FI71309B - REFRIGERATION FOR THERAPEUTIC USE OF THERAPEUTIC ACID 2, -DICHLORO-4'-PHENYL-OCH-4'-PHENYLTHIO-ALPHA- (N-IMIDAZOLYLMETHYL) - Google Patents

Description

"H rÄ, ADVERTISING 9-1700 B 11 UTLÄGGNINGSSKRIFT 7l309 (45> Patent IJ 12 1220 (51) Kv.lk.Vlnt.CI.- e 07 D 233/60 FINLAND - FINLAND (21) P # tenttlhakemus - P« entansökning 79 1 5 ^ + 3 (22) Application date - Ansöknlngsdag 1 ^ g ζ yg (H) (23) Starting date - Glltlghetsdag 15.05 79 (41) Has become public - Blivit offentlig ig ^ -j yg

National Board of Patents and Registration 144) Date of display and public publication—

Patent and registration authorities '' Ansökan utlagd och utl.skriften published 09.09.86 (86) Kv. application -! nt. ansökan (32) (33) (31) Claim claimed — Begärd priority 18.05.78 1 or ia-1taiien (IT) 235 ^ 6 A / 78 (71) Recordati SA, Chemical and Pharmaceutical Company, C.so S. Gottardo 5 ^, Chiasso, Switzerland-Switzerland (CH) (72) Dante Nardi, Milan, Elena Massarani, Milan, Alberto Tajana, Milan, Mario Veronese, Milan, I or ia-l or ien (IT) (7 * 0 Oy Kolster Ab (5 ^) Process for the preparation of therapeutically useful 2,4-dichloro-4'-phenyl- and β-phenylthio-OMN-imidazolylmethyl i) dibenzyl ethers - For the preparation of therapeutically useful compounds 2 , 4-dichloro-41-phenyl-och-phenyl-06- (N-imidazolylmethyl) dibenzyl ether

The invention relates to a process for the preparation of therapeutically useful 2,4-dichloro-4'-phenyl and -4'-phenylthio-N- (N-imidazolylmethyl) dibenzyl ethers of the formula I

C1-6-CH-O-CH2-R) -R

Cl CH0 "-N 1

2 V-N

wherein R is a phenyl or phenylthio group, and for the preparation of their pharmaceutically acceptable acid addition salts.

2 71309

Preferred pharmaceutically acceptable acid addition salts of Compounds I are those formed from both inorganic and organic acids such as hydrochloric, nitric, sulfuric, phosphoric, methanesulfonic, succinic, maleic, fumaric, citric and tartaric acids. These salts can be prepared by conventional methods, for example, by adding an equimolar amount of the desired acid to the base and then crystallizing the salt thus obtained from a suitable solvent.

The compounds I and their salts are of interest for their activity against fungi, yeasts and gram-positive aerobic and anaerobic bacteria. The effect is associated with low toxicity. The compounds of the invention have been compared to two well known fungicidal and bactericidal products, clotrimazole, i.e. ΖΓ (imidazol-1-yl) -2- (o-chlorophenyl) -2,2-diphenylmethane7, and miconazole.

Quite interesting was the comparison between the activity and toxicity of the compounds I and Miconazole, i.e. 2,2 ', 4,4'-tetrachloro-O1- (N-imidazolylmethyl) dibenzyl ether. In miconazole, both benzyl groups are substituted at the 2- and 4-positions by chlorine atoms, while in compounds I, the benzyl group which is unsubstituted is substituted at the 4-position by a phenyl or phenylthio group. This difference results in a significant reduction in toxicity, which is 3-4 times lower for these new products than miconazole, while the fungicidal and bactericidal effect is approximately the same. The compounds of the invention may be suitably used in human therapy for the topical treatment of skin diseases such as trichophytosis and candidiasis, as well as for the treatment of fungal infections caused by Staphylococci and streptococci. The compounds of the invention may be mixed with pharmaceutically acceptable diluents or carriers to form pharmaceutical compositions which may be in any suitable form, for example as powders, ointments, creams, suspensions and dispersions.

The results obtained in the biological experiments are shown in Tables I, II and III.

Table I shows the acute toxicity values (LD 2) of Compounds I and each reference substance. Toxicity at 71309 na mg / kg has been evaluated orally in mice by conventional methods.

Table II shows the minimum inhibitory concentrations (MICs) for Compounds I and each reference substance. MIC, minimum concentration, refers to the ability to inhibit the growth of multiple fungi and yeasts. MIC values have been determined according to the standard double broth dilution series method.

The MIC values of the test products and of each reference substance, which have been targeted to a number of gram-positive bacteria and determined according to the standard two-fold broth dilution series method, are shown in Table III.

The experimental conditions were as follows:

For fungi Medium: Sabouraud liquid, pH 5.7 (5 ml / tube)

Inoculum: A 10-day agar culture was washed with physiological saline containing 10% Tween 80, then filtered through a pellet and resuspended in physiological saline until the solution on a Coleman Jr II spectrophotometer at 650 nm gave 50% light transmittance (T) (0 , 1 ml spore suspension / ml). After filtration, a physiological solution of Aspergillus niger was prepared at a dilution of 1/10. 0.1 ml of this dilution formed an inoculum of 5 ml.

Temperature and incubation time: 25 ° C, 7 days.

For yeasts Medium: Sabouraud liquid, pH 5.7 (5 ml / tube)

Inoculum: Yeasts were grown in Saboraud fluid for 24 hours (Cryptococcus neoformans for 2 days). The cells were harvested by centrifugation at 6500 rpm and resuspended in physiological saline to give a suspension which gives a 50% light transmission (T) at 650 nm on a Coleman Jr II spectrophotometer.

0.1 ml of this suspension formed an inoculum of 5 ml.

Temperature and incubation time: 37 ° C, 48 hours.

For Gram-positive bacteria Medium: soy trypsin broth, pH 7.3 (5 ml / tube).

Inoculum: The day before the test, the microorganisms to be tested were inoculated into their respective media. After 18 days of incubation at 37 ° C, 0.1 ml of a 1: 100 dilution of a suspension of each strain in the broth was inoculated into 5 ml of medium containing 71309 test products in a dilution series of 0.009 to 160 mcg / ml.

Temperature and incubation time: 37 ° C, 18 hours.

Table I

Acute oral toxicity mg / kg (LD 50) in mice.

Formula I Formula I Miconazole Clotrimazole

I: R = C6H5 I: R-CgHgS

2400 3000 870 880

Table II

Fungicidal activity (MIC) in mcg / ml

Disease-causing Formula I Formula I Miconazole Clotrimating factor R = C6H5 R = CgH5S sol C.Albicans R 5 40 5 10 C.Albicans Griinenthal 5 40 5 10 C.Albicans 1040 10 80 20 40 C.Albicans 1041 20 80 20 40 C.Neoformans 0.312 0.156 0.078 0.625 T.Mentagroph.2538 0.156 0.156 0.078 0.078 T.Mentagroph.10148 0.625 0.625 0.625 1.25 T.Mentagroph.5865 1.25 1.25 0.31 0.62 T.Verrucosum 10 1 .25 2.5 1.25 T.Rubrum 2121 0.625 0.625 0.312 1.25 M.Canis 28 20 2.5 5 2.5 A.Niger 20 10 40 10 P.Chrysogenum 20 5 20 40 E.Floccosum 10 5 5 5 5 71 309

Table III

Bactericidal activity (MIC) in mcg / ml

Disease-inducing Formula I Formula I Miconazole Clotrimatrol

factor R = CgH ^ R = CgH ^ S

S.aureus SG 511 0.039 0.039 0.312 0.312 S.aureus 10 B 0.039 0.018 0.312 1.25

Str.hemolyt 8 21 0.156 0.156 1.25 2.5 B.subtilis 0.156 0.078 0.625 2.5

Cl.novyi <1.25 5.0 <1.25> 160

Str. Hemolyt 203 0.312 0.078 0.625 5

In the process, 1- (2 ', 4'-dichlorophenyl) -2- (N-imidazolyl) ethanol is condensed with halobenzyl of formula V

R —CH2X V

wherein R is as defined above and X is a chlorine or bromine atom, in a solvent, and that the resulting free base is, if desired, converted into a pharmaceutically acceptable acid addition salt.

As the solvent, an aromatic hydrocarbon, dimethylformamide, tetrahydrofuran, hexamethylphosphoramide and / or mixtures thereof can be used, but it has been found that condensation is more complete if dimethyl sulfoxide is used. This results in a higher yield and a cleaner product. In this case, it is not necessary to purify the base obtained from the condensation on a chromatography column, whereas this purification is necessary when using dimethylformamide or hexamethylphosphoramide as solvents. Simple filtration of the alkaline solution through a silica gel column 6 71309 is in fact able to retain small amounts of impurities. For compound I, where R is a phenyl group, even filtration is not necessary and crystallization of the nitrate results in a product which is sufficiently pure for pharmaceutical purposes. With the above-mentioned approbic solvents, an alkali metal hydride or amide capable of forming a salt with an hydroxy group of an ethanol derivative is usually used.

Alternatively, the condensation solvent may be an aliphatic alcohol having 3 to 6 carbon atoms, such as t-butanol, in which case the alkali metal hydride or amide should be replaced by an alkali metal alcoholate, for example potassium t-butylate.

Another very useful means is to add small amounts of potassium iodide as a catalyst before adding the halobenzyl derivative.

1- (2 ', 4'-dichlorophenyl) -2- (N-imidazolyl) ethanol can be prepared from 1-chloroacetyl-2,4-dichlorobenzene (Beilstein-Handbuch der Org. Chem. IV ° Edition, Volume 7, p. 28) by sensitizing the keto group using sodium borohydride and condensing with imidia-Sol. The reduction and condensation can be performed in either order. The combination can be performed in methanol and the condensation can be performed in dimethylformamide and methanol in the presence of sodium. Halo-benzyl compounds of general formula V are known. The preparation of 4-chloromethyl-bidenyl is described in Chem. Ber.66B, 1471, 1933 and the preparation of 1-bromomethyl-4-phenylthiobenzene is described in U.S. Patent No. 3,242,193.

The following Examples 1-4 illustrate the invention, while Examples A and B illustrate the preparation of the starting materials used in Examples 1-4.

Example A

1- (2 ', 4'-Dichlorophenyl) -2-chloroethanol 49.5 g of sodium borohydride were added slowly and in small portions to a suspension of 233 g of 1- (1'-hydroxy-2'-chloroethyl) -2 , 4-dichlorobenzene in 1 L of methanol with stirring at room temperature. The solution thus obtained was stirred at room temperature for a further two hours and then poured into 1 l of 5N hydrochloric acid, which was cooled on ice. After extraction with ethyl acetate or chloroform, the extract was washed with water, 1N sodium hydroxide, then again with water until neutral and finally with saturated sodium chloride solution. The extract was dried, the solvent was evaporated to give 220 g of an oil, the oil solidified on standing and the solid melted at 48-51 ° C.

Analysis for CoH_Cl_.0 C% H% Cl%

Calculated 42.61 3.13 47.17 Found 42.75 3.19 47.43

Example B

1- (2f, 4'-dichlorophenyl) -2- (N-imidazolyl) ethanol 30 g of sodium were added to a solution of 88.5 g of imidazole in 600 ml of methanol; the solvent was then evaporated. The residue was dissolved in 300 ml of dimethylformamide and heated to 115-120 ° C. To the solution thus obtained was added dropwise, with stirring, a solution of 225 g of 1- (21,4'-dichlorophenyl) -2-chloroethanol in 400 ml of dimethylformamide. The mixture was heated to 115-120 ° C and maintained at this temperature for 20 minutes, and after cooling to 40 ° C, 2500 ml of ice water was added with vigorous stirring. The product precipitated during stirring for about 2 hours, the supernatant was decanted off, another 2500 ml of water were added and after standing everything was filtered. The precipitate thus obtained was dried and crystallized from toluene. 170 g of the desired product were obtained, melting at 134-135 ° C.

Analysis for C ^ H ^ Cl ^ C ^ C% H% N% Cl *

Calculated 51.38 3.92 10.89 27.58 Found 51.62 3.80 10.73 27.76

Example 1 2,4-Dichloro-4'-phenylthio-O '- (N-imidazolylmethyl) dibenzyl ether (I: R = CcH_S)

Method I

A solution of 2.57 g of 1- (2 ', 41-dichlorophenyl) -2- (N-imidazolyl) ethanol prepared according to Example B in 10 ml of hexamethylphosphoramide was added dropwise to the suspension at 25 ° C. containing 0.52 g of sodium hydride (50% in oil) in 5 ml of hexamethylphosphoramide. When hydrogen evolution ceased, salt formation was completed by heating at 50 ° C for 1 hour. After cooling to 25 ° C, 2.58 g of 1-chloromethyl-4-phenylthiobenzene was added. The temperature was raised to 50 ° C 71309 and maintained at this temperature for 12 hours. After completion of the reaction, the mixture was poured into 200 ml of water, the product was extracted with diethyl ether, the solvent was evaporated and the residue was purified twice on a silica gel column using ethyl acetate as eluent, and the various fractions were tested by thin layer chromatography. The solvent was evaporated from the middle fraction to give 2.4 g of the desired base as a yellowish oil which gave a single spot on thin layer chromatography.

Analysis for C 24 H 22 N 2 O 2 OS C% HS NS CIS SS Calculated 63.30 4.44 6.13 15.57 7.04 Found 63.86 4.24 6.41 15.29 6.97.

Method II

0.66 g of sodium hydride (50% in oil) was added at 20-30 ° C under nitrogen to 3.86 g of 1- (21,4'-dichlorophenyl) -2- (N-imidiazolyl) ethanol In 15 ml of dimethyl sulfoxide (dried over calcium hydride).

The mixture was heated with stirring at 50-60 ° C until gas evolution ceased. After cooling to 20-25 ° C, 0.5 g of potassium iodide was added and then a solution of 3.51 g of 1-chloromethyl-4-phenylthiobenzene in 4 ml of dimethyl sulfoxide was added slowly dropwise. The mixture was stirred at 20-25 ° C until the addition of 1-chloromethyl-4-phenylthiobenzene was complete. The mixture was then poured into 150 ml of water and extracted with diethyl ether. To the ether solution was added, after drying over anhydrous sodium sulfate, an excess of 4N nitric acid solution in diethyl ether, the desired product precipitated as a nitrate, an oil which solidified on standing. After standing for 20 hours, the ethereal solution was decanted off and the residue was crystallized from ethanol. The incompletely pure nitrate thus obtained was dissolved in water and excess sodium carbonate was added to liberate the base, which was then extracted with ethyl acetate. The base obtained by filtration was purified on a silica gel column using ethyl acetate as eluent. The combined fractions containing the desired product were evaporated to dryness. The residue was dissolved in diethyl ether, reconverted to nitrate and crystallized from ethanol.

71309

Yield: 3.1 g of a white crystalline powder, melting at 134 ° C. Analysis for C24H2 () N2C120S .HNO3

CS HS N% CIS SS

Calculated 55.61 4.08 8.11 13.68 6.18 Found 55.32 4.08 8.16 13.56 6.32.

Example 2 2,4-Dichloro-4'-phenyl-N- (N-imidazolylmethyl) -dibenzyl ether (I: R = CgH4)

Method I

A mixture of 2.02 g of potassium t-butylate in 30 ml of t-butanol was prepared at 20-25 ° C under a nitrogen atmosphere. 3.86 g of 1- (2 ', 4'-dichlorophenyl) -2- (N-imidazolyl) ethanol prepared according to Example B were added. The solution was heated to reflux for 1 hour and then cooled to 20-25 ° C. 3.03 g of 4-chloromethyl-biphenyl was added and the solution was heated again under reflux for 5 hours. After cooling to 20-25 ° C, the solution was poured into water and the base was extracted with ethyl acetate. The extract was washed with diethyl ether and the solvent was evaporated. The residue was dissolved in diethyl ether (80 ml) and allowed to stand overnight. The insolubles were filtered off and the filtrate was treated with nitric acid dissolved in diethyl ether. An oil was obtained which solidified on standing. The residue comprising 2,4-dichloro-4'-phenyl-N- (N-imidazolylmethyl) dibenzyl ether nitrate was crystallized from ethanol or ethyl acetate. The product (4.3 g) was obtained, which was shown to be pure by thin layer chromatography and melted at 140-141 ° C.

Analysis for C24H20N2O2®.HNO2 C% H% N% CL%

Calculated 59.25 4.35 8.64 14.57 Found 59.17 4.14 8.61 14.46.

Method I (a)

To a solution of 0.37 g of sodium metal in 20 ml of n-propanol was added 4.1 g of 1- (21,4'-dichlorophenyl) -2- (N-imidazolyl) ethanol and the mixture was heated to reflux for two hours. hours with stirring. After cooling to room temperature, 0.5 g of potassium iodide and 3.25 g of 4-chloromethyldiphenyl were added with stirring, and the mixture was heated again under reflux. At the end of the reaction, the mixture was filtered off and washed with ethanol. The filtrate was evaporated to dryness and the residue was dissolved in diethyl ether. The insoluble residue was filtered off, and the clear ether solution thus obtained was treated with a mixture of nitric acid and diethyl ether. The 2,4-dichloro-4'-phenyl-N- (N-imidazolylmethyl) dibenzyl ether nitrate thus formed was crystallized from ethanol. A product (2.1 g) having the same properties as the product prepared according to the above procedure and having a single spot in thin-layer chromatography was obtained.

Method II

0.66 g of sodium hydride was added under a nitrogen atmosphere and at 20-25 ° C to a solution of 3.86 g of 1- (21,41-dichlorophenyl) -2- (N-imidazolyl) ethanol in 15 ml of dimethyl sulfoxide (dried over calcium hydride). The mixture was heated at 50-60 ° C until gas evolution ceased. The mixture was then cooled to 20-25 ° C, 0.5 g of potassium iodide was added and then a solution of 3.03 g of 4-chloromethylbiphenyl in 7 ml of dimethyl sulfoxide (dried over calcium hydride) was added dropwise. The mixture was stirred for about 20 hours at 20-25 ° C and then poured into water. The product was extracted with ethyl acetate and then treated as described in Method I to give 4.6 g.

Example 3 Salts of 2,4-dichloro-4'-phenylthio-dL- (N-imidazolylmethyl) -dibenzyl ether were prepared by reacting the free base, dissolved in ethanol, with an alcoholic solution of the desired acid and then crystallizing the salt thus obtained from a suitable solvent. . The free base was prepared according to Example 1, Method I, or liberated from its nitrate prepared according to Method II, treated with sodium carbonate, extracted with diethyl ether and evaporated.

Table IV shows the crystallization solvents, elemental analyzes, and melting points for some salts.

Table V lists the same information for some of the salts of 2,4-dichloro-4'-phenyl-N- (N-imidazolylmethyl) dibenzyl ether obtained using the methods described above.

n 71309 "Γ] σί in 'f' 'f m | r- σι οι ^ σι

LO ID in M O MO

* -P tH

o r- co o r-t v ro σ> m en o * - tH cv CV -H cv

CV tH tH tH M

^ O- O- 00 CO cv z cd Γ'- r- v —i m 't rj · tf · in <8 --- § 1-1¾¾ -f O cv σι co z co in co v r-i

V <t C · C

"£ 5 OI T-i rl CO ID

CJ V ID CV ^ CO

co oo v σι ’—i ___in_ut_in_in_in____ -h ϊ— * co cv σι in id id cv γ- 00 * ~ ~“ "id in —i O in

tH tH

CV tH ID O tH

H, ID H · CO CO 00

1-1 CJ I .H CV OI tH OJ

Q J CV f — I rH tH * H

-¾ -ö4 l σι O CO D D

3 2 | id σι O v o d 3 m TT m V n g ti ------- ^ Vs- O co σι O σι 2 co oj co m ή T rr t C ^ t— * m m co σι

-oi iO CO V co O

^ co co v σι cv lo lo_uo_in_in _ ro co m co in co tH in oj sf • tH «H rH tH <(¾ o I III i en o t-ι o- co tn co co r ^ in oj j rJ \ fH · —1 * “lw

•B

ε

: (0 I

PC ιό

-p · Η α · Η -Η -H -H

> 1 -P O rH Γ-l -H rH

<D o U 0 0 OO

d 2 oh cc SS

• H · Η o <H (0 (0 <0 C

h to O .p -p -P -P

MC MW MW

Ή.

t3 I -H

· Η I -H -P

^ P -H O C -P -H

H 2 O 4J MH d) C -P

o rH -P rH dj C -P

5 Λί m e-h cc e σο o e to -P m o e

C d) (ts Dl o MH MH

T3 H -PC -P> —I (0>, to eeieo ___ ^ KSSC & to En_ I - ^^ - 12--71309 ro ro cΨ cn co ui - ld in co co t-η c \ jo 1—1 <-ivmm co 0 - ....

n co co c \ j v

Oi i — 1 rL iH · —I

^ CO 10 --C V Ή z oo cm σι m o

> i in m V V V

P

4J

Q) —-- Ό> i „a o O σ> co e'en co cr> i-i co v a i - ....

__ ^ _ CO -St V V

O σ> m .π ον co o oo σι OJ V Oi co co

• ~ Z LO Id CO LO

LO co "c'-l r · ^ U") X - <o m c- co C V co cj

r-1 rJ CO - CO LO

CJ. ....

co ro ro rj r-i O] r-H - * —- 1 Ή> - -

O

C 'Γ- CO V

M -V: O co -: co m 3 z I -

^ 1 ίο co m v V

S _I-- E- <3 νϊ o LO cc r- co

jjj Tl I CD CM V CO LO

^ ^ V V c ro --—.......—

^ co cc LO in LO

co co ro oj in

• p. O

O - “

U O} ΙΓ O J T} · CO

O \ T. ‘3 O 10

o] σ> · $ o- O oj co

0 «- * CC - rH O

c i I i I in

rH Oi CD Oi rH

• O- O- OJ CO

_- ___

•B

-μ 1 4-) in cg 1 5

• P · 5 π! Ή · γ4 · Η * H

* P * r4 i-4 r-4 r-4 rH

39 £ S S B B

ö s 5 S S 3 S

Cd Cd W Cd Cd

^ I

• H -H I

P -H -H C 4) rl

O -P 4-> ffl (0 -P

cg rH 4-) -P uj CO 4-> h ^ s cg 3 s 2

Q O CO CO rH O CO

3 <44 φ Q 144 H

Ui Ό H 1-4 4J rH -P

έ1 Ui S§ tn 3 in

Claims (1)

Process for the preparation of therapeutically useful 2,4-dichloro-4'-phenyl and -4'-phenylthio-α- (N-imidazolylmethyl) dibenzyl ethers of the formula I 71309 Claim ci ^ ch-o-ch2 Cl CH _-- - N 1 1 -β in which R is a phenyl or phenylthio group, and for the preparation of their pharmaceutically acceptable acid addition salts, characterized in that 1- (2 ', 4'-dichlorophenyl) -2- (N-imidazolyl) ethanol is condensed with halobenzyl of the formula VR-CH 2 X v wherein R is as defined above and X is a chlorine or bromine atom, in a solvent, and the resulting free base is converted, if desired, into a pharmaceutically acceptable acid addition salt.