DK174358B1 - THERAPEUTIC AGENTS CONTAINING BENZHYDRYLTHIOMETHER DERIVATIVES - Google Patents

Description

in DK 174358 B1

The present invention relates to therapeutic agents containing as an active substance a benzhydrylthiomethane derivative which can be used in therapeutics because of its effect on the central nervous system and for certain purposes because of its immunostimulatory effect.

Some benzhydrylthiomethane derivatives used according to the invention have already been described.

Thus, FR-A-2385 discloses 2- (benzhydrythio) acetamide, EP-0 097,071 discloses 2- (4,4'-difluorobenzhydrylthio) acetamide, and EP-0 097,547 discloses 2- (4,4'-difluorobenzhydrylthio) droxamsyre. However, these compounds are only described as intermediates, and it was not clear that they would present interesting properties which enabled their use in the therapist in ka.

The present invention relates to agents containing as an active substance a benzhydrylthiomethane derivative of the formula

CH-S-CH2-Y II

Wherein Y is a group -CONH 2, -CO-NHOH or -CH = N-OH, and R 1 and R 2 are independently a hydrogen atom or a fluorine atom.

The compounds of formula II wherein Y is -COHH 2 can be prepared by reaction of ammonia to the chloride of the corresponding 2- (benzhydrylthio) acetic acid.

The compounds wherein Y is -CO-NHOH can be prepared by reaction of hydroxylamine hydrochloride with the methyl ester of the corresponding 2- (benzhydrylthio) acid.

The compounds wherein Y is -CH = N-OH can be prepared by reaction of hydroxylamine hydrochloride with the corresponding 2- (benzhydrylthio) acetaldehyde.

2 DK 174358 B1

The following examples illustrate the preparation of the compounds.

Example 1.

Preparation of 2- (4,41-difluorobenzhydrylthio) acetamide (code no. CRL 41334).

A solution of 26.46 g (0.08 mol) of 2- (4,4'-difluorobenzhydry-thi o) acetic acid is heated to reflux for 1 hour in 145 ml of benzene and 30 ml of thi onion 1 chl or id.

10

After evaporation under reduced pressure, the residue is taken up in 100 ml of ether and poured into a stirred mixture of 100 ml of 28% ammonia and 200 g of ice.

This is stirred for two hours, then the ether is poured off and the residue is washed with water, dried on Na 2 SO 4 and evaporated to dryness under reduced pressure. The residue is taken up in petroleum ether and separated.

After recrystallization in isopropyl ether, 28 g of amide is obtained (sue 1- 20 0 7 7 0 C, yield 58%).

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

25

Example 2.

Preparation of 2- (4,4'-difluorobenzyrohydriothio) acetohydroxamic acid (code no. CRL 42 335).

Dissolve 44.1 g (0.15 mol) of 2- (4,4'-difluorobenzhydry 11 h) acetic acid in 300 ml of methanol and add 1.5 ml of concentrated H 2 SO 4. After 4 hours at reflux, the methanol is evaporated, the residue is taken up in water and ether, washed with dilute bicarbonate, then with water, dried and evaporated under reduced pressure.

The oily residue is treated overnight at 20 ° C with a solution made up of 6.9 g (0.3 g atom) of sodium, 10.5 g 3 (0.15 mol), hydroxylamine hydrochloride and 480 ml of anhydrous methanol.

The oven is evaporated to dryness under reduced pressure, taken up in 500 5 mg of water, filtered on charcoal, acidified with 3N HCl, extracted with ether, washed with water, dried, evaporated and crystallized by petroleum ether.

28.8 g of 2- (4,4'-difluorobenzhydrylthio) acetohydroxamic acid (m.p. 76 ° C, yield 82%) are obtained.

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The compound is a pale pink powder soluble in ether, ethyl acetate, alcohols and insoluble in water.

Example 3

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Preparation of 2- (benzhydrylthio) acetamide (code no. CRL 41 05 5) 21 g (0.076 mol) of benzhydrylthioacetyl chloride in solution in 100 ml of methylene chloride are added dropwise with stirring to 40 ml of 20 ammonia and 40 ml of water. After stirring for 1 hour, the organic phase is decanted, washed with water and dried on Na 2 SO 4. The solvent is evaporated under reduced pressure, the residue is crystallized by isopropyl ether and recrystallized from ethyl acetate. The compound is obtained in a yield of 45%.

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It is a white powder which is soluble in alcohols, acetone, ethyl acetate and insoluble in water and isopropyl ether.

It melts at 107-108 ° C.

Example 4.

Preparation of 2- (benzhydrylthi) acetaleoxime (code no. CRL-40 956).

a) Preparation of 2- (benzhydrylthio) acetaldehyde.

20 g (0.1 mole) of diphenylmethanethiol are added to a solution of 3 g (0.13 g of atom) of sodium in 250 ml of ethanol and then added dropwise at reflux temperature over 1 hour 35 g (0.125 mole) bromoaceta 1dehyddi ethy1aceta 1. After 2 hours of refluxing, the alcohol is evaporated under reduced pressure, the residue is taken up in ether, washed three times with water, dried and filtered on charcoal. Evaporation of the ether under reduced pressure gives diethyl aceta 1 one of 2- (benzhydry 11 h o) aceta 1dehyde quantitatively.

The latter is hydrolyzed by heating for 2 hours on a water bath with 250 ml of 10% H2 SO4, extracted with ether, washed with water, dried, evaporated, crystallized by petroleum ether and recrystallized from isopropyl ether. 2- (Benzhydrylthio) acetaldehyde (mp 57-580 ° C) is obtained in 56% yield.

b) Preparation of 2- (benzhydrythi o) aceta 1doxim.

4.2 g (0.08 mole) of hydroxyl amine hydrochloride in solution in 15 ml of water is partially neutralized 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 are rapidly added at 20 ° C. This solution is stirred for 2 hours, the pH in one drops rapidly from 10 to 5 20 and the oxide precipitates. Add 20 ml of water, which is left in contact for 1 night, and then after separation, the residue is washed with water, dried and crystallized by 80% methanol. The compound is obtained in a total yield of 44%.

It is a white powder soluble in ether, ethyl acetate, methyl and ethyl alcohol.

It is insoluble in water and petroleum ether.

The equimolar mixture of "vision" and "ant in" forms has a melting point of 86-87 ° C.

The results of the pharmacological studies showing the properties of the compounds are set out below.

Neuropharmacological Study 35 1) Study of CRL 41 334

The compound CRL 41 334 in suspension in a solution of gum arabic was administered by intraperitoneal route in * 5 volumes of 20 ml / kg to mice (male NMRI.CER January) and in a volume of 5 ml. / kg for rats (male, CDj: SPRAGUE DAWLEY, Charles River).

5 I - Pre-toxicity (3 mice per dose).

128 mg / kg: unstable gait, exophthalmia, sedation, then arousal 2 hours after injection.

256 mg / kg: unstable gait, exophthalmia, convulsions (3/3) after 15 10 minutes, sedation followed by arousal 2 hours after injection. No mortality.

512 mg / kg: unstable gait, exophthalmia, cramps (3/3) for the first 10 minutes. Mortality (3/3) (1/3 in 15 minutes and 2/3 j 5 in 24 h for more).

II - Overall behavior and reactions.

Groups of 3 animals were observed before and then 15 minutes, 30 minutes, 1 hour, 2 hours, 3 hours and 24 hours after administration of the compound CRL 41 334.

1 - Mouse.

2, 8 and 32 mg / kg no clear changes in behavior and reactions.

25 128 mg / kg sedation for 30 minutes accompanied by unstable gait, reduction of respiratory rate, reactions, fear reaction and muscle tone. Hypothermia (-4.1 ° at 30 minutes for 1 hour).

30 2 - Rat.

1, A and 16 mg / kg of behavior, reactions, variation of pupil parameters and rectal temperature comparable to those in the control group.

64 mg / kg sedation for 15 minutes accompanied by dyspnea, reduction in response to touch and muscle tone.

35

This sedation is followed by an increase in the fear response (30 minutes) of reaction to touch and muscle tone for 3 hours.

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Mydriasis very moderate and short-lived after 1 hour.

5 III - Investigation of stereotyped movements.

Groups of 6 rats receive an injection of the compound CRL 41 334 or of amphetamine and are immediately placed in boxes of small dimensions (20 x 10 x 10 cm), with their stereotype behavior rated at 0 to 3 until the effect ceases.

In the two strongest doses used (64 and 128 mg / kg), the compound CRL 41 334 causes distinct stereotypes, the intensity of which is comparable to that obtained with ^ 5 i and 2 mg amphetamine, respectively.

However, the kinetics of action are different. The stereotypes induced by 2 mg / kg amphetamine reach their maximum of 60 to 90 minutes and last for 270 minutes, while the stereotypes introduced with 128 mg / kg of the compound CRi 41 334 reach their maximum wall of 120 to 150 minutes and last more than 420 minutes.

IV - The interaction with apomorphine.

1 - Mouse.

2 5

Groups of 6 mice receive CRL 41,334 half an hour before subcutaneous injection of apomorphine at a dose of i or 16 mg / kg.

a) Apomorphine 1 mg / kg b) Apomorphine 16 mg / kg 30 In the strongest dose (128 mg / kg) the compound causes CRL 41 334 hypothermia and aggravates the hypothermia induced by apomorphine (1 mg / kg).

The creation behavior and stereotype in the vein are not changed.

2 - Rats.

The compound CRL 41 334 is administered to groups of 6 rats half an hour before subcutaneous injection of 0.5 mg / kg apomorphine.

7 In the two strongest doses used (16 and 64 mg / kg), the compound reinforces the CRL 41 334 stereotypes introduced with apomorph in n.

5 V - Interaction with amphetamine.

Amphetamine (2 mg / kg) is injected along the intraperitoneal route to groups of 6 rats half an hour after administration of the compound CRL 41 334.

10 At doses of 16 but especially at doses of 64 mg / kg, the compound reinforces the CRL 41 334 amphetamine stereotypes.

VI - Saro work with reserpine.

Four hours after intraperitoneal injection of 2.5 mg / kg reserpine 15, groups of 12 mice receive the compound CRL 41 334.

1 - Effect on temperature 2 - Effect on ptosis.

2q In the strongest dose used (128 mg / kg), the compound exacerbates moderate reserpine hypothermia CRL 41 334. Ptosis that is already maximal does not change.

It should be noted that after 24 hours the compound CRL 41 334 (8, 32 or 128 mg / kg) seems to counteract reserpine hypothermia and 25 ptosi s.

VII - The interaction with oxotremorine.

The compound CRL 41 334 is administered to groups of 6 mice 30 minutes before intraperitoneal injection of 0.5 mg / kg oxotremorine.

1 - Effect on temperature.

In the strongest dose (128 mg / kg) the compound causes CRL 41 334 hypothermia and aggravates hypothermia introduced by oxotremor in n. 1 - Effect on shaking.

8 DK 174358 B1

Shaking caused by oxotremorin is not altered by compound CRL 41 334.

3 - Effect on peripheral shock symptoms.

5 At a dose of 128 mg / kg, the compound reduces CRL 41 334 defecation without altering salivation and tear formation.

VIII - Effect on the square test, distraction and electric shock.

1 o

The test is performed on groups of 10 mice half an hour after administration of the compound CRL 41 334,

The connection CRL 41 335 does not clearly change the number

Strsffsdv p3. SS30vP Qgn t) O V i Γ k * Γ Ί J (g Π0Ο £? Π t 0 P Γί1 2 3 bPV.'PGP 1 c λ r - 15 inefficiency.

In the strongest dose (128 mg / kg) used, the compound CRL 41 334 weakly counteracts the spasm, without altering the deadly effects of electric shock.

20 IX - Effect on spontaneous free movement 1 see.

1) Connection administered 10 minutes before entry into activity meter.

25 at doses of 32 and 64 mg / kg, compound CRL 41 334 causes an increase in the spontaneous free movement of mice. At a dose of 128 mg / kg, this effect is no longer apparent.

2) Connection administered 2 hours before entry to activity meter.

At a dose of 8 but in particular 32 and 128 mg / kg, the compound CRL 41 334 causes an increase in the spontaneous free movement of mice.

X - Effect on acresivity between groups, 2

After staying 3 weeks in each half of a cage separated by opaque partition, groups of 3 mice receive the compound CRL 41 334. Half an hour later, the two groups of mice in the same cage are brought together by removal of the partition. , and the number of matches occurring in 10 minutes is recorded. Half of the sample is performed with the usual mice (NMRI, C.E.R. January) and half on NMRI (Iffa Credo) mice.

5 In the two doses used (32 and 128 mg / kg), the compound CRL 41 334 reduces the number of matches.

* XI - Impact vis-a-vis behavior disturbed by various means.

10 1 - Movement reduced by accustomed to enclosure.

After 18 hours in the activity monitor, the mice (6 per dose, 12 controls) receive the compound CRL 41 334. The 15 are immediately reinserted in their respective enclosures and half an hour later their movement is recorded for 30 minutes.

Starting at a dose of 2 mg / kg, compound CRL 41 334 causes a distinct renewal of the movement activity of mice accustomed to its enclosure.

20 2 - Movement reduced by hypoxic aggression.

Half an hour after receiving the compound CRL 41 334, the mice (10 per dose, 20 controls) undergo an acute hypoxic oxygen deficiency (suppression of 60 mm Hg for 90 seconds held for 45 L t> seconds) and then placed in the activity meter, where their movement is recorded for 10 minutes.

Starting at a dose of 2 mg / kg, the compound causes CRL

41,334 an improvement in the movement recovery of mice whose motility had been suppressed after a short period in a reduced pressure enclosure. This improvement is significant for the two strongest of the doses used (23 and 128 mg / kg).

3 - Choking oxygen deficiency.

Groups of 20 mice receive the compound CRL 41,334 half an hour before intraperitoneal 1 administration of 32 mg / kg gallamine triiodide ethylate.

35 10 DK 174358 B1 In the strongest dose used (128 rag / kg), compound CRL 41 334 increases the period of death after an oxygen deficiency caused by a curative agent. The period of occurrence of seizures does not change.

5 XII - The interaction with barbital.

Half an hour after administration of the compound CRL 41 334, groups of 20 mice receive an intraperitoneal tonea 1 injection of barbital (220 mg / kg).

In doses of 8, 32 and 128 mg / kg, the compound CRL 41 334 causes a significant decrease in the duration of the barbiturate.

XIII - Impact on despair behavior.

1 5

Half an hour after receiving the compound CRL 41 334, groups of 6 mice (male, CD ^ Charles River) were placed in beaker filled with water to a height of 6 cm. The total duration of immobility between the second and sixth minutes after the immersion was recorded.

20 1 doses of 8, 32 and 128 mg / kg compound CRL 41 334 reduced the duration of immobility of mice placed in forced immersion.

XIV - Conclusion.

2 5 --------

The neuropsychopharmacological study of the compound CRL 41 334 shows-.

a stimulating effect on mice: hypermotility, distinct renewal of locomotor activity in mice accustomed to their enclosure, enhancement of locomotor recovery following hypoxia, antagonism to barbituric acid sleep, and reduction of despair hypomotility.

In a strong dose, a brief sedative effect is noted: sedation, hyporeactivity, hypoaggressiveness, hypothermia, exacerbation of hypothermia induced by reserpine and oxotremorine, moderate reduction of the anticonvulsant effect of an electric shock, and delay in the occurrence of death after oxygen. caused by curating agent. 2 hours after product administration, this sedative effect is replaced by a stimulating effect.

5

Furthermore, a strong dose is noted in the rat: stereotypes and a reinforcement of the stereotypes induced by apomorphine and amphetamine.

This stimulating effect seems to develop only after a clear latency of the order of at least 2 hours.

2) Investigation of CRL 41 055

The compound CRL 41 055 in suspension in a solution of gum arabic was administered by intraperitoneal route at a volume of 20 ml / kg in mice (male, NMRI, Evic Ceba).

Pre-toxicity (3 mice per dose).

64 mg / kg: arousal, stereotypes (snuff, erection) appear after 45 minutes and last more than 2 hours.

128 and 256 mg / kg: sedation for 45 minutes, decreased breathing rate, abdominal contractions followed by arousal, stereotypes (sniffing, erection) for more than 2 hours.

2 5

No mortality.

512 mg / kg: sedation for 45 minutes, diminished breathlessness, abdominal contractions, subconvulsive wakefulness followed by arousal, stereotypes (less evident than with the 30 weaker doses).

18 hours after injection, intermittent seizures in all mice interrupted (2/3) by stereotypes (toilette, jaw movements).

Mortality (1/3) after 48 hours.

35 1024 mg / kg: same symptoms.

Overall behavior and reactions.

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Groups of 3 animals were observed before and then 15 minutes, 30 minutes, 1 hour, 2 hours, 3 hours and 24 hours after administration of compound CRL 41055.

^ 2 mg / kg: no obvious changes in behavior and reactions.

8 and 32 mg / kg-. agitation for 2 hours.

128 mg / kg: sedation for 30 minutes followed by a late arousal (2 hours), lasting one hour with increase in response to 10 touch and fear reaction.

Effect on spontaneous free movement

Half an hour after receiving CRL 41 055, the mice (6

Dr. dose, 12 controllers) placed in an activity meter where their motility was recorded for 30 minutes.

At doses of 32 and 128 mg / kg, CRL 41 055 causes an increase in spontaneous mot in 1 in the mouse.

2o Movement reduced by accustomed to enclosure.

After staying for 18 hours in the activity meter, the mice (8 per dose, 12 controls) receive CRL 41 055. Immediately afterwards, they are placed back in their respective enclosures and half an hour later their movement is recorded for 30 minutes.

25 At doses of 39 and 128 mg / kg, CRL 41 055 causes a distinct renewal of the movement activity of mice accustomed to their enclosure.

3 q Conclusion.

CRL 41 055 shows a stimulating effect in mice: arousal, movements that appear to be stereotypes, hypermotion and distinct renewal of movement activity in mice that have become accustomed to their enclosures. Prior to this stimulation, a sedate often goes into the on-stage.

CRL 41 055 has been found to be useful in human therapeutics as an antidepressant at three doses of 50 mg a day.

13 DK 174358 B1 3. Investigation of CRL 40 956.

The compound CRL 40 956 in suspension in a solution of gum arabic was administered by intraperitoneal intraperine in a 5 volume of 20 ml / kg for mice (male, NMRI. Evic Ceba) and 5 ml / kg for rats (male, CD ^: SPRAGUE DAWLEY, Charles River).

Pre-toxicity (3 mice per dose).

1024 and 512 mg / kg-. abdominal cramps, dyspnoea, no mortality, 256 mg / kg: same symptoms.

Overall behavior and reactions.

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

1 - Mouse.

20 512 mg / kg: sedation for 2 to 3 hours with reduction of the response to touch and muscle tone for 1 to 3 hours.

Hypothermia (-2.9 °) for 3 hours.

25 128, 32 and β mg / kg: no obvious symptoms.

2 - Rats.

256 and 64 mg / kg: sedation for 3 hours with reduction of the response to touch and muscle tone.

30 16 and 4 mg / kg: behavior and reactions comparable to those in the control group.

The interaction with apomorphine.

The compound CRL 40 956 is administered to groups of 6 rats half an hour before subcutaneous injection of 0.5 mg / kg apomorphine.

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

14 DK 174358 B1

Effect ρά spontaneous free movement.

Half an hour after receiving CRL 40 956, the mice (6 per dose, 12 controls) are placed in an activity monitor where their motility is recorded for 30 minutes.

5 In the high dose (512 mg / kg), CRL 40 956 greatly reduces the spontaneous movement activity of mice.

Impact on aggression between groups.

10

After staying for 3 weeks in each half of a cage separated by an opaque partition, groups of 3 mice receive the compound CRL 40 956. Half an hour later, the two groups in the same cage are brought together by removal of the partition, easy matches that take place at 10 n> in utte r are not eres.

Ϊ a dose of 128 mg / kg compound CRL 40 956 reduces the number of matches.

Effect vis-à-vis suffocating oxygen deficiency.

20

Groups of 10 mice receive the compound CRL 40 956 half an hour before ntraper in tonea 1 administration of 32 mg / kg gallamine triiodethylate.

In a strong dose (512 mg / kg), the compound CRL 40 956 2 5 moderates the incidence of seizures and death after suffocating oxygen deficiency caused by a curative agent.

The interaction with barbital.

30 Half an hour after administration of the compound CRL 40 956, groups of 10 mice receive an intraperitoneal 1 injection of 200 mg / kg barbi.

In the strong dose (512 mg / kg), the compound CRL 40 956 reduces the duration of barbecue sleep.

35

Impact on despair behavior ·

Half an hour after receiving the compound CRL 40 956 groups of 10 mice (he, CDj, Charles River) are immersed in a small enclosure filled with water to a height of 6 cm. The total duration of immobility between the 2nd and the 6th minute after the immersion is recorded.

5 In the strong dose (512 mg / kg), the compound CRL 40 956 reduces the duration of despair immobility.

In conclusion, CRL 40 956 turns out to be a sedative.

Immunological examination.

10

Delayed hypersensitivity reaction ρά red & red blood cells

Following the method of Mi lier, Lagrange and Mackaness ("Immunopo-tentiation with BCG II modulation of the response to sheep red blood cells", Journal of the National Cancer Institute 1973, 15 51, 5, 1669 - 1676).

The compound to be tested is administered by the oral route to female mice of the strain OFi 3 days prior to immunization.

2Q The results are expressed as a percentage increase of the cushion.

25 30 35

Ex. % increase ___ Doses of product in mg / cow_ 5 in steps 0 0.1 1 10 100 ____ cushion (mm) _______ 16 DK 174358 B1 CRL 41 334 1 m 9.2 10.6 10.2 12.0 14.7 ie.sm ± 1.06 ± 1.42 ± 1.22 ± 2.16 ± 2.93

Ind. act i - 10 .....

__ v 1 ς 6 11 —______ 1 i 15 i ...... 1.10 _____ 1.29] 1.55 ™ 8.4 11.0 10.5 11.2 13.1 2 ie.s.m. ± 1.35 ± 1.77 ± 1.75 ± 1.67 ± 1.92 ind. act i - 1 5 L -] vj_te_t .............. J .................__ L_ 1_l.il! .. 1, 25 1 1.34 1.56 | CRL 41 335

Ex. % increase Doses of product in mq / kq 20. "1" in step 0.1 0.1 100 cushion (mm) __ "8.9 11.5 8.4 8.9 6.9 1 ie.sm ± 0.92 ± 1.75 ± 0.63 ± 2 , 41 ± 1.42 25 shares - __white ____ 1.29 ______ 0.94 1.00 0.77

Thus, it appears that the compound CRL 41 334 is in particular a product which, on the one hand, has an ant in depress iv effect on the central nervous system and on the other, an immune stimulating effect, which is so much more surprising than the compound. CRL 41 335 has no such immune effect 1 end effect.

The compound CRL 41 334 administered by the oral route in an amount of 50 mg four times a day has produced good results in humans in the treatment of depression in people suffering from burns.

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The therapeutic agents of the invention can be administered to humans or animals by the oral and rectal route.

They can be in the form of solid or semi-solid preparations. For example, mentioned are delayed release tablets, capsules, suppositories and forms 5.

In these agents, the active principle is generally mixed with one or more of the usual pharmaceutically acceptable excipients.

10

The amount of active principle administered, of course, depends on the patient being treated, the route of administration and the severity of the disease.

15 20 25 30 35

Claims (4)

1. Therapeutic agent containing as active substance a derivative of benzhydrythiomethane of formula II □ rK3 \ CH-S-CH2-V II in Krry where Y is a group -CONH 2, -CO-NHOH or -CH -OH, and R 1 and R 2 are independently a hydrogen or fluorine atom. 15 2. Therapeutic agent according to claim 1, characterized in that it contains as an active substance 2 ~ (4, 4 '- d in f'1 of benzhydrylthiojacetamide. The therapeutic agent of claim 1 wherein the active substance contains 2- (benzhydrylthio) - 20 a c e t a m i d. Therapeutic agent according to claim 1, characterized in that the soot active substance contains 2 - (benzhydry 11 h in o) -acetaldoxi m.