Novel polyamine derivatives novel polyamine derivatives

(74) Attorney General Tandbergs Patentkontor A / S, Oslo.

(3b) Priority Requested 03.02.37, US, No. 010380.

(54) DESCRIPTION OF THE INVENTION ANALOGUE IN THE PROCEDURE FOR THE PREPARATION OF NEW THERAPEUTIC ACTIVE POLYAM INDER1 WATER.

(57) Summary New compounds of formula

RHN (CH-) NH (CHJ NH (CH_n) NHR

L n 2 m 2 n and pharmaceutically acceptable salts thereof where n is an integer from 2 to 6, m is an integer from 3 to 12, and R is a C 1 -C 8 saturated or unsaturated hydrocarbyl radical or - - (Ar) -X wherein x is zero, one or two, Ar is phenyl or naphthyl, and X is H, C 1 -C 6 alkoxy, halogen, C 1 -C 2 alkyl, wherein R₁ is C 1 -C 6 alkyl, provided that R cannot be phenylmethyl when n is 2 and m is either 6 or 10, exhibit therapeutic activity. Preparation of the compound is described.

(56) Published publications BRD (DE) publication no. 2458222.

J.Med. Chem., Vol. 30 (1) 1987, pp. 201-204.

The present invention relates to the preparation of certain polyamine derivatives.

More particularly, the invention relates to an analogous process for the preparation of therapeutically active compounds of the formula

RHN (CH-) NH (CH_n) NH (CH 2) NHR n 2 m 2 and pharmaceutically acceptable salts thereof, wherein n is an integer from 2 to 6, m is an integer from 3 to 12 and R is a C or - (ΟΗ ^ Ιχ-ΑΓ-Χ wherein x is 0 or 1 and Ar is phenyl or ₁₅ naphthyl, and X is H or -SR wherein R is C-C. alkyl.

1 16

Where R is saturated hydrocarbyl, such compounds include straight-chain, branched or cyclized examples of alkyl radicals of up to 6 carbon atoms where t-butyl and cyclohexyl are preferred, and when R is an unsaturated hydrocarbyl moiety, such groups contain those radicals having one or two double bonds, and those having a triple bond which may be represented by such preferred radicals as -CH₂CH = CH₂, -CH₂CH₂CH = CH₂, -CH ^ CH, -CH₂CH = C = CH₂Optionally, such radicals may carry a phenyl or naphthyl group, such as e.g. OC = CHCH₂-. In the cases defined by the groups (CH₂)_n wherein n is 2 to 6, such moieties include straight-chain and branched-chain alkyl radicals of up to 6 carbon atoms, preferably ethylene, propylene and butylene as straight-chain alkylene groups, although these may be branched

JO chain groups. In the cases defined by the (Cl In the cases defined by - (CH₂) - (Ar) -X, it is preferred that x is 1 or 2, Ar is unsubstituted phenyl or naphthyl, but when substituted, it is preferred that the alkoxy radical is methoxy or ethoxy, the halogen is chlorine, the alkyl radical is methyl, ethyl or t -butyl and when the substituent is S (O)_nR-, preferably R ^ is methyl, ethyl or ⁵ t-butyl, and n is either 0, 1 or 2. In any particular compound defined by formula I, it is preferred that such compounds be symmetrical in structure. For example, it is preferred that the terminal R group for each individual compound is the same and that each (CH₂)_npart is the same.

¹⁰ In order to simplify the discussion and description of the application object, it is appropriate to use certain abbreviated forms to specify either generic or types of compounds. For example, the compound 1,18-bis - [(phenyl) methyl] 1,5,14,18-tetraazaoctadecane of the structure^1p och₂NH (CH₂) ₃NH (CH₂) _gNH. (CH₂) NHCH₂-Ø in its abbreviated version is shown as BnNH (3) NH (8) NH (3) NHBn, (Bn is benzyl), and may also be designated as a bis-benzyl 3-8-3 compound, wherein The nitrogen atoms, of course, must be understood. Where asymmetrical compounds are used, these will be indicated as, for example

N-benzyl-N'-phenethyl-3-8-4.

In general, the compounds of formula I can be prepared by chemical reactions analogous to those known in the art, the choice of any specific mode of preparation depending upon a number of factors. For example, the general availability and price of the reactants, the utility of certain generalized reactions to specific compounds, the presence of unsaturated hydrocarbyl groups, etc., are all factors fully understood by those skilled in the art and all of which contribute to the choice of synthesis in the preparation of any specific compound encompassed by the formula IN.

The analogous method of the invention is characterized in that

a) a suitable N-protected tetramine of the formula

HN (CH₂)_n-N- (CH₂)_m-N- (CH₂)_nNH

Pro Pro Pro Pro in which Pro is an N-protecting group, is reacted with a compound of formula

R¹-halide wherein R 1 is as defined for R in formula I, except that when R is - (CH 2) -Ar-X may not be 0, followed by cleavage ZX of the protecting group of the N-terminal alkylated product;

b) reductive alkylation of a compound of formula

H_, N (CH_J OH

2 n with a suitable aldehyde RUC ^ J ^ CHO wherein R 'is - (CH ^ J protected and mesylated product of the formula

R'N- (CH_) -OMS i ^{2 n}

Pro wherein Ms represents mesyl, with an N-protected diamine of the formula

Pro-NH (CH 2) NH-Pro m

followed by splitting of the protecting group,

c) N-aroylation of a partially N-protected tetramine of the formula

H_N- (CH) -NB

2 η n

-NB

2 η n wherein Bf represents benzyl is reacted with an aroyl chloride of the formula:

Ar-C-Cl followed by reduction and decomposition of the protecting group of the N-aroylated product,

d) for the preparation of a compound of formula 1 wherein ₁₀ R is - (CH₂)_x-Ar-X wherein x is 0, reaction of an N-protected compound of formula

Ar-N (CH_n) -NH '2 n iPro Pro with a suitable dihaloalkane of Formula I followed by cleavage of the protecting group, or

e) for the preparation of a compound of formula I wherein R is an unsaturated hydrocarbyl radical, reacting an N-protected diamine mesylate of the formula

MsO- (CH) -N- (CH_) -N- (CH_) -OMs n 2 m 2 n

Pro Pro wherein Ms is the mesyl group, with an N-protected amine of the formula R 1 -NH-Pro wherein is a suitable unsaturated hydrocarbyl radical,

Taking into account the foregoing, the following reaction schemes are illustrative of routes by which the compounds can be prepared.

Scheme A

<td>H₂N (CH₂)_mNH₂</td><td>EtOH</td>

<td> +</td><td>-------&gt; CN (CH₂ ) 2NH (CH2) mNH (CH2&gt; 2CN</td>

<td>h₂c = CHCN</td><td> 2</td>

2. + H₂ PtO₂

------------&gt;

HCl / AcOH

H2N (CH2) 3NH (CH2)_mNH (CH2) 3NH2 · 4HC1

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

<td>3 + OCOCO</td><td>• NaOH / H₂O_x</td>

<td>QQ</td><td>THF</td>

HN (CH2&gt; 3N- (CH2) mN- (CH2) 3NH

Boc Boc Boc Boc + R 'halide

Kt-BuO}

DMF

<img file="NO166362B_D0001.tif" />

BOC Boc Boc Boc + HC1 Et₂O

EtOH

R ¹ NH (CH₂ ) 3NH (CH₂ ) _mNH (CH₂ ) 3NHR ¹ -4HCL wherein R 'is as defined for R in formula I, except that when R is X - (Ar) - (CH₂)_χ, x cannot be zero, Boc is the tbutoxycarbonyl protecting group and Q is tert-butyl.

In the preceding five-step process, the initial step involves a specific N-alkylation intended to prepare compounds wherein n is 3 and which results in the reaction of a diamine (wherein m is as generically defined in formula I) with equivalents of acrylonitrile by heating the reactants, either in a suitable solvent or pure, according to standard conditions well known in the art. The resulting cyano derivatives (2) are chemically reduced by reaction with hydrogen in the presence of a catalyst (PtC 2) in a suitable solvent with 8 equivalents of hydrochloric or hydrobromic acid, to prepare the resulting hydrohalide salts according to standard procedures well known in the art. Of course, other reduction systems, such as reduction with lithium aluminum hydride, can also be used to prepare compounds of formula 3. After preparing these compounds, the hydrohalide salts are neutralized with base and the nitrogen atoms are protected, preferably with di-t-butyldicarbonate according to standard conditions. The tetra-N-protected amines (5) are alkylated with the appropriate alkyl compound or as alkyl halides (chlorine or bromine) by reaction in the presence of potassium butoxide according to standard alkylation procedures well known in the art. After alkylation, the N-protecting groups are removed by standard procedures, e.g. treatment with acid, preferably HCl, in the presence of a suitable solvent or solvent system, e.g. diethyl oxide in ethanol to give the desired products (6).

Alternatively, the compounds of formula 3 may be subjected to reductive alkylation using a suitable aldehyde (except that the aldehyde cannot carry an alkylthiosubstituent) and the reduction may be carried out by hydrogenation in the presence of PtCl according to well known procedures. This procedure does not require the protection of the nitrogen atoms of the intermediates. Where the desired final product carries an alkylthio group or an unsaturated hydrocarbyl radical on their terminal nitrogen atoms, the compounds 2 (° g of their homologs otherwise prepared, i.e. n is 2, 3 or 4) may be subjected to a reductive alkylation using a suitable aldehyde, but wherein the reduction is carried out with sodium cyanoborohydride according to standard techniques, nor does this reductive alkylation require protection of the nitrogen atoms by the intermediates.

A preferred method for preparing compounds of formula I wherein n is 4 (but which may also be used for those compounds wherein n is 2 to 6) and otherwise analogous to the compounds identified as (6) in Scheme A is following Scheme B.

Scheme B

H₂N (CH₂)_nOH

R (CH₂)_n&lt;CHO R'HN (CH₂)_nOH

--&gt;

PtO₂/ H₂ 8

N feskyttelse

<td>R'N (CH₂)_nOH</td><td></td><td>R ¹N- (CH₂) Noms</td><td></td>

<td> [</td><td>MsCI</td><td></td><td> --&gt; 5</td>

<td>BOC</td><td> -------------&gt;</td><td>Boc</td><td>NH (CH₂)_mNH</td>

<td></td><td>pyridine</td><td></td><td>1 i</td>

<td></td><td></td><td></td><td>Boc Boc</td>

<td>JL</td><td></td><td> 10</td><td></td>

wherein n is primarily 4 but may be 2 to 6, Boc is the tbutoxycarbonyl nitrogen protecting group (which is preferred, but which can be modified to any suitable N protecting group). R 'is X - (Ar) - (CH₂) where x is different from zero and Ar 'is an X-substituted alkyl or aryl group (as defined in formula I), n' is zero or a positive integer, and Ms is mesyl.

This reaction is started with reductive alkylation techniques using an amino alcohol (7) and a suitable aldehyde, to form R'-substituted amino alcohols (8) which are N-protected. The N-protected amino alcohols (9) are converted to their mesylates (10) under standard reaction conditions, for example, reaction with mesyl chloride in the presence of pyridine, preferably in the presence of a solvent such as ch₂ci₂.

The mesylate is subjected to alkylation with an N-protected diamine (ie, BocNH (CH-) NHBoc) using potassium-2 m butoxide in a solvent (DMF) using standard procedures. The tetra-N-protected tetramines (5) thus prepared are deprotected as indicated in Scheme A. The foregoing reductive alkylation, N-protection, mesylation, alkylation and deprotection utilize techniques and reaction conditions well known in the art.

In those cases where it is desirable to prepare compounds of formula I wherein n is 2, it is preferred to use Scheme C to obtain the necessary intermediates (13) which are subjected to the alkylation procedures discussed above in Reaction Scheme A.

Scheme C

H2NCH2CH2NH2 + Br (CH2) m^Br -&gt; H2N (CH2) 2NH (CH2)_mNH (CH2) 2NH2

12 13 wherein m is as defined in formula I.

The foregoing N-alkylation involves reacting a suitable dihaloalkane (12) with excess amounts (10x) of ethylenediamine (11) by heating the reactants to the reflux temperature in a suitable solvent, for example ethanol. Preferably, preparation of the desired end products bearing the R substituents on the terminal nitrogen atoms of the intermediates (13) can be carried out by reductive alkylation procedures using suitable aldehydes without N-protecting groups as alternatively discussed under Scheme A, or the intermediates (13) -protected, alkylated and deprotected by methods analogous to those depicted in steps 3, 4 and 5 of Scheme A.

A preferred method for preparing compounds in which Ar represents phenethyl (or naphthylethyl) (in particular, where n is 3 and m is 8) is reacting an aroyl chloride according to the method depicted in subsequent reaction scheme D.

Scheme D

<td></td><td>0 II</td>

<td>H₂N (CH₂) 3 ^ -Bn 0</td><td>II ØCH2CNH (CH2) 3N-Bn</td>

<td>(CH₂) 8 + ECH₂C-C1</td><td>(CH₂)₈__________________________in</td>

<td>H₂N (CH₂) 3Ν-ΒΠ</td><td>och₂CNH (CH₂) 3N-Bn II 0</td>

<td>Reduction</td><td>och₂CH₂NH (CH₂) 3N-Bn</td><td colspan="2">och₂CH₂NH (CH₂) 3NH</td>

<td> ------------&gt;</td><td> 1</td><td>H2</td><td> 1</td>

<td>16 LAH</td><td>(CH₂)8</td><td></td><td>-&gt; (CH₂) g</td>

<td></td><td> 1</td><td>Pd / C</td><td> 1</td>

<td></td><td>och₂CH₂NH (CH₂) 3Ν-ΒΠ</td><td></td><td>och₂CH₂NH (CH₂) 3NH</td>

wherein Bn is benzyl, O is phenyl and LAH is lithium aluminum hydride. As indicated above, the foregoing reaction is a preferred method of preparing a particular compound which results in N-alkylation of a partially protected intermediate (14) with an arylacetyl chloride (15) in the presence of triethylamine, using an inert solvent, under formation of an amide (16) which is chemically reduced, preferably with lithium aluminum hydride, and the resulting product (17) is catalytically debenzylated (H₂, Pd / C) to give the desired final product. These steps involve reaction techniques and procedures that are well known and understood in the art. Of course, the same reaction scheme can be used to prepare other compounds of formula I while adapting the technique to the usual precautions well understood by those skilled in the art.

Where Ar represents an aromatic group (X-phenyl or X-naphthyl) bonded directly to the terminal nitrogen atoms (i.e., x is zero), such compounds may be prepared according to the general reactions set forth in Scheme E. as follows:

Scheme E

<td> 5</td><td>ENT (CH2) 2^cn</td><td>LAH ENH (CH2&gt; 3NH2 ----&gt;</td>

<td></td><td></td><td>or</td>

<td></td><td></td><td>H2</td>

<td></td><td> 19</td><td> 20</td>

N protection -------------------&gt;

0N (CH2&gt; 3NH io II

Boc Boc

---------------&gt; NaH DMF

0-N (CHO) 3N-BOC II

Boc (CH2) m

0-N (CH₂) 3 ^ -Boc

IN

Boc

The foregoing reaction scheme illustrates the preparation of compounds wherein Ar is phenyl and the first step is a lithium aluminum hydride reduction performed according to the procedure published, for example, in (Bul. Soc. Chim. Fr., Part 2, 165-7 (1979)). Of course, this reaction can be extended to include naphthyl and X-substituted intermediates which are not adversely affected by the reaction conditions. Preferably, N protection is done with t-butoxycarbonyl protecting groups which are applied and removed according to standard techniques already discussed previously. The N-protected compounds are alkylated by reaction with a suitable dihaloalkane using well known standard procedures.

In those cases where it is desirable to prepare compounds of formula I containing an unsaturated hydrocarbyl group, i.e., compounds containing an acetylenic, allenic or allylic group, it is preferred to use the techniques shown in Reaction Scheme F as follows:

Reaction Scheme F

<td>BnNH (CH₂) mNHBn</td><td rowspan="2">Cl (CH₂) nOH ------------------_?nBnOH Needle</td><td>HO (CH₂) mN &lt;CH₂) mNtCH₂) 3OH in 1</td>

<td></td><td>Bn Bn</td>

<td> 23</td><td></td><td>_L4_</td>

<td>24 Debenzylate</td><td>HO (CH₂)_nNH (CH₂:</td><td> )_mNH (CH₂) 3OH</td>

---------------»

N-protected HO (CH₂)_nN (CH₂) mN (CH₂)_n0H MsCl -----------------------&gt; II -----------&gt;

Boc Boc CH₂C1₂

pyridine

MsO (CH2) N, N (CH₂) M ^ -tCH₂) nOMs R³ NHBoc loc Boc NaH, Na I

DMF 27

R3-N- (CH₂)_nN (CH₂) mN (CH₂) n³REF³ HCl / EtOH

Boc Boc Boc BOC

R3NH (CH₂)_nNH (CH₂) MNH (CH₂) _nNHR³ wherein R is a suitable unsaturated hydrocarbyl group, Bn is benzyl, MsCl is methanesulfonyl chloride, and Boc is the t-butoxycarbonyl protecting group.

In the foregoing reaction, a dibenzylated diamine (23) is N-alkylated by a simple displacement reaction to form compounds (24) which are then sequentially benzylated and

N-protected. These steps are performed according to well-known standard procedures. The resulting bis (hydroxy) aminoalkanes (26) are mesylated and the mesylates (27) are alkylated with two equivalents of an N-protected amine bearing a suitable unsaturated hydrocarbyl group, e.g. N- (t-butoxycarbonyl) -

2,3-butadienylamine. A tetraprotected tetramine (29) thus obtained is then easily deprotected to form the desired compounds (30).

In those cases where it is desirable to convert an alkylthio substituent to one of its higher oxidation states, the alkylthioether is treated with a peracid according to known conditions. Suitable oxidizing agents are 1 2 O 2 and NalO 2, but metachloroperoxybenzoic acid is preferred. In carrying out the oxidation, per. sulfinyl derivative 1 molar equivalent (per alkylthioether group) and 2 molar equivalents of the peracid to form the sulfonyl derivatives. The oxidations are carried out at temperatures of from 0 ° C to room temperature, in solvents which are not sensitive to oxidation. Preferred solvents are CF 2 Cl 2, CHCl 3, acetic acid and ethyl acetate.

Illustrative Examples for Reaction Scheme A

Example 1

1,18-bis - [(phenyl) methyl] -1,5,14,18-tetraazaoctadecan.4HC1

Step A. N, N'-bis- [2,2'-bis- (cyano) ethyl] -1,8-diaminooctane. 28.8 g (0.2 mole) of 1,8-diaminooctane was dissolved in 250 ml of EtOH. 27 ml (0.41 mol) of acrylonitrile was added and the mixture was gently refluxed overnight. The solvent was removed at reduced pressure. Analysis showed that the desired material was more than 95% pure.

Step B. 1,5,14,18-tetraazaoctadecane tetrahydrochloride.

50.0 g of the product of Example 1 was mixed with 2.0 g PtO₂, 133 ml of concentrated HCl and 600 ml of AcOH, and the resulting mixture was treated with H₂ at 3.2 kg / cm in a shake bottle until the hydrogen was no longer absorbed. The resulting mixture was filtered, the solvent was evaporated and the product was triturated with 1 liter of EtOH. The mixture was filtered and the product dried to give 51.6 g of the title compound, Rf 0.17 (silica gel plates eluted with 40% concentrated NH4 / CH2 OH).

Step C. 1,5,14,18-tetra- (t-butoxycarbony 1) -1,5,14,18tetraazaoctadecane. 28.0 g (0.069 mol) of the product of step B was treated with 10.99 g (0.274 mol) of NaOI-I in 120 ml of 1 When a homogeneous solution was obtained, 65.7 g (0.307 mol) of di-t-butyl dicarbonate in 750 ml of THF was added and the resulting mixture was stirred for 16 hours. The layers were separated, the aqueous layer was removed and washed twice with 500 ml of C ^ C C ·. eluted with 25% EtOAc / hexane to give 30.2 g of the desired product. Rf is 0.33 (silica gel plates eluted with 25% EtOAc / hexane). ί

Step D. 1,18-Bis - [(phenyl) methyl] -1,5,14,18-tetra (t-butoxycarbonyl) -1,5,14,18-tetraazaoctadecane. 20.0 g (0.03 mole) of the product of step C was dissolved in 30 ml of DMF and treated with 7.5 g (0.067 mole) of KtBuO and 7.96 ml (0.067 mole) of BnBr with stirring for 18 hours. The volatiles were evaporated (0.5 mm and 45 ° C) and the resulting residue was taken up in 1400 ml of EtOAc and washed twice with 500 ml of water. The organic layer was then dried (MgSO 4) and the solvent was evaporated in vacuo. Flash chromatography on silica gel eluting with 20% EtOAc / hexane gave 12.4 g (50%) of the desired product as a clear, viscous oil. Rf is 0.42 (silica gel plates eluted with 25% EtOAc / hexane).

Step E. 1,18-Bis - [(phenyl) methyl] -1,14,14,18-tetraaz aoctadecane.4HCl. 12.4 g (0.0147 mol) of the product of step D was dissolved in 14.7 ml of anhydrous EtOH and treated with 160 ml of 2N HCl in Et₂O with stirring overnight. Mix 166362 none was filtered, the filter cake was washed with Et₂O and dried to give 7.2 g of the desired compound, m.p. above 300 ° C. Rf is 0.24 (from silica gel eluted with 10% concentrated NH / CH2 OH).

^JJ

Example 1A

N, N'-bis- [3- (ethylamino) propyl] -1,7-heptanediamine

Steps A and B. 1,5,13,17-tetraazaheptadecane tetrahydrochloride. Prepare the title compound by the method of Israel et al., J ·. With. Chem. 710 (1964).

Step C. 1,5,13,17-tetra- (t-butoxycarbonyl) -1,5,14,18 tetraazaheptadencane. Combine 3.9 g (0.01 mole) of 1,5,13,17 tetraazaheptadecane tetrahydrochloride and 1.76 g (0.44 mole) of sodium hydroxide in 44 ml of water and stir until the mixture is homogeneous. Add to this mixture 9.6 g (0.044 mole) of di-tbutyl dicarbonate in 88 ml of THF and stir for 3 hours. Dilute the mixture with 300 ml of ethyl acetate (EtOAc) and separate the organic layer. Dry the organic layer over anhydrous MgSO4 and evaporate in vacuo to give a viscous oil. Purify the residue by flash chromatography (silica gel) and elute with 25% EtOAc / hexane to give 3.0 g of the title compound. Is 0.20 on silica gel plates eluted with 25% EtOAc / hexane.

Step D. 3,7,15,19-tetra- (t-butoxycarbonyl) -3,7,15,19 tetraazaheneicosane. Combine 3.0 g (0.0046 mol) of 1,5,13,17 tetra- (t-butoxycarbonyl) -1,5,14,18-tetraazaheptadecane and

0.45 g (0.011 mole) of sodium hydride (50% in oil) and stir the mixture until hydrogen evolution subsides. Add 0.9 ml of i-ο (0.011 mole) of ethyl iodide and stir the mixture for 18 hours. Evaporate DMF in vacuo and divide the residue between 600 ml of ethyl acetate and 200 ml of water. Separate the organic layer, dry the organic layer over anhydrous MgSO4 and evaporate in vacuo. Purify the residue by flash chromatography (silica gel) and elute with 20% EtOAc / hexane to give 1.68 g of the title compound. Rf is 0.5 on silica gel plates eluted with 25% EtOAc / hexane.

* Step E. N, N'-bis- [3- (ethylamino) propyl] -1,7 heptanediamine. Treat 1.68 g (0.0024 mol) of 3,7,15,19-tetra (t-butoxycarbonyl) -3,7,15,19-tetraazaheneicosane with 50 ml (1.0 N) HCl in methanol and stir over night. Filter among⁵ and recrystallize the title compound from methanol / water (20:80, v / v) to give 0.5 g of the title compound.

Rf is 0.39 on silica gel plates eluted with 40% ammonia (concentrated) in methanol; mp. 322-23 ° C under decomposition.

¹⁰ Example 2

1,18-bis- (butyl) -1,5,14,18-tetraazaoctadecane.4HCl

Step A. 1,18-Bis- (butyl) -1,5,14,18-tetra- (t-butoxycarbonyl) -1,5,14,18-tetraazaoctadecane. 3.5 g (0.0053 mole) of the product of Step C of Example 1 was combined with 2.7 g (0.024 mole) of KtBuO, 2.57 ml (0.024 mole) of 1-iodobutane in 10 ml of DMF, and the mixture was stirred for 18 hours. The volatiles were evaporated (0.5 mm at 45 ° C), the residue was dissolved in 500 ml of EtOAc, the organic layer was washed twice with 100 ml of water, and the organic layer was dried over MgSO 4.⁴the agents were evaporated and the residue was subjected to flash chromatography (silica gel eluted with 20% EtOAc / hexane) to give 1.32 g of the title compound. Rf is 0.36 (silica gel plates eluted with 20% EtOAc / hexane).

²⁵ Step B. 1.32 g (0.0017 mol) of the product of step A in this example was dissolved in 1.7 ml of EtOH and treated with 17 ml of 2N HCl in Et 2 O and the mixture was stirred overnight. The mixture was filtered and the precipitate was washed with Et 2 O, the washed material was recrystallized from isopropanol / water.

³⁰ The crystals were filtered and dried^Ρ2θ5 ^woe79 ° C and 0.1 mm) to give 0.62 g of the title compound, m.p. above 300 ° C. Rf is 0.47 (silica gel plates eluted with 20% concentrated NH 2 / CH 2 OH).

Example 2A

1,18- [(1-naphthyl) methyl] -1,5,14,18-tetraazoctadecanetetrahydrochloride hemihydrate

Step A. A solution of 1.7 g (9.6 mmol) of 1-chloromethyl166362 naphthalene in 5 ml of hexamethylphosphoric triamide (HMPA) was added to a mixture of 2.6 g, 5,14,18-tetra- (t-butoxycarbonyl) ) -

1,5,14,18-tetraazaoctadecane and 1.07 g of potassium t-butoxide in 50 ml of HMPA. The mixture was heated in an 80 ° C oil bath for 4 hours, poured into 300 ml of water and the aqueous mixture extracted with 2 x 300 ml of ethyl acetate. The combined extracts were extracted with 3 x 300 ml water and 300 ml saline. The organic layer was dried, evaporated and the residue was chromatographed (flash silica gel column and eluted with 4 to 1 toluene / ethyl acetate) to give 800 mg of 1,18-bis - [(1naphthyl) methyl) -1,5,14 , 18-tetra- (t-butoxycarbonyl) -1,5,14,18tetraazaoctadecan.

Step B. 800 mg of the product of step A was dissolved in 50 ml of methanol, excess HCl gas was added and the resulting mixture was stirred overnight at ambient temperature. The mixture was filtered, the solid was vacuum dried to give the desired product, m.p. 262-264 ° C.

Illustrative Examples for Reaction Scheme B

Example 3

1,20-bis - [(phenyl) methyl] -1,16,15,20-tetraazaeicosane.4HCl Step A. N, N'-bis- (t-butoxycarbonyl) -1,8-octanediamine

10.8 g (0.075 mole) of diaminooctane was dissolved in 200 ml and 100 ml of CH 2 OH, 32.7 g (0.156 mole) of di-t-butyldicarbonate was added and the mixture was stirred overnight. The mixture was evaporated in vacuo and the residue was crystallized from hexane to give 20.2 g of the desired compound, m.p. 96-97 ° C.

Step B. 4 - [[(Phenyl) methyl] amino] -butan-1-ol.

8.9 g (0.1 mole) of 4-amino-butan-1-ol were combined with 10.6 g (0.1 mole) of benzaldehyde, 100 ml of EtOH and 0.3 g of PtO hydrogenated at 3.2 kg / cm to H₂ was no longer recorded. The mixture was filtered, the solvent was evaporated in vacuo to give 17.7 g of the desired compound. Rf is 0.70 (eluted from silica gel with 10% concentrated nh₃/ ch₃OH).

Step C. 4- [N- (t-butoxycarbonyl) -N - [(phenyl) methyl] amino] -butan-1-ol. 17.7 g (0.1 mole) of the butanol from step B was combined with di-t-butyldicarbonate in 100 ml of CH 2 Cl 2 and the mixture was stirred overnight. The solvents were evaporated in vacuo and the residue was flash chromatographed and eluted from silica gel with 25% EtOAc / hexane to give the desired compound. Rf is 0.27 (silica gel plates eluted with 20% EtOAc / hexane).

Step D. 4- [N- (t-butoxycarbonyl) -N - [(phenyl) methyl] amino] -1-methanesulfonylbutane. A mixture containing

21.8 g (0.078 mol) of the product of step C, 250 ml of Cl 2 C 2 and 9.7 ml of pyridine (0.12 mol) were cooled in an ice bath and added dropwise (20 minutes) 6.65 ml (0.086 mole) of mesyl chloride in 6.6 ml of C ^ CC ^, and the mixture was allowed to warm to room temperature and stirred for 2 hours. The resulting mixture was poured into 200 mL of CH 7 g of the desired product. Rf is 0.36 (silica gel plates eluted with 25% EtOAc / hexane).

Step E. 1,20-Bis - [(phenyl) methyl) -1,6,15,20-tetra (t-butoxycarbonyl) -1,6,15,20-tetraazaeicosane. 5.16 g (0.015 mol) of the product of step A were mixed with 10.7 g (0.032 mol) of the product of step D, 3.92 g of Kt-BuO, 0.2 g of NaI and 60 ml of DMF, and the mixture was stirred for 72 hours at room temperature. The solvent was evaporated in vacuo, the residue was taken up in 600 ml of EtOAc and washed twice with 200 ml of water. The organic layer was dried (MgSO4), the solvents evaporated and the viscous residue flash chromatographed on silica gel eluting with 20% EtOAc / hexane to give the desired product. Rf is 0.22 (silica gel plates eluted with 20% EtOAc / hexane).

Step F. 1,20-bis - [(phenyl) -methyl] -1,6,15,20-tetraeicosane.4HCl. 4.7 g (0.0054 mol) of the product of step E was dissolved in 5 ml of EtOH and treated with 54 ml of 2N HCl in EtO2, the mixture was stirred overnight, filtered and the solid thus obtained was recrystallized from isopropanol / water. The mixture was cooled, filtered and the product dried (above?₂^Ο5 ^vECreduced pressure) to give the desired product, m.p. above 300 C. Rf is 0.47 (eluted from silica with 20% concentrated NH4 / CH2OH).

Illustrative Examples of Reaction Scheme C Example 4

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

4.75 g of 1,8-dibromooctane (0.017 mol) was combined with 20 ml of EtOH and 9.32 ml of ethylenediamine, and the mixture was refluxed overnight. The mixture was cooled and treated with 1.4 g of NaOH. The solvent was evaporated and the residue was triturated with 2 x 100 ml CH 2 Cl 2 and filtered. The filtrate was treated with 66.6 g of di-t-butyldicarbonate and the mixture was stirred overnight. The solvent was removed and the residue was subjected to flash chromatography and eluted with 25% EtOAc / hexane to give the desired product. Rf is 0.64 (eluted from silica gel with 50% EtOAc / hexane).

The foregoing may be bis-N-alkylated and the product deprotected by methods analogous to steps D and E of Example 1 to give the desired compounds of formula R'HN (CH ..) (CH ..) _ON (CH, J, NHR ', for example 1,16-bis - [(phenyl) 2 2 2 2 2 methyl] -1,4,13,16-tetraazahexadecane.4HCl.

Illustrative Example for Reaction Scheme D Example 5

1,18-bis - [(2-phenyl) ethyl] -1,5,14,18-tetraazaoctadecan.4HC1

Step A. 1,18-Bis - [[(phenyl) methyl] carbonyl] -5,14-bis [(phenyl) methyl ·] -1,14,14,18-tetraazaoctadecane. A solution of

2.2 g (5 mmol) of 5,14-bis - [(phenyl) methyl] -1,5,14,18-tetraazaoctadecane and 2 g (20 mmol) of triethylamine in 100 ml of chloroform were cooled on an ice bath. A solution of 2.3 g (15 mmol) of phenyl 16,6362 acetyl chloride in 10 ml of chloroform was added dropwise. The ice bath was removed and the mixture was stirred at ambient temperature for 18 hours. The reaction mixture was extracted with aqueous sodium bicarbonate, the organic layer was dried and evaporated. The residue was chromatographed on a flash silica gel column (ethyl acetate) to give 3 g of the desired product as a thick oil.

Step B. A solution of the product from step A in 150 THF was added dropwise to a suspension of 0.5 g LAH in 500 ml THF. The mixture was stirred for 48 hours at ambient temperature. Excess reducing agent was cleaved by dropwise addition of 1 ml of water, 1 ml of 15% NaOH and then 3 ml of water. The mixture was filtered and the filtrate was evaporated. The residue was taken up in 100 ml of ethanol and anhydrous HCl gas was added to convert the product_z1,18-bis - [(2-phenyl) -ethyl] -5,14-bis [(phenyl) -methyl] -1,15,14,18-tetraazaoctadecane, to its tetrahydrochloride salt. This product in 150 ml of ethanol was hydrogenated in the presence of Pearlman's catalyst (0.3 g) at 3.0 kg / cm 2 on a Parr hydrogenation apparatus for 24 hours. The catalyst was filtered off and the filtrate was evaporated. The residue was crystallized from 2-propanol to give the desired product 1,18-bis - [(2-phenyl) ethyl] -1,5,14,18-tetraazaoctadecanetetrahydrochloride salt hemihydrate, m.p. 228-231 ° C.

Alternative reductive alkylation procedures Example 6

1,14-bis - [(phenyl) methyl] -1,5,10,14-tetraazatetradecan.4HC1

2.02 g of sperm were combined with 2.13 ml of benzaldehyde, 40 ml of EtOH and 0.1 g of Pt0₉, the mixture was treated with H ^{2 2}(3.2 kg / cm) until H2 was no longer absorbed. The catalyst was removed by filtration, 100 mL of 1N HCl in EtOH was added, water was added until the solids dissolved, and isopropanol was added until the solution became pale. The solution was cooled and filtered and the resulting solid was dried to give 2.0 g of the desired product, m.p. above 29 ° C. Rf is 0.50 (eluted from silica gel with 20% concentrated NH₂ / CH₂OH).

Example 7

1,18-bis - [(4-methylthiophenyl) methyl] -1,5,14,18-tetraazaoctadecane. 4HC1

0.81 g of the product of B in Example 1 was combined with 0.05 g Na 2 CO 2, 0.25 g NaBH 3 CN and 0.53 ml, 4-methylthiobenzaldehyde in 100 ml CH 2 OH, and the mixture was stirred. overnight at room temperature. The reaction mixture was poured into 300 mL, washed with 100 mL of 1N NaOH and 100 mL of saturated NaCl, dried over MgSO4 and evaporated in vacuo. The crude product thus obtained was recrystallized from EtOAc, cooled, filtered and treated with 10 ml of EtOH and 30 ml of 2N HCl in Et₂O. The mixture was filtered and the solid thus obtained was dried to give 0.32 g of the desired product. R f is 0.58 (eluted from silica gel with 40% concentrated NH 2 / CH 2 OH).

Illustrative Example of Reaction Scheme E Example 8

1,18-bis- (phenyl) -1,5,14,18-tetraazaoctadecane

Step A. N-phenyl-N, N'-bis- (t-butoxycarbonyl) -propanediamine. 200 ml anhydrous Et₂O was cooled in an ice bath, and

8.74 g (0.23 mol) of lithium aluminum hydride was added. 14.6 g

3-Aniline Propionitrile in 50 ml Et₂O was added dropwise over 30 minutes, the ice bath was removed and the resulting mixture was refluxed overnight. In succession, 8.7 ml of water, 1.5 g of NaOH (in 10 ml of water) and 25 ml of water were added. The resulting precipitate was filtered, rinsed with 200 ml of Et₂O, and the solvent was removed in vacuo and the resulting N- (phenyl) -propanediamine was treated with 43.6 g of di-t-butyldicarbonate in 600 ml of CH₂C1₂After stirring overnight, the solvent was evaporated and the residue was subjected to flash chromatography from silica gel and eluted. with 17% EtOAc / hexane to give the desired compound. Rf is 0.50 (eluted from silica gel with 25% EtOAc / hexane).

Step Β. 1,18-bis- (phenyl) -1,5,14,18-tetra- (t-butoxy-carbonyl) -1,5,14,18-tetraazaoctadecane. A mixture containing 13.0 g of the product of step A, 3.70 g of diiodoctane and 4.14 g of potassium t-butoxide in 200 ml of DMF was stirred for 16 hours. The solvent was evaporated at 0.5 mm and 45 ° C, the residue was taken up in 800 ml of EtOAc. This solution was washed twice with 300 ml of water, dried (MgSO 4) and the solvent removed in vacuo. The viscous oil thus obtained was subjected to flash chromatography and eluted with 15% EtOAc from silica gel to give 5.7 g of the desired product.

Rf = 0.36 (eluted from silica gel with EtOAc / hexane). The N-boc protecting groups were removed according to the procedure described in step E of Example 1 to give the title compound with m.p. 264-267 ° C.

Illustrative Examples of Reaction Scheme F Example 9

1,18-Bis (2,3-butadienyl) -1,5,14,18-tetraazaoctadecantetrahydroklorid

Step A. N- (t-butoxycarbonyl) propargylamine.

g of propargylamine in 25 ml of CH was added dropwise to a stirred mixture of 99.18 g of di-t-butyl dicarbonate in 900 ml of Cf C After 2 hours, the solvent was removed in vacuo to give 70 g of the desired N-protected propargylamine.

Step B. N- (t-butoxycarbonyl) -2,3-butadienylamine.

A mixture containing 70 g of N- (t-butoxycarbonyl) propargylamine, 93.5 ml of 32% formaldehyde, 76.4 ml of diisopropylamine, 19.66 g of copper bromide and 860 ml of p-dioxane was refluxed for 12 hours. The resulting mixture was cooled and diluted with 3000 ml of Et 2 O, washed with 500 ml of water, 1000 ml of acetic acid, 500 ml of water (2 times), 200 ml of saturated sodium chloride and dried (MgSO 4) and evaporated in vacuo. Flash chromatography of the residue and elution from silica gel with 10% Et 2 O / hexane gave 40.8 g of the desired compound. Rf is 0.31 (eluted from silica gel with 10% EtOAc / hexane).

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Step C. N, N-bis - [(phenyl) methyl]] -, 8-diaminooctane.

14.4 g of diaminooctane was combined with 20.3 ml of benzaldehyde and 0.66 g of Pt 2 O in 100 ml of ethanol. The resulting mixture was treated with hydrogen at 3.2 kg / cm until the hydrogen uptake stopped. The mixture was filtered, the solvent was evaporated in vacuo and the remaining material was distilled to give 25.5 g of the desired product with b.p. 185-19 ° C at 0.1 mm.

Step D. 1,18-Bis (hydroxy) -5,14-bis - [(phenyl) methyl] -

5.14-diazaoctadecane. A mixture containing 25.5 g of the product of step C, 13.2 ml of 3-chloro-1-hydroxypropane, 50.4 g of Na₂CO₂ and 1.19 g of sodium iodide in 40 ml of n-butanol were refluxed for 18 hours. The mixture was cooled and poured into 700 ml of ethyl acetate, washed with water, dried over magnesium sulfate and the solvent removed in vacuo to give a residue which, by distillation, gave 30.0 g of the desired product with b.p. 25 ° -252 ° C at 0.1 mm.

Step Ε. 1,18-Bis (hydroxy) -5,14-diazaoctadecane.

A mixture containing 3.0 g of the product of step D, 30 ml

AcOH and 0.6 g of palladium oxide were hydrogenated at 3.2 kg / cm until the hydrogen uptake was stopped. The mixture was filtered and the solvent removed in vacuo to give 1.77 g of the desired product. Rf is 0.37 (eluted from silica gel with 10% concentrated NH4 / CH2OH).

Step F. 1,18-Bis- (hydroxy) -5,14-bis- (t-butoxycarbonyl) -

5.14-diazaoctadecane. A mixture containing 1.77 g of the product of step E, 2.97 g (0.0136 mol) of di-t-butyl dicarbonate, 3 ml of triethylamine and 50 ml of CH₂C1₂ was stirred overnight. The mixture was diluted with 200 ml of CH₂C1₂, washed with 200 ml of 0.5N HCl and then 100 ml of saturated NaCl, dried (over MgSOp, then the solvent was removed in vacuo. Flash chromatography of the residue and elution from silica gel with 75% EtOAc gave the desired product. Rf 0.29 (eluted from silica gel with 75% EtOAc / hexane).

Step G. 1,18-Bis- (methanesulfonyl) -5,14-bis- (t-butoxycarbonyl) -5,14-diazaoctadecane. A mixture containing 3.0 g of the product of step F, 3.3 ml of triethylamine and 70 ml of CH₂C1₂was cooled to 0 ° C. 1.22 ml of mesyl chloride in 10 ml of CH₂C1₂ was added dropwise and the resulting mixture was stirred at 0 ° C for 1.5 hours. The mixture was poured into 100 ml of CH₂C1₂, washed with 100 mL of 1N AcOH, 100 mL of water, 100 mL of saturated sodium bicarbonate, dried over MgSO 4, and the solvent removed in vacuo. Flash chromatography of the residue and elution from silica gel with 60% EtOAc / hexane gave 3.5 g of the desired product. Rf is 0.39.

Step Η. 1,18-Bis (2,3-butadienyl) -1,5,14,18-tetra (t-butoxycarbonyl) -1,5,14,18-tetraazaoctadecane. A mixture containing 3.5 g of the product of step G, 1.74 g of sodium iodide, 0.51 g of hexane washed sodium hydride (60% in oil) in 12 ml of DMF was mixed with 2.16 g of N- (t-butoxycarbonyl) -2,3-butanedienylamine (i.e., the product of step B), and the resulting mixture was allowed to stand for 2 hours. The solvent was removed in vacuo, 350 ml of ethyl acetate was added to the residue, washed with 50 ml of water (4 times), 100 ml of saturated sodium chloride and then dried over MgSO 4. The solvent was removed in vacuo and the residue was flash chromatographed from silica gel and eluted with 30% EtOAc / hexane to give 0.5 g of the desired product as a viscous oil. Rf is 0.39 (eluted from silica gel with 25% EtOAc / hexane).

Step I. 1,18-Bis- (2,3-butadienyl) -1,5,14,18-tetraazaoctadecane.4HCl. 0.5 g of the product of step H was dissolved in 2 ml of EtOH and treated with stirring with 10 ml of 2N HCl in Et₂O. The resulting mixture was stirred overnight, filtered and the solid dried in vacuo to give 0.22 g of the desired product, m.p. 283-284 ° C (dec.).

In their final application, the compounds have been found to be useful in the treatment of diseases caused by protozoal infections. In this final application, it has also been found that the use of concomitant therapy with either an ornithine decarboxylase inhibitor or an arginine decarboxylase inhibitor will improve the effectiveness of the treatment of the particular disease state being treated.

In general, compounds (I) are useful in the treatment of diseases caused by protozoal parasites living either intracellularly or extracellularly in a mammalian host to be treated. Included among such protozoa are parasites categorized into such genera as: Plasmodium (e.g., including such species as vivax, malariae, oval, falciparum, knowlesi, berghei, vinckei, chabaudi, gallinaceum and lophurae), Leishamania (e.g. species such as donovani, tropica, braziliensis and mexicana), Babesia (e.g. species such as bovis, rodhaini and microti), Trypanosoma (of the class stercoraria) (for example, such as cruzi, and it should be noted that this species is an example on a trypanosome that uses arginine decarboxylase in its polyamine metabolic pathway, and an accompanying therapy will therefore be with an ADC inhibitor), Toxoplasma (e.g., species such as gondii), and Theileria (e.g., parva). Protozoal parasites living outside the blood cells include Trypanosoma (of the class salivaria) (e.g. species such as rhodesiense, gambiense, brucei, evansi, equinum, equiperdum, congolense and vivax), Trichomonas (e.g. species such as vaginalis, fetus and gallinae), Entamoeba (e.g., such as histolytica and invadence), Penumocystis carini, Eimeria (e.g., tenella, necatrix and brunetti), Cryptosporidia, and Giardia (e.g., lamblia).

All of the foregoing protozoa are known to infect animals, and the particular diseases for which these protozoa are responsible are well known. Use of the compounds of formula I, with or without the concomitant use of suitable ornithine or arginine decarboxylase inhibitors in the treatment of diseases caused by the above protozoa, is within the scope of the invention. Important human disease states for which there is a need for new therapy are Amebiasis, Malaria, Leishmaniasis, Trypanosomiasis, Toxoplasmosis, as well as the so-called opportunistic infectious disease such as Penumocystic carinii.

As is known, malaria is the world's most important infection in terms of human suffering and death. Even today, there is a serious need for practical, effective and safe drugs to fight this protozoal infection, and despite significant advances in the treatment of malaria, the prevalence of malaria is steadily increasing, and strains of Plasmodium falciparum resistant to several drugs , is spreading steadily, and the degree of resistance to drugs of this very dangerous and prevalent plasmodial species is increasing. It has been stated that over 200 million people have malaria, and over one million deaths per year. Years are associated with malaria in Africa, and it is known that traveling to and from these regions is causing an increasing health problem. A particularly important feature of the invention is the use of the compounds of formula (I), effectively enhanced by concomitant treatment with an ornithine decarboxylase inhibitor, in the treatment of malaria, including, but not limited to, species such as vivax and oval malaria, falciparum malaria, (including cerebral malaria), malaria malaria and blackwater fever, and algal malaria.

Based on standard laboratory procedures (both in vitro and in vivo) well known for the assessment of compounds useful in the treatment of prozoal infections, as well as in comparison with known anti-protozoal agents (e.g. chloroquine), the compounds of Formula I effective in the treatment of protozoal diseases at doses of 1 to 100 mg per day. kg body weight per day. The preferred dose is 10 to 30 mg / kg when administered parenterally and 3 to 5 times when administered enterally. Preferably, the compound is administered in the initial phase of the treatment 3 times daily for approx. three days, followed by continuous treatment once per day. day until laboratory analyzes show a cure. The preferred mode of administration is intramuscular.

Suitable ornithine decarboxylase inhibitors are such compounds as α-difluoromethylornithine and α-monofluoromethylornithine, although other well-known ODC inhibitors may also be used. Suitable arginine decarboxylase inhibitors are mono-difluoromethylarginine compounds, and the ODC and ADC inhibitors are well known in the art. Of course, when a particular protozoa uses the arginine decarboxylase enzyme in its polyamine metabolic pathway, a companion therapy would use ADC inhibitors, and similarly, when the infecting protozoa uses ornithine decarboxylase enzyme in its polyamine metabolic pathway, include an ODC inhibitor. When used, the ODC and ADC inhibitors will be useful at a concentration of 50 to 500 mg / kg body weight per day. per day, generally extrapolated to 10-20 g per day. 70 kg patient. In the concomitant therapy, the inhibitors will be administered at the start of treatment with the novel polyamines of Formula I and will be administered to maintain a suitable blood level during the course of treatment. Depending on the condition of the patient to be treated as determined by the physician, the ODC or ADC inhibitors are preferably administered intravenously or as solutions suitable for drinking. In practice, the anti-protozoal polyamine and arginine or ornithine decarboxylase inhibitors will not be administered in a pharmaceutical composition. Rather, it is preferred to co-administer these components as separate entities. For example, it may be advantageous to administer the polyamine (I) in three equal doses, while it would be advantageous to administer the decarboxylase inhibitors twice daily. The important feature of treatment is that both drugs are used in conjunction with each other. The most effective way of treating diseases with polyamine (I) is with concomitant therapy with the appropriate decarboxylase inhibitor.

The suitable pharmaceutical formulations for enteral and parenteral administration may be prepared by well known methods in the art, such as in the preparation of sterile, physiologically acceptable solutions suitable for intramuscular injection.

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As is well known in the art in which generic classes of compounds are involved, certain subgeneric and certain specific compounds are more effective in their end uses than other members of the generic class. According to the invention, those compounds having a center alkyl chain of 6 to 9 carbon atoms are preferred, especially those having 6, 7 and 8 carbon atoms. Also preferred are those compounds wherein the alkylene chains which are on each side of the alkylene center chain are those having 2, 3 or 4 carbon atoms where 3 is the most preferred. Thus, the most preferred compounds are those in which the polyamine group has a 3-6-3-, a 3-7-3-, a 3-8-3-, a 3-9-3-, a 2-6-3-, a 2-7-2-, a 2-8-2-, a 2-9-2-, a 4-6-4-, a 4-7-4-, a 4-8-4- or a 4 -9-4-constitution. The preferred ones are 3-6-3, 3-7-3, 3-8-3 and 3-9-3 with 3-7-3 and 3-6-3 being the most preferred. The preferred terminal R group is benzyl, and the compound is preferably a bisbenzyl compound. In all cases, it has been shown that the symmetrical compounds are preferred. The most preferred is a bis-benzyl 3-7-3 compound. Where R is an alkyl radical, methyl or ethyl is preferred, or a bis-dimethyl or bis-diethyl.

The most preferred ornithine decarboxylase inhibitor is α-difluoromethylornithine, and the most preferred arginine decarboxylase inhibitor is α-difluoromethylarginine.

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