AP27A

AP27A - Substituted indoloquinoxalines.

Info

Publication number: AP27A
Application number: APAP/P/1987/000055A
Authority: AP
Inventors: Jan Olof Einar Bergman
Original assignee: Leif J I Lundblad
Priority date: 1986-01-21
Filing date: 1987-01-09
Publication date: 1988-12-02
Also published as: FI883373A0; WO1987004436A1; DK492587A; DK492587D0; KR880700801A; AU606047B2; RU1776259C; HU201937B; CA1303613C; US4990510A; AP8700055A0; EP0238459B1; JPH07107066B2; JPS63502588A; AU6899587A; ATE74920T1; ES2037105T3; FI883373A; HUT47937A; IL81209A

Abstract

Substituted indoquinoxalines of the general formula i wherein r1 represents one or several, preferably 1 to 4, similar or different substitutents in the positions 1-4 and/or 7-10, selected from halogen, preferably br, lower alkyl/alkkoxy group having not more than 4 carbon atoms, trifluoromethyl group, trichloromethyl group, x is a group -(ch2)n-r2 wherein r2 represents a nitrogen containing basic residue such as nh2,nhr4 or nr5r6 wherein r4, r5 and r6 independently are lower alkyl or cykloalkyl and n is an integer of from 1 to 4 and r3 represents hydrogen, lower alkyl/cykloalkyl group having not more than 4 crabon atoms, and physiologically acceptable addition products of the compounds with acids and halogen adducts, preferably adducts with iodine, iodine monochloride or iodine monobromide. Also methods for preparing said compounds by reacting a compound of the formula ii with a reactive compound containing the residue -chr3x, or by rearranging a compound the formula iii by heating are described. The novel indoloquinoxalines have antiviral effect and have effect against cancer.

Description

Substituted indoloquinoxalines

The present invention relates to novel substituted indoloquinoxalines of the general formula I

wherein represents one or several, preferably 1 to 4, similar or different substituents in the positions 1-4 and/or 7-10, selected from halogen, preferably Br, lower alkyl/alkoxy group having not more than 4 carbon atoms, trifluoromethyl group, trichloromethyl group;

X is a group ~(^CH2^n^_R2 ^w^^ere^^{n R}2 ^reP^{resents a} nitrogen containing basic residue such as NH^, NHR^ or NR^Rg wherein R₄, and Rg independently are lower alkyl or cykloalkyl and n is an integer of from 1 to 4 and .,

R^ represents hydrogen, lower alkyl/cykloalkyl group having not more than 4 carbon atoms, and the physiologically acceptable addition products of the compounds with acids and halogen adducts,preferably adducts with iodine, iodine monochloride or iodine monobromide.

The novel substituted indoloquinoxalines according to the present invention have a high antiviral effect and several of the compounds show a high anti-cancer effect.

A suitable group of compounds are compounds wherein X is

-(CH_n) -R_n wherein R_n is -N(CH,)_n and n is 1. z n z z j z

The invention also relates to methods for preparing the novel indoloquinoxalines.

The compounds can be prepared by reaction of 6H-indolo-(2,3-b) quinoxalines in 6-position with a reactive compound containing the residue -CHR^X. Suitable solvents in this context are e.g. dimethylsulphoxide, dimethylformamide and dimethylacetamide. Alkylation in 6-position is favored by low temperature during the reaction. Minor amounts of 5-alkylated 5H-indolo(2,3-b)-quinoxalines are formed as byproduct in said alkylation. Previously ethanol has been used as solvent in this type of alkylation, F.Knotz, Sci. Pharm, 39, 20 (1970), F.Knotz, W.Wendelin, Sci. Pharm., 43, 249 (1975). This is unsuitable since with solvents of this type and at the stated temperature a relatively unfavorable isomer ratio is obtained. The 5-alkylated substances have an essentially lower activity.

The 5-alkylated products can like unalkylated starting material, if any, be removed by chromatography for instance on silica gel with a suitable eluent, for instance methylene chloride/methanol. The starting materials can be prepared by condensation of isatines with o-phenylene diamines. If unsymmetrical o-phenylene diamine is used for this, which is necessary for for instance preparation of 1-, 2-, 3- or 4-mono substituted starting materials, generally isomeric mixtures are obtained which can be difficult to separate. These difficulties can be avoided by preparing 6H-indolo(2,3-b)quinoxalines by condensation of oxindoles with 2-nitrosotoluidines as is shown below. R can be hydrogen or a nitrogen containing basic alkyl residue. With aminoalkyl groups as substituent ftPO 0 0 0 2 7 the active compounds can be prepared in one stage.

Another way to avoid the difficulties is to use N-alkylated o-phenylene diamines whereby 5-alkylated 5H-indolo(2,3-b)cuinoxalines are obtained. Then the substituent in 5-position can. be removed by boiling with hydrogen bromide dissolved in acetic acid.

The 5-alkylated 5H-indolo(2,3-b)quinoxalines can be thermally rearranged to 6-alkylated 6H-indolo(2,3-b)quinoxalines. R can be an aminoalkyl group. Thus, by this rearrangement reaction there is an alternative preparation method for the active compounds .

The anions of indoloquinoxalines are easily alkylated by chloroacetonitrile whereby 6-cyanomethylindologuinoxalines are obtained which by a suitable reduction can be converted to

6-(2-aminoethyl)indoloquinoxalines. Compounds of this type can

The bases easily can be converted to physiologically acceptable addition compounds such as hydrochloride, oxalate, tartrate and halogen adducts, preferably adducts with iodine, iodine monochloride or iodine monobromide.

A very interesting group of compounds according to the present invention are the adducts between phosphonoalkanoic acids, such as phosphonoformic acid, phosphonoacetic acid, phosphonopropionic acid, phosphonoglycolic. acid and phosphonobutyric acid, and their O-acetyl derivatives, and the indoloquinoxalines of formula I. The last mentioned adducts are interesting since the components per se are antiviral and intervene in different ways in the life cycles of DNA virus. Thus, it is known that phosphonoacetic acid and phosphonoformic acid are active against herpes virus,cf. Applied Microbiology, Volume 26, No. 3, September 1973, Shipkowit2 et al, Suppression of Herpes Simplex Virus Infection by Phosphonoacetic Acid, p. 264-267; Biochemistry, Volume 15, No. 2, 1976,

S Leinbach et al, Mechanism of Pnosphonoacetate Inhibition of Herpesvirus-Induced DNA Polymerase, p. 426-430; Antiviral Research, I (19S1), A Larsson and B Oberg, Selective Inhibition of Herpesvirus DNA Synthesis by Foscarnet, p. 55-62;

J.gen. Virol., 45, (1979), Bo Sundquist and Bo Oberg Phosphonoformate Inhibits Reverse Transcriptase, p. 273-281.

Instead cf using isolated adcucts of the incolocuinoxalines

HP ο o 0 0 2 7

I with acids of course directly prepared mixtures of the indoloquinoxalines and the acids can be used.

The compounds according to the present invention have a high anti-cancer effect as can be seen from the following tests which show anti-tumor effects for the substituted indoloquinoxalines according to the present invention.

Knotz and coworkers in Sci. Pharm. 1975, 43 (4), pages 249-60 have described a number of indophenazine derivatives. The most closely related compounds described in said article differ^from the compounds according to the present invention as regards the ring substituents. Thus, the compounds of formula I in said article wherein R=H and which thus only have a substituent in the 6-position do not show a high activity. The compounds of formula I in said article wherein R-NC^ or COOH are completely devoid of activity.

tfnotz has shown that indoloquinoxalines have effect against certain bacteria which effect however is clearly lower than the effect shown by preparations used today. Knotz has not reported any studies of the effects of the described indoloquinoxalines neither against virus nor against tumor.

iowever, the compounds according to the present invention which lave the combination of suitable ring substituents and an propriate N-substi tuent. in 6-position as defined' in claim 1 assess a high antiviral activity and high anli-cancer ctivity.

ndoloquinoxalines without a nitrogen containing basic residue n 6-position have been described in the literature, cf. atagiri, K. et al., in Shionogi Kenkyosho Nempo 16 (1966) pp, 2-57, and 58-62. in said article a 6-acetylindoloquinoxaline

5a is described. However, the indologuinoxaiines without any nitrogen containing basic residue in 6-position did not show any effect against virus and/or cancer and the shown inhibition of phages was insignificant. The five indophenazines (i.e, indologuinoxaiines) described by Katagiri et al were all 1naotive in the cancer test. Sone of these indophenazines have been studied by the present inventors and were found inactive in all tests and fall outside of the definition of the active compounds according to the present invention.

In the following tests two substituted indologuinoxaiines according to the present invention were used, viz.

3196= 9-bromo-6(N,N-dimethyl ami noethyl)-6H-i ndolo( 2,3-b)guinoxali ne . HCl

B219= the 2,3-dimethyIderivative of the above compound.

Test system with cancer cells

Burkitt lymphoma cells were obtained from professor Georg Klein, Karolinska Institutet. HeHrl-cells contain Epstein-Barr /irus antigen on the surface of the cells while Raji cells contain the same antigen in the nucleus. The cells were grown

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in RPMI 1640 medium in the presence of 15% of fetal calf-serum, 2mM glutamine, 200 mg/1 penicillin and 200 mg/1 of streptomycine. L 1210 cells (mouse leukemia) were obtained from Dr, Miklos Deere, Rikshospitalet, Oslo. These cells were grown in RPMI 1640 medium with 10% horse serum.

Results

The results of the tests carried out are shown graphically in the enclosed figures 1, 2 and 3, wherein fig. 1 shows growth of 2 strains of Burkitt lymphoma cells in the presence of iododeoxyuridine (IDU) fig. 2 shows the effect of B196 against HeHrl-cells fig. 3 shows the effect of B196 against Raji-cells.

Fig. 1 shows that HeHrl-cells are sensitive to iododeoxyuridine (well known anti-herpes compound), while Raji-cells are not. Figures 2 and 3 show that both these cell types are rapidly killed in the presence of indoloquinoxaline derivative (B196). Also osteogenic sarcoma cells and giant tumor cells (strain T4) were inhibited to 90% in their growth after 6 days test with 1 mg/1.

Experiments with mice have also been carried out. LD^g intraperitoneally is of the order 1000 mg/kg and about 100 mg/kg intravenously. 20 mice were inocculated with 5 millions mouse ascites tumor cells intraperitoneally. 80 mg/kg of B219 were given daily for ten days. 2 mice died 22 days after treatment, mice survived the whole test period (3 months).

All controls were then dead.

In two experiments with ascites lymphoma cells in mice a survival time of 50.9 days was obtained in the first experiment and 45.6 days in the second. The survival time for the controls was 15.2 and 16.1 cays respectively.

When tearing at National Cancer Institute (USA) a statistically conf;r*ei effect w;rh mouse L1210 was ccrainec.

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Substituted indoloquinoxa1ines according to the present invention also have a high antiviral effect. In tests carried out at National Cancer Institute a strong effect against HTLV-3-virus in tissue culture has been shown with concentration as low as 1 mg/1. As can be seen from the following tests the indoloquinoxalines also have a high antiviral effect against herpes simplex virus of both type 1 and 2.

Test methods against virus

The virus strains were obtained from the National

Bacteriological Laboratory, Stockholm. Herpes simplex type 1 and 2 were passaged intracerebrally in mice.

For mouse experiments male mice 2-3 weeks old were used. Intracerebral infection was carried out by introducing the syringe 3 mm above the eye to a depth of approximately 3 mm. 0.03 ml virus suspension was injected above one eye and the substance above the other.

Control groups obtained on one hand injection with physiological saline and on the'Other hand virus suspension.

i

Tissue culture experiments took place with human amnion AV3-cells. The cells were grown in glass-tubes (12x100 mm) in Eagles medium containing Earles salt solution and 10% of calf-serum. Tubes with a full-grown homogenous layer of cells were infected with virus (0.15 ml) and the virus was allowed to adsorb to cells for 1 hour at 37°c. The tubes were treated with substance dissolved in medium 24 hours and 0 hour before infection. Medium containing test-substance was changed every second day throughout the experiment. In each test a series of concentrations of the test-substance was investigated simultaneously so that about 50% reduction in cytopatogenic effect could be established (ΕΟ^θ).

Results of the virus experiments

For all investigated synthetisized compounds was between

0.5-2 mc/1 in the tissue culture experiments. ___

L I 0 0 0 0 dV bad original

The results of the virus experiments carried out are also shown graphically in the enclosed figures 4 and 5 wherein fig. 4 shows effect of dialysis on the antiviral properties of

B196, fig. 5 shows inactivation of HSVl-virus after incubation with two different concentrations of B196 vs. time.

In the experiments shown in figure 4 various concentrations of B-196 were mixed with herpes simplex virus type 1 and injected intracerebrally into mice immediately at or after dialysis, at 4°C over night. No survivors were recorded in control groups.

Figure 4 shows a dramatically improvement of the survival rate if virus was mixed with B196 before injection. The survival rate increases from 0 to 100%. Even if the virus-substance mixture is subjected to dialysis before injection the effect is almost equally pronunced. This probably indicates a direct interaction between the substance and the virus particle. (Fig. 4) .

I

This belief is supported by experiments wherein virus and substance have been incubated together and injected in mice after different times.

In the experiments shown in figure 5 two concentrations of B-196 (75 and 150 ug/ml) were incubated for different times at 4°C with herpes simplex virus type 1 and the infectivity of the mixtures were determined by intracerebral injection in mice. Control virus was incubated at 4°C for the same period of time. No survivors were registered in these groups.

Fig. 5 clearly shows that virus slowly is inactivated in the presence of the substance.

Some results with other compounds according to the invention are given in Table 1.

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PAGINAL

The invention is further illustrated by means of the following examples;

Example 1

2,3-dimethy1-9-bromo-6H-indolo(2,3-b)guinoxaline

A mixture of 22.6 g 5-bromoisatin (0.1 mole), 15.0 g 4,5-dimethyl-o-phenylene diamine (0.11 mole) and 400 ml acetic acid is boiled under reflux for 4 hours. After cooling the product is sucked, washed with ethanol and dried. Yield 29 g, 89%. Recrystallisation from pyridine gives needle shaped crystals with melting point 319-322°C.

In the corresponding manner also the following related intermediates are prepared substitution mp °C

2-CH₃

2- OCH₃

3- OCH₃

3-COOH

9-F

9-Cl

9-Br

9-1

2.3- di-CH₃2-CH₃,9-Cl 2-CH₃,9-Br 9-CH₃

3.9- di-Cl

7.9- di-Cl

2.3- di-CH₃, 9-C1

2.3- di-Cl

2.3.9- tri-Cl

2,3,7,8-tet ra-Cl

2.3- diCl,7,8-diOCH₃

297-300

324-327

284-287

360

302-304

322-324

338-340

354-356

347-350

340-342

347-350

247-249

328-330

302-303

331-334

342-345

362-365

402-404

AP 0 0 0 0 2 7

/

Example 2

2-methyl-9-chloro-6H-indolo(2,3-b)quinoxaline

A solution of 1.68 g 5-cnloroindol (0.01 mole) and 1.50 g 2-nitroso-N-methyl-p-toluidine (0.01 mole), 10 ml acetic acid is boiled under reflux for 1 hour. Finally hydrogen bromide is introduced and the mixture is boiled for further 2 hours. After cooling it is poured into water. The dried reaction product is recrystallized from acetic acid.

Yield 2.0 g 75%, mp 340-343°C.

Example 3

2-methv1-9-bromo-6H-indolot 2,3-b)quinoxaline

A solution of 2.26 g 5-bromoisatin (0.01 mole) and 1.36 g 2-amino-N-methyl-p-toluidine (0.01 mole) in 15 ml acetic acid is boiled with reflux for 45 minutes. Finally hydrogen bromide is introduced and the mixture is boiled for further 2 hours. After cooling it is poured into water. The dried reaction product is recrystallized from acetic acid.

Yield 2.3 g, 74%, mp 347-350°C.

Example 4

9-methyl-6-(N,N-dimethylaminoethyl)-6H-indolo(2,3-b)guinoxal ine

5.38 g sodium hydride (0.22 mole) is added to a solution of 15.9 g of N,N-cimethylamino-2-chloroethan-HCi (0.11 mole) in 400 ml dimethyIformamide at 30°C under nitrogen and well stirred. After about 5 minutes 23.3 g 9-methyl-6H-indolo- (2,3-b)quinoxaline (0.10 mole) is added in portions. The mixture is stirred at 30°C until a clear solution is obtained. After 24 hours at 30°C the solvent is destilled off in vacuum at room temperature and the residue is purified by washing with water. The raw product is dried and dissolved in methyl acetate. When cooled down to -30°C orange yellow crystals are ofcrained. Yield 21.1 g, 70%, mp 118-120°C.

bad ORIGINS S

2,3-dimethyl-6-(N,N-dimethylaminoethyl)-6H-indolo(2,3-b)~ quinoxaline, mp 138-140°C

7,9-dichloro-6-(N,N-dimethylaminoethyl)-6H-indolo(2,3-b) quinoxaline, mp 187-189°C.

Example 5

9-chloro-6-(N,N-dimethylaminoethyl)-6H-indolo(2,3-b) quinoxaline-HCl

5.38 g sodium hydride (0.22 mole) are added to 600 ml dry dimethylsulphoxide, wherein 15.9 g N,N-dimethylamino-2-chloroethan-HCl (0.11 mole) has been dissolved at 30°C under nitrogen and a good stirring. After about 5 minutes 5.4 g 9-dichloro-6H-indclo(2,3-b)quinoxaline (0.10 mole) are added. The mixture is stirred at 40°C until a clear solution is obtained. Finally heating is carried out at 40°C for 3 hours. The solvent is destilled off in vacuum at room temperature and the residue is dissolved in 2% boiling hydrochloric acid. When cooled the hydrochloride precipitates.

In the corresponding manner the hydrochloride of the following compounds belonging to the general formula are prepared. Alternatively the raw dried base can be dissolved in acetone and the hydrochloride can be precipitated by introducing of hydrogen chloride.

AP 0 0 0 0 2 7

R^ substituent 2?3-di-CK₃,9-C1

2,3-di-CH₃,9-Br 2-CH₃,9-I

3.9- di-Cl 9-C1

7.9- di-Cl n(ch₃)₂ H n(ch₃)₂ H n(ch₃)₂ H n(ch₃)₂ H N(CH₃)₂ h N(CH₃)₂ h n mp °C

235-240

225-228

237-240

254-257

221-223

Example 6 , 3-di.T.ethyl-9-bromo-6-(N,N-dimethylaminoethyl)-6H-indolo(2,3-b)auinoxaline

To a supension of 5.38 g sodium hydride (0.22 mole) in 600 ml dry dimethylsulphoxide maintained at 35°C 32.6 g 2,3-dimethyl-9-bromo-6H-indolo(2,3-b)quinoxaline (0.10 mol) is added under nitrogen and with good stirring. 30 minutes after completed addition 15.9 g N,N-dimethylamino-2-chloroethane-HCl (0.11 * mole) are added at 35°C.

The mixture is stirred at 35°C for four hours and then the solvent is distilled off in vacuum at room temperature. The residue is dissolved after soaking out with water in methylacetate anc slowly cooled to -30°C whereby the base crystallizes out. Yield 22 g, 55%, mp. 146-147°c.

In the corresponding manner the following compounds are prepared :

2.3- dimethyl-9-bromo-6-(Ν,Ν-diisopropylaminoethyl)-6H-indolo(2,3-b)quinoxaline, mp. 121-123°C

I

9-fluor0-6-(Ν,Ν-dimetylaminoethyl)-6H-indolo( 2,3-b)quinoxaline, mp. 101-102’C

2.3- dimethyl-9-bromo-6-(morpholinoethyl)-6H-indolo{2,3-b)quinoxaline, mp. 208-210’C.

Example 7

7,9-dichloro-6-(N,N-dimethylaminoethyl)-6E-indolo(2,3-b).

au inoxaline-HCl

2.64 g sodium hydride (0.11 mole) are added to 300 ml dry dimethylsulphoxide wherein 7.95 g N,N-dimethylamino-2-chloroethane HC1 (0.055 mole) have been dissolved at 30’C under nitrogen and with good stirring. After about 5 minutes 14.4 g 7, 9-d ichloro-6H-indolo-( 2,3-b)quinoxaline (0.05 mole) is added. The mixture is stirred at 50°C until a clear solution is ocrained. Finally heating is carried out ar 60°C for 3 hours. The solvent is distilled off in vacuum at room temperature and - ' * ci c · — ~ ί c c i s 3 c v ⁰ in *ΐάπ^ρ·! 3,^e ” ~ ~ ξ ~ z >· * ^u BAD water. The mixture of the raw base then is subjected to chromatography on silica gel with methanol as eluent, the following being obtained in the order stated; a slight amount of non-reacted 7,9-dichloro-6H-indolo(2,3-b)quinoxaline,

7,9-dichloro-6-(N,N-dimethylaminoethyl)-6H-indolo(2,3-b)quinoxaline and finally 7,9-dichloro-5-(N,N-dimethylaminoethyl)-5H-indolo(2,3-b)quinoxaline (22%). The dried 6-substituted base is dissolved in acetone and the hydrochloride is precipitated by introducing hydrogen chloride. Yield 7'g, 39%, mp 254-257°C.

Example 5

9-methyl-6-(N,N-d imet bylaminoethy1-6H-indolo(2,3-b) ouinoxaline-KCl

5.06 g sodium (0.22 mole) is dissolved in 550 ml dry ethanol, 15.9 g N, N-dimethylamino-2-chloroethane-HCl (0.11 mole) is added followed by 23.3 g 9-methyl-6H-indolo(2,3-b)quinoxaline (0.10 mole). The mixture is boiled for 4 hours, cooled and filtered. The filtrate finally is concentrated in 2% warm hydrochloric acid. The hydrochloride is obtained as yellow crystals with slowly cooling. Yield 21 g, 62%, mp. 240-242°C.

Example 9

9-chloro-6-(N,N-dimethylaminoethyl)-6H-indolo(2,3-b)~ quinoxaline-HCl

5.06 g sodium (0.22 mole) is dissolved in 300 ml 2-methoxyethand , 15.9 g N,N-dimethy1amino-2-chloroethane-HCl (0.11 mole) is added followed by 25.4 g 9-chloro-6H-indolo(2,3-b) quinoxaline (0.10 mole). The mixture is heated for 3 hours at 100°C, cooled and filtered. The filtrate is concentrated and poured into water. The raw base obtained is filtered off and washed with water and subjected to chromatography on silica gel wish merr.and as eluent. The purified base is dissolved after drying ir. acecone. The hydrochlor i de is precipitated by slowly dropwise adding concentrated hydrochloric acid with stirring.

• - . . i ~ „ cc? - -, ? : . :

...... - f .. «-f . — - - - - - - .

L Z 0 0 0 0 dV

BAD ORIGINAL

Example 10

9-^ethv1-6-(N,N-dimethylaminoethy1)-6H-indolo(2,3-b)quinoxaline-ICl

3.04 g 9-methy1-6-{N,N-dimethylaminoethy1 )-6H-indolo( 2,3-b) quinoxaline (0.01 mole) is dissolved in 50 ml methanol. 1.62 g iodine chloride (0.01 mole) is added and the mixture is boiled 15 minutes. After cooling the crystals formed are sucked, 4.1 g; 87%, mp. 101-103°C.

In the corresponding manner the following compound is prepared:

2,3-dimethy1-6(N,N-dimethylaminoethy1)6H-indolo(2,3-b)cuinoxaline-ICl, mp. 1S3-1S5°C.

Example 11

9-methyl-5-(Ν,Ν-dimethylaminoethyl)-5H-indolo(2,3-b)quinoxaline

1.79 g (Ν,Ν-dimethylaminoethyl)-o-phenylenediamine (0.01 mole) and 1.61 g 5-methyl isatin (0.01 mole) are boiled 30 minutes in 25 ml acetic acid. The reaction solution is concentrated under reduced pressure and then is made alkaline with 5% sodium hydroxide solution. The precipitated base is washed, dried and recrystallised from methanol/water. Yield 2.2 g; 76%, mp. 150-152°C.

In the corresponding manner the following compounds are prepared :

9-bromo-5-(N,N-dimethylaminoethyl)-5H-indolo(2,3-b )qu inoxal ine, mp. 152-154°C.

7,9-dichloro-5-(N,N-dimethylaminoethyl)-5H-indolo(2,3-b)quinoxaline, mp. 180-182°C.

Example 12

9-r e t hy 1 - 6 - (N, N-c imet hy lami noet hy 1 ) - 6H-i ndolo ( 2,3-b )qu inoxal ine

3.0- c 9-methyl-5-(Ν,Ν-dimethylaminoethyl )-5K-indolo(2,3-b)q j i noxaline (0.01 mole) is heated to 250°C for 2 hours. The reassicr. mixture is dissolved in methanol and subjected to or. silica gel with :methanol as eluent wherebv

the following components are obtained in the order stated:

9-methy 1indolo(2,3-b)quinoxaline and

9-methy1-6-(N,N-dimethylaminoethyl)-6H-indolo(2,3-b)quinoxalinea nd finally a slight amount of non-reacted starting material. Yield 6-substituted compound 1.1 g; 36%, mp. 118-120°C.

In the corresponding manner the following compounds are prepared:

2,3-dimethyl-6-(N,N-dimethylaminoethyl)-6H-indolo(2,3-b)quinoxaline, mp. 138-140°C.

7,9-aichloro-6-(N, N-d imethy1aminoethyl)-6H-indolo(2,3-b)quinoxaline, mp. 187-189°C.

Typical compounds according to the invention

—1	—2	—3	n	smp °C
2-OCH₃	N(CH₃)₂	H	1	224-227 (HC1)
3-CH₃	N(CH₃)₂	H	1	248-250 (HC1)
3-NO₂	n(ch₃)₂	H	1	292-295 (HC1)
3-COOH	N(CH₃)₂ \|	H ,	1	302-305 (HC1)
3-OCH₃	N(CH₃)₂	H	1	228-230 (HC1)
9-CH₃	N(CH₃)₂	H	1	118-120 (base)
9-CH₃	N(CH₃)₂	H	1	251-253 (HC1)
9-OCH₃	n(ch₃)₂	H	1	231-234 (HC1)
9-F	N(CH₃)₂	H	1	101-102 (base)
9-F	N(CH₃)₂	H	1	238-240 (HC1)
9-C1	n(ch₃)₂	H	1	232-255 (HC1)
9-C1	N/CHiCH)^	H	1	290-295 (HC1)
9-Br	N(CH₃)₂	H	1	260-262 (HC1)
9-Br	N(CH₃)₂	H	1	224-227 (oxalate)
9-Br	N(CH₃)₂	H	1	207-211 (IC1)
9-Br	N(CH₃)₂	H	1	254-257 (HC1)
9-1	N(CH₃)₂	H	1	280-285 (oxalate)
9-1	n(ch₃)₂	H	1	204-208 (IC1)
2,3-di-CH₃	N(CH₃)₂	H	2	146-147 (base)
2,3-di-CH₃	N(CH₃)₂	H	1	138-140 (base)
2,3-d;-Cn₃	N{CH₃)₂	Π	1	234-236 (a'Cl)

AP000027 bad original Q

Typical compounds according to the invention (contn.)
*1	—2	—3	n	mp °C
2,3-di-CH₃	n(ch₃)₂	H	1	219-221	(oxalate)
2,3-di-CK₃	n(ch₃)₂	H	1	183-185	(IC1)
2,3-di-CH₃	nGTo	H	1	183-185	(base )
3,9-di-Cl	n(ch₃)₂	H	1	254-257	(HCl)
7,9-di-Cl	n(ch₃)₂	H	1	187-189	(base)
7,9-di-Cl	N(CH₃)₂	H	1	254-257	(HC1)
2-CH₃,9-Cl	n(ch₃)₂	H	1	123-125	(base)
2-CH₃,9-C1	nich₃)₂	H	1	230-233	(HC1)
2-CH₃,9-Br	N(CH₃)₂	H	1	.279-282	(oxalate)
2-C»₃,9-Br	n(ch₃)₂	H	1	217-220	(HCl)
3-Cl,9-Br	n(ch₃)₂	H	1	291-295	(oxalate)
2,3-di-CH₃,	9-C1 N(CH₃)₂	H	1	252-255	(HC1,
2,3-di-CH₃,	9-Br ί0Γθ	H	1	208-210	(base,
2,3-di-CH₃,	9-Br N(CH₃)₂	H	1	225-228 f	(HCl)
2,3-di-CH₃,	9-Br N/tiHCH-j)^	H	1	216-219	(HC1)
2,3,7,8-tetra-Cl N(CH₃)₂	H	1	282-284	(HCl)
2,3-diCl-7,	8-diMeO N(CH₃)₂	H	1	280-282	(HCl)
2, 3-diCH₃,7	-Br N(CH₃)₂	H	3	216-218	(HCl)
2,3-diCH₃r7	-Br N(CH₃)₂	H	4	212-214	(HCl)

Example 13

6-cyanomethyl-6H-indolo(2,3-b) quinoxaline'

Indoloquinoxaline (10.95 g) is dissolved in dry dimethylsulfoxide (150 ml) under nitrogen and sodium hydride (1.3 g) is added with good stirring at 35°C. After completed hydrogen gas generation (about 30 minutes) chloroacetonitrile (4.0 g) was dropped in with stirring at 20°C. After 1 day at this temperature water (20 ml) is added and crystals formed of pure 6-cvanomethyl indoloquinoxaline (8.9 g) are sucked and washed with methanol followed by water. The mother liquor is poured into a large amount of water and the precipitate formed is dried and purified by chromatography on silica gel with methylene chloride/methanol as eluent. In this manner further 3.0 g of 6-cyanomethyl-6H-indclo( 2,3-b) quinoxaline is obtained. Thus, the total yield is 11.9 g, 92%, mp. 231-232°C.

In the corresponding manner the following compounds are prepared

2.3- dichloro-6-cyanomethyl-6H-indolo( 2,3)quinoxaline, mp. 258-260°C.

2.3- dimethy 1-6-cyanomethyl-6H-indolo( 2,3-b)quinoxaline, mp. 244-246°C.

, 3,9-tr ichloro-6-cyanomethyl-6H-indolo(2,3-b)quinoxaline, mp. 296-298°C.

2.3- dimethy1-9-chloro-6-cyanomethyl-6H-indolo(2,3-b)quinoxaline, mp. 286-288°C.

Example 14

6-( 2-aminoethyl)-6H-indolo( 2,3-b)quinoxaline . HC1

6-cyanomethyl-6H-indolo(2,3-b)quinoxaline (2.58 g) is heated with reflux with lithium aluminum hydride (0.60 g) in tetrahydrofuran (40 ml) for 6 hours, and then water (2 ml) is cropped thereto cautiously. After stirring for 1 hour solid material is removed by separation and the rest is concentrated. The rest is dissolved in dry ether and the base is precipitated as hydrochloride by addition of hydrogen chloride 1.40 g, 47%, mp. 280-282°C.

Example 15

6-( 3-phtalimidopropvl)-6H-indolo(2,3-b)quinoxaline

Σndolocuinozaline is dissolved in dry dimethylsulfoxide (180 ml·) under nitrogen and sodium hydride (1.3 g) is added with good stirring at 33’C. After completed hydrogen gas generation (about 30 minutes) 3-chloropropylphtalimide (11.20 g) is added in portions at 20°C. After 1 day at this temperature the

APO 0 0 0 2 7 recrystallisation from acetonitrile is carried out 15.10 g, 74%, mp. 212-214°C.

Exemple 16

6-(3-aminopropy1)-6H-indolo(2,3-b)quinoxaline . HCl

6-(3-phtalimidopropyl)-6H-indolo(2,3-b)quinoxaline (4.06 g) is heated under reflux (4 hours) in ethanol (200 ml) wherein hydrazine (0.5 ml) has been dissolved. After warm filtration the filtrate is concentrated and dissolved in dry ethet and then hydrogen chloride is added. The precipitate formed is sucked and dried, 3.12 g, 57%, mp. 288-291°C.

Example 17

6-(N,N-dimethylaminoethyl)-2,3-dimethyl-6H-indolo(2,3-b)guinoxaline-phosphonoacetate

6-(N,N-dimethylaminoethyl )-2,3-dimethyl-6H-indolo(2,3-b)quinoxaline (318 mg, 0.001 mole) is dissolved in dioxane-ether (10 ml) and a solution of phosphonoacetic acid (140 mg, 0.001 mole) in dioxan-ether (10 ml) is added. After 2 hours at 25°C the adduct formed is sucked. Yield 412 mg (90%).

Exemple 18

6-(N,N-dimethylami noethyl )-2,3-dimethyl-6H-indolo(2,3-b)cu inoxaline-phosphonoformate

6-(N,N-dimethylaminoethyl)-6H-indolo-(2,3-b)quinoxaline . HCl (354.5 mg, 0.001 mole) is dissolved in water (50 ml) wherein acetic acid (2.0 g) and sodium acetate (2.3 g) have been dissolved. To this solution trisodiumphosphonoformate-hexahydrate (300.1 mg, 0.001 mole) in water (15 ml) is added. The solution thus obtained then directly is used for antiviral studies .

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Claims

1. Substituted indoloquinoxalines of the general formula I wherein R^ represents one or several, preferably 1 to 4, similar or different substituents in the positions 1-4 and/or

7-10, selected from halogen, preferably Br, lower alkyl/alkoxy group having not more than 4 carbon atoms, trifluoromethyl group, trichloromethyl group;

X is a group -(CH₂)_n-R₂ wherein R₂ represents a nitrogen containing basic residue such as NH₂, NHR^ or NR^Rg wherein R₄, R₅ and Rg independently are lower alkyl or cykloalkyl and n is an integer of from 1 to 4 and

2. Substituted indoloquinoxalines according to claim 1, characterized in that X is -(CH₂)_n~R₂ wherein R₂ is -N(CH₃)₂and n is 1.

3. Substituted indoloquinoxaline according to claim 1 or 2, characterized in that it is

9-bromo-6-(N,N-dimethylaminoethyl)-6H-indolo(2,3-b)qu inoxa1ine,

2,3-dimethy1-9-bromo-6-(N,N-dimethylaminoethyl)-6H-indolo(2,3-b)quinoxaline,

2, 3-d imet hy 1-6 - ( N, N-d ime t Ly 15.mi r.ce t hy 1 ) -6H - i r.dolo ( 2 , 3-b )BAO ORIGINAL quinoxaline,

2-methyl-9-chloro-6-(N,N-dimethylaminoethyl )-6H-indolo(2,3-b)quinoxaline, or

2-methy 1 -9-bromo-6-(Ν,Ν-dimethylaminoethy 1) - 6H-indolo( 2,3-b) qu inoxaline.

4. Substituted indoloquinoxaline according to any of the claims 1-3 characterized in that it is an adduct of an indoloquinoxaline of formula I and a phosphonoalkanoic acid.

5. Substituted indoloquinoxaline according to claim 4, characterized in that the phospnonoalkanoic acid is pbosphonoformic acid.

6. Substituted indoloquinoxaline according to claim 4, characterized in that the phosphonoalkanoic acid is phosphonoacetic acid.

I

7. Ά method for preparing substituted indoloquinoxalines of the general formula I

A Z 0 0 0 0 dV wherein represents one or several, preferably 1 to 4, similar or different substituents in the positions 1-4 and/or 7-10, selected from halogen, preferably Br, lower alkyl/alkoxy group having not more than 4 carbon atoms, trifluoromethyl group, trichloromethyl group;

X is a group “(^CH2^n~^R2 ^w^^ere^^{n R}2 ^reP^{resents a} nitrogen containing basic residue such as NI^, NHR₄ or NR^R^ wherein ?₄, R- and R- independently are lower alkyl or cykloaikyl and bad original n is an integer of from 1 to 4 and

P.^ represents hydrogen, lower alkyl/cykloalkyl group having not more than 4 carbon atoms, and the physiologically acceptable addition products of the compounds with acids and halogen adducts, preferably adducts with iodine, iodine monochlcride or iodine monobromide, characterized by

a) reacting a substituted quinoxaline of the general formula II wherein R^ is defined as above, with a reactive compound containing the residue -CHR^X, wherein R^ and X are defined as above, and if desired converting the compound obtained into the corresponding physiologically acceptable addition products with acids/halogen adducts, or

b) rearranging a substituted quinoxaline of the general formula III w n e r e 1 n R , R by heat inc and and X are defined as above, if desired converting the product obtained into ng phy.-i : Logically acceptable addition orcd-cts

BAD OPWSWW- with acids/halogen adducts.

8. An agent having effect against cancer characterized in that it as an active component contains a substituted indoloquinoxaline of the general formula wherein represents one or several, preferably 1 to 4, similar or different substituents in the positions 1-4 and/or 7-10, selected from halogen, preferably Br, lower alkyl/alkoxy group having not more than 4 carbon atoms, trifluoromethyl group, trichloromethyl group;

X is a group *'(^CH2^n^-R2 ^where*^{n R}2 ^reP^{resents a} nitrogen containing basic residue such as NH₂, NHR^ or.NR^Rg wherein R₄, Rg and Rg independently are lower alkyl or cykloalkyl and n is an integer of from 1 to 4 and

R^ represents hydrogen, lower alkyl/cykloalkyl group having not more than 4 carbon atoms, and the physiologically acceptable addition products of the compounds with acids and halogen adducts, preferably adducts with iodine, iodine monochloride or iodine monobromide.

AP 0 0 0 0 2 7

9. An antiviral agent characterized in that it as an active component contains a substituted indoloquinoxaline of the general formula wherein R^ represents one or several, preferably 1 to 4, similar or different substituents in the positions 1-4 and/or 7-10, selected from halogen, preferably Br, lower alkyl/alkoxy group having not more than 4 carbon atoms, trifluoromethyl group, trichloromethyl group;

X is a group ^-^^CH2^n^_R2 ^{wherein R}2 ^reP^{resents a} nitrogen containing basic residue such as NH₂, NHR^ or NR^Rg wherein R₄, R₅ and Rg independently are lower alkyl or cykloalkyl and n is an integer of from 1 to 4 and

10. An antiviral agent according to claim 9 active against DNA-virus characterized in that it consists of an adduct or a mixture of an indologuinoxaline with the formula I and a phosphonoalkanoic acid.

11. An antiviral agent according to claim 10 characterized in that the phosphonoalkanoic acid is phosphonoformic acid.

12. An =r.:ivi:si agenc according to claim. 10 characterized in that the phosphonoalkanoic acid is phosphonoacetic acid.