralstae O theon hic isnet or priority Pate and not tn conflict with the aritliCation but CA dcumnt efiingthe eneal tat 01the rt h~c Iinct cited to understand the Principle Or theory underlying the considered to be of particular relevance invention earlier document but publiahed on or otter the 1Iternational1 "X document ot particular relevance: the claimed Invention filing date cannot be conaidered novel ar cannot be consildered 1o document which may throw doubts on priority claimls) or involve an inventive stop which Is cited to establish the publication date of another document of particular relevance:the claimed invention citation or, other special man (as specified) cannot be considered to Involve an Inventive step when the document referring to an pof disclosure, use, exhibition or document is combined with one ot mote Other such docu. other means mnants. such combination being obvious to a person skilled document Published prior to the international filing date but 11n the arm. later than the pilority date claimed 'W document member Of the same patent famnily IV. CERTIFICATION Date 01 the Actual Completion of thet International Search Date of Mailing of this International Search Report 26 February 1990 (26.02.,90) 25 May 1990 (25.05.90) International Searching Authority Signature of Authorized officer European Patent Form PCTIISAI2iO isecond sheet) (Januarny 1315) -2- IS~us~a~u~letle~ PCT/FR 89/00569 tu. DocUMEnTo coMstaDIRI To sit RELvAmT (CownTuEO PROOm 713 BconO 514f *af ftan at OMWOM, Wiux k~mW". VANN W Od WNE USQSS 0WOM mo I to Clam NO of dianhydrogalactitol'in plas~ma by derivati- zation with sodium diethylditbiocarbamate", pages 375-382, see page 380, figures III, IV X Tetrahedron Letters, vol. 27, No. 35, 1986, 22 Pergamon Journals Limited, P. Rollin: "One-step stereospecific conversion of alcohols into dithiccarbamates: a smooth pathway for the introduction of a sulphur functionality", pages 4169-4170, see page 4169, table I X Chemical Abstracts, vol. 100, No. 14 37 2 April 1984 (Columbus, Ohio, US),I. Ymen: "Structure of 1lithiumn direthyldithiocarbamate tetrahydrate, Li[C H NS I.4H see page 592, abstract 112594mn, i Rct3 Cry~tallogr., Ser-t. C: Cryst. Struct. Commun. 1984, C40(1), 33-4 X Chemical Abstracts, vol. 100, No. 14, 2 April 1984, 37 (Columbus, C0io, US), I. Ymen: "Structure of lithium diethyldithiocarbamate trihydrate, Li[C HIONS 2 1 3H 2 011, see page 594, abstract 1126N Aca Crystallogr., Sect. C: Cryst. Struct. Commun. 1984, C40(2), 241-3 X J. Chem. Soc., Dalton Trans., 1973, 37 D.C. Bradley et al.: "Covalent compounds of quadrivalent transition metals. Part VI. Spectroscopic studies on titanium, vanadium, and zirconium diethyldithiocarbamates", pages 2228-2233, see page 2232, 6olumn 1, lines 40-43 X Chemical Abstract, vol. 91, No. 9, 27 August 1979, 37 (Columbus, Ohio, US), see page 192, abstract 70011g, JP, A, 794324 (TOKYO ORGANIC CHEMICAL INDUSTRIES, LTD) 18 April 1979, Chemical Abstracts, Tenth Collective Index, C H NSCarbamodithioic acid,dimethyl- lRt4iuXsalt A EP, A, 0179694 (INSTITUT MERIEUX) 30 April 1986 22-36 see page 7, line 23 page 9, line 33; claims 1,2,11,12 A EP, A, 0288871 (PROTER) 2 November 1988 22,26,35,36 see claims 1,10 Form PCT)ISAMOism enoo tit 4jauary IOUS) international Application No. PCT/FR 89/00569 FURTHER INFORMATION CONTINUED FROM THE SECOND SHEET IV.[:]OSERVATIONHS WHERE CERTAIN CLAIMS WERE FOUND UNSEARCI4ASLE' This International search report has not been established In respect of certain claims under Article 17(2) for the following reasons: *[)Claim because they rolat, to subject matter not required to be searched by this Authority, narmly:- ZM: Clim numbers because they relats to parts of the International application that do not comply with the piescribed require- meni,,to s~ch an extent that no meaningfl International search can be carried out, epecifically: clm nrniers.......becau"e they are depor~dent claima and are not drafted in accordanc with the second and thid settence of POT Rlule 6.41(a). VI.EJ ONSERVATIONS WHERE UNITY OF INVENTION IS LACKING'I This International Searching Authority found multiple Inventions In this International application as follows: 1. Claims 1-37. 2. Claims 38,39. As ali requirad additional search fees werhi msly paid by the applicant, this International search report covers all searchable clsime of the International 4pplication. L(3 As only some of the required additional search tees were timely paid by the applicant, this International search report covers only those claims of the International application for which foes were paid, specifically claimns:. &QNo required additional search fees were timely paid by the applicant Consequently, this international search report Is restricted to the Invention first mentioned In the clis: It Is covered by claim numbers; 1-37 4,E) Asalisearchabe claims could be searched without effort justifying en additional fee, the International Searching Authority did not Invite pament oi any additionai fee. flomarm on Protest 0 The additional search tees were accompanied by applicant's protest. [3 No protest accompanied the payment ot additional search tees. Form PCIIISAI2lO (supplemental sheet (January 1605) ANNEX TO THE INTERNATIONAL SEARCH REPORT ON INTERNATIONAL PATENT APPLICATION NO. FR 8900569 SA 32434 This annex lists the patent family members relating to the patent documents cited in the above-mentioned international search report. The memnbers sre as contained in the European Patent Office EDP file on 18/05/90 The European Patent Office is in no way liable for these particulars which are merely given for the purpose of inf~mnadon. citedl in search report EP-A- 0179694 Publication Patent family Publication memberQ') date 30-04-86 CA-A- EP-A, B WO-A- JP-A- JP-T- Th66058 0198878 8602639 61103827 62501069 02-05-86 20-02-90 29-10-86 09-05-86 22-05-86 30-04-87 EP-A- 0288871 02-11-88 None 'W For more details about this annex .1se Official Journal of the European Patent Oflke, No. 12182 RAPPORT DE RECHERCHE INTERNATIONALE Demandit interniatio~nal* N* PCT/FR 89 /00569 1. CLASSE MENT DE L'IiM4ENTION (61 otusiturs aymbotea do classification sont apolicabs. les indlipuet tous) Seion Is classification Internationals des brovo:, (CIS) ou A Is lois selon in classification national, at In CIS IB C 07 H 13/12, A 61 K 31/70, C 07 D 317/24, A 61 K 31/335, C 07 C 333/16, 333/18, 333/20, A 61 K_31/27 11, DOMAINES SUR LESQUELS LA RECH4ERCHE A PORTE Documentation minimal* consuitileI systie czLidesrilllionS, .Jio de clnaificationi C 07 H 13/00, C 07 D 31.7/00, C 07 C 333/00, CIB 5 A 61 K 31/00 Documentation eiiiulto suit* quo Is dociumetation minmmii dkns as mosurg ou do tots documerts font partis des domainha our losquala Is recherehi a Pon*i Ill. DOCUMENTS COMiVIDERVES COMME PERTIP4ENTS It Caltorle Identification der~ documents citts," avoc Indication, s4 niceasoiro. IN, dos reondicatioons dos Pasages Poflinoni 12 vi sgi 'I-eChemistry of the Thiol Group' partie 1, 1974, 6diteur Saul Patai, PubI4 par John Wiley Sons, (Londres, GB), pcges 210-211 voir pages 210-211 Recueji, volume 88, 1969, C.W. Plujgers et al.: "Synthesis and antifunga1 activity of some carbohydrate and amino acid derivatives of dimethyldithio- carbamic acid and of pyridine-2- thiol 1-oxide"l, pages 241-253 voir page 242, tableau 1, compos~s 1,2; page 246, tableau 3, ccmpos6s 1,2 Chemical $4 Pharmaceutical Bulletin, volume no. 3, mars 1967, 1-21 1-21, 24 1-21l,3,24 'A C610ha~s rnPOCIA10 do documents citla: 1t aA 2i documont difilissant I'itat gifliral de as techniqe~u, non cznsiO04 comma particuliiroment Ponlient 4' I a document anti"Aur, mas Publii A 1 data do dip~l Interna. tighal Pu aprk5 w.ie date Lai document Potivant ifflot on douto s&ir tine roendication do peioeite oti cilk o04i M~armtinat to dateat )Publication d'une auiteo citation o6 poir tine ragoon 904ciale (talls quIndiquio) a0it document is rollirtritl h ufn divulgation peate, A on tias, A tine exposition Cu toto* autreg moyans oP a documaint 0014~ avant ao date do d6061 international, irraim posteriouement a to date do OfMIeit eovendgquda eTa document ulttiour pubti# ovieloueoment AIs dots do d#0:t International ou A Is date do ptboritil at napovoneniant pas A ltat do to technique pfrinent, mam oil pour comoronidro Is Principe ou Is thAoria tonstuantI ase do linvontion oX a document Panicutlieirnant beirtinent; Vinveantin roveoi. quo. ne pout lite coflaidirde comma nauvoile o comma ImpIlItient tine actiVilt' inventive it Y a document particuiliiromeflt pertinent-, Vinventton rsoen. diQuis no pout dire considlrie comma implioqant. tri aiclie te inventive loranue IC document eat &&acil a un ou ptuaifurs suits$ documents do Mois nature, coltt COMMbi naioon tant tvidonta pbour tinie pelmonno du mnklia?. a4 a document qul faht parict do Is moms familie do bravots IV, CERTIFICATION Oat* it lerjuolle to recriorche lnternationotv a did otfectivement Octievoo 26 f6vrier 1990 Administration chargoe do Is rechercheo into nastionpe OFFICE EUROPEEN DES BREVETS Oata deoxohdition do PhAsent rapport do recrigrclto inlornationale Formijlaire PCTIISA1210 (derusuiil lou,1i Jinvwe A0110, Demand. International@ N* PCT/FR 89 /00569- 2- (SUITE DES RENSEIGNEMENTS INDIQUES SURt LA Ill. DOCUMENTS CONSIDjft9S COMME PERINENTS DEUXIME FEUILLE) Calteore 6 dm'iitcaein dompwnts aites, &a w kiaw~n. 06 nbo wNO ea rjacatxv,-, d" case"g peettlInts X. Studies on glycosyl N,N- dialkyldithiocarbamates", pages 255-263. voir l'article en entier XI Journal of Chromatography, volume 143, 1977, Elsevier Scientific Publishing Company, (Amsterdam, NL), ;.Munger et al.: "High-performance liquid chromatographic analysis of dianhydrogalactitol in plasma by derivatization with sodium diethyldithiocarbamate"l, pages 375-382 voir page 380, figures Ill, IV X Tetrahedron Letters, volume 27, no. 35, 2'2 1986, Pergaxnon Journals Limited, (GB), P. Rollin: "One-step stereospecific conversion of alcohols into dithiocarbamates: a smooth pathway for the introduction of a sulphur functionality", pages 4169-4170 voir page 4169,,tableau I X chemical Abstracts, volume 100, no. 14 37 2 avril 1984, (Columbus, Ohio, US), I. Ymen: "Structure of lithium dimethyldithiocarbamate tetrahydrate, LiCC3H6NS 2 ].4H 2 voir page 592, abr6g6 112594m, Acta Crysf-n.llogr., Sect. C: Cryst. Struct. Commun. 1984,, C40(l)i 33-4 X Chentical Abstracts, volume 100, no. 14, 37 2 avril 1984, (Columbus, Ohio, Wis), 1. Ymen: "Structure of lithium ditathyldithiocarbamate trihyd1rate, Li[:C 3 Hl 0 NS 2 J .3H 2 voir- Page 594, abr6ge 112628a, Acta Crystallogr., Sect. C: Cryst. Struct. Commun. 1984, C40(2), 241-3 X J. Chem. Soc., Dalton Trans., 1973, 37 Bradley et al.: "Covalent compounds of quadrivalent transition metals. Part VI. Spectroscopic studies on titanium, vanadium, and zirconium die thydithiocarbamates", pages 2228-22333 voir page 2232, colonne 1, lignes

40-43 Formulair. PCTIISA12I0 (Isulle addltionnali.) (Janvw. 19SS) Demand* International* N, PCT/FR 89 /00569-3 (SUITE DES RKN"StIOINT 5S INDIQUtS SUR LA Ill. DOCUMENTS~ CONSIDIR911 COMME PERlINENTS DEUXIIEME FEUILLE) Catigoris dtcabon Me. docurnents crie. avoc Wdca~on. wJ n .Me. j des rmndicatbo" d" Pasage Puew ftee X Chemical Abstract, volume 91, no.. 9, 37 27 aoat 1979, (Columbus, Ohio, US), voir page 192, abr6g6 70011g, JP, A, 794324 (TOKYO ORGANIC CHEMICAL INDUSTRIES, LTD) 18 avriJ. 1979, Chemical Abstracts, Tenth Collective Index, C 3 11 7 NS 2 Carbamodithioic acid, dimethyl- lithium salt A EP, A, 0179694 (INSTITUT MERIEUX) 22-36 avri. 1986 I voa:: page 7, ligrie 23 page 9, ligne 33; revendications 1,2,11,12 A EP, A, 0288871 (PROTER) 2.2,26,35, 2 novembre 1988" 36 voir revendications 1,10 FetmulaU.e PCTIISAMO1 (teullo additionnelle) (Janywe 1043) Demnands international* N, PCT/FR ec~/ 0056~9 SUITE DES RENSEIGNEMENTS INOIQU9S SUR LA DEUXItME MEILLE V. OBSERVATIONS LORSQUtIL A MT KS711illt QUE CERTAINIS REVENDICATIONS HE POUVAIINI PAS FAIR& L'OBJET D*UNE RECHIERCHE I Solon Particle 17.2) a) certalnes revandicatlona nWont pas fait Pobiot d'une roercho pour too motifs oulvanta: 1. Let ravendicationa nut rapportent A, cn objot. A l'igard duqusi Is gr~stnt. administration W'e pas libilgation do pro- C~dw 6 wl fecrico, A savoir: 2. Las revondicationa numirooa..... s uo apportent A des partiesa do Is demand* lntiornalionalt qui no Tomplacont pas le co~ditiols, preacrile dens une maure tolle qu'une recherche aignificative no pout Alto offecti'to, prieladment: 3, Los revandlcationa nuff*o sont des revendications cdpondantee at ne soot pas r6digdoa conformdniont Ala douitme et Aa oIstAsAme Wiuss do Ia rigle du PCT, VI. OBSERVATIONS LORSOU'lL Y A ABSENCE DIUNITI DE L'INVENTION L'ad ministration chargA. dolIn recherche Internationale a trouvA plusioura Inventions dana Is piriagnte demands Internationale, c'eat-&-dira: 1 .Revendications 1-37. 2 .Reveridications 38,39. 1. [3Comm. tout.. lee taxes additlonnell~a demandies ant Ot6 pay~eu dens lee ddilal, t@ prdaeont rapport dop recherche lnternatianalia ecuvre toute. Its revendicationo de Ia demand# Internationale pouvant fair* l'objet d'un. recherche, 2. QComm. souhment unoi portt des taxes addltk)ArnoWtI domandila. a Ott pay6o dans a oals1, It pr~soft rapport a recherchae Internatinale court. sousoettl caioea del revendications o It dornaxicd pour Bosqalos tee taxes ant Ott pays*&, c'est-dIro lo" roeondicationa: 3. A ucuno taxt additionnelle demandit na Ali payde dans lot dilula par It d~posant. En conaicluonce, I. prilsont rapport do recherche Internationale oat limit6 A llnvontlon montionndc an premier dons lot ravandicationa; al, eat couverte par lt toenoicattont num~roa: 1-37. 4,D Etant donn4 out louta* lei rovondicationa susoplibles do fair. Pobjet d'une recherchto o ouvalanl men*a florl particulior luatiflont une taxt additionnolle, Ilad mini strstio n charges do Is rech* ircho Internationale na soilicitA to paieonlont dasucunt laxe additionnoillo Remarque quant A la r~seive QLoa taco. additlonnolios do recherche itaiont accompagnis dun. r~terve du dipatant. Aucuno rserve nas 4t6 fel* Iota du polomont des taxes additionnelloa do rechche. Formulairo PCTIISA/2IO (loulle supplimontaire (Janvae 'lO6t) ANNEXE AU RAPPORT DE RECHERCH-E INTERNATIONALE RELATIF A LA DEMANDE IN'U6aNATIONALE NO. FR 8900569 SA 32434 La presente annexe indique los membres de Is famnile de brevets vitatifs auw documents brevets cites dans le rapport de recherche internationale visi ci-dessus. Losdits inembres sont Contenus au fichier informatique de l'Office europien des brevets i Ia date du 18105190 Les ronseignements lournis sont donnes a titre indicatif et n'engagent pas Is responsabiliti de l'Offico etaropien des brevets. Document brevet citi Date do IMembre(s) de Is Date do au rapport do recherche publicato fainlle de brevet(s) publication EP-A- 0179694 30-04-86 FR-A,B 2572284 02-05-86 CA-A- 1266058 20-02-90 EP-A, B 0198878 29-10-86 WQ-A- 8602639 09-05-86 JP-A- 61103827 22-05-86 JP-T- 62501069 30-04-87 EP-A- 0288871 02-11-88 Aucun Pour tout renseignement concenan conte annexe voir Journal Officiel do l'OMco europher des brevets, No.12/82