IE60234B1 - Therapeutic compositions containing benzhydrylthiomethane derivatives - Google Patents

Abstract

1. Claims for the Contracting States BE, CH, DE, FR, GB, IT, LI, LU, NL, SE Therapeutic compositions containing as active principle a derivative of benzhydrylthiomethane having the Formula : see diagramm : EP0258134,P10,F2 where Y denotes a -CONH2 , -CO-NHOH or -CH=N-OH group, and R1 and R2 denote a hydrogen or fluorine atom independently of one another. 1. Claims for the Contracting States AT, GR, SP A process for the preparation of a therapeutic composition, characterized in that a pharmaceutically acceptable form is given to a derivative of benzhydrylthiomethane having the Formula : see diagramm : EP0258134,P10,F3 where Y denotes a -CONH2 , -CO-NHOH or -CH=N-OH group, and R1 and R2 denote a hydrogen or fluorine atom independently of one another.

Description

Ths present invention is concerned with therapeutic compositions containing as active principle a benzhydrylthiomethsne derivative which can be used In therapeutics because of their activity on the central nervous system and for certain purposes because of their immuno-stisaulating activity.

A certain number of benzhydrylthiomethan© derivatives which are used in the present invention have already been described.

Thus , FR-A-2 385 693 describes 2-(benzhydrylthio)acetamide, Patent Specification No. 12.77/33 describes 2-(4,4’-difluorobenzhydrylthio)acetamide thio) acetohydroxamic acid. However, these compounds have only been described as intermediate products and it was not evident that they offer interesting properties permitting their use in therapeutics.

Tho subject of tho present invention is compounds containing as active principle a benzhydryIthiomethane derivative with the formula R CH-S-CHn-Y II in which ϊ represents a group -CONKj, -C0-NH0H or -CH - N-OH, and and represent independently of each other a hydrogen or a fluorine atom.

The compounds with the formula IX in which Ϊ = -CONHj can be prepared by the reaction of ammonia on th® chloride of the corresponding 2-(benzhydrylthio)acetie acid.

The compounds in which T - -GQ-KHOH can be prepared by the reaction of hydroxylamine hydrochloride on ths methyl ester of the corresponding 2-(benzhydrylthio) acid. f The compounds in which ϊ - -CH=K~0H can be prepared by the reaction of hydroxylaminehydrochloride on the corresponding 2-(benzhydrylthio)acetaldehyde, The following examples illustrate the preparation of the compounds.

Example 1 Preparation of 2-(4,48-difluorobenzhydrylthio)acetamide (code No. CRL 41334 ) A solution of 26.40 g (0.08 mole)) of 2-(4,.4!-dif luor obenzhydrylthio)acetic acid is heated to reflux for one hour in 145 ml of benzene and 30 ml of thionyl chloride.

After evaporating under reduced pressure, the residue is taken up with 100 ml of ether and poured into an agitated mixture of 100 ml of 28% ammonia and 200 g of ice.

This is agitated for 2 hours, then the ether is poured off and the remainder is washed with water, dried on NajSO^, and evaporated to dryness under reduced pressure. The residue is taken up with petroleum ether and separated.

After re-crystallisation in isopropyl ether, there is obtained : g of amide. (fflP”inst yield + 68 %).

The compound is a white powder, insoluble in petroleum ether, soluble in ether, ethyl acetate and the alcohols. Its solubility in water is less than 0.1 %.

Example 2 Preparation of 2-(4,4g-difluorobenshydrylthio)acetohydroxamic acid (code No. CRL 41 335) 44.1 g (0.15 mole) of 2-(4,41-dlfluorobenshydrylthio)acetic acid is dissolved in 300 ml of methanol and 1.5 ml of concentrated H2S0^ is added to it. After 4 hours at reflux. the methanol is evaporated, the residue is taken up with water and ether, washed with dilute bicarbonate, then with water» dried and evaporated under reduced pressure.

The oily residue is treated for one night at 20°C with a solution obtained with 6.9 g (0.3 g.atom) of sodium, 10„5 g (0.15 mole) of hydroxylamine hydrochloride end 480 ml of anhydrous methanol.

It Is then evaporated to dryness under reduced pressure, taken up with 600 mg of water, filtered on charcoal, acidified, with 3N HCl, extracted with ether, washed with water, dried, evaporated and crystallized from petroleum ether. 28.8 g of 2-(4:,4'-difluorobenzhydrylthio)acetohydroxamic acid is recovered. (m.p. insj._ = 76° C: Yield = 62 %)» The compound is a slightly pink powder: soluble in ether, ethyl J acetate, alcohols: insoluble in water» ♦ Example 3 Preparation of 2-(benzhydrylthio)acetamide, (code No. CRL-41055). g (0.076 mole) of benzzhydrylthioaeetyl chloride, in solution in 100 ml of methylene chloride, is added drop by drop under agitation to 40 ml of ammonia and 40 ml of water. After agitation for one hour, the organic phase is decanted, washed with water and dried on NajSO^.

The solvent is evaporated under reduced pressure, the residue is crystallized froa isopropyl ether and recrystallized from ethyl acetate. The compound is obtained with a yield of 45 %» It is a white powder, soluble in alcohols, acetone, ethyl acetate: insoluble in water, isopropyl ether.

It melts at 107 - 108°C.

Example 4 Preparation of 2-(benzhydrylthio)acetaldoxime (Code H°. CRL-40^956 ) a) Preparation of 2-(benzhydrylthio)acetaldehyde. g (0.1 mole) of diphsnylmethanethiol is added to a solution of g (0.13 g.atom) of sodium in 250 ml of ethanol, then, over one hour, g (0.125 mole) of bromoacetaldehyde diethylacetai is added drop by drop at reflux. After 2 hours at reflux, the alcohol is evaporated off under reduced pressure, the residue is taken up with ether, washed 3 times with water, dried and filtered on charcoal. By evaporating th© ether under reduced pressure, the diethylacetal of the 2-(benzhydrylthio )acetaldehyde is obtained quantitatively.

The latter is hydrolysed by heating for 2 hours on a water-bath with 250 ml of 10 a H2S0z, extracted with ether, washed with water, dried, evaporated, crystallized from petroleum ether, and recrystal- ? lized from isopropyl ether. 2«(benzhydrylthio)acetaldehyde (ω»ρ. 5758°) is obtained with a yield of 56 %. (. > b) Preparation of 2-(benzhydrylthio)acetaldoxime. 4.2 g (0.08 mole) of hydroxylamine hydrochloride in solution in 15 al of water is partially neutralised with 3.2 g (0.04 mole) of sodium bicarbonate. 9.6 g (0,04 mole) of 2-(benzhydrylthio)acetaldehyde in solution in 50 ml of methanol is added rapidly at 20° C. This solution is agitated for 2 hours, the pH falls rapidly from 10 towards 5 and the oxime precipitates. 20 ml of water is addedleft in contact for one night, then, after separation, the residue is washed with water, dried and crystallized from 80 % methanol. The compound is obtained with an overall yield of 44 Z.

This is a white powder, soluble in ether, ethyl acetate, methyl and ethyl alcohols.

Insoluble in water and petroleum ether.

The equimolecular mixture of the '’syn” and anti” forms has a melting point of 86-87° C.

The results of the pharmacological studies which make clear the properties of the compounds are given hereafter.

Neuyopsycho-pharmacologieal study 1) Study of CRL 41 334 The compound CRL 41 334, in suspension in a solution of gum arabic, was administered by intraperitoneal route at a volume of 20 ml/kg to mice (male, NMRI.C.E.R. January) and at a volume of 5 ml/kg to rats (mala, GD-j: SPRAGUE DAWLEY, Charles River).

I - PRE-TOXICITY ( 3 mice per dose). - 128 mg/kg : unsteady walk, exophthalmia, sedation, then excitation 2 hours after injection. 256 mg/kg ; unsteady walk, exophthalmia, convulsions (3/3) at 15 minutes, sedation followed by excitation 2 hours after the injection. No mortality. - 512 mg/kg : unsteady walk, exophthalomia, convulsions (3/3) during the first 10 minutes. Mortality (3/3) (1/3 in 15 minutes and 2/3 in 24 hours).

II - OVERALL BEHAVIOUR AMD REACTIONS Groups of 3 animals ware observed before, then 15 minutes, 30 minutess, 1 hour. 2 hours, 3 hours and 24 hours after the administration of the compound CRL 41 334» 1 ~ Mice 2, 8, and 32 mg/kg ~ no clear modifications to behaviour and reactions. 128 mg/kg ~ sedation for 30 minutes accompanied by unsteady walk, reduction of the respiratory frequency, of the reactions, of th© fear reaction, and of the muscular tons» Hypothermia (-4-10 at 30 minutes, lasting 1 hour).

~ Rat 1, 4, and 16 mg/kg - behaviour, reactions, variation of th© pupillary diameter and rsctal temperature comparable to those of ths contract group» mg/kg ~ sedation during 15 minutes accompanied by dyspnoea, reduction of reaction to touching aad of muscular tone. - this sedation is followed by an increase of fear reaction (30 minutes), of reaction to touching and of muscular tone for 3 hours. - mydriasis, very moderate and short, at 1 hour.

III - INVESTIGATION OF STEREOTYPE MQVQISNTS Groups of ό rats receive an injection of the compound CRL 41 334 or of amphetamine, and are then immediately placed in boxes of small dimensions (20 x 10 x 10 cm) where their stereotype behaviour is marked at 0 to 3 until the extinction of the effect.

At the two strongest doses used (04 and 128 mg/kg), the compound CRL 41 334 causes clear stereotypies the intensity of which is comparable to that obtained respectively ’with 1 and 2 mg of amphetamine.

The kinetics of the action, however,, are different. The stereotypies induced by 2 mg/kg of amphetamine reach their maximum in to 90 minutes and last 270 minutes, while the stereotypies introduced by 128 mg/kg of the compound CRL 41 334 reach their maximum in 120 - 15Ο minutes and last more than 420 minutes.

IV _ INTERACTION WITH APOMORPHINE - Mice Groups of 6 mice receive the CRL 41 334 compound half-anhour before sub-cutaneous injection of apomorphine at a dose of 1 or 16 mg/kg,. a) Apomorphine, 1 mg/kg b) Apomorphine 16 mg/kg At the strongest dosage used (128 mg/kg) the compound CRL 41 334 brings on hypothermia and aggravates th© hypothermia induced by apomorphine (1 mg/kg)The behaviours of verticalisation and of stereotypies are not modified» - Rats , The compound CRL 41 334 is administered to groups of 6 rats half-an-hour before ths sub-cutaneous injection of 0,.5 mg/kg of apomorphine., At the two strongest doses used (16 and 64 mg/kg), the compound CRL 41 334 strengthens the stereotypies induced by apomorphine.

V - INTERACTION WITH AMPHETAMINE Amphetamine (2 mg/kg) is injected by intraperitoneal route to groups of 6 rats h&lf~an~hour after the administration of the compound CRL 41 334At doses of 16, but particularly at 64 mg/kg, the compound CRL 41 334 strengthens the amphetamine stereotypies.

VI - INTERACTION WITH RESERPINE Four hours after Intraperitoneal injection of 2.5 mg/kg of reserpine, groups of 12 mice receive the CRLsf 41 334 compound.

- Action on the temperature.

- Action on ptosis.

At the strongest dose used (128 mlg/kg)„ the compound CRL 41 334 moderately aggravates reserpinic hypothermia. Ptosis, already maximal, is not modified.

It should be noted that at 24 hours, the compound CRL 4-1 334 (8, 32 or 128 mg/kg) seems to oppose the reserpinic hypothermia and ptosis.

VII - INTERACTION WITH OXOTREMORINE The compound CRL 41 334 is administered to groups of ό mice half~an~hour before the intraperitoneal injection of 0.5 mg/kg of oxotremorine.

- Action on temperature.

At the strongest dos© used (128 mg/kg), the compound CRL 41 334 brings on hypothermia and aggravates hypothermia induced by oxotremorine.

- Action on trembling Trembling caused by oxotremorine is not modified by the compound CRL 41 334« - Action on peripheral cholinergic symptoms.

At a dose of 128 mg/kg, ths compound CRL 41 334 reduces defecation without modifying salivation and lacrimation.

VIII - ACTION ON,THE FOUR-PLATE- TEST, TRACTION AND ELECTRIC.SHOCK.

The test is carried out on groups of 10 mice, half-an-hour after the administration of th® compound CRL 41 334.

The compound CRL 41 335 does not clearly modify the number of passages punished. It does not cause any major movement incapacity.

At the strongest dos© used (128 mg/kg), the compound CRL 41 334 weakly opposes the convulsive effects without modifying the lethal effects of the electric shock.

IS - ACTION ON SPONTANEOUS FREE MOVEMENT. r (_ 1) Compound administered 10 minutes before passage into activity matey?,, At doses of 32 and 64 mg/kg, the compound CRL 41 334 brings on an increase of the spontaneous free movement of the mouse; at a dos© of 128 mg/kg, this effect does not further appear. 2) Compound administered 2 hours before passage into activity meter At the dose of 8, but particularly at 32 and 128 mg/kg, the compound CRL 4,1 334 brings on an increrass in the spontaneous free movement of the mouse.

X - ACTION ON INTER-GROUP AGGRESSIVENESS After staying for 3 weeks in each of the halves of a cage separated by an opaque partition, groups of 3 mica receive the compound CRL 41 334. Half-&n-hour later, the two groups of the same cage are put together by removal of the partition, and the number of fights which take place in 10 minutes Is noted. Half of ths test is carried out on the usual mice (NMRI, C.E.R. January) and half on the NMRI (Iffa Credo) mice.

At ths two doses used (32 and 128 mg/kg), th© compound CRL 41 334 reduces the number of fights.

XI - ACTION VIS-A-VIS BEHAVIOUR DISTURBED BI VARIOUS AGENTS - Motility reduced by habituation to th© enclosure After remaining for 18 hours in ths activity meters,, the mice (ό per dose, 12 controls) receive ths compound CRL 41 334- They are immediately replaced in their respective enclosures and, half-an-hour later, their motility is recorded for 30 minutes.

Starting at a dos© of 2 mg/kg, the compound CRL 41 334 brings on a clear renewal of the motor activity in ths mouse habituated to its enclosure.

- Motility reduced by hypoxic aggression Half-an-hour after having received the compound CRL 41 334. the mice (10 per dose, 20 controls) are submitted to an acute hypobaric anoxia (depression of 60 mmHg in 90 seconds, held for 45 seconds), then they ar© placed in the activity meter where their motility Is recorded for 10 minutes.

Starting at a dose of 2 mg/kg, the compound CRL 41 334 brings on an improvement in the motor recovery in the mouse of which the Ο motility had been depressed following a brief period in an enclosure with reduced pressure. This improvement is considerable for the two strongest doses used. (32 and 128 mg/kg).

- Asphyxiating anoxia Groups of 20 sice receive the compound CRL 11 334 half-an- | hour before the intraperitoneal administration of 32 mg/kg of gallamine triiodoethylate At the strongest doss utilised (128 mg/kg). the compound CRL 41 334 increases the period for the appearance of death following an anoxia caused by a curarizing agent. Ths period for the appearance of convulsions is not modified.

Ill - INTERACTION WITH BARBITAL Half-an-hour after the administration of the compound CRL 41 334, groups of 20 mice receive an intraperitoneal injection of barbital (220 mg/kg).

At doses of 8, 32 and 128 mg/kg, the compound CRL 41 334 brings about a considerable decrease in the duration of barbituric sleep.

XIII - ACTION ON BEHAVIOURAL DESPAIR Half-sn-feour after having received the compound CRL 41 334, groups of 6 mice (males. CD^ „ Charles River) were placed in a beaker filled with water to a height of ό cm.. The total duration of immobility between the 2nd and the 6th minute following the immersion was noted.

At doses of 8. 32 and 128 mg/kg, the compound CRL 41 334 reduced the duration of immobility of the mice placed in forced immersion. r IIV - CONCLUSION V The neuropsycho pharmacological study of the compound CRL 41 334 shows s - a stimulating effect in the mouse ϊ hypermotility, clear ί 1 renewal of motor activity in the mouse habituated to its enclosure, improvement in the motor recovery after hypoxia, antagonism to barbituric sleep and reduction of despair*3 hypomotility At a strong dose, a brief sedative effect is noted : sedation, hypo-reactivity, hypo-aggressivity, hypothermia, aggravation of hypothermia induced by reserpine and oxotremorine, moderate reduction of the convulsivating effect of an electric shock and of the delay in the appearance of death following anoxia caused by a curarising agent. Two hours after the administration of th® product, this sedative effect is replaced by a stimulating effect.

Further, in the rat, there is noted with a strong dose : stereotypies and a poientiallz&tion of the stereotypies induced by apomorphine and amphetamine.

This stimulating effect seems to develop only after· a considerable latency period of the order of at least 2 hours. 2) Study of CRL 41 055 Tbe compound CRL 41 055» in suspension in a solution of gum arable, was administered by intraperitoneal route at a volume of 20 ml/kg to mice (male, NMRI» Evie Ceba).

Pre-toxicity ( 3 mice per dose). - 64 mg/kg : excitation, stereotypies (sniffing, standing-up) appearing after 45 minutes and lasting more than 2 hours. - 128 and 256 mg/kg : sedation for 45 minutes, respiratory frequency diminished, abdominal contractions followed by excitation, stereotypies (sniffing, standing-up) for more than 2 hours.

Io mortality. - 512 mg/kg ; sedation for 45 minutes, respiratory frequency diminished, abdominal contractions, sub-convulsive titubations followed by excitation, stereotypies (less clear than with the weaker doses). hours after injection, intermittent convulsions present in all the mice, interrupted (2/3) by stereotypies (toilet, jaw movements).

Mortality (1/3) after 48 hours. - 1024 mg/kg : the same symptoms.

Overall behaviour and reactions Groups of 3 animals were observed before, then 15 minutes, 30 minutes, 1 hour, 2 hours. 3 hours, and 24 hours after the administration of the compound CRL 41 055» - 2 mg/kg : no clear modifications of behaviour and reactions» - 8 and 32 mg/kg : excitation for 2 hours. - 128 mg/kg : sedation for 30 minutes giving way to a late excitation (2 hours) lasting 1 hour with increase in reaction to touching and of the fear reaction.

Action on spontaneous free movement.

Half-an-hour after having received the CRL 41 055, the mice (6 per dose, 12 controls) were placed in an activity meter where their motility is recorded during 30 minutes.

The CRL 41 055 at doses of 32 and 128 mg/kg brings about an increase in spontaneous motility in the mouse, immersion» Motility reduced hy habituation to the enclosure.

After remaining for 18 hours in the activity meters, the mice (8 per dose, 12 controls) receive CL 41 055» They are immediately replaced in their respective enclosures and, half-an-hour later, their motility is recorded for 30 minutes» CRL 41 055 at doses of 32 and 128 mg/kg causes a clear renewal of the motor activity in the mouse habituated to its enclosure.

CONCLUSION CRL 41 055 presents a stimulating effect in the mouse : excitation, movements appearing to be stereotypies, hypermotility, and clear renewal of motor activity in the mouse habituated to its . enclosure. This stimulation is sometimes preceded by a phase of sedation. r The CRL 41 055 has proved to be useful in human therapeutics as an / anti-depressant, at a dose of three capsules of 50 mg per day. > 3 3) Study of CRL 40 956 . The compound CRL 40 956, in suspension in a solution of gum arabic, wag administered by intraperitoneal route at a volume of 20 / ml/kg to mice (male. NMRI. Evic Ceba) and at 5 ml/kg to rats (male.

CD-p Sprague Davley, Charles River).

Pre-toxicity (3 mice per dose). 1024 and 512 mg/kg : abdominal cramps, dyspnoea, no mortality. 256 mg/kg : same symptoms.

Overall behaviour and reactions Groups of 3 animals were observed before, then 15 minutes, 30 minutes, 1 hour, 2 hours, 3 hours and 24 hours after the administration of the compound CRL 40 956.

- Mice 512 mg/kg ; sedation for 2 to 3 hours, with reduction of the reaction to touching and of the muscular tone for 1 to 3 hours. Hypothermia (-2.9°) for 3 hours. 128, 32 and 8 mg/kg ; no clear symptoms.

- Rats 256 and 64 mg/kg : sedation for 3 hours with reduction of the reaction to touching and of the muscular tone. and 4 mg/kg : behaviour and reactions comparable to those of the control group.

Interaction with apomorphlne The CRL 40 956 compound is administered to groups of 6 rats half25 an-hour before sub-cutaneous injection of 0.5 mg/kg of apomorphlne.

At doses of 64 and 256 mg/kg, the CRL 40 956 very moderately reduces the intensity of the stereotypies induced by the apomorphlne.

Action on spontaneous free movement.

/ Half-an-hour after having received the CRL 40 956, the mice (6 per dose, 12 controls) are placed in an activity meter where their motility is recorded for 3θ minutes. 4.

At the strong dose (512 mg/kg), CRL 40 956 strongly reduces the spontaneous motor activity of the mice.

Action on inter-group aggressiveness After staying for 3 weeks in each of the halves of a cage separated by an opaque partition, groups of 3 mice receive the compound CRL 40 950. Half-an-hour later, the two groups of the same cage are put together by removal of the partition, and the number of fights which take place in 10 minutes is noted.

At a dose of 128 mg/kg, the compound CRL 40 956 reduces the number of fights.

Action vis-a-vis asphyxiating anoxia Groups of 10 mice receive ths compound CRL 40 956 half-an- hour before tha intraperitoneal administration of 32 mg/kg of gallamine triiodoetbylate At a strong dose (512 mg/kg), the compound CRL 40 956 moderately retards the appearance of convulsions and of death following asphyxiating anoxia caused by a curarising agent.

Interaction with barbital Half-an-hour after the administration of the compound CRL 40 956 groups of 10 mice receive an intraperitoneal injection of 200 mg/kg of barbital.

At th© strong dose (512 mg/kg) the compound CRL 40 956 reduces the duration of barbituric sleep.

Action on behavioural despair18 Half-an-hour after having received th® compound CRL 40 956, groups of 10 mice (males, GD-j, Charles River) are plunged into a small enclosure filled with water to a height of 6 cm. The total duration of immobility between the 2nd and the 6th minute following the immersion is noted.

At the strong dose (512 mlg/kg), the compound CRL 40 956 reduces the duration of immobility of despair In conclusion, the CRL 40 956 proves to be a sedative.

IMMUNOLOGICAL STUDY Retarded hypersensitivity reaction to sheep red blood cells According to the method of Miller, Lagrange and Mackaness (Immunopotentiation with BCG II modulation of the response to sheep red blood calls. Journal of the National Cancer Institute 1973, 51.. . 1669 - 1076), The compound to be tested is administered by oral route to female mice of the OF-j strain, three days before immunisation.

The results lare expressed in percentage increase of the plantar pad, Compound CRL 41 334 Compound CRL 41 335 Ό x ba X increase in pad (ma) Doses of the product in mg/kg 0 0.1 1 10 100 m 8.9 11,5 8.4 8,9 6.9 1 'Τ’θ 15 S » ul 1! 4- 0.92 4- 1.75 + 0.63 + 2.41 j- 1.42 Ind. act- 30 ivity 1 OO 1 Q> tw X 0.94 1.00 0.77 je It thus appears that the compound CRL 41 334 is presented particularly as a product having on the one hand an anti-depressive action at the central nervous system end on the other hand an immuno-stimulating action which is all the more surprising since the compound CRL 41 335 has no such immuno-stiaulating action. ( The compound CRL 41 334 administered by oral route at the rate of 50 mg four times per day has given good results in man in the treatment of depression in persons suffering from bums.

The therapeutic compositions according to the invention can be 10 administered to man or to animals by oral and rectal route.

They can be in the form of solid or semi-solid preparations. For example, there can bs cited tablets, capsules, suppositories, as well as delayed forms.

Ia these compositions, ths active principle is generally mixed 15 with one or more of the usual pharmaceutically acceptable excipients well known to an, expert.

The quantity of active principle administered obviously depends on the patient under treatment, oa the administration route and on the severity of the illness.

Claims (3)

1., Therapeutic composition containing as active principle a derivative of beazhydrylthioaethan® with the formula CH-S-CH-,-Υ II in which 1 represents a group -CONH·?, -CO-NHOH or -CH=N-OH and P- ( and R 2 represent independently of each other a hydrogen or a fluorine atom.

2. Therapeutic composition according to Claim 1, containing as active principle 2-(4,4.’-difluorobenzhydrylthio) acetamide.

3. Therapeutic composition according to Claim 1, containing as active principle 2-(benzhydrylthio)acetamide4- Therapeutic composition according to Claim 1, containing as active principle 2-(benzhydrylthio)acetaldoxime5. A therapeutic composition according to Claim 1, 15 substantially as hereinbefore described with particular reference to the accompanying Examples.