EP1001936A1 - Bis -acridinecarboxamide having anti-tumor activity - Google Patents

Abstract

The present invention relates to bis-acridinecarboxamides and the derivatives thereof substituted with aminoalkyl chains, have evidenced a remarkable antitumor activity, in particular in colon tumors.

Description

B S-ACR DIMECARBQXAMIDES HAVING AftTTITUMQR ACTIVITY

The present invention relates to bis- acridinecarboxamides and the derivatives thereof substituted with aminoalkyl chains.

Said compounds have evidenced a remarkable antitumor activity, in particular in colon tumors. PRIOR ART

The treatment of solid tumors is a still unresolved problem of the antitumor therapy. The search for new molecules able to cure solid tumors is hindered by a number of factors, such as the development, by the tumor cells, of resistance to many antitumor drugs (Multi-Drug Resistance, MDR) and the difficulty for the medicament to reach the tumor mass which is scarcely vascularized.

A number of compounds which are at present in preclinical or clinical development or which are already used in the antitumor therapy, such as mitoxantrone or doxorubicine, belong to the wide family of the DNA intercalating molecules. Although these molecules share a tri- o tetra-cyclic planar structure, they belong to well distinct classes, having frequently different chemicc-physical and biological properties.

The nature of the substituents and of the aromatic rings in the backbone is critical since often small structure changes give the analogue compounds an altogether different biological behaviour. It is in fact known, for example, that a change from anthracenediones to the corresponding benzo[g]isoquinolin-5 ,10-diones causes a decrease in cardiotoxicity, while maintaining an antitumor activity identical to, if not better than, that of the carbocyclic analogues (Krapcho, P., WO

92/15300: Krapcho et al . , US 5,587,382).

However, the molecules presently used in therapy induce the cited multi-drug resistance, limiting therefore their effective use in a long-term therapy.

Another known class of DNA intercalating agents comprises acridines [Atwell G.J. et al . , J. Med. Chem.,

30, 564-9 (1987); Bagulβy B.C. et al., Cancer Chemother. and Pharmacol., 36, 244-8 (1995); Atwell G.J. et al., J. Med. Chem., 27, 1481-5 (1984); Rewcastle G.W. et al . , J.

Med. Chem., 29, 472-7 (1986); Denny W.A. et al . , J. Med.

Chem., 30, 658-663 (1987); Wakelin .P.G. et al . , J.

Med. Chem., 30, 855-861 (1987)].

Examples of bis-imidazoacridones with antitumor activity are known in literature [Cholody W.M. et al.,

J. Med. Chem., 38, 3043-52 (1995); Hernandez L. et al . ,

Cancer Res., 55, 2338-45 (1995)]. However, even within the same class of molecules, as the authors themselves suggest, small structural modifications in the bifunctional agents can dramatically affect their activity and selectivity.

We have now found a novel class of bis-acridine derivatives which show a marked antitumor activity, predominantly in solid tumors such as the colon tumor, compared with the compounds of the prior art. In particular, the novel bifunctional derivatives are distinguished in that they have acridine, acridone, pyrazole-acridone, pyrimido-acridone and pyrazole- pyrimidoacridone systems bearing alkylamino side chains at the para position to the polyamino chain linking the two units . DISCLOSURE OF THE INVENTION

The present invention relates to compounds of general formula (I):

A-NH-Z-NH-A' wherein:

Z is a (C2-C ) alkylene chain or a group of formula (CH₂)_n-N(B)-(CH₂)_q-N(B^,)-(CH₂)m-(CH₂)_n-N(B)-X-(CH₂)_m wherein: n and are independently an integer from 2 to 4,

B is hydrogen or (C-^-C^-alkyl,

X is a single bond or a group of formula

-(CH₂)_p-N(B' )- wherein: q is an integer from 2 to 4 and B' is hydrogen or (C₁-C₄) alkyl

A and A' , which are the same or different, are selected from:

in which:

R is hydrogen, or one or more substituents which are the same or different, selected from straight or branched (C^-Cg)alkyl, -OH, -NH , (C^-C^)alkoxy , mono- and di-(C₁-C^)alkylamino, chlorine, bromine, iodine and fluorine, nitro, mono- and poly-fluoro( C.,-C₄)alkyl, (C^-C₄ )alkylsulfonyl, (C^-C₄ )alkylaminosulfonyl;

Y is a -(CH )_D~T group; p is the integer 2, 3 or 4; - T is selected from -NH , -OH, mono- or di-(C^-C₄)- alkylamino, hydroxyethylamino , N-morpholinyl , N-pi- perazinyl, N-thiomorpholinyl , N-pyrrolidinyl, N-pipe- ridinyl, possible stereoisomers or stereomeric mixtures and salts thereof with pharmaceutically acceptable acids.

A second object of the present invention is a process for the preparation of the compounds of formula

(I).

A further object of the present invention are pharmaceutical formulations containing an effective dose of at least one compound of formula ( I ) in admixture with pharmaceutically acceptable excipients, as well as the use of said compounds of formula (I) and of the related pharmaceutical formulations as antitumor agents. Preferred compounds of formula (I) are those in which n and m are the integer 3.

Particularly preferred compounds are those in which A = A' is a group of formula (b), (c) or (e).

Most preferred compounds are those in which E is a methyl group. PREPARATION OF THE COMPOUNDS OF THE INVENTION

The compounds of formula (I) in which A s A' is a group of formula (b) (in the following defined compounds

(I)-(b); the compounds of formula (I)-(a), (I)-(c), (I)- (d), (I)-(e) and (I)-(f)) will be defined analogously) can be prepared according to the synthetic scheme reported in Figure 1, which involves the following synthetic steps:

(1) reaction of two equivalents of acridone of formula (III) with an equivalent of polyamine of formula (II) to give a compound of formula (IV). Said reaction can be carried out in the presence of a condensing agent, such as dicyclohexylcarbodiimide, or after activation of the carboxylic group of the intermediate of formula (III), for example via imidazolide (by reaction with 1,1'- carbonyldii idazole) , mixed anhydride (by reaction with ethyl chloroformate in the presence of triethylamine) or N-hydroxysuccinimidoester (by reaction with N- hydroxysuccinimide in the presence of morpholinoethyl isonitrile) .

(2) reaction of the dimer obtained at step (1) with at least 2 equivalents of an amine of formula Y-NH in a solvent such as ethoxyethanol, in the presence of a base such as triethylamine and at a temperature up to 150 "C to give compounds of formula (I)-(b).

Compounds of formula (I)-(d) can be obtained according to the synthetic scheme reported in Figure 2, which comprises reacting a compound of formula (IV) with at least 2 equivalents of hydrazine of formula Y-NH-NH in a solvent such as ethoxyethanol, in the presence of a base such as triethylamine and at a temperature up to 150°C.

The compounds of formula (I)-(c) can be prepared according to the synthetic scheme reported in Figure 3, which comprises reacting at least two equivalents of the intermediate of formula (V) with an equivalent of the polyamine of formula (II) in a solvent such as ethoxyethanol, in the presence of a base such as triethylamine and at a temperature up to 150°C.

The compounds of formula (I)-(a) can be prepared according to the synthetic scheme reported in Figure 4, by reducing the corresponding compounds of formula (I)- (c) with aluminum amalgam in ethanol/water .

The compounds of formula (I)-(e) can be prepared according to the synthetic scheme reported in Figure 5, which comprises the following synthesis steps:

(3) reaction of an intermediate of formula (V) with phosgene in a solvent and in the presence of a base, such as triethylamine, preferably at temperatures ranging from 0°C to room temperature, to give an intermediate of formula (VI);

(4) reaction of at least two equivalents of an intermediate of formula (VI) with an equivalent of the polyamine of formula (II) in a solvent such as ethoxyethanol, in the presence of a base such as triethylamine and at a temperature up to 150°C, to give the compounds of formula (I)-(e).

The compounds of formula (I)-(f) can be prepared by reduction of the compounds of formula (I)-(b) with aluminum amalgam in ethanol/water. The compounds of formula (I) in which A and A' are different are prepared by a reaction sequence involving a first condensation between, for instance, a compound of formula A-W wherein A is as defined above and W is a halogen atom or a carboxy-activating group, and a diamine of formula NH₂-Z-NH₂, wherein Z is as defined above, followed by a second reaction between the product resulting from the first reaction and a compound of formula A'-W, wherein A' and W are as defined above.

The first reaction is usually carried out using an at least 1:2 excess of the diamine compound in a solvent such as 2-ethoxyethanol, ethanol, methanol, ethyl acetate, dimethylsulfoxide, dimethylformamide .

The intermediates of formula (III) and (V) can be prepared according to the synthetic scheme reported in Figure 6, which comprises the following synthetic steps: (5) condensation of a 2-halocarboxylic acid with 5- chloro-2-carboxy aniline (or the corresponding methyl carboxylate ) in the presence of copper or of the salts thereof and in basic medium. Examples of said reactions are the use of copper acetate in the presence of a tertiary amine or the use of copper and of an alkali metal carbonate. The reaction is preferably carried out in a solvent, such as an alcohol or a dipolar aprotic solvent. Preferred solvents are N-methylpyrrolidone and isopentyl alcohol. (6) cyclization of the intermediate obtained in (5) to give a iso eric mixture of l-chloro-9-oxo-9 ,10- dihydroacridine-4-carboxylic and 6-chloro-9-oxo-9 ,10- dihydroacridine-4-carboxylic acids (or of the corresponding methyl carboxylates) in an about 2 : 1 ratio. The cyclization is carried out in the presence of condensing agents such as polyphosphoric acid or phosphorous oxychloride, the latter preferably in the presence of a solvent such as an aromatic hydrocarbon.

(7) basic hydrolysis of any methyl carboxylates obtained in (6) to give the corresponding acids of formula (III ) . '

(8) condensation of the acid derivative obtained in (6) or (7) with a suitable amine of formula H N-Y, to give the intermediates of formula (V). Said reaction can be carried out in the presence of condensing agents such as carbodiimides or after activation of the carboxylic group, for example by formation of the imidazolide (by reaction with carbonyldii idazole ) , of the acid chloride (by reaction with thionyl chloride) or of the hydroxysuccinimido ester (by reaction with N-hydroxy- succinimide in the presence of morpholinoethylisonitrile) .

When the group Y present on the amines or hydrazines used in the synthesis described above contains i its turn primary and secondary amino groups or hydroxy groups, these will be protected before the condensation reaction, by conventional reactions with protective groups. After the condensation reaction, said protective groups will be removed.

Examples of protection and deprotection reactions of primary and secondary amines- protective groups and of alcohols- protective groups are described in Green, T.W., Wuts, P.G.M., "Protective Groups in Organic Synthesis", second edition, John Wiley & Sons, 1991.

The polyamines of formula (II) are compounds widely known and described for example in the already cited Cholody W.M. et al . , J. Med. Chem., 38, 3043-52 (1995) and Hernandez L. et al., Cancer Res., 55, 2338-45

(1995), which are herein incorporated by reference.

The resulting compounds of formula (I) can be trasformed into other compounds of formula (I) which are also included within the meanings of formula (I) by deprotection reactions of the protective groups, such as transformation of compounds of formula (I) with R = methoxyl into compounds with R = hydroxyl by cleavage reaction in acidic conditions (48% hydrobromic acid), or for example, when R is a nitro group, by reduction to amine and subsequent optional functionalization of the amine via reductive alkylation or by reaction with an alkylsulfonyl chloride.

The compounds of formula (I) can also be salified by reaction of the free base with pharmaceutically acceptable acids, preferably with hydrochloric acid. BIOLOGICAL ACTIVITY OF THE COMPOUNDS OF THΞ INVENTION

The compounds of the invention were tested for their antitumor activity on the human HT 29 cell line of colon adenocarcinoma. The cells were incubated for 144 hours with the test compound, then cytotoxicity was evaluated using the "MTT Assay" (Mosman, T. "Rapid Colorimetric Assay for Cellular Growth and Survival: Application to Proliferation and Cytotoxicity Assay"; J. Immunol. Methods, (1983), 65, 66-63; Green, L.M., "Rapid Colorimetric Assay for Cell Viability; Application to the Quantitation of Cytotoxic and Growth Inhibitory Lymphokines", J. Immunol. Methods, (1984), 70, 257-268). The results are expressed as IC₅₀ (μg/ml), i.e. the concentration of the test compound causing the death of 50% of the cell population. The data concerning some representative compounds of the invention are reported in Table.

Table - Cytotoxic activity of the compounds of the invention on the human HT 29 cell line of colon adenocarcinoma '

A-NH-( CH₂ )_n-NMe- ( CH₂ )_m~NH-A'

R n = m Y A = A' IC₅₀ (μg/ml)

7-OMe 3 -(CH₂)₂NMe_? (b) 0.00073

7-N0₂ 3 -(CH₂)₂NMe₂ (c) <0.0001

H 3 -(CH₂)₂NMe_? (c) 0.00037

H 3 -(CH₂)₂NMe₂ (d) 0.001

H 3 -(CH₂)₂NMe₂ (e) <0.0001

H 3 -(CH₂)₂NMe₂ (a) 0.038

mitoxantrone 0.005

The data reported in Table clearly show that the compounds of the invention have an activity comparable to, if not higher than, mitoxantrone.

The compounds of formula (I), when administered to man and animals bearing tumors susceptible of treatment with DNA intercalating agents, at dosages ranging from 1 mg to 1200 mg per square metre of body area, are capable of inducing the regression of the above mentioned tumors .

The effective dosage of the compounds of the invention can be determined by the expert clinician with conventional, known methods. The relationship between the dosages used for animals of various species and those for humans (on the basis of mg/m2 of body area) is described by Freirich,

E.J. et al., Cancer Chemother. Rep., 50, n.4, 219-244,

May 1966.

Tumors which can be treated with the compounds of the present invention are those sensitive to the therapy with DNA intercalating agents.

In particular, colon tumor can advantageously be treated.

The pharmaceutical compositions containing the compounds of formula (I) are comprised in the invention.

These pharmaceutical compositions can contain any amounts of compounds of formula (I) capable of exerting in mammals an antitumor activity against tumors sensitive to the therapy with DNA intercalating agents. The pharmaceutical compositions can contain, in addition to at least one compound of formula (I), pharmaceutically compatible excipients, so as to allow the administration through any route, such as the oral, parenteral, intravenous, intradermal, subcutaneous or topical routes, in the liquid or solid form.

An administration route for the compounds of formula (I) is the oral one. Oral compositions will usually include an inert diluent or an edible carrier.

They can be included in gelatin capsules or compressed into tablets. Other forms for the oral administration are capsules, pills, elixirs, suspensions or syrups.

Tablets, pills, capsules and similar compositions can contain the following ingredients (in addition to the active ingredient): a ligand, such as microcrystalcell lineulose, gum tragacanth, or gelatin; an excipient such as starch or lactose; a disintegration agent such as alginic acid, primogel, maize starch, and the like; a lubricant such as magnesium stearate; a fluidifier such as colloidal silicon dioxide; a sweetening agent such as sucrose or saccharin or a flavour such as mint essence, methyl salicylate or orange flavour. When the selected composition is in the form of capsules, it can contain in addition a liquid carrier such as a fatty oil. Other compositions can contain various materials which modify their physical state, for example coating agents (for tablets and pills) such as sugar or shellac. The materials used in the preparation of the compositions will be pharmaceutically pure and non toxic at the used dosages.

For the preparation of pharmaceutical compositions for the parenteral administration, the active ingredient can be incorporated in solutions or suspensions, which can also include the following components: a sterile diluent such as water for injections, physiological saline, oils, polyethylene glycols, glycerin, propylene glycol or other synthetic solvents; antibacterials such as benzyl alcohol; antioxidizers such as ascorbic acid or sodium bisulfite; chelating agents such as ethylenediamino tetraacetic acid; buffers such as acetates, citrates or phosphates and agents for adjusting the tonicity of the solution, such as sodium chloride or dextrose. The parenteral preparation can be included in glass or plastic ampoules, mono-dose syringes or vials.

The invention is further described by the following examples.

Preparation 1 - l-Chloro-7-nitro-9~oxo-9 , 10- dihydroacridine-4-carboxylic acid

2-Amino-4-chlorobenzoic acid (7 g), 2-chloro-5- nitrobenzoic acid (7 g), potassium carbonate (7 g) and copper powder (0.25 g) are suspended in 150 ml of isopentyl alcohol. After refluxing the reaction mixture under stirring for about 20 minutes, an orange-red mass precipitates. The reaction is continued for 4-5 hours, then 150 ml of a 1M potassium carbonate aqueous solution are added and insolubles are filtered off while hot. The aqueous phase is separated, acidified to pH 5 with 2M hydrochloric acid to precipitate the product, which is filtered, suspended in boiling methanol (100 ml), filtered, resuspended in boiling water (200 ml), filtered again and washed with ethanol. The resulting diacid is dissolved in xylene (55 ml), added with 55 ml of phosphorous oxychloride and refluxed for 2 hours. After cooling, the precipitate is filtered, suspended in boiling methanol, filtered, washed with ethyl ether and dried to give 5.8 g of a mixture of l-chloro-7-nitro-9-oxo-9 ,10-dihydroacridine- 4-carboxylic acid and 6-chloro-2-nitro-9-oxo-9,10- dihydroacridine-4-carboxylic acid in an about 2 : 1 ratio.

Preparation 2 - 2-( 5-Chloro-2-methoxycarbonyl- phenylamino)-5-methoxybenzoic acid

A suspension of methyl 2-amino-4-chlorobenzoate (0.8 g), 2-bromo-5-methoxybenzoic acid (1 g) and copper(II) acetate onohydrate (0.86 g) in 5 ml of N- methylpyrrolidone and 10 ml of N,N-diisopropylethylamine is heated at 160°C under stirring for 6 hours. After cooling, dilution with water and acidification to pH 2, a solid separates, which is recovered by filtration and dried. The solid is then suspended under stirring in ethyl ether and filtered to give 0.75 g of product, m.p.

176-177°C (crystallized from methanol). ¹H-NMR in' d6-DMSO: 3.80 ppm (s, 3H ) ; 3.88 ppm (s,

3H); 6.87 ppm (d, 1H); 7.03-7.21 ppm (m, 2H); 7.33-7.57 ppm (m, 2H); 7.89 ppm (d, 1H); 10.42 ppm (s, 1H); 11.29 ppm (br s, 1H) .

Preparation 3 - Methyl l-chloro-7-methoxy-9-oxo- 9 , 10-dihydroacridine-4-carboxylate

1 g of 2-( 5-chloro-2-methoxycarbonylphenylamino)-5- ethoxybenzoic acid is refluxed v/ith PPE (40 g) in 50 ml of chloroform until the whole solid is dissolved. Chloroform is evaporated off to obtain an oil which is heated to 100°C for one hour. The mixture is carefully diluted v/ith 5 ml of methanol and 10 ml of water, then extracted with chloroform (3 x 20 ml). The combined organic extracts are dried and the residue is purified by flash-chromatography on silica gel (eluent chloroform/benzene 3 : 2) to give 0.58 of product, m.p. 219-220°C.

^^■H-NMR in CDC1₃: 3.92 ppm (s, 3H ) ; 4.01 ppm (s, 3H); 7.20 ppm (d, 1H); 7.25-7.33 ppm (m, 2H); 7.81 ppm ( , 1H); 8.24 ppm (d, 1H); 12.10 ppm (s, 1H). Preparation 4 - Methyl l-chloro-6 ,7-dimethoxy-9- oxo-9 , 10-dihγdroacridine-4-carboxylate .

A suspension of methyl 2-amino-4-chlorobenzoate (0.7 g, 3.83 mmol), commercially available 2-bromo-4,5- dimethoxybenzoic acid (1 g, 3.83 mmol), and Cu(OAc)₂«H₂0 (0.77 g, 3.83 mmol) in l-methyl-2-pyrrolidinone (5 L) and JV^-V-diisopropylethylamine (10 mL) is stirred for 8 h at 160^βC. After cooling, dilution with water, and acidification to pH 2, the collected precipitate is stirred with hot MeOH, then with Et₂0, filtered and dried. The solid obtained (0.32 g) is refluxed with PPΞ (15 g) in CHCI3 (25 mL) until all solid is dissolved.

The reflux condenser is removed, and CHCI3 is evaporated off, to give an oil v/hich is heated for 1 h at 100°C.

After cooling, the mixture is diluted cautiously with

MeOH (5 mL) and water (50 mL) and then extracted with CHCI3 (3 x 20 mL). The concentrated chloroform extracts are chromatographed on silica gel using CHCl₃/CgHg (9:1 v/v) to yield the title product (0.19 g, 63.3%): C₁₇H₁₄C1N0₅; m.p. 277-278°C; NMR (CDCI3) δ 3.95-4.00 (m, 6H, 2 x O-CH3), 6.70 (ε, Hi, ar ) , 7.18 (d, 1H, ar ) , 7.75 (s, 1H, ar), 8.20 (d, 1H, ar ) , 12.04 (br s, 1H, 10- H, ex).

Preparation 5 - l-Chloro-7-methoxy-9-oxo-9 ,10- dihydroacridine-4-carboxylic acid

A suspension of methyl l-chloro-7-methoxy-9-oxo- 9,10-dihydroacridine-4-carboxylate (1 g) in 100 ml of ethanol and 100 ml of 2M sodium hydroxide is refluxed for 30 minutes. The reaction mixture is acidified with 4M hydrochloric acid and stirred at room temperature for 20 minutes. The precipitate is recovered by filtration, washed with water, methanol and ethyl ether to give 0.72 g of pure product, m.p. 340-341°C.

¹H-NMR in d6-DMS0: 3.89 ppm (s, 3H ) ; 7.28 ppm (d, 1H); 7.43 ppm (m, 1H); 7.59 ppm (d, 1H); 7.75 ppm (d, 1H); 8.30 ppm (d, 1H); 12.35 ppm (s, 1H ) . Preparation 6 - l-Chloro-6 , 7-dimethoxy-9-oxo-9,10- dihydroacridine-4-carboxylie acid The ester prepared in preparation 4 (0.7 g, 2.01 mmol) is suspended in ΞtOH (70 L) and 2 M NaOH (70 mL ) and heated under reflux for 30 min. After cooling, the resulting mixture is acidified with 4 M HCl and stirred at room temperature for 20 min; the precipitate is collected and washed v/ith water, MeOH, and Ξt₂0 to give the crude title compound (0.5 g, 74.6%) that is used for the subsequent step without further purification.

Preparation 7 - 4-[N-[3-(dimethylamino)propyl] ]- carboxamido-l-chloro-9-oxo-9,10-dihydroacridine

A mixture of 1 g of l-chloro-9-oxo-9 ,10- dihydroacridine-4-carboxylic acid (obtained according to Rewcastle et al., Synthesis, 1985, 220-2) and 1.1 g of 1 ,1 ' -carbonyldiimidazole in 10 ml of dimethylforma ide is stirred, if necessary with heating, until complete dissolution. The mixture is cooled to 10°C and 1.02 g of 3-dimethylaminopropylamine are added. After 15 minutes at room temperature, the mixture is partitioned between chloroform and 1M sodium carbonate. The organic phase is worked up to give the product which solidifies by treatment with ethyl ether (0.86 g), m.p. 150-151°C.

¹H-NMR in CDC1₃: 1.85 ppm (m, 2H ) ; 2.38 ppm (s, &H); 2.63 ppm (t, 2H); 3.60 ppm (q, 2H); 7.16 ppm (d, 1H); 7.22-7.38 ppm (m, 2H); 7.59-7.69 ppm (m, 2H); 8.40 ppm (dd, 1H); 8.57 ppm (s, 1H); 13.14 ppm (s, 1H).

The following compounds are prepared analogously: 4-[N-[2-(dimethylamino)ethyl] ]carboxamido-l- chloro-7-methoxy-9-oxo-9 ,10-dihydroacridine, m.p. 170- 172 °C, iH- MR in CDCI3: 2.43 ppm (s, 6H); 2.76 ppm (t, 2H); 3.65 ppm (q, 2H ) ; 3.93 ppm (s, 3H); 7.19 ppm (d, 1H); 7.25-7.34 ppm (m, 2H ) ; 7.62 ppm (br ε, 1H); 7.78- 7.89 ppm (m, 2H); 12.77 ppm (s, 1H);

4-[N-[2-( tert-butoxycarbonylamino)ethyl] ]car- boxyamido-l-chloro-9-oxo-9,10-dihydroacridine, m.p. 212-

213 °C, -NMR in dδ-DMSO: 1.42 ppm (s, 9H); 3.25 ppm (t, 2H); 3.38 ppm (t, 2H); 7.01 ppm (t, 1H); 7.28-

7.42 ppm (m, 2H); 7.60-7.82 ppm (m, 2H); 8.08-8.25 ppm (m, 2H); 9.02 ppm (t, 1H); 12.80 ppm (s,

1H).

Preparation 8 - 4-{iV-[2-(Dimethylamino)ethyl]- carbamoyl}-l-chloro-6 , 7-dimethoxy-9-oxo-9 , 10-dihydroa- cridine : mp 221-222°C; ¹H NMR (CDC1₃) δ 2.35 (s, 6H, 2 x CH₃), 2.60 (t, 2H, CH₂), 3.50-3.60 (m, 2H, CH₂ ) , 4.00- 4.01 (m, 6H, 2 x O-CH3 ) , 6.71 (s, 1H, ar ) , 7.18 (d, 1H, ar), 7.30 (br ε, 1H, CO-NH, ex), 7.68 (d, 1H, ar ) , 7.80 (s, 1H, ar), 12.72 (br s, 1H, 10-H, ex).

Preparation 9 - 4-[N-[2-(dimethylamino )ethyl] ]- carboxyamido-l-chloro-7-nitro-9-oxo-9 , 10-dihydroacridine hydrochloride

An isomeric mixture of l-chloro-7-nitro-9-oxo-9,10- dihydroacridine-4-carboxylic acid and 6-chloro-2-nitro- 9-oxo-9,10-dihydroacridine-4-carboxylic acid (1 g of mixture; preparation 1) and anhydrous triethylamine (0.314 g) in 30 ml of chloroform, cooled to 0"C and stirred, is added drop by drop with a solution of ethyl chloroformate (0.34 g) in 20 ml of chloroform. After 1 hour under stirring at room temperature, the mixture is cooled to 0°C and added with N,N-dimethylethylendiamine (0.32 g), then kept at room temperature and under stirring overnight. The precipitated solid is filtered and v/ashed with methanol, chloroform and finally with ethyl ether, to give 0.58 g of product, m.p. 295-297°C (crystallized from methanol). iH-NMR in d6-DMSO: 2.88 ppm (s, 6H); 3.34 ppm (t,

2H); 3.75 ppm (q, 2H); 7.33 ppm (d, IH); 7.90 ppm (d,

IH); 8.30 ppm (d, IH); 8.49 ppm (dd, IH); 8.93 ppm (d, IH); 9.47 ppm (t, IH); 10.03 ppm (br s, IH); 11.03 ppm

(br m, IH) .

Preparation 10 - 6-chloro-2-[3-(dimethylamino)pro- pyl]pyrimido[5, 6 ,l-de]acridine-l , 3 ,7-dione

A mixture of l-chloro-4-[N-[3-(dimethylamino)pro- pyl] ]carboxamido-9-oxo-9,10-dihydroacridine (0.33 g) in 20 ml of chloroform and 1 ml of triethylamine is added drop by drop with a solution of phosgene (2 ml of 20% solution in toluene) in 10 ml of chloroform, keeping the temperature at 0°C. When the addition is completed, the mixture is stirred at room temperature for 20 minutes, then the reaction mixture is partitioned between chloroform and IM sodium carbonate. The organic phase is worked up to give a residue which is chromatographed on a silica gel column (eluent chloroform/methanol 19 : 1), to give 280 mg of product, m.p. 212-214°C (dec. ) .

¹H-NMR in CDC1₃: 2.31-2.51 ppm (m, 2H); 2.85 ppm (s, 6H); 3.20 ppm (t, 2H); 4.35 ppm (t, 2H); 7.47-7.80 ppm (m, 3H); 8.28-8.60 ppm (m, 3H). Preparation 11 - 6-chloro-2-[2-(dimethylamino)- ethyl]-9-methoxy-l,3,7-trioxo[lH,2H,3H,7H,12H]pyri- mido[5, 6 ,1 ,-de]acridine

A solution of 4-{N-[2-(dimethylamino)ethyl] }- carboxamiido-l-chloro-7-methoxy-9-oxo-9 , 10-dihydroacri- dine (700 mg) and Et₃ (2ml) in CHC1₃ (10 ml), is added drop by drop with C0C1 (20% in toluene, 3.7 ml) in CHC1₃ (10 ml) at 0^oC. When the addition is completed, the mixture is stirred at room temperature for 20 minutes. The reaction mixture is partitioned between

CHC1₃ and IM Na₂C0₃. The organic phase is worked up to give a residue which is suspended in ethyl ether (20 ml), stirred for 10 minutes, then filtered to obtain 620 mg of a solid, which is slightly impure by TLC and is used as it is for the subsequent reactions, but is transformed into hydrochloride for characterization: m.p. > 300°C (crystallized from EtOH); ¹H-NMR (DMSO-dg) 2.90 ppm (s, 6H), 3.43-3.56 ppm (m, 2H), 3.95 ppm (s, 3H), 4.42 ppm (t, 2H), 7.50 ppm (d, IH), 7.64 ppm (s, IH), 7.78 ppm (d, IH), 8.43 ppm (d, IH), 8.54 ppm (d, IH), 10.16 ppm (br s, IH). Preparation 12 - 6-Chloro-2-[2-(dimethylamino)- ethyl] -9 , 10-dimethoxy-pyrimido[ 5 , 6 , 1-de] acridine-1 ,3,7- trione

To a solution of 4-{N-[2-(dimethylami- no )ethyl ] carbamoyl }-l-chloro-6 , 7-dimethoxy-9-oxo-9 ,10- dihydroacridine (0.2 g , 0.49 mmol) in CHC1₃ (10 mL ) and anhydrous triethylamine (0.5 mL) C0C1₂ is added (20% in toluene, 1.26 mL, 2.52 mmol) in CHC1₃ (10 mL ) dropwise under stirring at 0°C. The resulting mixture is stirred for 30 min at room temperature and then partitioned between CHC1₃ and aqueous 1 M a₂C0₃. The organic layer is worked up to give a residue that is purified by column chromatography on silica gel using CHC1₃/CH₃0H (19:1 v/v) to give the title product (0.14 g, 66.7%): C₂₁H₂₀ClN₃O₅; m.p. 214-215°C; ^ NMR (CDCI3) δ 2.35 (s, 6H, 2 x CH₃), 2.70 (t, 2H, CH₂ ) , 4.00-4.03 (m, 6H, 2 x O-CH3), 4.34 (t, 2H, CH₂), 7.57 (d, IH, ar ) , 7.74 (s, IH , ar ) , 8 . 15 ( s , IH , ar ) , 8 . 40 ( d , IH , ar ) .

Preparation 13 - JY4-[2-(Dimethylamino)ethyl]-l-( {3- [ ( 3-aminopropyl)- (methyl )amino]propyl}-amino )-9-oxo- 9 , 10-dihydro-4-acri-dinecarboxamide A suspension of l-chloro-N-4-[2-(dime- thylamino )ethyl]-9-oxo-9 , 10-dihydro-4-acridinecarboxa- ide, disclosed in J. Med. Chem., 1995, 38, 3282-3286 (0.3 g, 0.97 mmol), 3 , 3 ' -diamino-JV-methyldipropylamine (0.32 mL, 1.96 mmol) and triethylamine (0.5 L ) in 2- ethoxyethanol (10 mL) is stirred at 80°C for 24 h. After cooling the mixture is partitioned between CHCI3 and aqueous 1 M a₂Cθ3. The organic layer is worked up to give a residue that is purified by column flash- chromatography on silica gel using CHCl₃/MeOH (1:1, v/v) and 32% aqueous NH₃ (20 mL for 1 L of eluent) to give the title product as an oil (0.22 g, 50%): C25^H36^N6°2^; NMR (CDCI3) δ 1.48-1.65 (m, 2H, CH₂ ) , 1.72-1.92 (m, 2H, CH₂), 2.15 (s, 3H, CH3), 2.22 (s, 6H, 2 x CH3 ) , 2.28-2.52 (m, 6H, 3 x CH₂ ) , 2.68 (t, 2H, CH, ) , 3.15-3.28 (m, 2H, CH₂), 3.40-3.51 (m, 2H , CH₂ ) , 6.10 (d, IH, ar ) , 7.00 (br t, IH, CO-NH, ex), 7.15 (t, IH , ar ) , 7.28 (d, IH, ar), 7.50 (d, IH, ar), 7.60 (t, IH, ar ) , 8.25 (d, IH, ar), 10.87 (br t, IH, ex), 13.39 (br s, IH, ex).

Example 1 - 1 ,7-Bis(l-chloro-9-oxo-9 ,10-dihydro- acridine-4-carbonyl)-4-methyl-l ,4, 7-triazaheptane l-Chloro-9-oxo-9 , 10-dihydroacridine-4-carboxylic acid (500 mg) is suspended in CHCI3 (20 ml), added with Et₃N (0.26 ml), cooled to 0°C, then added drop by drop with ClCOOEt (0.26 ml) in CHCI3 (10 ml). The mixture is stirred at room temperature for 1 hour, cooled again to 0°C, added with N²-methyldiethylenetriamine (0.12 ml) and stirred at room temperature for 4 hours. The resulting precipitate is filtered and treated with boiling MeOH (20 ml), to obtain 230 mg of product, m.p.

> 300°C; -NMR (DMSO-dg) 2.80-3.11 ppm (m, 7H), 3.53- 3.75 ppm (m, 4H), 7.09 ppm (d, 2H), 7.35 ppm (d, 2H),

7.52 ppm (t, 2H), 8.00-8.11 ppm (m, 4H), 9.20 ppm (br t,

2H), 9.68 ppm (br s, IH), 12.50 ppm (s, 2H).

The following compounds are prepared analogously:

1 , 9-Bis ( l-chloro-9-oxo-9 , 10-dihydroacridine-4-car- bonyl)-5-methyl-l,5,9-triazanonane, m.p. 245-247°C (dec); iH- MR (DMSO-dg) 1.92-2.18 ppm ( , 4H), 2.80 ppm (ε, 3H), 3.04-3.30 ppm (m, 4H), 3.40-3.51 ppm (m, 4H), 7.20-7.34 ppm (m, 4H), 7.55 ppm (d, 2H), 7.70 ppm (t, 2H), 8.08-8.21 ppm (m, 4H), 9.28 ppm (br t, 2H), 10.21 ppm (br s, IH), 12.80 ppm (s, 2H);

1 , 7-Bis ( l-chloro-7-methoxy-9-oxo-9 , 10-dihydroacri- dine-4-carbonyl )-4-methyl-l , 4 , 7-triazaheptane ( after completion of the reaction, the mixture is partitioned between CHCI3 and a₂Cθ3 IM. The organic phase is worked up to give a residue which is chromatographed on a silica gel column eluted with CHCl₃/MeOH (9:1)), m.p. 155-156°C; ¹H-NMR (DMSO-dg) 2.35 ppm (s, 3H), 2.60-2.72 ppm ( , 4H), 3.40-3.56 ppm ( , 4H), 3.79 ppm (s, 6H), 7.00 ppm (d, 2H), 7.24 ppm (d, 2H ) , 7.38-7.56 ppm (m, 4H), 7.87 ppm (d, 2H), 8.78 ppm (br t, 2H), 12.54 ppm (s, 2H);

1 , 9-Bis(l-chloro-7-methoxy-9-oxo-9 , 10-dihydroacri- dine-4-carbonyl )-5-methyl-l , 5 , 9-triazanonane ( after completion of the reaction, the mixture is partitioned between CHCI3 and Na₂C0₃ IM. The organic phase is worked up to give a residue which is chromatographed on a silica gel column eluted with CHCl₃/MeOH (4:1)), m.p.

158-159°C; ^- MR (CDClg) 1.90-2.09 ppm (m, 4H), 2.48 ppm (s, 3H), 2.69-2.89 ppm (m, 4H), 3.49-3.67 ppm (m,

4H), 3.88 ppm (s, 6H), 6.81 ppm (d, 2H), 7.15-7.24 ppm ( , 4H), 7.57-7.66 ppm (m, 4H), 8.20 ppm (br t, 2H),

12.44 ppm (s, 2H) .

JV4- ( 3-{ [ ( l-Chloro-9-oxo-9 , 10-dihydro-4-acridinyl )- carbonyl] amino}propyl )-l-chloro-9-oxo-9 , 10-dihydro-4- acridinecarboxamide: 0.42 g, 97.6%): m.p. >300°C; NMR (DMSO-dg) δ 1.90-2.10 (m, 2H, CH₂), 2.91-3.09 (m, 4H, 2 x CH₂ ) , 7.12-7.28 (m, 4H, ar), 7.48 (d, 2H, ar), 7.70 (t, 2H, ar), 8.02-8.40 (m, 6H, 4 ar + 2 x CO-NH, ex), 15.65 (s, 2H, ex) . JV4-{2-[{2-[(2-{[(l-Chloro-9-oxo-9,10-dihydro-4- acridinyl ) carbonyl ]amino}-ethyl ) (methyl )amino]ethyl}- (methyl )amino]ethyl}-l-chloro-9-oxo-9 , 10-dihydro-4- acridinecarboxamide

Example 2 - 1 ,7-Bis[l-( 2-(dimethylamino)ethyl )- amino-9-oxo-9 , 10-dihydroacridine-4-carbonyl]-4-methyl- 1,4,7-triazaheptane (compound (I)-(b))

The bis-derivative 1 , 7-Bis ( l-chloro-9-oxo-9 ,10- dihydroacridine-4-carbonyl )-4-methyl-l , 4 , 7-triazaheptane hydrochloride (150 mg) is refluxed in 2- (dimethylamino)ethylamine (2 ml) for 1 hour, then cooled to room temperature and the reaction mixture is partitioned between CHCI3 and IM a₂C0₃. The organic phase is worked up to give a residue which is suspended in ethyl ether (10 ml) to give the product, pure by TLC (130 mg), m.p. 116-117°C; hydrochloride m.p. 238-240°C; -H-NMR (CDCI3) 2.31 ppm (s, 12H), 2.50 ppm (s, 3H), 2.60 ppm (t, 4H), 2.77-2.87 ppm (m, 4H), 3.54-3.65 ppm (m,

4H), 5.80 ppm (d, 2H), 6.78 ppm (br t, 2H), 7.10-7.21 ppm (m, 4H), 7.41-7.52 ppm ( , 4H), 8.28 ppm (d, 2H),

10.85 ppm (br t, 2H), 13.18 ppm (s, 2H). The following compounds are prepared analogously:

1 , 9-Bis[1- ( 2- (dimethyla ino )ethyl ) amino-9-oxo-9 , 10- dihydroacridine-4-carbonyl]-5-methyl-l , 5 , 9-triazanonane , m.p. 119-120°C; hydrochloride m.p. 226-228°C (dec); -

NMR (CDC1₃) 1.77-1.99 ppm (m, 4H), 2.28 ppm (s, 12H), 2.38 ppm (s, 3H), 2.51-2.68 ppm ( , 8H), 3.15-3.31 ppm

(m, 4H), 3.46-3.62 ppm (m, 4H), 6.05 ppm (d, 2H), 7.10- 7.26 ppm (m, 4H), 7.40-7.61 (m, 6H), 8.30 ppm (d, 2H), 10.91 ppm (br t, 2H), 12.36 ppm (s, 2H);

1 , 7-Bis [ 1- ( 2- ( dimethylamino ) ethyl ) ammo-7-methoxy- 9-oxo-9 , 10-dihydroacridine-4-carbonyl ] -4-methyl-l ,4,7- triazaheptane (the organic phase is worked up to give a residue which is flash-chromatographed on a silica gel column eluted with CHCl₃/Me0H/38% H₃ (1:1:0.01) to give the product), m.p. 217-218°C; hydrochloride m.p. 270- 273°C (dec); ifi- M (CDC1₃) 2.28 ppm (s, 12H), 2.40 ppm (s, 3H), 2.54 ppm (t, 4H), 2.72 ppm (t, 4H), 3.00-3.10 ppm (m, 4H), 3.50-3.63 ppm (m, 4H ) , 3.88 ppm (s, 3H), 5.72 ppm (d, 2H), 6.58 ppm (br t, 2H), 7.10-7.20 ppm (m, 4H), 7.41 ppm (d, 2H), 7.68 ppm (s, 2H), 10.90 ppm (br t, 2H), 13.10 ppm (s, 2H);

1 , 9-Bis [1- ( 2- ( dimethylamino ) ethyl ) amino-7-methoxy- 9-OXO-9 , 10-dihydroacridine-4-carbonyl ]-5-methyl-l ,5,9- triazanonane (the organic phase is worked up to give a residue which is flash-chromatographed on a silica gel column eluted with CHCl₃/Me0H/38% NH₃ (1:1:0.01) ), m.p. 143-144°C; hydrochloride m.p. 250-252°C (dec); iH- M (CDCI3) 1.82-1.88 (m, 4H), 2.28 ppm (s, 12H), 2.34 ppm

(s, 3H), 2.53-2.61 ppm (m, 8H), 3.20-3.28 ppm (m, 4H),

3.48-3.56 ppm (m, 4H), 3.86 ppm (s, 3H), 6.01 ppm (d,

2H), 7.13-7.21 ppm (m, 4H), 7.42 ppm (br t, 2H), 7.49 ppm (d, 2H), ^'7.69 ppm (s, 2H), 10.96 ppm (br t, 2H),

13.38 ppm (ε, 2H) .

JV4-( 3-{ [ (l-{ [2-(Dimethylamino)ethyl]amino}-9-oxo-

9, 10-dihydro-4-acridinyl)-carbonyl]amino}propyl)-l-{ [2-

( dimethylamino)ethyl]amino}-9-oxo-9, 10-dihydro-4-acri- dinecarboxamide: (0.11 g, 47.8%): m.p. 220-221"C; hydrochloride m.p. 198-200°C; NMR (DMSO-dg) δ 2.11- 2.31 (m, 14H, 4 x CH3 + CH₂ ) , 2.47-2.62 (m, 6H, 3 x CH₂), 2.63-2.75 (m, 6H, 3 x CH₂ ) , 6.20 (d, 2H, ar ) , 7.19 (t, 2H, ar), 7.40 (d, 2H, ar ) , 7.60 (t, 2H, ar ) , 8.09- 8.21 (m, 4H, ar), 10.41 (br t, 2H, CO-NH, ex), 15.80 (br s, 2H, ex) .

JV4-{ 2- [{2-C(2-{[( l-{[2- (Dimethylamino )ethyl ]amino}- 9-OXO-9 , 10-dihydro-4-acridinyl )carbonyl ] amino}ethyl ) ( methyl )amino]ethyl} (methyl )amino] ethyl}-l-{ [ 2- ( dimethyla- mino)ethyl] amino}-9-oxo-9, 10-dihydro-4-acridinecarboxa- mide: mp 268-270°C; NMR (DMSO-dg) δ 2.91 (s, 12H, 4 x CH₃), 3.00 (s, 6H 2 x CH3 ) , 3.25-3.59 (m, 8H, 4 x CH₂), 3.61-4.00 (m, 12H, 6 x CH₂ ) , 6.52 (d, 2H, ar ) , 7.23 (t, 2H, ar), 7.45 (d, 2H, ar ) , 7.64 (t, 2H, ar ) , 8.12 (d, 2H, ar ) , 8.30 (d, 2H, ar ) , 9.14 (br s, 2H, ex), 10.90 (br s, 2H, ex), 13.12 (br s, 2H, ex), 13.40 (br s, 2H, ex), 13.60 (br s, 2H, ex).

Example 3 - 1, 7-Bis[l-( 2- (dimethylamino)ethyl )- amino-7-hydroxy-9-oxo-9,10-dihydroacridine-4-carbonyl]- 4-methyl-l,4,7-triazaheptane

1 , 7-Bis [1-( 2- ( dimethylamino ) ethyl ) amino-7-methoxy- 9-OXO-9 , 10-dihydroacridine-4-carbonyl]-4-methyl-l ,4,7- triazaheptane(130 mg) is refluxed in 48% HBr (3 ml) for

1 hour, then cooled to room temperature and the reaction mixture is partitioned between CHC1₃ and IM Na₂C0₃. The organic phase is worked up to give a residue which is flash-chromatographed on a silica gel column eluted with

CHCl₃/MeOH/38% NH₃ (1:1:0.01) to give the product (80 mg), m.p. 162-163°C; hydrochloride m.p. 262-264°C (dec); iH-NMR (DMSO-dg) 2.23 (s, 12H), 2.34 ppm (s, 3H), 2.51- 2.74 ppm (m, 8H), 3.17-3.22 ppm (m, 4H), 3.42-3.46 ppm (m, 4H), 6.05 ppm (d, 2H), 7.16 ppm (d, 2H), 7.41-7.50 ppm (m, 4H), 7.89 ppm (d, 2H), 8.22 ppm (br t, 2H), 9.65 ppm (br s, 2H), 10.83 ppm (br t, 2H), 13.62 ppm (s, 2H). The following compound is prepared analogously: 1 , 9-Bis[l-( 2- (dimethylamino ) ethyl ) amino-7-hydroxy- 9-oxo-9 , 10-dihydroacridine-4-carbonyl ]-5-methyl-l ,5,9- triazanonane, m.p. 221-222°C; hydrochloride m.p. 250- 251°C; !H-NMR (DMSO-dg) 1.69-1.76 ppm ( , 4H), 2.19-2.21 ppm (m, 15H), 2.40 ppm (t, 4H), 2.52-2.56 ppm (m, 4H), 3.26-3.46 ppm (m, 8H), 6.18 ppm (d, 2H), 7.19 ppm (d, 2H), 7.40-7.50 ppm (m, 4H), 7.95 ppm (d, 2H), 8.48 ppm (br t, 2H), 9.69 ppm (br s, 2H), 10.83 ppm (br t, 2H), 13.70 ppm (s, 2H) .

Example 4 - 1 ,7-Bis[l-( 2- (dimethylamino ) ethyl )- aminoacridine-4-carbonyl]-4-methyl-l ,4,7-triazaheptane (compound ( I )-( f ) )

1 , 7-Bis [ 1- ( 2- ( dimethylamino )ethyl ) amino-9-oxo-9 , 10- dihydroacridine-4-carbonyl]-4-methyl-l , 4 , 7-triazaheptane trihydrochloride (350 mg) is refluxed in EtOH/H₂0 (3:1, 30 ml). Al pellets (350 mg) placed for 1 minute in a HgCl₂ ethanol εolution (1300 mg in 25 ml of ΞtOH) are added during 30 minutes. When the addition is completed, the mixture is refluxed for 30 minutes, then filtered while hot washing with EtOH (20 ml), diluted with H₂0

(50 ml) and added with FeCl₃ (1200 mg). The reaction mixture is partitioned between CHCI3 and IM Na₂C0₃. The organic phase is worked up to give a residue which is flash-chromatographed on a silica gel column eluted with

CHC13/MeOH/38% NH3 (1:1:0.01) to give the product (40 mg), m.p. 196-197^βC; hydrochloride m.p. 248-249°C (dec); iH-NMR (CDCI3) 2.46 ppm (s, 12H), 2.62 ppm (s, 3H),

2.75-2.90 ppm (m, 4H), 2.90-3.06 ppm (m, 4H), 3.30-3.44 ppm (m, 4H), 3.81-3.97 ppm (m, 4H), 5.74 ppm (br t, 2H),

6.27 ppm (d, 2H), 7.07 ppm (t, 2H), 7.32 ppm (t, 2H),

7.51 ppm (d, 2H), 7.71 ppm (d, 2H), 8.32 ppm (s, 2H), 8.50 ppm (d, 2H), 11.60 ppm (br s, 2H).

The following compound is prepared analogously: 1 , 9-Bis [1- ( 2- ( imethylamino ) ethyl ) aminoacridine-4- carbonyl]-5-methyl-l, 5 , 9-triazanonane , m.p. 166-167°C; hydrochloride m.p. 240-242°C (dec); ¹H-NMR (CDCI3) 2.01- 2.10 ppm (m, 4H), 2.29-2.50 ppm (m, 15H), 2.72-2.88 ppm (m, 8H), 3.31-3.46 ppm (m, 4H), 3.60-3.73 ppm (m, 4H), 6.14 ppm (br t, 2H), 6.44 ppm (d, 2H), 7.27 ppm (t, 2H), 7.60 ppm (t, 2H), 7.74 ppm (d, 2H), 7.90 ppm (d, 2H), 8.71 ppm (d, 2H), 8.88 ppm (s, 2H), 11.71 ppm (br s, 2H).

Example 5 - 1 , 7-Bis[2- (2- (dimethylamino )ethyl )- pyrazole[3,4, 5-cZ acridine-5-carbonyl]-4-methyl-l ,4,7- triazaheptane (compound (I)-(d))

1 , 7-Bis ( l-chloro-9-oxo-9 , lO-dihydroacridine-4- carbonyl)-4-methyl-l, 4, 7-triazaheptane monohydrochloride (270 mg) and 2- ( dimethylamino )ethylhydrazine (420 mg) are suspended in 2-ethoxyethanol (10 ml) at 120°C for 1 hour . The mixture is cooled to room temperature and partitioned between CHCI3 and IM Na₂C0₃. The organic phase is worked up to give an oily residue which is flash-chromatographed on a silica gel column eluted with

CHCl₃/MeOH/38% NH3 (1:1:0.02) to give the product as an oil, pure by TLC (70 mg), which is transformed into the hydrochloride by conventional methods for characterization: m.p. trihydrochloride 230-231°C; ^H- NMR (DMSO-dg) 2.88 ppm (s, 12H), 3.01 ppm (s, 3H), 3.32- 4.00 ppm (m, 12H), 4.50-4.78 ppm (m, 4H), 6.64 ppm (d, 2H), 7.10 ppm (t, 2H), 7.30 ppm (t, 2H), 7.44 ppm (d, 2H), 7.68-7.91 ppm (m, 4H), 8.82 ppm (br t, 2H), 10.40- 10.65 ppm (m, 3H), 11.00 ppm (br ε, 2H). The following compoundε are prepared analogously: 1, 9-Bis[2- (2- (dimethylamino) ethyl )pyrazole[ 3,4,5- &l]acridine-5-carbonyl]-5-methyl-l , 5 , 9-triazanonane (after chromatography the product is obtained as a pure solid residue, which is characterized as the free base), m.p. 159-160°C; hydrochloride m.p. 200-202°C (dec); ¹E- NMR (CDCI3) 1.82-1.97 ppm (m, 4H), 2.27 ppm (s, 12H), 2.37 ppm (s, 3H), 2.53-2.71 ppm ( , 4H), 2.77 ppm (t, 4H), 3.50-3.64 ppm (m, 4H), 4.29 ppm (t, 4H), 6.36 ppm (d, 2H), 6.97-7.12 ppm (m, 4H), 7.15-7.35 ppm (m, 6H), 7.91 ppm (d, 2H), 10.52 ppm (br ε, 2H);

1 , 7-3is [ 2- ( 2- ( dimethylamino ) ethyl )-9-methoxypyrazo- le[ 3,4, 5-;Z]acridine-5-carbonyl]-4-methyl-l, 4, 7-triazaheptane trihydrochloride, m.p. 245-247°C; ¹H-NMR (DMSO-dg) 2.86 ppm (s, 12H), 3.00 ppm (s, 3H), 3.30-3.75 ppm (m, 12H), 3.84 ppm (s, 6H) , 4.60-4.80 ppm (m, 4H), 6.56 ppm (d, 2H), 6.92 ppm (d, 2H), 7.20 ppm (ε, 2H), 7.40 ppm (d, 2H), 7.78 ppm (d, 2H), 8.71 ppm (br t, 2H),

10.30-10.54 ppm (m, 4H), 10.88 ppm (br s, IH);

1 , 9-Bis [2- ( 2- ( imethylamino )ethyl )-9-methoxypyra- zole [3,4, 5- kl]acridine-5-carbonyl ]-5-methyl-l , 5 , 9-tria- zanonane trihydrochloride, m.p. 204-205°C; ^-H-NMR (DMSO- dg) 1.90-2.15 ppm (m, 4H), 2.68-2.92 ppm (m, 15H), 3.00- 3.50 ppm (m, 8H), 3.51-3.70 ppm (m, 4H), 3.80 ppm (s, 6H), 4.69-4.85 ppm (m, 4H), 6.81 ppm (d, 2H), 6.97 ppm (d, 2H), 7.26 ppm (s, 2H), 7.52 ppm (d, 2H), 7.84 ppm (d, 2H), 8.56 ppm (br t, 2H), 10.69 ppm (br s, 2H), 10.83 ppm (br s, IH), 11.15 ppm (br s, 2H).

Example 6 - 1 ,7-Bis[2-( 2- (dimethylamino )ethyl)-9- hydroxypyrazole[3,4, 5- kl acridine-5-carbonyl]-4-methy1- 1,4, 7-triazaheptane 1 ,7-Bi [2-(2- (dimethylamino )ethyl )-9-methoxypyrazo- le[3 ,4, 5- kl] acridine-5-carbonyl]-4-methyl-l, 4, 7-triazaheptane trihydrochloride (200 mg) is refluxed in 48% HBr (3 ml) for 1 hour; the mixture is cooled to room temperature and partitioned between CHCI3 and IM a₂Cθ3- The organic phaεe is worked up to give a residue which is flash-chromatographed on a silica gel column eluted with CHCl₃/MeOH/38% NH3 (1:1:0.01) to give the product (40 mg), m.p. 282-283°C; hydrochloride m.p. 255-257°C (dec); ¹H-NMR (DMSO-dg) 2.18 ppm (s, 12H), 2.34 ppm (ε, 3H), 2.55-2.77 ppm (m, 8H), 3.30-3.50 ppm (m, 4H), 4.33 ppm (t, 4H), 6.54 ppm (d, 2H), 6.80 ppm (d, 2H), 7.18 ppm (ε, 2H), 7.36 ppm (d, 2H), 7.56 ppm (d, 2H), 8.03 ppm (br t, 2H), 9.45 ppm (br s, 2H), 10.56 ppm (s, 2H). The following compound is prepared analogously: 1 , 9-Bis [2- ( 2- ( dimethylamino )ethyl ) -9-hydroxypyrazo- le[3,4, 5-&1] acridine-5-carbonyl]-5-methyl-l, 5, 9-triaza- nonane, m.p. 239-240°C; hydrochloride m.p. 260-261^βC;

^-H-NMR (DMSO-d₆) 1.67-1.84 ppm (m, 4H), 2.20 ppm (s,

12H), 2.25 ppm (s, 3H), 2.39-2.52 ppm (m, 4H), 2.74 (t,

4H), 3.23-3.42 ppm (m, 4H), 4.36 ppm (t, 4H), 6.63 ppm (d, 2H), 6.81 ppm (d, 2H), 7.18 ppm (ε, 2H), 7.37 ppm

(d, 2H), 7.62 ppm (d, 2H), 8.21 ppm (br t, 2H), 9.45 ppm

(s, 2H) , 10.58 ppm (s, 2H).

Example 7 - 1, 7-Bis[4- (N-( 2-(dimethylamino)ethyl) )- carboxamido-9-oxo-9,l0-dihydroacridine-l-yl]-4-methy1- 1 ,4,7-triazaheptane (compound (I)-(c))

4-{N-[2- (Dimethylamino ) ethyl] }carboxamido-l-chloro- 9-oxo-9,10-dihydroacridine (500 mg), N²- methyldiethylenetriamine (0.1 ml) and Et₃N (0.5 ml) are placed in 2-ethoxyethanol (10 ml) at 120 °C, under stirring until the starting product disappearε. The reaction mixture iε cooled to room temperature and partitioned between CHC1₃ and IM Na₂C0₃. The organic phase is worked up to give a residue which is flash- chro atographed on a silica gel column eluted with CHCl₃/MeOH/38% H3 (1:1:0.01) to give the product (170 mg) as solid pure by TLC, m.p. 135-136°C; hydrochloride m.p. 258-260°C (dec); ^-NMR (CDCI3) 2.32 ppm (s, 12H), 2.48 ppm (s, 3H), 2.59 ppm (t, 4H), 2.83 ppm (t, 4H), 3.34-3.44 ppm (m, 4H), 3.45-3.55 ppm (m, 4H), 6.16 ppm (d, 2H), 6.88 ppm (br t, 2H), 7.10 ppm (t, 2H), 7.25 ppm (d, 2H), 7.49-7.60 ppm ( , 4H), 8.20 ppm (d, 2H), 10.97 ppm (br t, 2H), 13.20 ppm (s, 2H).

The following compounds are prepared analogously: 1 , 9-Bis C4-( - ( 2- ( dimethylamino )ethyl ) )carboxamido- 9-OXO-9 ,10-dihydro-acridine-l-yl] -5-methyl-l, 5, -triaza- nonane, m.p. 112-113°C; hydrochloride m.p. 235- 236°C (dec); iH-NMR (CDCI3) 1.90-2.07 ppm (m,

4H), 2.30 ppm (s, 12H), 2.40 ppm (s, 3H), 2.59 ppm

(t, 4H), 2.70 ppm (t, 4H), 3.24-3.39 ppm (m, 4H), 3.50-

3.67 ppm ( , 4H), 6.08 ppm (d, 2H), 7.05-7.20 ppm ( , 4H), 7.28 ppm (d, 2H), 7.50 ppm (t, 2H), 7.62 ppm (d,

2H), 8.28 ppm (d, 2H), 10.89 ppm (br t, 2H), 13.27 ppm

(s, 2H);

1 , 7-Bis[4-(N- ( 2- (dimethylamino)ethyl ) )carboxamido-

7-methoxy-9-oxo-9,10-dihydroacridine-l-yl]-4-methyl- 1,4,7-triazaheρtane, m.p. 160-162°C; hydrochloride m.p. 220-222 °C (dec); ^-H-NMR (CDC1₃) 2.34 ppm (ε, 12H), 2.53 ppm (ε, 3H), 2.60 ppm (t, 4H), 2.87 ppm (t, 4H), 3.29-3.45 ppm (m, 4H), 3.46-3.60 ppm (m, 4H), 3.88 ppm (ε, 3H), 6.04 ppm (d, 2H), 6.85 ppm (br t, 2H), 7.15-7.26 ppm (m, 4H), 7.48 ppm (d, 2H), 7.59 ppm (s, 2H), 11.02 ppm (br t, 2H), 13.10 ppm (s, 2H);

1 , 9-Bis [4- ( N- ( 2- ( dimethylamino ) ethyl ) ) carboxamido- 7-methoxy-9-oxo-9,10-dihydroacridine-l-yl]-5-methyl- 1, 5,9-triazanonane, m.p. 160-161°C; hydrochloride m.p. 230-233°C (dec); ^-H-NMR (CDC1₃) 1.90-2.10 ppm (m, 4H), 2.20-2.38 ppm (m, 15H), 2.52-2.68 ppm (m, 8H), 3.18-3.34 ppm (m, 4H), 3.45-3.63 ppm (m, 4H), 3.90 ppm (s, 3H), 5.92 ppm (d, 2H), 6.90 ppm (br t, 2H), 7.06-7.28 ppm (m, 4H), 7.48 ppm (d, 2H), 7.61 ppm (s, 2H), 10.90 ppm (br t, 2H), 13.17 ppm (ε, 2H);

1 , 9-Bis [4- ( N- ( 2- ( imethylamino )ethyl ) ) carboxamido- 7-nitro-9-oxo-9 , 10-dihydroacridine-l-yl]-5-methyl-l ,5,9- triazanonane, m.p. 152-153°C; hydrochloride m.p. 255- 257°C (dec); ÷i-H- MR (CDCI3) 1.84-2.04 ppm (m, 4H), 2.51 ppm (s, 15H), 2.48-2.70 ppm ( , 8H), 3.20-3.37 ppm (m, 4H), 3.40-3.55 ppm (m, 4H), 6.10 ppm (d, 2H), 6.89 ppm (br t, 2H), 7.15 ppm (d, 2H), 7.42 ppm (d, 2H), 8.21 ppm (d, 2H), 8.91 ppm (s, 2H), 10.57 ppm (br t, 2H),

13.58 ppm (br s, 2H). Example 8 - 1, 7-Bis[4-(N-( 2- ( dimethylamino )ethyl ) )- carboxamido~7-hydroxy-9-oxo-9,10-dihydroacridine-l-yl]-

4-methyl-l , 4 , 7-triazaheptane

1 , 7-Bis[4-(N-( 2-(dimethylamino )ethyl ) )carboxamido-

7-methoxy-9-oxo-9,10-dihydroacridine-l-yl]-4-methyl- 1,4,7-triazaheptane (280 mg) is refluxed in 48% HBr (3 mi) for 1 hour. The reaction mixture is cooled to room temperature and partitioned between CHC1₃ and IM Na₂C0₃. The organic phase is worked up to give a residue which is flash-chromatographed on a silica gel column eluted with CHCl₃/MeOH/38% NH₃ (1:1:0.01) to give the product (210 mg), m.p. 190-192'C; hydrochloride m.p. 254-255"C; iH- MR (DMSO-d₆) 2.25 ppm (s, 12H), 2.38-2.50 ppm (m, 7H), 2.75-2.93 ppm (m, 4H), 3.32-3.77 ppm (m, 8H) , 6.31 ppm (d, 2H), 7.10 ppm (d, 2H), 7.41-7.54 ppm (m, 4H), 7.98 ppm (d, 2H), 8.30 ppm (br t, 2H), 9.64 ppm (br s, 2H), 10.96 ppm (br t, 2H), 13.69 ppm (s, 2H).

The following compound is prepared analogously: 1 , 9-Bis [4- ( -( 2- ( dimethylamino ) ethyl ) )carboxamido- 7-hydroxy-9-oxo-9,10-dihydroacridine-l-yl]-5-methyl- 1,5 ,9-triazanonane, m.p. 186-188°C; hydrochloride m.p. 270-271°C; ^-H-NMR (DMSO-dg) 1.80-1.90 ppm (m, 4H), 2.22 ppm (s, 15H), 2.42-2.51 ppm (m, 8H), 3.34-3.44 ppm (m, 8H), 6.21 ppm (d, 2H), 7.21 ppm (d, 2H), 7.42-7.51 ppm (m, 4H), 7.94 ppm (d, 2H), 8.29 ppm (br t, 2H), 9.62 ppm (br s, 2H), 10.91 ppm (br t, 2H), 13.67 ppm (s, 2H).

Example 9 - 1, 7-Bis[4-(N-( 2- (dimethylamino)ethyl ) )- carboxamidoacridine-1-yl]-4-methyl-l, 4, 7-triazaheptane

(compound ( I )-( a) ) .

1 , 7-Bis [4- ( N-( 2- ( dimethylamino )ethyl ) )carboxamido-

9-OXO-9 , 10-dihydroacridine-l-yl ]-4-methyl-l , 4 , 7-tria- zaheptane trihydrochloride (190 mg) is refluxed in

EtOH/H₂0 (3:1, 15 ml). Al pellets (190 mg) placed for 1 minute in a HgCl₂ ethanol solution (760 mg in 18 ml of

EtOH) are added during 30 minutes (mg). When the addition is completed, the mixture is refluxed for 30 minutes, the filtered while hot, washing with ΞtOH (20 ml), diluted with H₂0 (50 ml) and added with FeCl₃ (600 mg). The reaction mixture is partitioned between CHCI3 and IM Na Cθ . The organic phase is worked up to give a residue which is flash-chromatographed on a silica gel column eluted with CHCl₃/MeOH/38% NH3 (1:1:0.01) to give the product (90 mg), m.p. 138-139°C; hydrochloride m.p. 255-257°C (dec); ^-NMR (CDCI3) 2.45 ppm (s, 12H), 2.57 ppm (s, 3H), 2.70 ppm (t, 4H), 2.90-3.02 ppm (m, 4H), 3.40-3.53 ppm (m, 4H), 3.68-3.82 ppm ( , 4H), 5.80 ppm (br t, 2H), 6.43 ppm (d, 2H), 6.97-7.17 ppm (m, 4H), 7.58 ppm (t, 2H), 7.78 ppm (d, 2H), 8.28 ppm (s, 2H), 8.73 ppm (d, 2H), 11.65 ppm (br t, 2H).

The following compound is prepared analogously:

1 , 9-Bis[4- (N- ( 2- (dimethylamino ) thyl ) )carboxamido- acridine-l-yl]-5-methyl-l, 5 ,9-triazanonane, m.p. 130- 131°C; hydrochloride m.p. 250-252°C (dec); ¹H-NMR (CDCI3) 2.02-2.16 ppm (m, 4H), 2.44 ppm (s, 12H), 2.54 ppm (s, 3H), 2.65-2.87 ppm (m, 8H), 3.29-3.44 ppm (m, 4H), 3.69-3.82 ppm (m, 4H), 6.26 ppm (d, 2H), 6.85 ppm (br t, 2H), 7.35 ppm (t, 2H), 7.67 ppm (t, 2H), 7.78 ppm (d, 2H), 8.02 ppm (d, 2H), 8.68 ppm (d, 2H), 8.74 ppm ( s , 2H ) , 11 . 81 ppm ( s , 2H ) .

Example 10 - 1,7-Bis[2- (2-(dimethylamino)ethyl )-

1,3, 7-trioxo- [IH, 2H , 3H, 7H, 12H]pyrimido[ 5 , 6 , 1-de]acridi- ne-6-yl]-4-methyl-l,4,7-triazaheptane (compound (I)-(e)) 6-Chloro-2-[2- (dimethylamino )ethyl]-1,3, 7-trioxo-

[lH,2H,3H,7H,12H]pyrimido[5,6,l-de]acridine (300 mg; the preparation is described in Antonini, I. et al., J. Med.

Chem., 1995, 38, 3282-6), N²-methyldiethylenetriamine

(0.06 ml) and Et₃ (0.5 ml) are placed in 2- ethoxyethanol (10 ml) at 80°C, under stirring until the starting product disappears. The reaction mixture is cooled to room temperature and partitioned between CHCI3 and IM Na CU3. The organic phase is worked up to give a residue which is chromatographed on a silica gel column eluted v/ith CHCl₃/MeOH (4:1) to give the product (190 mg) as solid pure BY TLC, m.p. 183-184°C; hydrochloride m.p. 275-277°C (dec); -NMR (CDCI3) 2.42 ppm (s, 12H), 2.58 ppm (s, 3H), 2.71 ppm (t, 4H), 2.86 ppm (t, 4H), 3.20-3.30 ppm (m, 4H), 4.25 ppm (t, 4H), 6.60 ppm (d, 2H), 7.16 ppm (t, 2H), 7.52 ppm (t, 2H), 7.91 ppm (d, 2H), 8.08 ppm (d, 2H), 8.52 ppm (d, 2H), 10.90 ppm (br t, 2H).

The following compounds are prepared analogously: 1 , 9-Bis [2- ( 2- ( dimethylamino )ethyl ) -1 , 3 , 7-trioxo- [lH,2H,3H,7H,12H]pyrimido[5, 6, 1-de]acridine-6-yl]-5-methyl-l, 5, 9-triazanonane, m.p. 143-144'C; hydrochloride m.p. 259-260°C (dec); ¹H-NMR (CDCI3) 1.90-2.04 ppm (m, 4H), 2.32 ppm (s, 3H), 2.40 ppm (ε, 12H), 2.60 ppm (t, 4H), 2.71 ppm (t, 4H), 3.28-3.40 ppm (m, 4H), 4.28 ppm (t, 4H), 6.44 ppm (d, 2H), 7.28 ppm (t, 2H), 7.52 ppm (t, 2H), 7.96 ppm (d, 2H), 8.20 ppm (d, 2H), 8.74 ppm ( d , 2H ) , 10 . 75 ppm ( br t , 2H ) ;

1 , 7-Bis [2- ( 2- ( dimethylamino )ethyl )-9-methoxy-l ,3,7- trioxo- [IH, 2H , 3H, 7H, 12H]pyrimido[ 5 , 6 , 1-de]acridine-6- yl]-4-methyl-l,4,7-triazaheptane, m.p. 200-202°C; hydrochloride m.p. 253-254°C (dec); ¹H-NMR (CDCI3) 2.40 ppm (s, 12H), 2.60 ppm (s, 3H), 2.70 ppm (t, 4H), 2.85 ppm (t, 4H), 3.34-3.46 ppm (m, 4H), 3.71 ppm (s, 3H),

4.25 ppm (t, 4H), 6.54 ppm (d, 2H), 7.08 ppm (d, 2H),

7.26 ppm (s, 2H), 8.10 ppm (d, 2H), 8.52 ppm (d, 2H), 10.94 ppm (br t, 2H);

1 , 9-Bis[2-( 2-(dimethylamino ) thyl )-9-methoxy-l ,3,7- trioxo-[lH,2H,3H,7H,12H]pyrimido[5,6,l-de]acridine-6- yl]-5-methγl-l,5,9-triazanonane, m.p. 183-185°C; hydrochloride m.p. 258-259°C (dec); ¹H-NMR (CDC1₃) 1.90- 2.10 ppm (m, 4H), 2.31 ppm (s, 3H), 2.40 ppm (s, 12H), 2.56-2.78 ppm (m, 8H), 3.25-3.42 ppm ( , 4H), 3.85 ppm (s, 3H), 4.27 ppm (t, 4H), 6.40 ppm (d, 2H), 7.05 ppm (d, 2H), 7.58 ppm (ε, 2H), 7.98 ppm (d, 2H), 8.60 ppm (d, 2H), 10.77 ppm (br t, 2H). Example 11 - 1 , 9-Biε[2-( 2- (dimethylamino )ethyl )-9- nitro-l,3,7-trioxo-[lH,2H,3H,7H,12H]pyrimido[5,6,l-de]- acridine-6-yl]-5-methyl-l , 5 , 9-triazanonane

A solution of 1 ,9-Bis[4-(N-( 2- (dimethylamino )- ethyl) )carboxamido-7-nitro-9-oxo-9 ,10-dihydroacridine-l- yl]-5-methyl-l,5,9-triazanonane(230 mg) and EtgN (0.75 ml) in CHCI3 (10 ml), is added drop by drop with C0C1₂ (20% in toluene, 0.7 ml ) in CHCI3 (10 ml) at 0°C. When the addition is completed the mixture is stirred at room temperature for 20 minutes. The reaction mixture is partitioned between CHCI3 and IM a₂C0₃. The organic phase is worked up to give a residue which is chromatographed eluting with CHCl₃/MeOH (1:1) to give the product (140 mg) as solid pure by TLC, m.p. 148-

149^βC; hydrochloride m.p. 262-263°C (dec); iH-NMR

(CDClg) 1.90-2.10 ppm (m, 4H), 2.35 ppm (s, 15H), 2.60- 2.75 ppm ( , 8H), 3.32-3.48 ppm (m, 4H), 4.25 ppm (t,

4H), 6.50 ppm (d, 2H), 7.98 ppm (d, 2H), 8.23-8.33 ppm

( , 4H), 8.79 ppm (d, 2H), 8.96 ppm (ε, 2H), 10.65 ppm

(br t, 2H) .

Example 12 - 1 ,7-Bis[2- ( 2- (dimethylamino)ethyl)-9- hydroxy-l,3,7-trioxo-[lH,2H,3H,7H,12H]pyrimido[5,6,l- de]acridine-6-yl]-4-methyl-l , 4 , 7-triazaheptane

1 , 7-Bis [ 2- ( 2- ( dimethylamino )ethyl )-9-methoxy-l ,3,7- trioxo-[lH,2H,3H,7H,12H]pyrimido[5,6,l-de]acridine-6- yl]-4-methyl-l,4,7-triazaheptane(150 mg) is refluxed in 48% HBr (3 ml) for 1 hour. The reaction mixture is cooled to room temperature and partitioned between CHC1₃ and IM Na₂C0₃. The organic phase is worked up to give a residue which is flash-chromatographed on a silica gel column eluted with CHCl₃/MeOH/38% NH3 (1:1:0.01) to give the product (140 mg) as solid pure by TLC, m.p. 279- 280°C; hydrochloride m.p. 270-272°C (dec); iH-NMR (DMSO- dg) 2.31 ppm (s, 12H), 2.50 ppm (s, 3H), 2,63-2.81 ppm (m, 8H), 3.39-3.54 ppm (m, 4H), 4.02-4.20 ppm ( , 4H), 6.69 ppm (s, 2H), 6.79 ppm (d, 2H), 6.90 ppm (d, 2H), 8.01 ppm (d, 2H), 8.30 ppm (d, 2H), 10.40 ppm (br ε, 2H) , 10.74 ppm (br t, 2H).

The following compound iε prepared analogously: 1 , 9-Bis [2- ( 2- ( imethylamino ) ethyl ) -9-hydroxy-l ,3,7- trioxo-[IH , 2H, 3H, 7H, 12H]pyrimido[5,6, 1-de] acridine-6- yl]-5-methyl-l,5,9-triazanonane, m.p. 263-264°C; hydrochloride m.p. 265-267°C (dec); ^- MR (DMSO-dg) 1.80-1.97 ppm (m, 4H), 2.25-2.42 ppm (m, 15H), 2.56-2.74 ppm ( , 8H), 3.21-3.35 ppm (m, 4H), 4.05-4.20 ppm (m,

4H), 6.50 ppm (d, 2H), 7.05 ppm (d, 2H), 7.44 ppm (s,

2H), 7.76 ppm (d, 2H), 8.46 ppm (d, 2H), 10.01 ppm (br s, 2H), 10.66 ppm (br t, 2H).

Example 13 - l,9-Bis{2-[2-(dimethylamino)ethyl]-

9 , 10-dimethoxypyrimido [5,6, 1-de] -acridin-6-yl}-5-methyl-

1,5, 9-triazanonane

A suspension of pyrimidoacridine from preparation 10 (0.35 g, 0.814 mmol), 3 , 3 ' -diamino-JV- methyldipropylamine (0.071 mL, 0.41 mmol) and triethylamine (0.5 mL ) in 2-ethoxyethanol (10 mL ) is stirred at 80°C for 2 h. After cooling, the mixture is partitioned between CHCI3 and aqueous 1 M a₂C0₃. The organic layer is worked up to give a residue that is purified by column chromatography on silica gel using CHCI3/CH3OH (9:1 v/v). The residue obtained by evaporation of the fractions containing the desired product is further purified by stirring in Et 0. Filtration of the mixture gives the pure title compound (0.08 g, 21.0%): C_4gH_{57 g}O₁₀; m.p. 190-191°C; hydrochloride m.p. 240-242°C (dec); NMR (CDCI3) δ 1.90-2.10 (m, 4H, 2 x CH₂ ) , 2.30 (s, 3H, CH3 ) , 2.35 (s, 12H, 4 x CH₃), 2.58-2.76 ( , 8H, 4 x CH₂ ) ,. 3.24-3.40 (m, 4H, 2 x CH₂), 3.93 (s, 6H, 2 x 0-CH₃ ) , 4.00 ((s, 6H, 2 x 0-CH₃), 4.29 (t, 4H, 2 x CH₂ ) , 6.43 (d, 2H , ar ) , 7.58 (s, 2H, ar), 7.97 (d, 2H, ar ) , 8.40 (s, 2H, ar ) , 10.85 (br ε, 2H, ex) .

Example 14 - 2-[2-(Dimethylamino)ethyl]-6-{ [3-( {2- [2- ( imethylamino ) ethyl ]-1 , 3 , 7-trioxo-2 , 3-dihydro-lff, 1H- pyrimido [5,6, 1-de]acridin-6-yl }amino )propyl ] amino}-2 , 3- dihydro-1#, 7Jϊ-pyrimido[5,6, 1-de]acridine-1 , 3 , 7-trione

A suspension of 2-[2-(dimethylamino)ethyl]-6- chloro-2 , 3-dihydro-liJ, 7#-pyrimido [5,6, l-de]acridine-

1,3,7-trione, disclosed in J. Med. Chem. 1995, 38, 3282- 3286 (0.2 g, 0.54 mmol), 1 , 3-diaminopropane (0.023 mL ,

0.27 mmol) and triethylamine (0.5 mL) in 2-ethoxyethanol

(10 mL) is stirred at 80°C for 2 h. After cooling, the mixture is partitioned between CHC1₃ and aqueous 1 M

Na C0₃. The organic layer is worked up to give a reεidue that iε purified by column chromatography on εilica gel using CHC1₃/CH₃0H (9:1 v/v) to give the pure title compound (0.1 g, 50%): ^c4i^H40^N8°6^{; m}-^p- 222-224°C; hydrochloride m.p. 273-274°C; NMR (CDC1₃) δ 2.30 (t, 2H, CH₂), 2.45 (s, 12H, 4 x CH₃ ) , 2.70 (t, 4H, 2 x CH₂ ) , 3.56-3.69 (m, 4H, 2 x CH₂ ) , 4.30 (t, 4H, 2 x CH₂ ) , 6.75 (d, 2H, ar), 7.41 (t, 2H, ar ) , 7.68 (t, 2H, ar ) , 8.21- 8.30 (m, 4H, ar ) , 8.70 (d, 2H, ar ) , 11.00 (br t, 2H, ex ) .

The following compounds are prepared analogously: 2- [2- (Dimethylamino)ethyl] -6-{[6- ({2- [2- (dimethylamino )ethyl]-1 , 3 , 7-trioxo-2 , 3-dihydro-l#, 7 -pyrimi- do[ 5 , 6 , 1-de]acridin-6-yl }amino )hexyl ] amino }-2 , 3-dihydro- 1H, 7#-pyrimido[5,6 , 1-de]acridine-1 , 3 ,7-trione: 67.7%; C₄₄H₄₆N₈G₅; rn.p. 275-276°C; hydrochloride m.p. 239- 241°C; NMR (CDClg) δ 1.57-1.73 (m, 4H, 2 x CH₂ ) , 1.74-1.98 (m, 4H, 2 x CH₂ ) , 2.40 (s, 12H, 4 x CH₃), 2.71 (t, 4H, 2 x CH₂), 3.31-3.37 (m, 4H , 2 x CH₂ ) , 4.31 (t, 4H, 2 x CH₂), 6.68 (d, 2H, ar ) , 7.41 (t, 2H, ar ) , 7.68 (t, 2H, ar), 8.20 (d, 2H, ar ) , 8.30 (d, 2H, ar ) , 8.71 (d, 2H, ar), 10.90 (br t, 2H, ex).

2-[2-(Dimethylamino)ethyl]-6-{[8-({2-[2-(dimethyla- mino )ethyl]-1 , 3 , 7-trioxo-2 , 3-dihydro-l.ff, 7iϊ-pyrimi- do[5 , 6 , 1- e]acridin-6-yl}amino )octyl]amino}-2 , 3-dihydro- 1H, 7ώ-pyrimido [5,6, 1-de]acridine-1 , 3 , 7-trione : 63.6%; ^C46^H50^N8°6^{; m,p}- 242-243°C; hydrochloride m.p. 185- 187°C; iH NMR (CDCI3) 5 1.41-1.62 (m, 8H, 4 x CH₂ ) , 1.63-1.90 (m, 4H, 2 x CH₂ ) , 2.35 (s, 12H, 4 x CH3 ) , 2.68 (t, 4H, 2 x CH₂), 3.30-3.46 (m, 4H, 2 x CH₂ ) , 4.30 (t, 4H, 2 x CH₂), 6.68 (d, 2H, ar ) , 7.42 (t, 2H, ar ) , 7.65 (t, 2H, ar), 8.22 (d, 2H, ar), 8.35 (d, 2H, ar ) , 8.71 (d, 2H, ar), 10.87 (br t, 2H, ex).

2- [ 2-(Dimethylamino ) ethyl]-6-{ [ 12- ( { 2- [2- ( dimethylamino )ethyl]-l , 3 , 7-trioxo-2 , 3-dihydro-l#, 7i?-pyrimi- do[ 5 , 6 , 1-de]acridin-6-yl}amino ) odecyl] amino}-2 , 3-di- hydro-liϊ, 7iϊ-pyrimido[5,6, 1-de]acridine-1 , 3 , 7-trione : 80.8%; C₅₀H₅₈NgO₆; m.p. 187-189°C; hydrochloride m.p. 254-256°C; ^-H NMR (CDCI3) δ 1.31-1.59 (m, 14H, 7 x CH₂), 1.64-1.90 (m, 6H, 3 x CH₂ ) , 2.38 (s, 12H, 4 x CH3 ) , 2.70 (t, 4H, 2 x CH₂), 3.29-3.45 (m, 4H, 2 x CH₂ ) , 4.30 (t, 4H, 2 x CH₂), 6.68 (d, 2H, ar ) , 7.46 (t, 2H, ar ) , 7.69 (t, 2H, ar), 8.22 (d, 2H, ar ) , 8.40 (d, 2H, ar ) , 8.71 (d, 2H, ar), 10.87 (br t, 2H, ex).

2- [2- (Dimethylamino)ethyl] -6- ({2- [{2- [[2- ({2- [2- ( dimethylamino)ethyl]-1,3, 7-trioxo-2 , 3-dihydro-l#, 1 H- pyrimido[5,6, 1-de] acridin-6-yl}amino ) ethyl ]- (methyl )- amino]ethyl} (methyl )amino]ethyl}amino)-2 , 3-dihydro-

1H, 7#-pyrimido- [5,6, 1-de]acridine-1 , 3 , 7-trione : 23.5%; ^C46^H52^N10°6^{; m,p}° 235-23δ°C; hydrochloride m.p. >300^QC; ¹H NMR (CDCI3) δ 2.24-2.61 (m, 18H, 6 x CH3), 2.62-2.74 (m, 8H, 4 x CH₂), 2.76-3.00 (m, 4H, 2 x CH₂ ) , 3.15-3.31 (m, 4H, 2 x CH₂), 4.30 (br t, 4H, 2 x CH₂ ) , 5.93 (d, 2H, ar), 7.34-7.59 (m, 4H, ar ) , 7.70 (t, 2K, ar ) , 8.28 (d, 2H , ar ) , 8 . 71 ( d , 2H, ar ) , 10 . 77 ( br s , 2H , ex ) .

Example 15 - NA- [2- (Dimethylamino )ethyl]-l-( {3-[ [3-

( {2-[2- (dimethylamino )ethyl]-1,3, 7-trioxo-2,3-dihydro-

1H, 7#-pyrimido[5,6, 1-de]acridin-6-yl}amino )propyl] (me- thyl)amino]propyl}amino)-9-oxo-9,10-dihydro-4-acridine- carboxamide

A suspension of the compound of preparation 11

(0.21 g, 0.46 mmol), pyrimidoacridine used in Example 14

(0.34 g, 0.92 mmol) and triethylamine (0.5 mL ) in 2- ethoxyethanol (10 mL) is stirred at 80 °C for 3 h. After cooling the mixture is partitioned between CHCI3 and aqueous 1 M a₂C0₃. The organic layer is worked up to give a residue that is purified by column flash- chromatography on silica gel using CHC^/MeOH (1:1, v/v) and 32% aqueous NH3 (4 mL for 1 L of eluent). The residue obtained by evaporation of the fractions containing the desired product is further purified by stirring in Ξt₂0. Filtration of the mixture gives the pure title product (0.3 g, 83.3%): C₄₄H_{51 9}0₅; m.p. 160- 162°C; hydrochloride m.p. 233-235°C; ¹H NMR (CDCI3) δ 1.85-2.04 (m, 4H, 2 x CH₂ ) , 2.23-2.40 ( , 15H, 5 x CH3 ) , 2.53-2.73 (m, 8H, 4 x CH₂ ) , 3.20-3.40 (m, 4H, 2 x CH₂ ) , 3.43-3.55 (m, 2H, CH₂ ) , 4.30 (t, 2H, CH₂ ) , 6.00 (d, IH, ar), 6.45 (d, IH, ar), 6.78 (br t, IH, ex), 7.05-7.15 (m, 2H, ar), 7.30-7.54 (m, 3H, ar ) , 7.97 (d, IH, ar), 8.12-8.24 (m, 2H, ar ) , 8.62 (d, IH, ar ) , 10.72 (br s, IH, ex), 10.84 (br s, IH, ex), 13.11 (s, IH, ex).

Claims

1. Compounds of general formula (I):

A-NH-Z-NH-A' wherein:

Z is a (C₂-C₄) alkylene chain or a group of formula (CH₂)_n-N(B)-(CH₂)-N(B' )-(CH₂ )m- (CH₂ )_n-N(B)-X-(CH₂ )_m wherein: n and m are independently an integer from 2 to 4,

B is hydrogen or (C^-C )-alkyl,

X is a single bond or a group of formula

-(CH₂)_p-N(B' )- wherein: q is an integer from 2 to 4 and B' is hydrogen or (C₁-C₄) alkyl

A and A' , which are the same or different, are selected from:

(d) (a) (f) in which :

R is hydrogen, or one or more substituents which are the same or different, selected from straight or branched (C₁-Cg)alkyl, -OH, -NH₂, (C₁~C₄)alkoxy, mono- and di-(C₁-C₄)alkylamino, chlorine, bromine, iodine and fluorine, nitro, mono- and poly-fluoro(C^- C₄)alkyl, (C₁-C₄ )alkylsulfonyl, (C₁-C₄)alkylaminosulfo- nyl;

Y is a -(CH₂) -T group; - p is the integer 2, 3 or 4;

T i╬╡ selected from -NH , -OH, mono- or di-(C₁~C₄)- alkylamino, hydroxyethylamino, N-morpholinyl, N-pi- perazinyl, N-thiomorpholinyl, N-pyrrolidin╬│l, N-pipe- ridinyl, possible stereoisomers or stereomeric mixtures and salts thereof with pharmaceutically acceptable acids .

2. Compounds according to claim 1, in which n and m are the integer 3.

3. Compounds according to claim 1 or 2, in which A = A¹ is a group of formula (b), (c) or (e).

4. Compounds according to claims 1 to 3, in which B is a methyl group.

5. Compounds according to claims 1 to 4, selected from: - 1 ,7-Bis(l-chloro-9-oxo-9,10-dihydro-acridine-4-car- bonyl ) -4-methyl-l , 4 , 7-triazaheptane;

1 , 9-Bis( l-chloro-9-oxo-9 , 10-dihydroacridine-4-car- bonyl ) -5-methyl-l , 5 , 9-triazanonane;

1 , 7-Bis( l-chloro-7-methoxy-9-oxo-9 , 10-dihydroacri- dine-4-carbonyl ) -4-methyl-l , 4 , 7-triazaheptane;

1 , 9-Bis ( l-chloro~7-methoxy-9-oxo-9 , 10-dihydroacri- dine-4-carbonyl )-5-methyl-l , 5 , 9-triazanonane; iV4-( 3-{ [ ( l-chloro-9-oxo-9 , 10-dihydro-4-acridinyl )- carbonyl]amino}propyl ) -l-chloro-9-oxo-9 , 10-dihydro-4- acridinecarboxamide; - iV4-{2-[{2-[(2-{[(l-chloro-9-oxo-9,10-dihydro-4-a- cridinyl )carbonyl]amino}-ethyl ) (methyl ) amino]ethyl}- (methyl ) amino]ethyl}-l-chloro-9-oxo-9 , 10-dihydro-4-acridinecarboxamide;

1 , 7-Bis [1- ( 2- ( dimethylamino )ethyl ) -amino-9-oxo- 9, 10-dihydroacridine-4-carbonyl] -4-methyl-l ,4, 7-triazaheptane;

1, 9-Bis[l-( 2-(dimethylamino )ethyl )amino-9-oxo-9, 10- dihydroacridine-4-carbonyl]-5-methyl-l , 5 ,9-triazanonane;

1 , 7-Bis [ 1- ( 2-( dimethylamino )ethyl ) amino-7-methoxy- 9-oxo-9,10-dihydroacridine-4-carbonyl]-4-methyl-l, 4,7- triazaheptane;

1, 9-Bis[l-( 2- (dimethylamino)ethyl )amino-7-methoxy- 9-oxo-9 , 10-dihydroacridine-4-carbonyl]-5-methyl-l ,5,9- triazanonane; - JV4- (3-{ [(!-{[ 2- (dimethylamino )ethyl]amino}-9-oxo- 9, 10-dihydro-4-acridinyl)-carbonyl]amino}propyl)-l-{ [2- (dimethylamino)ethyl]amino}-9-oxo-9,10-dihydro-4-acri- dinecarboxamide;

N4- {2- [{2- [(2-{[( l-{[2- (dimethylamino ) ethyl]amino}- 9-oxo-9, 10-dihydro-4-acridinyl)carbonyl] amino}ethyl) (methyl ) amino]ethyl} ( methyl )amino] ethyl }-l-{ [2-(dimethylamino )ethyl]amino}-9-oxo-9 , 10-dihydro-4-acridinecarboxa- mide;

1 , 7-Bis [1- ( 2- ( imethylamino )ethyl )-amino-7-hydroxy- 9-oxo-9 , 10-dihydroacridine-4-carbonyl ] -4-methyl-l ,4,7- triazaheptane; 1 , 9-Bis [1- ( 2- ( imethylamino )ethyl )amino-7-hydroxy-

9-OXO-9 , 10-dihydroacridine-4-carbonyl]-5-methyl-l ,5,9- triazanonane;

1 , 7-Bis [1- ( 2- ( dimethylamino )ethyl )-aminoacridine-4- carbonyl]-4-methyl-l , 4 , 7-triazaheptane;

1 , 9-Bis[1-( 2-( dimethylamino)ethyl ) aminoacridine-4- carbonyl]-5-methyl-l , 5 , 9-triazanonane;

1 , 7-Bis[2- ( 2- ( dimethylamino )ethyl ) -pyrazole[3,4,5- kl]acridine-5-carbonyl]-4-methyl-l , 4 , 7-triazaheptane; - l,9-Bis[2-(2-(dimethylamino)ethyl)pyrazole[3,4,5- A"I]acridine-5-carbonyl]-5-methyl-l , 5 , 9-triazanonane;

1 , 7-Bis[2-( 2-( dimethylamino )ethyl ) -9-methoxypyra- zole[3,4, 5 -kl^~ acridine-5-carbonyl]-4-methyl-l, 4, 7-triazaheptane trihydrochloride; - l,9-Bis[2-(2-(dimethylamino)ethyl)-9-methoxypyra- zole [ 3 , 4 , 5-kl]acridine-5-carbonyl ]-5-methyl-l , 5 , 9-tria- zanonane trihydrochloride;

1 , 7-Bis [2- ( 2- ( dimethylamino ) ethyl ) -9-hydroxypyrazo- le[3 ,4, 5 -kl ^~ acridine-5-carbonyl]-4-methyl-l, 4, 7-triaza- heptane;

1 , 9-Bis [2-( 2- ( dimethylamino ) ethyl ) -9-hydroxypyra- zole [3,4, 5- kl]acridine-5-carbonyl]-5-methyl-l , 5 , 9-tria- zanonane;

1 , 7-Bis[4-(N-( 2-( imethylamino)ethyl ) )-carboxamido- 9-oxo-9,10-dihydroacridine-l-yl]-4-methyl-l, 4, 7-triazaheptane;

1 , 9-Bis [4- ( -( 2- (dimethylamino )ethyl ) )carboxamido- 9-OXO-9 , 10-dihydroacridine-l-yl]-5-methyl-l , 5 , 9-triaza- nonane; - 1, 7-Bis[4- (N-( 2- (dimethylamino)ethyl ) )carboxamido- 7-methoxy-9-oxo-9,10-dihydroacridine-l-yl]-4-methyl- 1,4, 7-triazaheptane;

1 , 9-Bis[4-( - ( 2-( dimethylamino )ethyl ) )carboxamido- 7-methoxy-9-oxo-9,10-dihydroacridine-l-yl]-5-methyl- 1,5, 9-triazanonane; - l,9-Bis[4-(N-(2-(dimeth╬│lamino)ethyl) )carboxamido- 7-nitr╬▒-9-oxo-9,10-dihydroacridine-l-yl]-5-methyl-l, 5, 9- triazanonane;

1 , 7-Bis[4-(N-( 2-(dimethylamino)ethyl) )-carboxamido- 7-hydroxy-9-oxo-9 ,10-dihydroacridine-l-yl]-4-methyl- 1,4,7-triazaheptane; l,9-Bi╬╡[4-(N-(2-(dimethylamino)ethyl) )carboxamido- 7-hydroxy-9-oxo-9,10-dihydroacridine-l-yl]-5-methyl- 1 , 5,9-triazanonane;

1 , 7-Bi╬╡ [4- (N- ( 2- ( dimethylamino )ethyl ) )-carboxamido- acridine-l-yl]-4-methyl-l , 4 , 7-triazaheptane;

1 , 9-Bis[4-( - ( 2- ( dimethylamino )ethyl ) )carboxa i- doacridine-l-yl]-5-methyl-l , 5 , 9-triazanonane;

1 , 7-Bi [2- ( 2- (dimethylamino ) ethyl )-1 , 3 , 7-trioxo- [1H,2H,3H,7H, 12H]pyrimido[5,6,1-de]acridine-6-yl] -4- methyl-1, 4, 7-triazaheptane; l,9-Bi╬╡[2-(2-(dimethylamino)ethyl)-l,3,7-trioxo- [1H , 2H , 3H , 7H, 12H]pyrimido[5,6, 1-de]acridine-6-yl]-5- methyl-1, 5,9-triazanonane;

1, 7-Bis[2- (2-(dimethylamino )ethyl )-9-methoxy-l, 3,7- trioxo-[lH,2H,3H,7H,12H]pyrimido[5,6,l-de]acridine-6- yl ]-4-methyl-l , 4 , 7-triazaheptane;

1 , 9-Bis [2- ( 2- (dimethylamino )ethyl ) -9-methoxy-l ,3,7- trioxo- [IH, 2H , 3H, 7H, 12H]pyrimido[5,6, 1-de]acridine-6- yl]-5-methyl-l, 5,9-triazanonane; - l,9-Bis[2-(2-(dimethylamino)ethyl)-9-nitro-l,3,7- trioxo-[lH,2H,3H,7H,12H]pyrimido[5,6,l-de]-acridine-6- yl]-5-methyl-l , 5 , 9-triazanonane;

1 , 7-Bis[2- ( 2- ( imethylamino )ethyl )-9-hydroxy-l ,3,7- trioxo-[lH,2H,3H,7H,12H]pyrimido[5,6,l-de]acridine-6- yl]-4-methyl-l,4,7-triazaheptane; - l,9-Bis[2-(2-(dimethylamino)ethyl)-9-hydroxy-l,3,7- trioxo- [IH,2H, 3H, 7H, 12H]pyrimido[5,6, 1-de]acridine-6- yl]-5-methyl-l , 5 , 9-triazanonane;

1 , 9-Bis{2-[2- (dimethylamino )ethyl]-9 , 10-dimethoxy- pyrimido[5,6, 1-de]-acridin-6-yl}-5-methyl-l , 5 , 9-triaza- nonane;

2-[2-(dimethylamino)ethyl]-6-{[3-({2-[2-(dimethyla- mino )ethyl]-1 , 3 , 7-trioxo-2 , 3-dihydro-l#, 7#-pyrimido- [5,6, 1-de]acridin-6-yl}amino )proρyl]amino}-2 , 3-dihydro- 1H, 7 H-pγτimido[5,6, 1-de]acridine-1 , 3 , 7-trione; - 2-[2-(dimethylamino)ethyl]-6-{[6-({2-[2-(dimethyla- mino ) ethyl]-l , 3 , 7-trioxo-2 , 3-dihydro-l , 7#-pyrimido- [5,6, 1-de]acridin-6-yl}amino )hexyl]amino}-2 , 3-dihydro- 1H, 7i?-pyrimido[ 5 , 6 , l-de]acridine-l , 3 , 7-trione;

2-[2-(dimethylamino )ethyl]-6-{ [8-( {2-[2-(dimethyla- mino)ethyl]-1,3, 7-trioxo-2,3-dihydro-li7, 7J-pyrimido-

[5,6, 1-de] acridin-6-y1}amino )octyl] amino}-2 , 3-dihydro- 1H, 7H- YTimido[5,6, 1-de]acridine-1 , 3 , 7-trione;

2-[2-(dimethγlamino)ethyl]-6-{[12-({2-[2-(dime- thylamino ) ethyl]-1 , 3 , 7-trioxo-2 , 3-dihydro-lif, 7irT-pyrimi- do[5, 6, 1-de] acridin-6-yl}amino)dodecyl] amino}-2 , 3-di- hydro-l#, 7ϊ-pyrimido[ 5 , 6 , 1-de]acridine-1 , 3 , 7-trione;

2-[2-(dimethylamino )ethyl]-6- ({2-[{2-[[2-({2-[2- ( dimethylamino)ethyl]-1,3, 7-trioxo-2, 3-dihydro-l#, 1H- pyrimido[5, 6, 1-de]acridin-6-yl}amino) ethyl]- (methyl)- amino]ethyl} (methyl ) amino]ethyl}amino )-2 , 3-dihydro- 1H, 7H-pyrimido-[5,6, 1-de]acridine-1 , 3 , 7-trione; N4-[2-( dimethylamino )ethyl]-l-({3-[[3-({2-[2-( imethylamino )ethyl]-l , 3 , 7-trioxo-2 , 3-dihydro-lJϊ, 7#-pyrimi- do[5,6,l-de]acridin-6-yl}amino)propyl] (methyl)amino]pro- pyl}amino )-9-oxo-9 , 10-dihydro-4-acridinecarboxamide .

6. The use of the compounds of claims 1 to 5 for the preparation of a medicament having antitumor activity.

7. The use according to claim 6, in which the tumor to treat is the colon tumor.

8. Pharmaceutical formulations containing an effective dosage of at least one compound according to claims 1 to 5 in admixture with pharmaceutically acceptable excipients.