Novel polyamine derivatives novel polyamine derivatives

The present invention relates to certain polyamine derivatives, the intermediate compounds and methods used in their preparation and their use in the treatment of diseases caused by infestation with a large number of parasitic animals belonging to the Protozoa Phylum.

More specifically, the present invention relates to the treatment of a large number of diseases caused by parasitic animals belonging to the Phylum Protozoa that infest warm-blooded animals, the treatment being the administration to the infected host with a protozoan effective amount of a derivative. polyamine of general formula

RHN (CH₂)_noNH (CH₂)_niNH (CH₂)_noNHR

I

<td>at</td><td>what</td><td></td><td></td>

<td>no</td><td>represents</td><td>one</td><td>integer from 2 to 6;</td>

<td>m</td><td>represents</td><td>one</td><td>integer from 3 to 12; and</td>

<td>R</td><td>represents</td><td>one</td><td>hydrocarbyl group eventually</td>

or a group of formula - (CH₂)_x- (Ar) -X where x represents zero or the integer 1 or 2, Ar represents a phenyl or naphthyl group and X represents a hydrogen or halogen atom, an alkoxy group

<img file="PT86687B_D0001.tif" />

Cj _θ or C1-6 alkyl₄ or a group of general formula -S (O)_XWherein R 1 is a C 1-6 alkyl group or a pharmaceutically acceptable salt thereof. The above compounds of formula I may be administered to the infected host individually or in combination with an ornithine or arginine decarboxyase inhibitor.

When in the compounds of formula I above, R represents a saturated hydrocarbyl group, these compounds have straight chain, branched or cyclic alkyl groups of up to 6 carbon atoms, preferably tertiary butyl groups. and cyclohexyl; when R represents an unsaturated hydrocarbyl group they contain radicals having one or two double bonds or one triple bond, preferably groups of formula -CH₂CH = CH₂ , -CH₂CH₂CH = CH₂, -CH₂C = CH, -CH₂CH = C = CH₂· These groups may optionally have a phenyl or naphthyl radical such as a group of formula CH = CHCH₂- · In the case of groups of general formulas (CH₂)_nS where n represents an integer from 2 to 6, these include straight or branched chain alkyl groups of up to 6 carbon atoms such as, for example, straight chain alkylenic groups, preferably ethylene, propylene and butylene or chain groups. branched. When the groups of general formula (CH₂)_m include straight chain or ramyl alkylenic groups. with up to 12 carbon atoms, preferred groups contain

5,6,7,8 or 9 carbon atoms and exhibit linear chains. No grju powder of general formula - (CH₂)_x- (Ar) -X, x preferably represents number 1 or 2 and Ar represents a substituted phenyl or naphthyl group but when Ar represents a substituted phenyl or naphthyl group X represents an alkoxy group, preferably a methoxy group or ·³L, ethoxy, a halogen atom, preferably a chlorine atom or an alkyl group, preferably a tert-methyl, ethyl or butyl group. and when X represents a group of formula (R) R 1 represents preferably a methyl, ethyl or tertiary butyl group. and n represents zero or the number 1 or 2. The compounds of formula I should preferably be symmetrical. For example, in each of the compounds of formula I the terminal groups represented by R should preferably be the same as the groups of formula (CH₂)_no·

In order to facilitate discussion and description of the concepts according to the present invention it is convenient to use certain abbreviated forms with respect to both generic compounds and compound classes. For example, the 1,18-bis- (phenyl) -methylJ-1,5,14,18-tetraazaoctadecane compound of structure 0 CH₂NH (CH₂)₃NH (CH₂)_gNH (CH₂)₃NHCH₂0 could be represented, in its abbreviated version, by BηH (3) NH (8) NH (3) N representandoB, representing B_noa benzyl group and may also be referred to as a bis-benzyl 3-3-8-3 compound, of course, nitrogen atoms. If non-symmetrical compounds are used they could be referred to as, for example, N-benzyl-N'-phenethyl-3-8-4.

In general, the compounds of formula I may be prepared by chemical reactions analogous to well known conventional reactions, the choice of any particular preparation process being dependent upon a large number of factors. For example, in general, the ease of acquisition and cost of reagents, the applicability of certain general reactions to specific compounds, the presence of unsaturated hydrocarbic groups, and the like, are all natural factors to those skilled in the art. in the art which contributes to the choice of the synthesis method used in the preparation of any specific compound within general formula I.

Accordingly, the following reaction schemes illustrate the steps for preparing the compounds according to the present invention.

Reaction Scheme A

H₂N (CH₂)_mNH₂

H₂C = CHCN + H₂ Pt0₂

HCl / AcOH

R¹ halide

HCl Et₂O

ItOH

EtOH

---&gt; CN (CH₂)₂NH (CH₂)_mNH (CH₂)₂CN

H₂N (CH₂) 3NH (CH₂)_mNH (CH₂)₃NH₂.4HCl

NaOH / H_?O

THF

Kt-BuQ ----&gt;

DMF

R'NH (CH₂)₃NH (CH₂)_niNH (CH₂)₃NHR '. Wherein R 'represents one of the groups represented above by the symbol R in general formula I except that when R' represents a group +

-5po of general formula X- (Ar) - (CH₂) x cannot represent zero; Boc represents a t-butoxycarbonyl protecting group and Q represents a tertiary butyl group.

In the above 5-phase process the initial phase requires a specific N-alkylation to produce compounds wherein n represents the integer 3, which is achieved by reacting, under heating and under conventional conditions, a diamine in which is not as defined above in general formula I with 2 equivalents of acetonitrile, possibly in an appropriate solvent. The resulting cyan derivatives of the general formula are then chemically reduced with hydrogen in the presence of platinum dioxide as a catalyst and in an appropriate solvent containing 8 equivalents of hydrochloric acid or hydrobromic acid to obtain the corresponding hydrohalogenated salts, using conventional techniques. Other reduction techniques may also be used, such as using lithium aluminum hydride to obtain compounds of formula 3. Continuing the preparation of the above compounds under conventional conditions, the hydrohalogenated salts are neutralized. using bases and protecting the nitrogen atoms preferably using tert-dibutyl dicarbonate. N-protected tetraamines of formula 5 are alkylated using the appropriate alkyl groups or alkyl halides wherein the halogen atom represents a chlorine or bromine atom in the presence of potassium butoxide and using alkylation techniques. conventional. After this alkylation reaction the nitrogen protecting groups are removed by conventional techniques such as, for example, treatment with an acid, preferably

<img file="PT86687B_D0002.tif" />

hydrochloric acid in the presence of a suitable solvent or solvent system such as diethyl ether in ethanol to give the desired compounds of formula 6.

Alternatively, the compounds of general formula 3 may be subjected to reductive alkylation using an appropriate aldehyde which, as a sole requirement, may not include, as a substituent, an alkylthio group; using conventional techniques this reduction is accomplished by hydrogenation in the presence of platinum oxide. This technique does not require the protection of nitrogen atoms of intermediate compounds. Thus, when the final compounds are intended to contain an alkylthio group or an unsaturated hydrocarbyl group next to their terminal nitrogen atoms, the compounds of formula 3 or their otherwise prepared homologues should be subjected, i.e. compounds of general formulas 2,3 or 4, to reductive alkylation using an appropriate aldehyde although such reductive alkylation again does not require protection of the nitrogen atoms of the intermediate compounds when reduction is carried out with sodium cyanoborohydride by conventional techniques.

reaction scheme B represents a preferred technique for the preparation of compounds of formula I wherein n represents the integer 4, but which may also be applied in the preparation of compounds wherein n represents an integer from 2 to 6, these compounds being obtained by another process, analogues of the compounds of formula 6 obtained in reaction scheme A cited above.

<img file="PT86687B_D0003.tif" />

Reaction Scheme B

H₂N (CH₂)_no0H

<td>R (CH₂)_no'CHO</td><td>R¹ HN (CH₂)_no0H</td><td>Protection of</td>

<td>PtO₉/H₉</td><td> 8</td><td rowspan="2">nitrogen atom, ------------------------------</td>

<td>cc</td><td></td>

R'N (CH₂)_no0H

Mouth

Ms Cl · · ~ ΰ ~ - pin dina

R'N- (CH₂)_noOMs

5OC

3oc Boc where n is fundamentally the number 4, although it may represent any integer from 2 to 6; BoC represents a nitrogen protecting group, preferably a t-tuboxy carbonyl group which may be substituted by another group which is suitable for protecting nitrogen atoms; R 'represents a group of general formula^x - (A r) - (CH ₂) _χ wherein x is not zero and Ar 'is an X-substituted alkyl or aryl group as defined above in general formula I, n is zero or a positive integer and Ms is a mesyl group.

This reaction is initiated by reductive alkylation techniques using an amino alcohol of formula 7 and an appropriate allyl to obtain amino alcohols.¹-substituted for. general formula 8 carrying protected nitrogen atoms. The N-protected amino alcohols of formula 9 are converted to their mesylates of formula 10 using conventional techniques such as by reaction with mesyl chloride in the presence of pyridine, preferably in the presence of a dissolve.

<img file="PT86687B_D0004.tif" />

such as, for example, methylene chloride.

By conventional techniques the mesylate corresponding to alkylation is reacted with an N-protected amine such as BocNH (CH₂)_mNHBoC, and using tert-butoxide. of potassium in dimethylformamide. Using the techniques described in Reaction Scheme A, the protecting groups of the resulting N-protected tetraamines of general formula 5 are removed. All reactions cited prior to reductive alkylation, N-protection, mesylation, alkylation and removal of the protecting groups use conventional reaction techniques and conditions.

Thus, when optionally preparing compounds of general formula I wherein n is number 2, reaction scheme C is preferably used to obtain the necessary intermediate compounds of general formula 13 which will be subjected to the same techniques. alkylation presented above in Reaction Scheme A.

I

Reaction scheme C

H₂NCH₂CH₂NH₂ + Br (CH₂)_mBr H₂N (CH₂)₂NH (CH₂)_mNH (CH₂)₂NH₂

12 13 who has the meaning defined above in general formula I.

The above N-alkylation reaction requires reaction of an appropriate dihalogenoalkane of formula 12 with an excess (10x) amount of ethylenediamine of formula 11 by heating the reagents to reflux temperatures in a suitable solvent as for example ethanol. THE

<img file="PT86687B_D0005.tif" />

The preparation of the desired final compounds having substituents R at the terminal nitrogen atoms of the intermediate compounds of formula 13 may preferably be carried out by means of reductive alkylation techniques using appropriate aldehydes which do not exhibit atom protecting groups. nitrogen, alternative technique shown in Reaction Scheme A or N-protecting, alkylating and removing the protecting groups of the intermediate compounds of formula 13 by similar techniques to those described in steps 3, 4 and 5 of Reaction Scheme A.

A preferred method for preparing compounds wherein Ar represents a phenethyl or naphthylethyl group, especially those wherein n represents the integer 3 and n represents the integer 8, is the reaction of an aroyl chloride according to the method described in Scheme. reaction D.

<td>Scheme</td><td>reactionary</td><td>D</td>

<td>H₂n (ch₂]</td><td>KN-Bn³I</td><td> 0 ||</td>

<td></td><td>(CH) 8 +</td><td>0CH₂C-C1</td>

<td>H₂n (ch₂]</td><td> 1 1 ₃ N - B n</td><td></td>

<td> 14</td><td></td><td> 15</td>

II

0CH₂CNH (CH₂)₃N-Bn (Ç^H₂)8

0CH_?CNH (CH_?M-Bn

II o

<td>Reduction</td><td>0CH_?CH_?NH (CH_?)_?N-Bn</td><td>0CH₂CH_?NH (CH₂) 3NH</td>

<td> 16 ^LAH</td><td>(CH₂)8</td><td>H „1</td>

<td></td><td>0CH₂CH₂NH (CH₂)₃N-Bn</td><td>^{Pd / C} 0CH₂CH₂NH (CH₂)₃NH</td>

<img file="PT86687B_D0006.tif" />

wherein Bn represents a benzyl group, 0 represents a phenyl group and LAH represents aluminum hydride and TiO. As stated above, the above reaction is the preferred method for preparing special compounds which require the N-alkylation of a partially protected intermediate of formula 14 by reaction with arylacetyl chloride of formula 15 in the presence of triethylamine and in the presence of triethylamine. an inert solvent, yielding a chemically reducing amide of formula 16, preferably using aluminum hydride and iTtio, Thereafter, the resulting product of formula 17 is subjected to a di-benzylation reaction in the presence of hydrogen and palladium on charcoal as catalyst to give the desired end product. In these reaction steps conventional techniques are used. It is evident that this same reaction scheme can be used in the preparation of others with compounds of general formula I being well known to those skilled in the art the adoption of the art and the warnings involved.

Thus, when Ar represents an aromatic group, ie a phenyl or naphthyl group, (X-phenyl or X-naphthyl), bonded directly to the terminal nitrogen atoms, which means that x represents zero, these compounds may be prepared. according to the following general reactions of Reaction Scheme E.

Reaction scheme E

0NH (CH₂)₂CN LAH 0NH (CH₂)₃NH₂

----&gt;

or

Protection of nitrogen atoms ------------------_&gt;

-110N (CH₂)₃NH

Boc Bo

NaH

DMF

0-Ν (CH₂) gN-Boc jt

Boc (CH₉) ^v cm I i 0-N (CH₂) gN-Boc

I Boc £ 2

In the reaction scheme cited above which represents the preparation of compounds wherein Ar represents a phenyl group, the first step is a reduction which is carried out with lithium aluminum hydride according to techniques published in journals such as, for example, Bul. Soc. Chim. Br., Part 2, 165-167 (1979). Of course, this reaction can be extended by allowing the inclusion of naphthyl and X-substituted intermediates provided that the reaction conditions do not influence them unfavorably. For the protection of nitrogen atoms, t-butoxycarbonyl groups are preferably used which are attached and eliminated by conventional techniques cited above in the present invention. By conventional techniques, the N-protected compounds are alkylated by reacting them with an appropriate dihalogen and alkane.

Thus, when preparing compounds of formula I having an unsaturated hydrocarbyl group, i.e. compounds having acetylenic, allenic or allyl groups, the techniques of reaction Scheme F are preferably used.

<img file="PT86687B_D0007.tif" />

Reaction scheme F

BnNH (CH₂)_mNHBn

Cl (CH₂)_no0H nBnOH

Nal

H0 (CH₂)_niN (CH₂)_frogN- (CH₂)₃0H

Bn Bn

Unbenzi1 Action_

N atom protection,

2'n

Ms Cl

CH₂C1₂

Pi ri di na

MsO (CH₂)_noN- (CH₂)_mN- (CH₂)_noOMs

Boc Boc

<img file="PT86687B_D0008.tif" />

R-NHBoc&gt;

NaH, Nal

DMF

HCl / EtOH

<img file="PT86687B_D0009.tif" />

R₃NH (CH₂)_laughNH (CH₂)_niNH (CH₂)_noNHR₃ where R₃ represents an appropriate unsaturated hydrocarbyl group, Bn represents a benzyl group, MsCl represents methanesulfonyl chloride and Boc represents the t-butoxy carbonyl protecting group.

In the above reaction scheme N-alkylate one diamine containing two benzene groups of general formula 23 by a single displacement reaction yielding compounds of general formula 24 in which benzyl groups are first removed and then protect nitrogen atoms. These reaction steps are carried out according to well known techniques. Bis- (hydroxy) -aminoalkanes of formula 26 are obtained which upon a mesylation reaction give rise to mesylates of formula 27 which are alkylated using 2 equivalents of an N-protected amine having an appropriate unsaturated hydrocarbyl group such as for example, N- (t-butoxycarbonyl) -2,3-butadieneylamine. A tetramine of 4 protected nitrogen atoms of general formula 29 is obtained from which the protecting groups are readily removed to yield the desired compounds of general formula 30.

Thus, when one optionally converts one alkylthio substitute into another corresponding to its highest oxidation state, the alkylthio is treated with a peracid according to conventional conditions. Hydrogen peroxide and sodium periodate are suitable oxidizing agents but acj is preferred. of m-chloroperoxybenzoic acid. When by oxidation a derj is obtained. sulfinyl ester 1 molar equivalent of peracid per alkylthioether group is used; when 2 molar equivalents of peracid are used, sulfonyl derivatives are obtained. These oxidation reactions are carried out at a temperature of from about 0 ° C to room temperature in unsolvent solvents. likely to oxidize. Methylene chloride, chloroform, acetic acid and ethyl acetate are the preferred solvents.

<img file="PT86687B_D0010.tif" />

Examples illustrating Reaction Scheme A

Example 1

1,18-bi-sL (phenyl) -methyl-1-5,15,18-tetraazaoctadecane tetrahydrochloride

Phase A. N, N ¹-bis-e 2,2'-bis (cyano) ethyl] -1,8-dimethyloctane. Dissolve 28.8 g (0.2 mol) of 1,8-diaminooctane in 250 ml of ethyl alcohol. Add 27 ml (0.41 mole) of acrylonitrile and gently reflux the reaction mixture overnight. Discard solvent under reduced pressure. Analytically it was observed that the desired compound was greater than 95% pure.

Phase B. 1,5,14,18-tetraazaoctadecane tetrahydrochloride. Mix 50.0 g of the compound prepared in Example 1 with 2.0 g of platinum oxide, 133 ml of concentrated hydrochloric acid and 600 ml of acetic acid, treat the resulting mixture with hydrogen at about 28.5 Kg / cm ( 451bas / sq. In.) On a shaker to saturation. Filter the resulting reaction mixture, evaporate the solvent and triturate the resulting compound with one liter of ethyl alcohol. Filter and dry the compound. 51.6 g of the title compound were chromatographed by TLC (thin layer chromatography) using silica gel plates and a mixture of 40% concentrated ammonium solution and methyl alcohol as eluting agent. Rf 0.17.

Step C. 1,5,14,18-tetra (t-butoxycarbonyl) -1,2,14,18-tetraazaoctadecane. Treat 28.0 g (0.069 mol) of the compound obtained in reaction phase B with 10.99 g (0.274 mol) of hydroxy.

-1 5 ^ -, i /

<img file="PT86687B_D0011.tif" />

of sodium in 120 ml of water until a homogeneous solution is obtained. Add 65.7 g (0.307 mole) of tert-dibutyl dicarbonate. in 750 ml tetrahydrofuran and stir the resulting reaction mixture for 16 hours. Separate the phases, discard and wash the aqueous phase with 2x500 ml of methylene chloride. Combine the organic phases and dry over magnesium sulfate. Filter, evaporate the solvents in vacuo and flash chromatograph the residue on silica gel and elute 25% ethyl acetate / hexane. 30.2 g of the title compound were chromatographed using silica gel plates and eluting with 25% ethyl acetate / hexane. Rf 0.33.

Step D. 1,18-Bis- (Phenyl) -methyl-1,1,14,18-tetra- (t-butoxycarbonyl) -1,2,14,18-tetraazaoctadecane. Dissolve 20.0 g (0.03 mole) of the product prepared in reaction phase C in 30 ml of dimethylformamide and treat with 7.5 g (0.067 mole) of tert-butoxide. of potassium and 7.97 ml (0.067 mole) of benzyl bromide with stirring for 18 hours. Evaporate volatile liquids in a vacuum (0.5 mm) at 45 ° C, treat the resulting residue with 1400 ml of ethyl acetate and wash it with 2x500 ml of water. The organic phase is then dried over magnesium sulfate and the solvent evaporated in vacuo. Flash chromatography on silica gel using 20% ​​ethyl acetate / hexane as eluting agent provided 12.4 g (50% yield) of the desired compound as a clear viscous oil. This oil was chromatographed using silica gel plates and a 25% ethyl acetate / hexane mixture as eluting agent. Rf. 0.42.

<img file="PT86687B_D0012.tif" />

Phase E. 1,18-BisZ (phenyl) methyl -1,5,14,18-tetraazaoctadecane tetrachloridate. Dissolve 12.4 g (0.0147 mol) of the compound prepared in step D in 14.7 ml of anhydrous ethyl alcohol and treat with 160 ml of 2N hydrochloric acid in diethyl ether with stirring overnight. Filter, wash the resulting filter cake with diethyl ether and dry. 7.2 g of the expected compound are obtained. Mp 300 ° C. The resulting compound was chromatographed using silica gel and a mixture of a concentrated 10% ammonia / methyl alcohol solution as eluting agent. Rf. 0.24.

<img file="PT86687B_D0013.tif" />

Example IA

Ν, Ν¹-Bis [3- (ethylamino) propyl] -1,7-heptanediamine

Phases A and B: 1,5,13,17-tetraazaheptadecane tetrachlorhydrate

The title compound is prepared according to the method of Israel et al., J. Med. Chem. 7, 710 (1964).

Phase C 1,5,13,17-tetra- (butoxy tert-carbonyl) -1,5,14,18 Combine 3.9 g (0.01 mole) of 1,5,13,17-tetrachloride tetraazaheptadecane with 1.76 g (0.44 mole) of sodium hydroxide in 44 ml of water and stir until homogeneous. To this mixture is added 9.6 g (0.044 mole) of tert-dibutyl dicarbonate. in 88 ml of THF and stir for 3 hours. The mixture is diluted with 300 mL of ethyl acetate (EtOAc) and the organic layer is separated. The organic layer is dried over anhydrous MgSO4 and evaporated in vacuo to give a viscous oil. Purifies itself

16-residue by flash chromatography (silica gel) eluting with 25% EtOAc / hexane to give 3.0 g of the title compound. The Rf value is 0.20 on silica gel plates eluted with 25% EtOAc / hexane.

Step D: 3,7,15,19-tetra- (butoxy tert-carbonyl) -3,7.15,

19-tetraazaheneicosane

3.0 g (0.0046 mol) of 1,5,13,17-tetra- (butoxy tert-carbonyl) -1,5,14,18-tetraazaheptadecane and 0.45 g (0.011 mol) are combined. of sodium hydride (50% in oil) in 9 ml of DMF and the mixture is stirred until hydrogen evolution ceases. Ethyl iodide (0.9 ml, 0.011 mol) is added and the mixture is stirred for 18 hours. Evaporate the DMF in vacuo and partition the residue between ethyl acetate (600 mL) and water (200 mL). The organic layer is separated, dried over anhydrous MgSO4 and evaporated in vacuo. The residue is purified by flash chromatography (silica gel) eluting with 20% EtOAc / hexane to give 1.68 g of the title compound. The Rf value is 0.5 after elution on silica gel plates using 25% EtOAc / hexane as eluent.

Step E: Ν, Ν'-BisZ * 3- (ethylamino) -propyl-1,7-heptanediamine

1.68 g (0.0024 mol) of 3,7,15,19-tetra- (butoxy tert-carbonyl) -3,7,15,19-tetraazaheneicosane is treated with HCl in methanol (50 ml, 1 mL). , ON) and stir overnight. The mixture is filtered and the title compound is recrystallized from methanol / water (20:80, v / v) to give 0.5 g of the title compound. The value of R ^ is

16-Β equal to 0.39 on silica gel plates eluted with 40% ammonia (concentrate) in methanol; Mp 322 ° -323 ° C with degradation.

Example 2 1,18-bis (butyl) -1,5,14,18-tetraazaoctadecane tetrahydrochloride

Step A. 1,18-Bis (butyl) -1,5,14,18-tetra (t-butoxycarbonyl) -1,5,14,18-tetraazaoctadecane. Mix 3.5 g (0.0053 mol) of the product prepared in reaction phase C of Example 1, 2.7 g (0.024 mol) of tert-butoxide. of potassium, 2.57 ml (0.024 mol) of 1-iodobutane in 10 ml dimethylformamide and leave the resulting mixture to stir for 18 hours. Evaporate volatile liquids in vacuo (0.5 mm) at 45 ° C, dissolve the resulting residue in 500 ml of ethyl acetate, wash the organic phase with 2x100 ml of water and dry over magnesium sulfate. Evaporate off the solvents and subject the residue to flash chromatography on silica gel using a 20% ethyl acetate / hexane mixture as eluting agent. 1.32 g of the title compound are obtained and chromatographed on silica gel plates using a% ethyl acetate / hexane mixture as eluting agent. Rf. 0.36.

Phase B. Dissolve 1.32 g (0.0017 mol) of the product prepared in reaction step A of this example in 1.7 ml of ethyl alcohol, treat with 17 ml of 2N hydrochloric acid in diethyl ether and stir the mixture. reaction overnight. Filter and wash the resulting precipitate with diethyl ether, recrystallize the washed material from the isopropanol / water mixture. Filter and dry the crystals in vacuo (0.1 mm) over phosphorus pentoxide at 79 ° C. 0.62 g of the title compound is obtained. Mp) 300 ° C. The compound was chromatographed using silica gel plates and a concentrated 20% ammonia / acetic acid solution as eluting agent. Rf. 0.47.

Example 2A 1,18-Γ (1-naphthyl) -methyl 1J-1,5 tetrahydrochloride hemihydrate,

14,18-tetraazoctadecane

Step A. To a mixture of 2.6 g of 1,15,18-tetra- (t-butoxycarbonyl) -1,1,14,18-tetraazaoctadecane and 1.07g of tert-butoxide. of potassium in 50 ml hexamethylphosphosphoriamide (HMPA) add a solution of 1.7 g (9.6 mmol) of 1-chloromethylnaphthalene in 5 ml hexamethphosphoriamide triamide (HMPA). In an oil bath at 80 ° C for 4 hours, pour into 300 ml of water and extract the aqueous mixture with 2x300 ml of ethyl acetate. The extracts are combined and extracted with 3x300 ml of water and 300 ml of a concentrated sodium chloride solution. The organic phase is dried, evaporated and the resulting residue is chromatographed using a silica gel flash chromatographic column and a toluene / ethyl acetate mixture as eluting agent.

<img file="PT86687B_D0014.tif" />

(4: 1). 800 mg of 1,18-bis- (1-naphthyl) methyl) -1,5,14, are obtained.

18-tetra- (t-butoxycarbonyl) -1,1,14,18-tetraazaoctadecane.

Step B. 800 mg of the product prepared in reaction phase A is dissolved in 50 ml of methanol, excess gas hydrochloric acid is added and the resulting reaction mixture is stirred overnight at room temperature. The resulting solids are filtered off and dried under vacuum to give the title product. Mp 262 ° -264 ° C.

Examples illustrating Reaction Scheme B

Example 3 1,20-bis-sf (phenyl) -methyl 1J1,16,15,20-tetraaza Phase A tetrahydrochloride.¹bis (t-butoxycarbonyl) -1,8-octanediamine

Dissolve 10.8 g (0.075 mole) of diaminooctane in 200 ml of methylene chloride and 100 ml of methyl alcohol, add 32.7 g (0.156 mole) of tert-dibutyl dicarbonate. and stir the resulting mixture overnight. Evaporate in vacuo and crystallize the resulting residue in hexane. 20.2 g of the desired compound are obtained. Mp 96 ° -97 ° C.

Stage B. 4- /? Z- (phenyl) -methyl-amino-7-butanol-1. Mix

8.9 g (0.1 mol) 4-amino-butanol-1, 10.6 g (0.1 mol) benzaldehyde, 100 ml ethyl alcohol and 0.3 g platinum oxide and saturate the mixture hydrogen at 28.5 kg / cm (45 lbs./sq. in.). Filter and evaporate the solvent in a vacuum.

17.7 g of the desired compound is chromatographed on silica gel using a mixture of a concentrated 10% ammonia / acetic acid solution as eluting agent.

Step C. 4- [N- (t-Butoxycarbonyl) -Nf (phenyl) methyl] amino] butanol-1. Mix 17.7 g (0.1 mol) of the butanolic compound prepared in reaction step B mentioned above and tert-dibutyl dicarbonate. in 100 ml of methylene chloride and stir the resulting mixture overnight. Evaporate the solvents in vacuo and chromatograph the resulting residue by chromatography. rapid elution using a 25% ethyl acetate / hexane mixture as eluting agent. The desired compound is obtained which is chromatographed on silica gel plates using a 20% ethyl acetate / hexane mixture as eluting agent. Rf 0.27.

Phase D. 4- [N- (t-butoxycarbonyl) -N-zS (phenyl) -methyl] -amino-7-1-methanesulfonyl butane. Cool in an ice bath a mixture containing 21.8 g (0.078 mol) of the product obtained in reaction step C above, 250 ml of methylene chloride and

9.7 ml (0.12 mol) of pyridine. Add dropwise for 20 minutes 6.65 ml (0.086 mol) of mesyl chloride in 6.6 ml methylene chloride. Stirring the reaction mixture for 2 hours store it at room temperature until it is at this temperature. Pour the resulting mixture into 200 ml of methylene chloride, wash with 500 ml of 0.5N hydrochloric acid, saturate with sodium hydrogen carbonate, dry over magnesium sulfate, evaporate in vacuo and chromatograph using flash silica gel chromatography. and as eluting agent a 25% ethyl acetate / hexane mixture. 10.7 g of the expected product are obtained by chromatography on silica gel plates using a 25% ethyl acetate / hexane mixture as eluting agent. Rf. 0.36.

Step E. 1,20-Bis (phenyl) -methyl 17-1,6,15,20-tetra- (t-butoxycarbonyl) -1,6,15,20-tetraazaeicosane Mix 5.16 g (0.015 mol) of the product prepared in reaction phase A and 10.7 g (0.032 mole) of the product prepared in reaction phase D, 3.92 g of tert-butoxide. of potassium, 0.2 g of sodium iodide and 60 ml of dimethylformamide and stir the resulting mixture for 72 hours at room temperature. Separate the solvent by evaporation in vacuo, treat the resulting residue with 600 mL of ethyl acetate and wash with 2x200 mL of water. Dry the organic phase over magnesium sulfate, evaporate the solvents and chromatograph the resulting viscous residue by flash chromatography using silica gel and a 20% ethyl acetate / hexane mixture as eluting agent. The desired product is obtained by chromatography on silica gel plates using a 20% ethyl acetate / hexane mixture as eluting agent. Rf. 0.22.

Phase F. tetrachloride 1,20-bi sZ- (phenyl) -methyl 1J-1,

6,15,20-tetraeicosane Dissolve 4.7 g (0.0054 mol) of the product prepared in reaction phase E in 5 mL of ethyl alcohol and treat with 54 mL of 2N hydrochloric acid in ethyl ether; Stir the reaction mixture overnight, filter and recrystallize from the mixture. isopropanol / water mixture the resulting solid products. Cool, filter and dry the product over phosphorus pentoxide at reduced pressure. The desired product is obtained which is chromatographed on silica using a mixture of eluents.

Of a concentrated 20% ammonia / methyl alcohol solution. MP&gt; 300 ° C. ; Rf 0.47.

Examples illustrating reaction scheme C

Example 4

1,4,13,16-tetra- (t-butoxycarbonyl) -1,1,13,16-tetraazahexadecane

Mix 4.75 g (0.017 mol) of 1,8-dibromooctane, 20 ml of ethyl alcohol and 9.32 ml of ethylenediamine and heat the resulting mixture to reflux overnight. Cool and treat with 1.4 g of sodium hydroxide. Evaporate the solvent and triturate the resulting residue with 2 x 100 ml methylene chloride. Filter Treat the filtrate with 66.6 g of tert-dibutyl dicarbonate. and stir the reaction mixture overnight. Discard the solvent and subject the residue to flash chromatography using a 25% ethyl acetate / hexane mixture as eluting agent. The desired product is obtained which is chromatographed on silica gel using a 50% ethyl acetate / hexane mixture as eluting agent. Rf 0.64.

The above product can be bis-N-alkylated and the protecting groups are then removed by methods analogous to those used in reaction steps D and E of Example 1 to give the desired compounds of formula RΉN (CH₂)₂(CH₂)_gN (CH₂) HR 'as, for example, 1,16-bis-methyl, 4,13,16-tetraazahexadecane tetrahydrochloride.

-22Examples illustrating Reaction Scheme D

Example 5 1,18-bis / (2-phenyl) -ethyl-1,5,14,8-tetraaza octadecane tetrachloride

Step A. 1,18-Bisf / (phenyl) -methyl 17-carbonyl-5,14-bis [(phenyl) -methyl-1,5,14,18-tetraazaoctadecane. Using an ice bath, a solution of 5.14-bis [(phenyl) -methyl 17 -1,5,14,18-tetraazaoctadecane and 2 g (20 mmol) of triethylamine was ice-cold. in 100 ml chloroform. 2.3 g (15 mmol) of a phenylacetyl chloride solution and 10 ml of chloroform were added dropwise. The ice bath was removed and the reaction stirred for 18 hours at room temperature. The reaction mixture was extracted with aqueous sodium hydrogen carbonate solution, the organic phase was dried and evaporated. The resulting residue was chromatographed using a flash column of silica gel and ethyl acetate as eluting agent. 3 g of the expected product were obtained as a thick oil.

Step B. To a solution of 0.5 g lithium aluminum hydride in 500 ml tetrahydrofuran was added dropwise a solution of the product prepared in reaction phase A in 150 ml tetrahydrofuran. The reaction mixture was stirred for 48 hours at room temperature. Excess reducing agent was decomposed by adding dropwise 1 ml water, 1 ml 15% sodium hydroxide and then 3 ml water. The reaction mixture was filtered and the filtrate was evaporated. The remaining residue was treated with ethanol (100 ml) and anhydrous hydrogen chloride was added as a gas, converting 1,18-bis (2-phenyl) -ethyl 1.7-5.14 -bis- / (phenyl) -methyl-1,1,14,18-tetraazaoctadecane in the corresponding tetrachlorhydrate. Using a Parr hydrogenation apparatus, the above product was hydrogenated for 24 hours at a pressure of 43 psig in 150 ml ethanol and in the presence of 0.3 g Pearlman catalyst. The catalyst was filtered off and the filtrate was evaporated. The resulting residue was crystallized from propanol-2 to give 1,1-bis / (2-phenyl) -ethyl (M), 5,14,18-tetraazaocta (t) -trachlorhydrate hemihydrate. . Dean. Mp 228 ° -231 ° C.

Alternative Reduction Alkylation Techniques

Example 6 1,14-bis / ((phenyl)) -methyl-1, 5,10,14-tetraazatetradecane tetrachloride

Mix 2.02 g spermine, 2.13 ml benzaldehyde, 40 ml ethyl alcohol and 0.1 g Platinum oxide and saturate the resulting mixture with hydrogen at 45 lbs./sq. in.) until the absorption of hydrogen ceases.

Filter off the catalyst, add 100 ml of 1N hydrochloric acid in ethanol and water until the solids dissolve. Add isopropanol until a cloudy solution is obtained. Cool and filter and dry the resulting solid. 20 g of the desired compound are obtained. Mp / 290 ° C. This compound is chromatographed using silica gel and as eluting agent a mixture of a concentrated solution of 20% ammonia and methyl alcohol. Rf 0.50.

Example 7 1,18-bis (4-methylthiophenyl) -methyl-1,1,1,1,18-tetraazaoctadecane tetrahydrochloride

Mix 0.81 g of the product prepared in the reaction phase.

Example 1 B, 0.05 g sodium carbonate, 0.25 g NaBH4 CN and 0.53 ml 4-methylthiobenzaldehyde in 100 ml methyl alcohol and stir the resulting mixture overnight at room temperature. Pour the reaction mixture into 300 ml of methylene chloride. ►

Wash with 100 ml of 1 N sodium hydroxide and 100 ml of saturated sodium chloride solution, dry over magnesium sulfate and evaporate in vacuo. Recrystallize the resulting product from ethyl acetate, cool, filter and treat with 10 mL of ethyl alcohol and 30 mL of 2N hydrochloric acid in ethyl ether. Filter and dry the resulting solid to yield 0.32 g of the expected product, which is chromatographed using silica gel and a mixture of a concentrated solution of ammonia 40% methyl alcohol as eluting agent. Rf 0.58.

Example illustrating Reaction Scheme E

Example 8 1,18-bis (phenyl) -1,1,14,18-tetraazaoctadecane

Stage A. N-Phen1-N, N¹bis (t-butoxycarbonyl) propanediamine Using an ice bath, cool 200 ml anhydrous ethyl ether and add 8.74 g (0.23 mol) lithium aluminum hydride. Add dropwise over 30 minutes 14.6 g of 3-anionopropionitrile in 50 ml of ethyl ether, remove the ice bath and heat the resulting mixture to reflux over

<img file="PT86687B_D0015.tif" />

night. In the order given, add 8.7 ml of water, 1.5 g of sodium hydroxide in 10 ml of water and 25 ml of water. Filter off the resulting precipitate, wash with 200 ml ether and evaporate the solvent in vacuo. Treat the resulting N- (phenyl) -propanediamine with 43.6 g of tert-dibutyl dicarbonate. in 600 ml of methylene chloride. Stir overnight, evaporate the solvent and evaporate the resulting residue using silica gel and eluting with a mixture of 17% ethyl acetate / hexane. The desired compound is obtained which is chromatographed using silica gel and a 25% ethyl acetate / hexane mixture as eluting agent. Rf. 0.50.

Phase B. 1,18-bis (phenyl) -1,5,14,18-tetra- (t-butoxycarbonyl) -1,1,14,18-tetraazaoctadecane. Stir for about 16 hours a mixture containing 13 g of the product prepared in reaction phase A, 3.70 g of diiodooctane and 4.14 g of tert-butoxide. of potassium in 200 ml of dimethyl formamide. Evaporate the solvent in vacuo (0.5 mm) at 45 ° C and treat the resulting residue with 800 ml of ethyl acetate. Wash with 2x300 ml of water, dry over magnesium sulfate and discard the dissolve in vacuo. Subject the resulting viscous oil to flash chromatography using silica gel and eluting as 15% ethyl acetate. 5.7 g of the desired compound is obtained which is chromatographed using silica gel and eluting with a mixture of ethyl acetate / hexane. Rf. 0.36. Using the technique described in reaction step E of Example 1 remove the nitrogen protecting groups, i.e. t-butoxycarbonyl groups, to give the title compound thereof

-26 // example. Mp 264-267 ° C.

Examples illustrating reaction scheme F

Example 9 1,18-bis (2,3-butadienyl) -1,5,1,18-tetraaza-octadecane tetrahydrochloride

Step A. N- (t-Butoxycarbonyl) propargylamine. A stirred solution of 99.18 g of tert-dibutyl dicarbonate. In 900 ml of methylene chloride was added dropwise 25 g of propargamine in 25 ml of methylene chloride. After 2 hours, separate the solvent by evaporation in vacuo to give 70 mg of desired N-protected propargylamine.

Step B. N- (t-butoxycarbonyl) -2,3-butadienylamine. Heat at reflux for 12 hours a mixture containing 70 g of N- (t-butoxycarbonyl) -propargamine, 93.5 ml of a solution of 32% formaldehyde, 76.4 ml diisopropylamine, 19.66 g cuprous bromide and 860 ml p-dioxane. Cool and dilute the resulting mixture with 3000 mL of ethyl ether, wash with 500 mL of water, 1000 mL of acetic acid, 2x500 mL of water and 200 mL of a saturated sodium chloride solution, dry over magnesium sulfate and evaporate. in the void. Flash chromatography of the resulting residue using silica gel and eluting with a 10% ethyl ether / hexane mixture affords 40.8 g of the desired compound. This compound is chromatographed using silica gel and a 10% ethyl acetate / hexane mixture as eluting agent. Rf. 0.31.

Phase C. N, N-bi s / T (phenyl) methyl-1,8-di-aminoctane. Mix 14.4 g diaminooctane, 20.3 ml benzaldehyde and 0.66 g platinum oxide in 100 ml ethanol. Treat mix results

- with hydrogen at 28.5 kg / cm (45 lbs./sq. in.) until saturation. Filter, evaporate the solvent in vacuo and distill the prepared compound. 25.5 g of the expected product are obtained, eg 185 ° -190 ° C at 0.1 mm.

Stage D. 1,18-bi s (hydroxy) -5,14-bi sf (phenyl) -methyl J-5,

14-diazaoctadecane. Reflux for 18 hours a mixture containing 25.5 g of the product prepared in reaction phase C,

13.2 ml of 3-chloro-1-hydroxypropane, 50.4 g of sodium carbonate and 1.19 g of sodium iodide in 40 ml of n-butanol. Cool this reaction mixture and pour over 700 ml of ethyl acetate, wash with water, dry over magnesium sulfate and remove the solvent in vacuo to give a residue which, after distillation, affords 30.0 g of the desired product, eg 25 ° -252 ° C ( 0.1 mm).

Phase E. 1,18-bis (hydroxy) -5,14-diazaoctadecane. Saturate with hydrogen at 28.5 kg / cm (45 lbs./sq. In.) A mixture containing 3.0 g of the product prepared in reaction phase D, 30 ml of acetic acid and 0.6 g of palladium oxide. . Filtration and removal of the solvent in vacuo affords 1.77 g of the desired product which is chromatographed using silica gel and a concentrated 10% ammonia / methyl alcohol solution as eluting agent.

Rf. 0.37.

-28..X⁴/

Fa^if F. 1,18-bis (hydroxy) -5,14-bis- (t-butoxycarbonyl) -5,14-diazaoctadecane. Stir overnight a mixture containing 1.77 g of the product prepared in reaction phase E, 2.97 g (0.0136 mole) of tertiary dibutyl dicarbonide, 3 ml of triethyl amine and 50 ml of methylene chloride. Dilute the mixture with 200 ml methylene chloride, wash with 200 ml 0.5 N hydrochloric acid and then with 100 ml saturated sodium chloride solution, dry over magnesium sulfate and evaporate the solvent in vacuo. Subject the resulting residue to flash chromatography using silica gel and 75% ethyl acetate as an eluting agent. The desired product is obtained which is chromatographed on silica gel using a 75% ethyl acetate / hexane mixture as eluting agent. Rf. 0.29.

Step G. 1,18-bis (methanesulfonyl) -5,14-bis (t-butoxycarbonyl) -5,14-diazaoctadecane. Cool to 0 ° C a mixture containing 3.0 g of the product prepared in reaction phase F, 3.3 ml of triethylamine and 70 ml of methylene chloride. Add dropwise 1.22 ml of mesyl chloride in 10 ml of methylene chloride and stir the resulting mixture at 0 ° C for 90 minutes. Pour the mixture into 100 ml methylene chloride, wash with 100 ml 1N acetic acid, 100 ml water, 100 ml saturated sodium bicarbonate solution, dry over magnesium sulfate and remove solvent in vacuo. . Flash chromatography of the resulting residue using silica gel and eluting with a 60% ethyl acetate / hexane mixture affords 3.5 g of the desired compound. Rf 0.39.

<img file="PT86687B_D0016.tif" />

Phase Η. 1,18-bis (2,3-butadienyl) -1,1,14,18-tetra- (t-butoxycarbonyl) -1,5,14,18-tetraazaoctadeGano. Add a mixture containing 3.5 g of the compound prepared in reaction phase G.

1.74 g sodium iodide, 0.51 g hexane washed sodium hydride (60% in Oil) in 12 ml dimethylformamide with 2.16 g N- (t-butoxycarbonyl) -2,3-butanedienylamine that is, the compound prepared in reaction phase B and leave the resulting mixture to stand for 2 hours. Evaporate the solvent in a vacuum, add)! After 350 ml of ethyl acetate are added to the resulting residue, wash with

4x50 ml of water and 100 ml of a saturated sodium chloride solution and dry over magnesium sulfate. Discard the solvents in vacuo and flash chromatograph the resulting residue using silica gel and a 30% ethyl acetate / hexane mixture as eluting agent. 0.5 g of the desired compound are obtained as a viscous oil which is chromatographed using silica gel and a 25% ethyl acetate / hexane mixture as eluting agent.

)

Step I. 1,18-Bis (2,3-butadienyl) -1,5,14,18-tetraazaoctadecane tetrahydrochloride. Dissolve 0.5 g of the product prepared in reaction H in 2 ml of ethyl alcohol and continuously stirring treat the resulting mixture with 10 ml of 2N hydrochloric acid in ethyl ether. Stir the resulting mixture overnight, filter and dry the resulting solids in vacuo. 0.22g of the desired compound is obtained. Mp 283-284 ° C (decomposed).

Using the abbreviated form to designate the compounds included in the general formula

NHR (CH₂)_nNH (CH₂)_mNH (CH₂)_noNHR

In which the groups represented by the symbol R are terminal groups and the groups positioned between those symbols R are polyamino groups, it is observed that the following specific compounds are readily prepared using the above techniques and conventional chemical principles which allow to perform the necessary modifications:

<td>Polyamine Groups</td><td>R Terminal Groups</td>

<td> 3-8-3</td><td>bis-meti 1</td>

<td> 3-8-3</td><td>bi s-eti1</td>

<td> 3-8-3</td><td>bi s-propi1</td>

<td> 3-8-3</td><td>bis - buti 1</td>

<td> 3-8-3</td><td>bi st-buti1</td>

<td> 3-8-3</td><td>bis - feni 1</td>

<td> 3-8-3</td><td>bi s-naphthi1</td>

<td> 3-8-3</td><td>bis- (phenyl) methyl 1]</td>

<td> 3-8-3</td><td>bi s- | F (phenyl) -ethyl 1]</td>

<td> 3-8-3</td><td>bi s - [(naphthyl 1) -methyl 1 /</td>

<td> 3-8-3</td><td>bios- (* -naphthyl) -eti</td>

<td> 3-8-3</td><td>bis - £ (4 - chlorophenyl) - me tilj</td>

<td> 3-8-3</td><td>bi sf (4-hydroxyphenyl) -methyl 1 /</td>

<td> 3-8-3</td><td>bis- (4-methoxyphenyl) methyl</td>

<td> 3-8-3</td><td>bis-f (4-methylphenyl) methyl</td>

<td> 3-8-3</td><td>bis - (4-methylthio) methyl</td>

<td> 3-8-3</td><td>bis- (4-methyl Isulphinyl) methyl /</td>

<td> 3-8-3</td><td>bi sf (4-methylsulfonyl) -methyl 1 /</td>

<td> 3-8-3</td><td>bis - (a ce ti 1 eni 1)</td>

<td> 3-8-3</td><td>bis s- (2,3-butadienyl)</td>

<td> 3-8-3</td><td>bis-ali1</td>

<td> 3-8-3</td><td>bis-aleni1</td>

31 as well as its analogs 2-8-2, 4-8-4, 5-8-5 and 6-8-6 and those in which the polyamine group represents a specific group as shown in the following table:

Polyamine Group

<td> 2-4-2</td><td> 3-4-3</td><td> 4-4-4</td><td> 5-4-5</td><td> 6-4-6</td>

<td> 2-5-2</td><td> 3-5-3</td><td> 4-5-4</td><td> 5-5-5</td><td> 6-5-6</td>

<td> 2-6-2</td><td> 3-6-3</td><td> 4-6-4</td><td> 5-6-5</td><td> 6-6-6</td>

<td> 2-7-2</td><td> 3-7-3</td><td> 4-7-4</td><td> 5-7-5</td><td> 6-7-6</td>

<td> 2-9-2</td><td> 3-9-3</td><td> 4-9-4</td><td> 5-9-5</td><td> 6-9-6</td>

<td> 2-10-2</td><td> 3-10-3</td><td> 4-10-4</td><td> 5-10-5</td><td> 6-10-6</td>

<td> 2-11-2</td><td> 3-11-3</td><td> 4-11-4</td><td> 5-11-5</td><td> 6-11-6</td>

<td> 2-12-2</td><td> 3-12-3</td><td> 4-12-4</td><td> 5-12-5</td><td> 6-12-6</td>

The compounds of the present invention find application in the treatment of protozoan diseases. The combination of these compounds with an ornithine decarboxyase or arginine decarboxyase inhibitor has been found to allow greater efficacy in the treatment of the specific disease being treated.

In general, the compounds of formula I according to the present invention are used in the treatment of diseases caused by parasitic protozoa installed intracellularly or extracellularly in host mammals to be treated.

Parasitic animals belonging to the Protozoa Phylum include the genera Plasmodium (which includes, for example, vivax, malariae, ovale, falciparum, knowlesi, berghei, vinckei, chabaudi, gallinaceum and lophurae), heishamania

(Which includes, for example, the Donovani, tropica, brazi-liensis and Mexican species), Babesia (which includes, for example, bovis, rodhaini and microti), Trypanosoma (of the stercoraria class) (including, for example, cruzi species, noting that this species is an example of Trypanosoma which uses arginine-decarboxyase in its polyamine metabolic cycle and that combination therapy with a arginine-decarboxy ase), Toxoplasma (which includes, for example, the species gondii) and Theileria (which includes, for example, the silly species). Parasitic protozoa that live outside the blood cells are, for example, the salivary class Trypanosoma protozoa (which includes, for example, the Rhodesian, Gambian, Brucei, Evansi, equinum, equiper dum, Congolese and vivax species), Trichomonas (which includes, for example, the species faginalis, fetus and gallinae), Entamoeba (which includes, for example, histolytica and invadens), Pneumocystis carini, Eimeria (which includes, for example, tenella, necatriz and brunetti), Cryptosporidia and Giardia (which includes, for example, the lamblia species).

All of the above protozoa are known to infect animals and the diseases for which these protozoa are responsible are well known. Thus one of the objects according to the present invention is to use the compounds of formula I, either alone or in combination with appropriate ornithine or arginine decarboxylase inhibitors, in the treatment of diseases caused by the protozoa mentioned above. The main human diseases requiring new therapy are amoebiasis, malaria, eishmaniasis, trypanosomiasis, toxoplasmosis, as well as diseases caused by so-called infections.

-33L opportunists as provoked by the protozoan Pneumocystic cari ni i.

As is well known, malaria is still the most important infection worldwide in terms of human suffering and death. Even today there is a desperate need for clinically tested, effective and safe chemicals to beat this protozoan infection because, despite advances in the treatment of malaria, its transmission is progressing, the most common species of Plasrnodium falciparum. dangerous and predominant species of plasmodia, resistant to a large number of compounds propagate and increase their degree of resistance. In fact, more than 200 million people are said to suffer from malaria and in Africa alone more than one million deaths per year are associated with malaria, and travel to and from these regions is known to be present. Endemic diseases are a major health problem. Thus, a particularly important aspect in accordance with the present invention is the use of the compounds of formula I, the efficacy of which is enhanced by association with an ornithine decarboxyase inhibitor in the treatment of the following types of malaria other than 1 prime examples of the application of this vivax malaria, ovale malaria, falciparum malaria (including cerebral malaria), malaria malariae and hemoglubinurica fever (blackwater boil) and algid malaria (algid malaria).

Based on conventional in vivo and in vitro laboratory assays well known in the evaluation of compounds useful in the treatment of protozoan infections

As well as by comparison with agents which exhibit action against protozoa such as chloroquine, the compounds of formula I have been found to be effective in treating protozoan diseases when administered at doses of from about 1 to about 100 mg / kg body weight per day. When administering these compounds parenterally, a dose of about 10 to 30 mg / kg and a dose of 3 to 5 times higher when administered enterally is preferred. In the initial stages of treatment these compounds are preferably administered 3 times per day for about 3 days, and treatment is continued once daily until laboratory analysis demonstrates cure. The preferred route for administration is intramuscular administration.

Suitable ornithine decarboxyase inhibitors are, for example, Q-difluoromethyl ornithine and α-monofluoromethyl ornithine, although other well known ornithine decarboxyase inhibitors may also be used. Suitable inhibitors of arginine decarboxyase are, for example, mono- and difluoro methyl-arginine; These ornithine decarboxyase and arginine decarboxyase inhibitors are known and may be purchased by those skilled in the art. It is therefore evident that when a given protozoan uses the enzyme arginine decarboxylase in its polyamine metabolic cycle, the compounds of the present invention associated with arginine decarboxylase inhibitors should be administered; Similarly, when the protozoan causing the infection utilizes the enzyme ornithine decarboxyase in its metabolic cycle of the polyamines, the compounds according to the present invention should be administered.

<img file="PT86687B_D0017.tif" />

associated with an ornithine decarboxyase inhibitor. These ornithine decarboxylase and arginine decarboxylase inhibitors are used in doses ranging from about 50 to 500 mg / kg body weight and per day, generally extrapolated to about 10-20 g by weight. patients with about 70 kg. In this combination therapy these inhibitors may be administered at the beginning of treatment with the new polyamines of general formula I or may be administered throughout the treatment allowing an appropriate blood level to be maintained. Ornithine decarboxylase or arginine decarboxylase inhibitors may preferably be administered intravenously or in the form of drinkable solutions depending on the condition of the patient and the diagnosis of the physician. In practice, the anti-protozoan agent, in this case a polyamine and the arginine or ornithine decarboxylase inhibitors, are not administered in a single pharmaceutical composition. It is even preferred to co-administer these components as separate stations. For example, it may be preferable to administer the polyamine of formula I in three equal doses and the decarboxyase inhibitor (s) twice daily. 0 An important aspect of treatment is to use both medicines together. The most effective method for treating these diseases is to administer a polyamine of formula I together with the appropriate decarboxylase inhibitor.

Using conventional techniques suitable pharmaceutical compositions for enteral and parenteral administration may be prepared, such as, for example, physiologically acceptable sterile solutions suitable for intramuscular injection.

As is well known in pharmaceutical research, when clinical practice involves the use of a generic class of compounds, some subclasses and some specific compounds are more efficient than the other members of the class. In accordance with the present invention, compounds having a central alkylene chain of 6 to 9 carbon atoms, especially those having 6.7 and 8 carbon atoms, are preferred. Preferred are also those compounds wherein the alkylene chains positioned at each end of the Central alkylenic chain carry 2,3 or 4 carbon atoms, the most preferred having 3 atoms. Thus, preferred compounds are those wherein the polyamine group exhibits a 3-6-3, 3-7-3, 3-8-3, 3-9-3, 2-6-3, 2-7-2 constitution. , 2-8-2, 2-9-2, 4-6-4, 4-7-4, 4-8-4 or 4-9-4. Preferred constitutions are 3-6-3, 3-7-3, 3-8-3 and 3-9-3, most preferred being 3-7-3 and 3-6-3. As the terminal group represented by R, a benzyl group is preferred, the compound preferably being a bis-benzyl compound. In all circumstances it has been shown that symmetrical compounds are preferred. Most preferred are bis-benzyl-3-7-3. Thus, when R represents an alkyl group, a methyl or ethyl or bis-dimethyl or bis-diethyl group is preferred.

As an inhibitor of ornithine decarboxyase, we prefer β-difluoromethyl ornithine, and as an inhibitor of arginine decarboxylase, Q-difluoromethyl arginine is preferred.