CS233445B1 - Substituted 4-anilino1,3-dimethyl-1h-pyrazole/3,4-b(3,4-b)quinolines and method of theier preparation - Google Patents

Abstract

The invention relates to substituted 4-anilino-1,3-dimethyl-1H-pyrazolo [3,4-b] of Ohinolines of Formula I HN Ί CH, (I) CH3 wherein R 1 is hydroxy, methoxy, propyloxy, cetyloxy, ethoxycarbonylmethoxy, dimethylamino, carboxy or phenyl and R2 is hydrogen, or R 1 is hydrogen and R 2 is methoxy, or R 1 and R 2 are methoxy, or R 1 is acetamido-and R 2 is methyl, or R 1 and R 2 are taken together methylenedioxy. These new, yet unlabeled substances show antiviral activity activity against the influenza A2-Honkong virus against encephalomyocardial virus and against vaccinia virus in mice.

Description

The present invention relates to substituted 4-enino-1,3-dimethyl-1H-pyrazolo [3,4-b] quinolines of formula I

wherein R ¹ is hydroxy, methoxy, propyloxy, cetyloxy, ethoxycarbonylmethoxy. dimethylamino, carboxy or phenyloxy; and

R ² is hydrogen or R is hydrogen and R ² is methoxy, or R and R ² is

2 * 2 * methoxy, or R is acetamido and R is methyl, or R and R together form methylenedioxy. The invention also relates to a process for the preparation of substituted 4-anilino-1,3-dimethyl-1H-pyrazolo [3,4-b] cholineines of formula (1).

. These novel substances, which have not been described yet, exhibit anti-viral activity against A2-Honkong influenza virus, against encephalomyocarditis virus and against vaccinia virus in mice. *

The compounds of the formula I according to the invention can be prepared by reaction of the known 4-chloro-1,3-dimethyl-1H-pyrazolo [3,4-b] quinoline Jstein R. 0, Biel J. H, J. Med.

Chem. 13, 153 O973j] with the corresponding aniline derivative in phenol at 80 ° C to 120 ° C, preferably at 100 ° C. Compounds of formula 1 wherein R 1 is propyloxy or ethoxy and R ² is hydrogen may be prepared by alkylation of the compound of the invention.

A compound of formula I wherein R is hydroxy and R is hydrogen, the corresponding alkyl bromide in acetone in the presence of anhydrous potassium carbonate and preferably in the presence of potassium iodide.

Said Compounds of the invention belong to the class of anti-viral agents. The compounds were evaluated in the form of hydrochlorides against A2-Honkong raft and against encephalomyocarditis virus (EMC) in mice. For some compounds, efficacy against vaccinia virus in mice was also evaluated.

Evaluation of efficacy against A2-Honkong influenza virus in mice:

Female white SPF mice (Velaz) weighing 10 to 11 g were used. The virus was administered intranasally by light ether anesthesia mice. The amount of LD40 virus was determined in a separate experiment and was calculated according to REED and MUENCH / Reed L. J., Muench H., Amer. J. Hyg. 27, 493 (1938)]. Seven mice were used for each virus concentration. Untreated mice died of pneumonia according to the concentration used, blaming them 3 to 10 days after infection. Test substances were suspended using mono-oleic acid ester with tripolyethylene glycol anhydride sorbitol (Tween 80). Animals were administered per os by gavage, or were administered subcutaneously into the skin lash on the back in a volume of 0.2 ml each. Mygi were treated orally or subcutaneously twice daily for 5 days (1 day before and 4 days after infection). The valuable dose was x 150 mg / kg. Efficacy was assessed by extension, mean survival compared to untreated control. ,

After oral administration, the compounds of formula (I) wherein R ¹ is methoxy and R ² is hydrogen (extends the survival time of mice against 5 virus by 50 56) were active, wherein:

R ^{1 is} hydroxy and R ² is a hydrogen atom (extends the mouse survival time against 5 ^LD 5q virus by 66 Se and against 50 LC-q virus by 15%) and where R ¹ and R ² together form a methyl endioxy group (extends the survival time of mice against 50.LDgQ virus by 25%).

Compounds of Formula I having activity against the A2-Honkong influenza virus in mice following subcutaneous administration are summarized in Table 1 and are reported as% survival in mean survival.

Table I

R ¹ R ² 0.5 ld ₅₀ % 5 LD ₅₀ 50 LD ₅₀ % OCH ₃ H 59 - 46 ₃ ) ₂ H - - 105 COOH H - 35 49 OH H - 46.5 70 OC3H7 H - 61 - ^OC 16 ^H 33 H - 63 - -o-ch ₂ -o- - 44 - OCH ₂ C00C ₂ H ₅ H - 75 H och ₃ 60 60 0 OCH ₃ och ₃ 49 44 0 ^C 6 ^H 5 Η 21 37.5 0

Explanatory notes to Table 1:

-...... has not been tested

Evaluation against EMC in mice:

*

Aemics of white SPF mice (Velaz) weighing 10 to 11 g were used. The virus was injected sublcutaneously in a volume of 0.2 ml into the fold of the back skin. Untreated mice died according to the virus concentration used 3 to 7 days after infection. Subcutaneous treatment was performed with four doses of 100 mg / kg each 28, 22, 2 hours before and 2 hours after infection. Oral treatment was performed 24 hours before infection with a single dose of 400 mg / kg. The determination of the amount of virus used and the evaluation of the efficacy of the substances were carried out as in the evaluation of the efficacy against the influenza virus A2-Hong Kong.

The efficacy of the compounds of formula I against EMC virus in mice following oral (p. O.) And subcutaneous (a. P.) Administration of these compounds is shown in Table II in increasing the mean survival time of mice in% after application of 5 LD ^ q virus.

Table II

R ¹ R ² Mr. O · (%) Se C * W OH H 0 66 oc ₃ h ₇ H 32 33 ^OC 16 ^H 33 H 47 83 -o-ch ₂ -o- 47 95 OCH ₂ C00C ₂ H ₅ H 50 100 ALIGN! NHC0CH ₃ ch ₃ 0 40 ^C 6 «5 H 0 36

233445 4

Evaluation against vaccinia virus in mice:

Vaccinia virus as a lyophilized vaccine Sevac (ÚSOL Prague). The vaccine contains 5x10? units of virus per ml. Female white SPF mice (Velaz) weighing 15 g were used. The virus was injected intravenously in mice with a volume of 0.1 ml of 1.25 x 10 6 units of virus, causing approximately 100 necrotic forests on the tail in untreated mice. 6 to 6 days after infection. For easier counting of forests, the tails were stained with a solution containing 1% fluoreacein and 0.5% methylene blue, which stained necrosis tissue. After 14 days, the skin forests healed spontaneously and the symptoms disappeared. The treatment was performed in the same manner as the evaluation of efficacy against EMC virus. Efficacy was evaluated by percent reduction of necrotic forest compared to untreated control.

Activity against vaccinia virus in mice was determined for compounds of formula I wherein R ¹ is methoxy or or dimethylamino and R ² is hydrogen. Compound 1 of formula I wherein R is dimethylamino and R is hydrogen reduces the number of forests after oral administration by 82% and by subcutaneous administration by 74%. '♦

As can be seen from the above data, the compounds of formula I are active against A2-Honkong virus, EMC virus and vaccinia virus in mice. The preparation process is simple and provides the desired products in satisfactory yields. The following examples illustrate the details. The examples given are merely illustrative and not limiting.

He did

4- (4-methoxyanilino) -1,3-dimethyl-1H-pyrazolo [3,4-b] quinoline

A mixture of 2.32 g of 4-chloro-1,3-dimethyl-1H-pyrazolo [3,4-b] quinoline (10 mmol), 1.35 g of 4-methoxyaniline (11 mmol) and 15 g of phenol was stirred for 1 hour at 100 ° C.

The reaction mixture was poured into 100 ml of 2M sodium hydroxide solution, the insoluble material was aspirated, washed with water and then dissolved in 200 ml of 10% acetic acid while boiling. The filtrate, after filtering off the insoluble matter, was cooled and basified with 3M sodium hydroxide solution. The precipitate was filtered off with suction, washed with water and recrystallized from 3.0% aqueous dimethylformamide. 2.2 g (69.2%) of m.p. 154.4-155.4 ° C. After dissolution in ethanol, the compound was converted to the hydrochloride by conventional methods.

201.1 - 202.4 ° C.

»

Example 2

4- (4-H.methylaminoanilino) -1,3-dimethyl-1H-pyrazolo [3,4-b] quinoline

Stas 1.16 g of 4-chloro-1,3-dimethyl-1H-pyrazolo [3,4-b] quinoline (5 mmol), 0.75 g of 4-dimethylaminoaniline (5.5 mmol) and 7 g of phenol were 1 After stirring at 100 ° C for 1 hour. After diluting with 25 ml of ethanol, the solution was precipitated with ether, the precipitate was washed with ether and dissolved in water acidified with a few drops of hydrochloric acid. The insoluble matter was filtered off and the filtrate was basified with 2M sodium hydroxide solution to pH 10. The precipitate was filtered off with suction and washed with water. After dissolution in ethanol, the blue solution was passed through a column of 100 g silica gel (Kieselgel 60 Merck). The yellow column solution was evaporated to dryness to give 1.33 g (80.3%) of m.p. 79.6 ° C to 81.3 ° C. The residue was dissolved in hot in 10 ml of ethanol and acidified with a few drops of ethanolic hydrogen chloride. The crystals precipitated upon cooling were aspirated and recrystallized from ethanol. 1.52 g (75.2%) of m.p. 237.9 ° C to 239.7 ° C.

Example 3

4- (4-Hydroxyanilino) -1,3-dimethyl-1H-pyrazolo [3,4-b] quinoline hydrochloride

A mixture of 2.32 g of 4-chloro-1,3-dimethyl-1H-pyrazolo [3,4-b] quinoline (10 maol), 1.21 g of 4-amino phenol (11 mmol) and 15 g of phenol was stirred for 1 hour. After diluting with 50 ml of ethanol, the mixture was precipitated with ether. The insoluble material was aspirated, washed with ether and recrystallized from ethanol. Yield: 3.1 g (91.0%), mp 346 ° ^{C to} 352 ° C (dec.).

Example 4

4- (4-Ethoxycarbonylmethoxyanilino) -1,3-dimethyl-1H-pyrazolo [3,4- b] quinoline hydrochloride

A mixture of 2.78 g of 4-chloro-1,3-dimethyl-1H-pyrazolo [3,4-b] quinoline (12 maol), 2.55 g of 4-ethoxycarbonylmethoxyaniline (13 mmol) and 20 g of phenol was stirred for 2 hours at 100 ° C. In the same manner as in Example 3, 2.95 g (57.6%) of m.p. 199.2 ° C to 203.8 ° C.

Example 5

4- (4-Carboxyanilino) -1,3-dimethyl-1H-pyrazolo [3,4-b] quinoline hydrochloride.

A mixture of 2.32 g of 4-chloro-1,3-dimethyl-1H-pyrazolo [3,4-b] quinoline (10 mmol), 1.65 g of 4-aminobenzoic acid (12 mmol) and 15 g of phenol was 8 hours stirred at 100 ° C. After dilution with 50 ml of ethanol, the mixture was precipitated with ether. The insoluble material was aspirated, washed with ether and boiled in acetone. 2.7 g (73.2%) of m.p. 308 ° C to 310 ° C.

Example6

4- (4-Biphenylamino) -1,3-dimethyl-1H-pyrazolo [3,4-b] quinoline hydrochloride

A mixture of 1.85 g of 4-chloro-1,3-dimethyl-1H-pyrazolo [3,4-b] quinoline (8 mmol), 1.49 g of 4-aminobiphenyl (8.8 mmol) and 10 g of phenol was 4 g. stirred at 100 ° C for 1 hour.

The working-up procedure described in Example 3 gave 1.8 g (56.1%) of m.p.

207.3 ° C to 211.8 ° C.

Example 7

4- (3-Methoxyanilino) -1,3-dimethyl-1H-pyrazolo [3,4-b] quinoline hydrochloride

A mixture of 0.46 g of 4-chloro-1,3-dimethyl-1H-pyrazolo [3,4-b] quinoline (2 mmol), 0.26 g of 3-methoxyaniline (2.1 mmol) and 3 g of phenol was was stirred at 100 ° C for 2 hours.

In the same manner as in Example 3, 0.44% of the substance was obtained. 151.0-154.7 ° C.

Example 8

4- (3,4-Dimethoxyanilino) -1,3-dimethyl-1H-pyrazolo [3,4-b] quinoline hydrochloride

A mixture of 1.74 g of 4-chloro-1,3-dimethyl-1H-pyrazolo [3,4-b] quinoline (7; 5 maol), 1.26 g of 3,4-dimethoxyaniline (8.25 mmol), g of phenol was stirred at 100 ° C for 2 hours. In the same manner as in Example 3, 1.82 g (63.0%) of m.p. 208.6 ° C to 218.7 ° C (dec.).

Example 9

1,3-Dimethyl-4- (3,4-methylenedioicyanilino) -1H-pyrazolo [3,4-b] quinoline hydrochloride

A mixture of 2.32 g of 4-chloro-1,3-dlmethyl-1H-pyrazolo [3,4-b] quinoline (10 mmol), 1.51 g of 3,4-methylenedioxyaniline (11 mmol) and 15 g of phenol was stirred 1 hour at 100 ° C. In the same manner as in Example 3, 3.1 g (84.0%) of m.p. 251.0 ° C to 252.8 ° C.

He did

4- (4-Acetamido-3-methylanilino) -1,3-dimethyl-1H-pyrazolo [3,4-b] quinoline hydrochloride

A mixture of 2.32 g of 4-chloro-1,3-dimethyl-1H-pyrazolo [3,4-b] quinoline (10 mmol), 1.8 g of 5-amino-2-acetamido toluene (11 mmol) and 15 g. The phenol was stirred at 100 ° C for 1 hour. In the same manner as in Example 3, 2.32 g (58.6%) of m.p. 238.6 ° C to 241.8 ° C.

They did

1,3-Dimethyl-4- (4-propoxyanilino) -1H-pyrazolo [3,4-b] quinoline hydrochloride

To a suspension of 1.7 g of 4- (4-hydroxyanilino) -1,3-dimethyl-1H-pyrazolo [3,4-b] quinoline hydrochloride (5 mmol) in 150 ml of dry acetone was added 2.05 g of anhydrous potassium carbonate and 0.15 g of potassium iodide and 10.7 g of propyl bromide (5.5 mmol). The mixture was refluxed under stirring for 80 hours. After evaporation to dryness, the residue was extracted with chloroform after diluting with 25 ml of water. The extract was shaken with 5% sodium hydroxide solution and dried over magnesium sulfate. The filtrate after evaporation to dryness was dissolved in 2-propanol and acidified with hydrochloric acid. The crystals precipitated upon cooling were aspirated and recrystallized from ethanol. 0.97 g (50.7%) was obtained. 199 ° C to 202.6 ° C.

Example 12

4- (4-Cetyloxyanilino) -1,3-dimethyl-1H-pyrazolo [3,4-b] chiholine hydrochloride

From 1.7 g of 4- (4-hydroxyanilino) -1,3-dimethyl-1H-pyrazolo [3,4-b] quinoline hydrochloride (5 mmol) and 1.7 g of cetyl bromide (5.5 mmol), as described in Example 11, 1.15 g (40.6%) of m.p. 170.1 ° C to 176.5 ° C. Obtained by ether precipitation from solution in ethyl acetate.

Claims (3)

object of the invention Substituted 4-anilino-1,3-dimethyl-1H-pyrazolo [3,4-b] quinolines of general formula (I); (I) wherein R ¹ is methoxy, hydroxy, propyloxy, cetyloxy, ethoxycarbonylmethoxyacin, dimethylamino, carboxy or phenyl and R ² is hydrogen, Or wherein R is hydrogen and R is methoxy, or wherein both R and R are methoxy, or wherein R is acetamido and R is methyl, or wherein R and R together form ethylenedioxy. 2. Process for preparing compounds according to claim 1 of formula I wherein R is methoxy, hydroxy, ethoxykarbonylmethoxyskupina, dimethylamino, carboxy or phenyl slňipina and ^R2 is hydrogen, or wherein R ¹ is hydrogen and R ² is methoxy, or R ¹ R ² is methoxy, or wherein R ¹ is acetamido and R ² is methyl, or wherein R and R of the appellation are methylenedioxy, wherein 4-chloro-1,3-dimethyl-1H-pyrazolo [3,4] b) The quinoline is heated with an additional aniline derivative, preferably with a 0.1 molar excess, in phenol to a temperature of 80 ° C to 120 ° C, preferably to a temperature of 100 ° C. 3 Process for preparing compounds according to claim 1 of formula I wherein R ¹ is a propoxy or cetyloxyskupina and R ² is hydrogen, characterized in that a compound of vsorce I wherein R ¹ is hydroxy and R ² is hydrogen, alkylating the corresponding alkylbroid, wherein alkyl is propyl or cetyl, preferably in a 0.1 molar excess, in acetone in the presence of anhydrous potassium carbonate, preferably in the presence of potassium iodide.