FR2826006A1 - Pyrimidine acyclonucleoside derivatives are antiviral agents, especially suitable for the treatment of HIV infections - Google Patents

Abstract

Pyrimidine derivatives (I), and their salts are new. Pyrimidine derivatives (I), and their salts are new. n = 3; R<1> = ethyl or isopropyl; R<2>, R2'' = H, 1-3C alkyl or halogen; one of R<3> and R<4> = H and the other = OH, or OR<5>; R<5> = 2-7C acyl, 1-6C alkyl amino carbonyl, aryl 1-6C alkyl amino carbonyl, (optionally substituted on the aryl), aryl carbonyl (optionally substituted on the aryl), or heteroaryl amino carbonyl. Independent claims are included for compositions containing (I) as active ingredient.

Description

<Desc / Clms Page number 1>

PYRIMIDINIC ACYCLONUCLEOSIDE DERIVATIVE, ITS PROCESS
PREPARATION AND USE Technical Field The present invention relates to a pyrimidine acyclonucleoside derivative active as a non-nucleoside inhibitor of HIV-1 reverse transcriptase, its method of preparation and its use.

STATE OF THE ART The current trend in the treatment of AIDS consists in resorting to combination therapy no longer including antiproteases, the side effects of which are bothersome and the modalities of taking the drug restrictive for the patient.

The recommended combination therapies therefore include one or two nucleoside inhibitors with which one or two non-nucleoside inhibitors are associated [G. J. Moyle, Infect. Med. 17 (6) 442-455 (2000)].

It is therefore important to develop new allosteric reverse transcriptase inhibitors with high specific activity and low toxicity.

étant t'E) \/) MR ! NE ayant la Formule A suivante :

Ces inhibiteurs allostériques de la transcriptase inverse à structure d'acyclonucléosides agissent à des concentrations nanomolaires et ils ont le désavantage de sélectionner très rapidement des mutants résistants. Allosteric inhibitors of reverse transcriptase with an acyclonucleoside structure are described, for example, in patent applications EP-A-0 449 726, WO-A-97/00432, WO-A-97/00309, WO-A-96 / 16675, WO-A-95/18109, EP-A-0 631 783, EP-A-0 420763, the best known example

being t'E) \ /) MR! BN having the following Formula A:

These allosteric reverse transcriptase inhibitors with an acyclonucleoside structure act at nanomolar concentrations and they have the disadvantage of selecting resistant mutants very quickly.

<Desc / Clms Page number 2>

qu'un reste N-hydroxyuréido fixé en o sur le radical porté par N-1 d'une thymine se comportait comme une pseudo-nucléobase contractant dans le cristal deux liaisons hydrogène (par son groupe CONH2) avec la nucléobase d'une autre molécule tandis que le groupe N-OH fonctionnait comme donneur de liaison hydrogène vis-àvis de l'atome d'oxygène d'un alcool. On the other hand, it has been shown by JMJ Tronchet, M. Zsély, M. Iznaden, F. BarbalatRey, M. Geoffroy & G. Bernardinelli, Carbohydr. Lett. 2,101-108 (1996) in connection with the following compound of Formula B:

that an N-hydroxyureido residue fixed in o on the radical carried by N-1 of a thymine behaved like a pseudo-nucleobase contracting in the crystal two hydrogen bonds (through its CONH2 group) with the nucleobase of another molecule while the N-OH group functioned as a hydrogen bond donor to the oxygen atom of an alcohol.

traités par l'enzyme. The N-hydroxyureido group could therefore constitute an interesting candidate for contracting new bonds with the allosteric site of reverse transcriptase and possibly outside it with nucleobases of nucleic acids.

treated with the enzyme.

In addition, from the point of view of the cytotoxicity of the products, the N-hydroxyureido group could also prove to be a good candidate because a structure-activity study of acyclonucleosides active against HIV-1 had shown that a hydrophilic group attached to N -1 on the nucleobase should decrease the cytotoxicity of the compound [J. M. J. Tronchet, M. Grigorov, N. Dolatshahi, F. Moriaud & J. Weber, Eur. J. Med. Chem., 32, 279-299 (1997)].

Thus, by introducing an N-hydroxyureido group in the co position of the chain fixed at N-1 of a pyrimidine substituted at C-4 and C-5, it was possible to improve the anti-HIV-1 activity of the already analogous products. known.

<Desc / Clms Page number 3>

[J. M. J. Tronchet, M. Iznaden, & N. Laroze, Carbohydr. Lett. 2, 313-320 (1997)] s'est révélé très modérément actif (cul50 70 nM).

However, the N-hydroxyureido moiety was not found to be sufficient to provide activity since the first compound prepared, namely the compound having Formula B above, was found to be completely inactive against HIV-1 and the second compound prepared. , namely the compound having the following Formula C

[JMJ Tronchet, M. Iznaden, & N. Laroze, Carbohydr. Lett. 2, 313-320 (1997)] has been shown to be very moderately active (70 nM cell).

On the basis of these facts, the present inventors continued their research and found with surprise that by lengthening the chain located between the nucleobase and the N-hydroxyureido group of a carbon atom to pass from a chain to two atoms of carbon to a chain with three carbon atoms, it was possible to obtain an acyclonucleoside which was non-toxic and active against HIV-1 at concentrations much lower than those achieved hitherto with similar products already known, while the same approval carried out on the analogue of the compound of Formula C bearing a hydroxyl group in place of the N-hydroxyureido group [cf. N. Larose, University of Geneva, Thesis No. 3212 (2000)] significantly reduced activity.

The present invention has been completed on the basis of these results.

<Desc / Clms Page number 4>

dans laquelle : - n est égal à 3 ; - R1 représente un groupe isopropyl ; - chacun des groupes R2 représente indépendamment l'un de l'autre un groupe méthyle ; - le groupe R3 représente un groupe -ORs, où RI est un groupe pivaloyl et R4 représente un atome d'hydrogène, ou un sel pharmaceutiquement acceptable de celui-ci. Disclosure of the Invention The subject of the present invention is a compound represented by the following general Formula 1:

in which: - n is equal to 3; - R1 represents an isopropyl group; - Each of the R 2 groups represents, independently of one another, a methyl group; the group R3 represents a group -ORs, where RI is a pivaloyl group and R4 represents a hydrogen atom, or a pharmaceutically acceptable salt thereof.

A subject of the present invention is also: the compound of Formula 1 as defined above for use as a medicament; - the compound of Formula 1 as defined above for use as an antiviral agent, in particular as an anti-HIV-1 agent; - a process for preparing the compound of Formula 1 as defined above; - a pharmaceutical composition containing as active ingredient at least the compound of Formula 1 as defined above; a pharmaceutical composition containing an effective amount as antiviral of the compound as defined above; and - the use of the compound of Formula 1 as defined above for the manufacture of an anti-HIV-1 drug;

<Desc / Clms Page number 5>

it being understood that by “compound as defined above”, it is also necessary to understand a pharmaceutically acceptable salt thereof.

Detailed description of the invention.

dans laquelle : - n est égal à 3 ; - R1 représente un groupe isopropyl ; - chacun des groupes R2 représente indépendamment l'un de l'autre un groupe méthyle ;

- le groupe R3 représente un groupe-OR5, où R5 est un groupe pivaloyle et R4 représente un atome d'hydrogène, à savoir le 1- (3-N1-pivaloyloxyuréidopropyloxyméthyl)-6- (3, 5-diméthylbenzyl)-5- isopropyluracile. The compound of the present invention is represented by the following General Formula 1:

in which: - n is equal to 3; - R1 represents an isopropyl group; - Each of the R 2 groups represents, independently of one another, a methyl group;

- the group R3 represents a group -OR5, where R5 is a pivaloyl group and R4 represents a hydrogen atom, namely 1- (3-N1-pivaloyloxyuréidopropyloxymethyl) -6- (3, 5-dimethylbenzyl) -5- isopropyluracil.

This compound of the present invention may be in the form of conventional pharmaceutically acceptable salts thereof.

The compound of the present invention can be prepared according to the following reaction scheme in which n, R ', R2, R3, R4 and R5 have the same meanings as defined above.

<Desc / Clms Page number 6>

R2 R2 )-\ Etape a) \- COOEt + RIe HBrC00Et 1) Zn/solvant \/Ri iv R2 III 2) acide p2 II Etape b) thiourée dans milieu alcoolique basique 0 2 0 JLRt Etape c) ! i Rl acide organique AR2 acide organique, 32 H dilué H VI V Etape d) 1) glycosidation avec MeCOOCH20 (CH2) nOCOMe (n=3) VII 2) solvolyse 9 2 0 - ? 1 R JL R Ri R QH-R2--------. --R 0 I ?--0 1j", R 0-1 réaction de l~ o 1 Mitsunobuavec o) (CHOH PhOCONHOCOOPh) t (CH ; PhON IX 1 X OCOOPh Etape f) aminolyse Yé- y 0 1 R2 )... R2 : s) YYA - R2 0 N 0, 1, 1, 1 2-K 0*'N 0 2 0-1 o-acylation o-J 1 1 (CH2) nOH (CH2) n N-OR5 HO--N '-NH2 ONH2 0 Ib la Reaction scheme

R2 R2) - \ Step a) \ - COOEt + RIe HBrC00Et 1) Zn / solvent \ / Ri iv R2 III 2) acid p2 II Step b) thiourea in basic alcoholic medium 0 2 0 JLRt Step c)! i Rl organic acid AR2 organic acid, 32 H diluted H VI V Step d) 1) glycosidation with MeCOOCH20 (CH2) nOCOMe (n = 3) VII 2) solvolysis 9 2 0 -? 1 R JL R Ri R QH-R2 --------. --R 0 I? - 0 1j ", R 0-1 reaction of l ~ o 1 Mitsunobuavec o) (CHOH PhOCONHOCOOPh) t (CH; PhON IX 1 X OCOOPh Step f) aminolysis Yé y 0 1 R2). .. R2: s) YYA - R2 0 N 0, 1, 1, 1 2-K 0 * 'N 0 2 0-1 o-acylation oJ 1 1 (CH2) nOH (CH2) n N-OR5 HO-- N '-NH2 ONH2 0 Ib la

<Desc / Clms Page number 7>

The nucleobases represented by Formulas V and VI above are known and they are usually prepared by electrophilic substitution [cf., p. ex. Y. S. Lee & Y. H. Kim, Synth. Common. 29 (9) 1503-17 (1999)].

However, in the process of the present invention, the pyrimidine ring of the modified nucleobases of Formulas V and VI was constructed by condensation according to the principle of the technique described in S. M. Hannick & Y. Kishi, J. Org. Chem., 48, 3833-3835 (1983).

Thus, the first step of the process, called step a), consists in reacting a phenylethanenitrile of Formula II with a compound of Formula H), in the presence of Zn in an appropriate solvent, for example an ether such as tetrahydrofuran, at a appropriate temperature, for example at reflux, then treating the resulting mixture with an acid, for example hydrochloric acid, to obtain the compound of Formula IV.

Then, in a step b), this compound of Formula IV is reacted with thiourea in a basic alcoholic medium, for example in a sodium / ethanol (Na / EtOH) medium, at an appropriate temperature, for example at reflux. , to obtain thiouracil of Formula V.

The thiouracil of Formula V is then converted in a step c) into uracil of Formula VI by treatment with an appropriate dilute organic acid, for example 10% chloroacetic acid.

The attachment of the chain to N1 of the nucleobase of Formula VI is carried out in step d) by a conventional glycosidation reaction with a compound of Formula VII in which n is equal to 3, for example in the presence of hexamethyldisilazane (HMDS) , chlorotrimethylsilane (TMSCI) and tin tetrachloride (SnCl4) followed by solvolysis of the ester obtained, for example in a methanol / triethylamine (MeOH / EtsN) medium, to provide the alcohol of Formula IX.

The alcohol of Formula IX is then subjected to a Mitsunobu reaction to replace its-OH group with a nitrogen group which will lead to the N-hydroxyureido group.

<Desc / Clms Page number 8>

Thus, treatment of the alcohol of Formula IX under the conditions of a conventional Mitsunobu reaction using N, O-bis (phenoxycarbonyl) hydroxylamine as a nucleophile (step e) provides the compound of Formula X which, after conventional aminolysis (step f), for example with ammonia or an amide in a suitable solvent, results in the compound of Formula Ia which bears a terminal N1-hydroxyureido group, namely a compound represented by Formula 1 in which R3 represents a group -OH and R4 represents a hydrogen atom.

The compound of Formula Ia can then be converted by an O-acylation reaction (step g) into the compound of Formula Ib, namely the compound of Formula 1 of the present invention in which R3 represents a group -OR and R represents a group. hydrogen atom, as follows.

The compound of Formula la is then subjected to a conventional O-acylation reaction, for example with an acylating agent of general formula X-CO-R6 (where R6 is a Me3C group, X possibly being for example a halogen atom or an R6COO- group, without X being limited thereto), to lead to the monoacyl compound of Formula lb of the present invention in which R5 is a Me3CCO group, more lipophilic than its precursor la and in which the N- group OH is protected against oxidation.

The compound of the present invention can further be converted in a conventional manner into pharmaceutically acceptable salts thereof.

In order to demonstrate the effectiveness of the compound of the present invention as an antiviral agent, and in particular as an anti-HIV-1 agent, in vitro tests were carried out according to the following principle and procedure.

Principle
The aim of the screening is to evaluate the antiretroviral activity of the compound of the present invention and of the comparative compounds on the HIV-1 lai strain cultured on peripheral blood mononuclear cells (PBMC).

<Desc / Clms Page number 9>

For this, the compounds to be tested were incubated for seven days (duration of the anti-HIV test) with the PBMCs in the exponential growth phase.

The capacity of the compounds to inhibit viral replication was then measured in the culture supernatants by assaying the “Reverse Transcriptase” (RT) activity using the RetroSys TR kit (INNOVAGEN)).

The concentration of the test compound making it possible to inhibit viral replication by 50% was determined. This is respectively the Iso inhibition concentration.

In parallel with the screening, and under the same conditions, a test for the cytotoxicity of the compounds to be tested was carried out.

This test was revealed by MTT (3- [4, 5-dimethylthiazol-2-yl] -2, 5-diphenyltetrazolium bromide) after 7 days of culture [J. G. Park et al., Cancer Res. 47 (22), 5875-5879 (1987)].

This test made it possible to determine the concentration of product which reduces the growth and viability of PBMCs (CCgo) by 50%.

The therapeutic index (TI) of the compound was calculated as follows: tTso = (CCgo / Ctgo).

Procedure The PBMCs were isolated as described in Ulmer et al., Immunobiology, 166 (3), 238-250 (1984) from a healthy donor by a Ficoll gradient (Nicomed Pharma AS).

PBMCs were activated by phytohemagglutinin for three days and then cultured in a 96-well plate, in basal medium (RPMI) supplemented with recombinant human interleukin-2.

Throughout the culture, the cells were maintained at 37 ° C., in an atmosphere saturated with humidity, and under 5% CO2.

The PBMCs were pre-treated for one hour in the presence of the compounds to be tested at different concentrations, then infected with 100 TCID50 of HIV-1-Lai [S. Wain-Hobson et al., Science 17,252 (5008): 961-965 (1991)].

<Desc / Clms Page number 10>

Three days after infection with HIV-1, half of the culture medium was renewed.

On day 7, the supernatants were removed and frozen at -20oC.

Viral replication was then measured by assaying the “Reverse Transcriptase” (RT) activity in the culture supernatants using the RetroSys kit (Innovagen).

The RT contained in the supernatant synthesizes, in the presence of bromodeoxyuridine triphosphate (BrdUTP), a DNA strand complementary to a matrix fixed at the bottom of the wells of a 96-well plate. The incorporation of BrdU is quantified by the binding of an anti-BrdU antibody conjugated to alkaline phosphatase. The activity of bound alkaline phosphatase, measured by colorimetry, is proportional to the activity of RT in the culture supernatant.

The results obtained result from at least two separate experiments carried out in triplicate.

Composé Comp. 1 : 1- (2-Nl-hydroxyuréidoéthyloxyméthyl)-6- (3, 5-diméthylbenzyl)-5- éthyluracile Composé Comp. 2 : 1- (4-Nl-hydroxyuréidobutyloxyméthyl)-6- (3, 5-diméthylbenzyl)-5- éthyluracile Composé Comp. 3 : 1- (2-N1-hydroxyuréidoéthyloxyméthyl)-6-benzyl-5-éthyluracile Composé Comp. 4 : 1-éthyloxyméthyl-6-benzyl-5-isopropyluracile (Composé de Formule A) Le composé de l'invention et les Composés Comp. 1, Comp. 2 et Comp. 3 testés ont été synthétisés comme indiqué dans les Exemples et le composé Comp. 4 a été synthétisé comme décrit dans la littérature [M. Baba, H. Tanaka, T. Miyasaka, S. Compound of the invention 1- (3-N3-pivaloyloxyuréidopropyloxymethyl) -6- (3, 5-dimethylbenzyl) -5-ethyluracil Comparative compounds tested:

Compound Comp. 1: 1- (2-Nl-hydroxyureidoethyloxymethyl) -6- (3, 5-dimethylbenzyl) -5-ethyluracil Compound Comp. 2: 1- (4-Nl-hydroxyureidobutyloxymethyl) -6- (3, 5-dimethylbenzyl) -5-ethyluracil Compound Comp. 3: 1- (2-N1-hydroxyureidoethyloxymethyl) -6-benzyl-5-ethyluracil Compound Comp. 4: 1-ethyloxymethyl-6-benzyl-5-isopropyluracil (Compound of Formula A) The compound of the invention and Compounds Comp. 1, Comp. 2 and Comp. 3 tested were synthesized as indicated in the Examples and the compound Comp. 4 was synthesized as described in the literature [M. Baba, H. Tanaka, T. Miyasaka, S.

<Desc / Clms Page number 11>

Yuseda, M. Ubasawa, R. T. Walker & E. De Clerq, Nucleosides Nucleotides, 14,575-583 (1995)].

Results of the tests The results of the tests resulting from the assay of the “Reverse Transcriptase” (RT) activity are reported in Table 1 below which shows the 50% inhibition concentrations (IC50) against HIV-Lai in nM. , the cytotoxic concentrations at 50% (CC50) in nM and the therapeutic index iT5o (CCso / Ciso) of the Compound of the present invention and of the comparative compounds Comp. 1,2, 3 and 4.

Table 1

<tb>
<tb> R1 <SEP> R2 <SEP> n <SEP> R3 <SEP> R4 <SEP> CI50 <SEP> CC50 <SEP> IT50
<tb> (nM) <SEP> (nM)
<tb> Compound <SEP> of <SEP> iPr <SEP> Me <SEP> 3 <SEP> OOCCMe3 <SEP> H <SEP> 1x10-6 <SEP> 6. <SEP> 5x104 <SEP> 6. <SEP > 5x101
<tb> the invention
<tb> Compounds
<tb> Comparisons
<tb> Comp. <SEP> 1 <SEP> And <SEP> Me <SEP> 2 <SEP> OH <SEP> H <SEP> 0.92 <SEP>> S <SEP> x <SEP> 1 <SEP> OS <SEP>> 1, <SEP> 1x106
<tb> Comp <SEP> 2 <SEP> And <SEP> Me <SEP> 4 <SEP> H <SEP> H <SEP> 2.3 <SEP>> 0.5 <SEP> x <SEP> 105 <SEP>> 0, <SEP> 25 <SEP> x <SEP> 105
<tb> Comp. <SEP> 3 <SEP> Et <SEP> H <SEP> 2 <SEP> OH <SEP> H <SEP> 70 <SEP>> 2,5 <SEP> x <SEP> 105 <SEP>> 3, <SEP> 5x103
<tb> (Formula <SEP> C)
<tb> Comp. <SEP> 4 <SEP> 2 <SEP>> 3 <SEP> x <SEP> 1 <SEP> 04 <SEP>> 1, <SEP> S <SEP> X104
<tb> (Formula <SEP> A)
<tb>

<Desc / Clms Page number 12>

As can be seen in Table 1, the compound of the present invention has been shown to be a very active antiviral against HIV-1, for which the spectacular and unpredictable activity, namely 1000 activity, can be observed. 000 times higher than that of its congeners or its competitors.

The present invention therefore provides a novel acyclonucleoside as an antiviral agent, in particular acting as a non-nucleoside inhibitor of the reverse transcriptase of HIV-1, non-toxic, and which can be active at femtomolar concentrations.

Thus, the compound of! the present invention or a pharmaceutically acceptable salt thereof can be used as an active ingredient in a pharmaceutical composition, and it can be used either alone or in admixture with other active ingredients, especially in a pharmaceutical composition for use as a drug as part of a combination therapy.

The present invention therefore also provides a pharmaceutical composition containing as an active ingredient at least the compound of the present invention or a pharmaceutically acceptable salt thereof.

The amount of the compound of the present invention or of its pharmaceutically acceptable salt contained in the composition will depend in particular on the weight, age and condition of the patient as well as on the effectiveness of the compound.

The pharmaceutical composition of the present invention may be in an orally or systemically administrable form, and it may contain any suitable pharmaceutically acceptable carrier or excipient.

5 un groupe OR, à savoir un groupe pivaloyloxy. The compound of the present invention is a compound of Formula 1 wherein R3 is

An OR group, namely a pivaloyloxy group.

Such a compound where R3 is an OR group may in particular be advantageously used in a depot formula.

The compound of the present invention in which R5 is a pivaloyl group is particularly advantageous because it is more lipophilic than its precursor and the group

<Desc / Clms Page number 13>

N-OH is protected against oxidation. In addition, it can be very slowly deacylated in the blood.

The use of the branched Rs group is also advantageous because it makes it possible to delay the hydrolysis of the ester group by the esterases.

Thanks to the present invention, it has been possible to improve the binding to the receptor, namely the allosteric site of the reverse transcriptase of HIV-1, which manifests itself in a lower inhibitory concentration, which implies a better selectivity. , a lower preparation cost and a lower propensity to select resistant mutants.

The following examples are intended to illustrate the present invention. However, they should in no way be considered as limiting the scope of the present invention.

<Desc / Clms Page number 14>

EXAMPLES The starting materials, reagents and solvents used in the following syntheses are all commercial products from Fluka (Buchs, Switzerland), Merck (Darmstadt, Germany), or Aldrich (Buchs, Switzerland), unless anything else is specified.

Synthèse du 1- (3-Nl-pivaloyloxyuréidopropyloxyméthyl)-6- (3, 5-diméthylbenzyl)- 5-isopropyluracile de la présente invention 1. Synthèse du 1- (3--hydroxyuréidopropy) oxyméthy !)-6- (3, 5-diméthy) benzy !)- 5-isopropyluracile (Composé de Formule I, dans lequel n=3, R1 est un groupe isopropyl, chaque R2 est un groupe méthyle, R3 est un groupe OH et R4 est un atome d'hydrogène). Example 1

Synthesis of 1- (3-Nl-pivaloyloxyuréidopropyloxymethyl) -6- (3, 5-dimethylbenzyl) - 5-isopropyluracil of the present invention 1. Synthesis of 1- (3 - hydroxyureidopropy) oxymethyl!) - 6- (3, 5-dimethyl) benzy!) - 5-isopropyluracil (Compound of Formula I, wherein n = 3, R1 is an isopropyl group, each R2 is a methyl group, R3 is an OH group and R4 is a hydrogen atom) .

A. Synthesis of 6- (3, 5-dimethylbenzyl) -5-isopropyl-uracil (Compound of Formula VI according to Reaction Scheme 1 in which R1 is an isopropyl group and each R2 is a methyl group) To an acid solution 2-Bromo-3-methylbutanoic (3g, 16.5mmol) in ethanol (200ml), concentrated sulfuric acid (4ml) was added. After 36 h of boiling at reflux, the reaction medium, brought to room temperature, was neutralized with a saturated aqueous solution of sodium carbonate. After distillation of the ethanol, the reaction medium was extracted with dichloromethane (100 ml). The organic phase was then dried over magnesium sulfate and the solvent was evaporated to give ethyl 2-bromo-3-methylbutanoate (1.91 g, 55%).

A suspension of zinc powder (31.5 g, 0.48 mol) in tetrahydrofuran (300 ml) was brought to the boil under reflux, then a few drops of ethyl 2-bromo-3-methylbutanoate were added. to initiate the reaction. After 45 min at reflux with magnetic stirring, there was added 3, 5-dimethylphenylethanenitrile (13.2 g, 91 mol), then dropwise, the remainder of ethyl 2-bromo-3-methylbutanoate (at

<Desc / Clms Page number 15>

total 19.1 g, 91 mmol). The boiling was maintained for 15 min, the mixture was then cooled, then tetrahydrofuran (500 ml) and 50% aqueous potassium carbonate solution (100 ml) were added thereto. After 45 min of vigorous stirring, the organic phase was separated by decantation and the aqueous phase was washed with tetrahydrofuran (2 x 100 ml). The combined organic phases were treated with a 10% aqueous hydrochloric acid solution (300 ml) for 45 min. After removing the tetrahydrofuran by distillation under reduced pressure, the residue was taken up in dichloromethane (300 ml), and the organic phase was washed with saturated sodium monohydrogencarbonate solution (100 ml), dried over magnesium sulfate and concentrated. Distillation of the residue (134 ° C, 10-1 mmHg) provided ethyl 3-methyl-2 (3, 5-dimethylphenylacetyl) - butanoate (13.2 g, 52%).

Metallic sodium (23.8 g, 1,034 moles) was reacted with anhydrous methanol (500 mL). To the clear solution obtained were added thiourea (54.35 g, 714 mmol) and ethyl 3-methyl-2 (3, 5-dimethylphenylacetyl) butanoate (13.14 g, 47.6 moles) ). The reaction medium was maintained at reflux for 6 h, then concentrated under vacuum at 40.50 C. To the residue obtained, concentrated hydrochloric acid (100 ml) was added then the solution was brought to pH 4 by acetic acid. The obtained 6-dimethylbenzyl-5-isopropyl-2-thiouracil was dissolved in a 10% aqueous solution of chloroacetic acid (200 ml) and the solution was kept under reflux for 24 h then cooled to room temperature and the precipitate. obtained was filtered off and washed with cold ethanol then ether, then dried under vacuum at 40 C to provide 6- (3, 5-dimethylbenzyl (-5-isopropyluracil (7 g, 54%) .

Melting point: 213-214 C.

B. Synthesis of 1-acetoxy-3-acetoxymethoxypropane (Compound of Formula VII according to Reaction Scheme 1, in which n = 3) To a mixture of 1,3-dioxane (3 ml, 0.035 mmol) and acetic anhydride ( 3.3 ml, 0.035 mmol) brought to 0 oC, a drop of concentrated sulfuric acid was added. The mixture was then stirred for 14 h at 20 C, added with sodium acetate (2 g) and

<Desc / Clms Page number 16>

filtré. Le résidu distillé sous vide (120 C, 16 mmHg) fournissait le 1-acétoxy-3acétoxyméthoxypropane (3, 34 g, 50 %) sous forme d'une huile incolore.

filtered. The residue distilled in vacuo (120 C, 16 mmHg) provided 1-acetoxy-3acetoxymethoxypropane (3.34 g, 50%) as a colorless oil.

1H-NMR (200 MHz, CDC! S): 8 1.90 (quint., 2H, J = 6Hz, CH2-CH-CH2), 2.04 (s, 3H, OAc), 2.09 (s, 3H, OAc), 3.69 (t, 2H, J = 6Hz, OCH2-CH2-CH2-OAc), 4.44 (t, 2H, J = 6Hz, OCH2CH2CH20Ac), 5.22 (s, 2H, AcOCHO ).

IR (KBr): Vmax 2994 (vc-H), 1706 and 1682 (vc = o), 1225, 1074 cm-1.

C. Synthesis of 1- (3-N1-hydroxyureidopropyloxymethyl) -6- (3,5-dimethylbenzyl) -
5-isopropyluracil A a solution of 6- (3, S-dimethylbenzyl) -S-isopropyluracil (1, S g, 5, 5 mmol) obtained in step A above and of 1-acetoxy-3-acetoxymethoxypropane ( 2.1 g, 11 mmol) obtained in step B above in ethanenitrile (60 ml), hexanemethyldisilazane (1.78 g, 11 mmol) and chlorotrimethylsilane (1, 2 g, 11 moles). After 15 min at 20 ° C, a solution of 1v tin chloride (2.87g, 11mmol) in ethanenitrile (15ml) was added dropwise. After 14 h of stirring at 20 ° C, dichloromethane (100 ml) and saturated aqueous sodium hydrogencarbonate solution (50 ml) were added. The organic phase was then washed with a saturated aqueous solution of sodium hydrogencarbonate (30 ml) then with a saturated solution of sodium chloride (30 ml), and dried over magnesium sulfate, then concentrated. The obtained residue was subjected to flash column chromatography (petroleum ether / ethyl acetate 1: 1, RF = 0.41) to provide 1-acetoxypropyloxymethyl) -6- (3,5-dimethylbenzyl) -5isopropyluracil (1.73 g, 75%) as a viscous oil.

(dichlorométhane/méthanol 99, 5 : 0. 5) pour fournir le 1- (3-hydroxypropyloxyméthyl) -6- (3, 5-diméthylbenzyl)-5-isopropyluracile (1,43 g, 93 %). A une solution de cet alcool (1,43 g, 3,96 mmoles), de N, O- (diphénoxycarbonyl) -hydroxylamine (1, 2 g, 4,36 mmoles) et de triphénylphosphine (1, 25 g, 4,76 mmoles) dans du tétrahydrofurane This acetylated derivative (1.7g, 4.05mmol) was dissolved in methanol (40ml) and thereto were added triethylamine (5ml) and water (5ml). After 48 h of stirring at 20 ° C, the reaction was complete (TLC). The reaction medium was concentrated, the remains of volatiles were removed by coevaporation with toluene and the residue was subjected to flash column chromatography.

(dichloromethane / methanol 99.5: 0.5) to provide 1- (3-hydroxypropyloxymethyl) -6- (3, 5-dimethylbenzyl) -5-isopropyluracil (1.43 g, 93%). Has a solution of this alcohol (1.43 g, 3.96 mmol), of N, O- (diphenoxycarbonyl) -hydroxylamine (1.2 g, 4.36 mmol) and of triphenylphosphine (1.25 g, 4, 76 mmol) in tetrahydrofuran

<Desc / Clms Page number 17>

(30 ml), a solution of diisopropyl azodicarboxylate (0.97 g, 4.76 mmol) in tetrahydrofuran (5 ml) was added dropwise at 0 ° C. The reaction medium was concentrated and then subjected to to flash column chromatography (methanol / chloroform 0.3: 9.7) to provide 6- (3,5-dimethylbenzyl) - 1- [3- (N-phenoxycarbonyl-N-phenoxycarbonyloxyamino) propyloxymethyl] -5- isopropyluracil (1.8 g, 79%) as a solid, Melting point: 59-61 C.

1 H-NMR (200 MHz, CDC! S): 8 1, 23 (d, 6H, J = 6.5Hz, CH-Me2), 2.03 (quint, 2H, J = 6Hz, CHs-CH-CHz ), 2.29 (s, 6H, MePh), 2.82 (sept., 1H, CHMe2), 3.75 (t, 2H, OCH-CHz-CHz-N), 3.95 (t, 2H, NCH2CH2CH20), 4.09 (s, 2H, CHAr), 5.12 (s, 2H, NCH O),
6.70 (s, 2H, Hortho-benzyl), 6.90 (s, 1H, Hpara-benzyl), 7.10-7.48 (m, 10.2PhO),
8.1 (bs, 1H, NH).

IR (CH2CI2): vmax 3382 (vN-H), 3050-2870 (vC-H), 1806, 1741,1707 and 1682 (vc = o) cm-1.

A solution of the latter compound (1.8 g, 3.13 mmol) in methanol saturated with ammonia was stirred at 20 ° C. until disappearance of the starting product (reaction followed by TLC). The residue was subjected to flash column chromatography (methanol / ethyl acetate 1: 9) to provide the desired 1- (3-N1-hydroxyureidopropyloxymethyl) -6- (3,5-dimethylbenzyl) -5-isopropyluracil ( 0.74 g, 60%) as a solid.

Melting point: 88-90 C.

6, 70 (s, 2H, Hortho-benzyle), 6, 90 (s, 1 H, Hpara-benzyle), 9, 00 et 9, 87 (2 s !, 3H, NH). 1 H-NMR (200 MHz, CDCl3): 5 1.29 (d, 6H, J = 6.5Hz, CHMe),
1.89 (quint, 2H, J = 6Hz, CH2-CH2-CH2), 2.28 (s, 6H, MePh),
2.88 (sept, 1H, CHMe2), 3.62 (t, 2H, OCH2-CH2-CH2-N),
3.68 (t, 2H, NCHCH2CH2O), 4.09 (s, 2H, CHzAr), 5.11 (s, 2H, NCHzO),

6.70 (s, 2H, Hortho-benzyl), 6.90 (s, 1H, Hpara-benzyl), 9.00 and 9.87 (2 s!, 3H, NH).

IR (CH2CI2): Vmax 3S30 (vO-H), 3430, 3372 and 3190 (VN-H), 3000-2850 (vc-H), 1681, 5 (vc = o) cm '.

<Desc / Clms Page number 18>

(Me3CCOO) et R4 est un atome d'hydrogène). 2. Synthesis of 1- (3-N1-pivaloyloxyureidopropyloxymethyl) -6- (3, 5- dimethylbenzyl) -5-isopropyl uraci le (Compound of the present invention of Formula i, wherein n = 3, R1 is isopropyl group , each R2 is a methyl group, R3 is a pivaloyloxy group

(Me3CCOO) and R4 is a hydrogen atom).

To a solution of 1- (3 - hydroxyureidopropytoxymethyl)) - 6- (3, 5-dimethyl! Benzy)) - 5- isopropyluracil obtained in Example 1-C above (134 mg, 0.32 mmol) in pyridine (1.5 ml), 46 mg (0.38 mmol) of pivaloyl chloride was added. After 14 h stirring at 24 ° C, the pyridine was coevaporated off with toluene (2 x 3ml) and the residue was taken up in dichloromethane (10ml). The organic phase was then washed with a saturated aqueous solution of sodium hydrogencarbonate (10 ml). The aqueous phase was extracted with dichloromethane (3 x 10 ml) and the organic phases, combined, washed with an aqueous solution of sodium hydrogencarbonate (10 mL), a saturated aqueous solution of sodium chloride (10 mL), (sodium sulfate) leave on distillation of the solvent 180 mg of crude 1- (3-N1-pivaloyloxyuréidopropyloxymethyl) -6- (3,5-dimethylbenzyl) - 5-isopropyl uracil which was subjected to "flash" chromatography on a column of silica gel (dichloromethane / methanol 24: 1) to provide 127 mg (78%) of the desired 1- (3-N1-pivaloyloxyuréidopropyloxymethyl) -6- (3,5-dimethylbenzyl) -5isopropyluracil as a solid.

Melting point 68-69 C; RF 0.22 (dichloromethane / methanot 24: 1).

UV (MeOH) nm (s): 210 (17500) and 268 (9420).

1H NMR (200 MHz, CDOs): 8 1.27 (d, 6H, J = 6.5Hz, CHMe2), 1.29 (s, 9H, CMe3),
1.80 (quint., 2H, J = 6Hz, CH2-CH2-CH2), 2.28 (s, 6H, Me2Ph),
2.86 (seven, 1H, CHMe2), 3.62 (t, 2H, OCH2CH2CH2N),
3.72 (t, 2H, OCH2CH2CH2N), 4.08 (s, 2H, CH2Ar), 5.09 (s, 2H, NCH20),
6.69 (s, 2H, Hortho-benzyl), 6.89 (s, 1H, Hpara-benzyl), 8.66 (s el, 1H NH).

13C NMR (50 MHz, CDCts): ô 20.4 (Me2Ar), 21.3 (Me2CH), 26, 8 (NCH2CH2CH2-0), 27.0 (Me3C), 28.4 (Me2CH), 33.3 (CH2Ar), 38.5 (Me3C), 46.6 (NCH2-CH2CH2), 66.4 (OCH2CH2CH2N), 73.1 (NCH20), 119.6 (C-5), 125.0 (Cortho-Ar ),

<Desc / Clms Page number 19>

128, 8 (Cpara-Ar), 135, 1 (Cipso-Ar), 138, 8 (Cméta-Ar), 148, 8 (C-2), 151, 8 (C-6), 158, 8 (CONH2), 162, 3 (C-4), et 175, 7 (COCMe3).

128, 8 (Cpara-Ar), 135, 1 (Cipso-Ar), 138, 8 (Cméta-Ar), 148, 8 (C-2), 151, 8 (C-6), 158, 8 (CONH2 ), 162, 3 (C-4), and 175, 7 (COCMe3).

IR (CH2Cb): vmax 3535 (VN-H), 3430 and 3386 (VNH2), 3052 (asymmetric vc-H),
1779 (vc = o ester), 1705.1684 (vc = o CON) cm '.

MS m / z (%): 355 (4, B-CH2-OCH2CH2CN +), 301 (2, B-CHsO), 285 (3, B-CH2 +),
273 (13, BH2 +), 272 (62, BH +), 271 (5, B +), 257 (100, BH ± Me).

Elemental analysis for C26H38N406 (502, 62), 0.5 CH2Cl2):
Calculated: C, 58.39; H, 7.21; N, 10.28.

Found: C, 58.14; H, 7.20; N, 10.13.

Example 2 Synthesis of 1- (2-N1-hydroxyureidoethoxymethyl) -6- (3,5-dimethylbenzyl) -5- ethyluracil (Compound Comp. 1) (Comparative compound of Formula 1 in which n = 2, R1 = ethyl, R2 = methyl, R = OH, R = H).

A. Synthesis of 1- (2-acetoxyethoxymethyt) -6- (3, 5-dimethy) benzy)) - 5-ethy! urac!) e 6- (3, 5-dimethylbenzyl) -5-ethyluracil (see Example 3) (516 mg, 2 mmol) and 1-acetoxy-2-acetoxymethoxyethane [A. Rosowski, S. H. Kim. M. Wick, J. Med. Chem.

(1981) 24, 1177-81]) (704 mg, 4 mmol) were suspended, under a nitrogen atmosphere, in acetonitrile (20 ml). HMDS (hexamethyldisilazane) (646 mg, 4 mmol) and TMSCl (chlorotrimethylsilane) (435 mg, 4 mmol) were added and stirred for 10 min. A solution of tin chloride (625 mg, 2.4 mmol) in acetonitrile (5 ml) was then added dropwise over 10 min. After 14 h of stirring, dichloromethane (50 ml) and saturated aqueous sodium hydrogencarbonate solution (50 ml) were added. Extracted with dichloromethane (3 x 50 mL). The combined organic phases were washed with a saturated aqueous solution of sodium hydrogencarbonate (25 ml), with water (25 ml), then with a saturated aqueous solution of sodium chloride. The organic phase was dried (sodium sulfate), concentrated, then subjected to flash column chromatography (petroleum ether / ethyl acetate 3: 2) to

<Desc / Clms Page number 20>

provide 6- (3,5-dimethylbenzyl) -5-ethyluracil (690 mg, 92%) as a white solid.

Melting point: 126.0-127.0 C RF 0.28 (hexane / ethyl acetate 3: 2).

1H NMR (CDOs, 200 MHz): 8.10 (t, 3H, J = 7.25 Hz, CHzCHs), 2.08 (s, 3H, Ac),
2.30 (s, 6H, MePh), 2.69 (q, 2H, cCH2CH3), 3.80 (m, 2H, AcOCH2CH2O),
4.08 (s, 2H, CH2Ph), 4.20 (m, 2H, AcOCH2), 5.18 (s, 2H, OCH2N),
6.71 (s, 2H, Ho-benzyl), 6.90 (s, 1H, Hpara-benzyl), 8.25 (s el. 1H, NH).

IR (KBr): 3030 (VC-H arom.), 2950 (aliphatic vC-H), 1740 and 1705 (vc = o) cm- '.

UV (methanol) vamx (v) 266 nm (8180).

MS (m / z (%)) 374 (4, M +), 255 (16, M. + - AcOCH2CH2O), 119 (11, M '+ - base),
87 (100, AcOCH2CH2 +).

Elemental analysis for C20H26N2O5 (374, 44):
Calculated: C, 64, 16; H, 7.00; N, 7.48.

Found: C, 63.88; H, 7.10; N, 7.37.

B. 6- (3, 5.dimethylbenzyl) -5-ethyl-2- (2-hydroxyethoxymethyl) uracil A solution of 1- (2-acetoxyethoxymethyl) -6- (3,5-dimethylbenzyl) -5-ethyluracil (690 mg, 1.9 mmol) in an 8: 1: 1 methanol / triethylamine / water mixture was stirred for 20 h at 20 C. The solvents were removed by evaporation in vacuo and then by coevaporation with toluene. The solid obtained, recrystallized (methanol) provided 390 mg of 6- (3,5-dimethylbenzyl-5-ethyl-1- (2-hydroxyethoxymethyl) uracil. The mother liquors of crystallization concentrated, subjected to flash column chromatography on Silica gel (dichloromethane / methanoI19: 1) provided an additional 80 mg sample Total yield 470 mg (77%) of a white solid.

1H RMN (CD30D, 200 MHz) : 8 1, 01 (t, 3 H, J = 7, 5 Hz, CH2CHs), 2, 25 (s, 6 H, Me2Ph), 2, 42 (q, 2 H, CHCHs), 3, 60 (m, 4 H, CH2CH2), 4, 12 (s, 2 H, CH2Ph). 5, 14 (s, 2 H, OCH2N), 6, 78 (s él. 2 H, Horlho-benzyle), 6, 90 (s él., 1 H, H-para-benzyle). Melting point: 178.0-179.0 C; RF 0.12 (petroleum ether / ethyl acetate 9: 1)

1H NMR (CD30D, 200 MHz): 8 1.01 (t, 3H, J = 7.5Hz, CH2CHs), 2.25 (s, 6H, Me2Ph), 2.42 (q, 2H, CHCHs), 3.60 (m, 4H, CH2CH2), 4.12 (s, 2H, CH2Ph). 5.44 (s, 2H, OCH2N), 6.78 (s el 2H, Horlho-benzyl), 6.90 (s el, 1H, H-para-benzyl).

<Desc / Clms Page number 21>

IR (KBr): 3380 (VO-H), 3020 (VC-H arom.), 1708 (vc = o) cm '.

UV (methanol) vmax (v) 207 (41500), 268 (15400).

MS (m / z (%)): 332 (2, M +), 258 (100, BH +).

Elemental analysis for C18H24N2O4 (332, 40):
Calculated: C, 65.04; H, 7.28; N, 8.43.

Found: C, 64, 93; H, 7.30; N, 8.36).

C. 1- (2-N1-hydroxyureidoethoxymethyl) -6- (3,5-dimethylbenzyl) -5-ethyluracil To a solution of 6- (3,5-dimethylbenzyl) -5-ethyl-1- (2-hydroxyethoxymethyl) uracil (0.2 g, 0.6 mmol), N, O-bis (phenoxycarbonyl) hydroxylamine (0.18 g, 0.66 mmol) and triphenylphosphine (0.19 g, 0.66 mmol) in tetrahydrofuran (6 mL), DIAD (diisopropylazodicarboxylate) (145 mg, 0.71 mmol) was added dropwise at 0 ° C. The reaction medium, concentrated, was subjected to two chromatographies on a flash column of silica gel successively, the first using as eluent the dichloromethane / methanol mixture 49: 1 and the second, the ethyl acetate / petroleum ether mixture 2: 3. There was thus obtained 6- (2, 3-dimethylbenzyl) -5-ethyl-1- (2-N, O-bis (phenoxycarbonyl) hydroxyaminoethoxymethyl) uracil (0.30 g, 85%) of which 0.15 g (0.25 mmol) was dissolved in a saturated methanolic solution of ammonia (10 ml) and stirred for 24 h at 10 C. The concentrated reaction medium, subjected to flash column chromatography on silica gel (dichloromethane / methanoI19 : 1) provided an oil which, taken up in ether, provided 1- (2-N1-hydroxyureidoethoxymethyl) -6- (3,5-dimethylbenzyl) -5-ethyluracil (36.6 mg, 36%) as a white solid.

1H RMN (CDCb, 200 MHz) : 81, 09 (t, 3 H, J = 7, 5 Hz, CHzCHs), 2, 29 (s, 6 H, MePh), 2, 48 (q, 2 H, CHzCH3), 3, 22-3, 90 (m, 4 H, CH2CH2), 4, 07 (s, 2 H, CHPh), 5, 10 (s, 2 H, OCH2N), 6, 70 (s, 2 H, H-benzyle), 6, 90 (s, 1 H, Hpara-benzyle),
9,76 (s él., 1 H, NH). Melting point: 147 - 148 C RF 0.18 (dichloromethane / methanol 9: 1)

1H NMR (CDCb, 200 MHz): 81.09 (t, 3H, J = 7.5Hz, CHzCHs), 2.29 (s, 6H, MePh), 2.48 (q, 2H, CHzCH3 ), 3, 22-3, 90 (m, 4H, CH2CH2), 4, 07 (s, 2H, CHPh), 5, 10 (s, 2H, OCH2N), 6, 70 (s, 2H , H-benzyl), 6.90 (s, 1H, Hpara-benzyl),
9.76 (s el, 1H, NH).

IR (KBr): 3498,3235 and 3190 (VOH and VNH), 1703,1699 and 1688 (vc = o) cm-1.

MS (m / z (%)) 345 (1, M '+ - NH2CHO), 327 (6, M. + - NH2COH - H2O),
271 (100, M. + - NH2CONOHCH2CH2O), 258 (73, BH +).

<Desc / Clms Page number 22>

Elemental analysis for C19H26N40s. 1 / 2H2O (339, 45):
Calculated: C, 57.13; H, 6.81; N, 14.03.

Found. C, 56.85; H, 6.63; N, 13.57.

Example 3 Synthesis of 1- (4-N1-hydroxyureidobutyloxymethyl) -6- (3,5-dimethylbenzyl) -5- ethyluracil (Compound Comp. 2) (Comparative compound of Formula 1 in which n = 4, R1 = ethyl, R2 = methyl, R3 = OH, R4 = H).

A. 4-Hydroxybutyl benzoate To a solution of 1,4-butanediol (18 ml, 0.2 mmol) in pyridine (16 ml), was added slowly at 0 ° C., under a nitrogen atmosphere, chloride benzoyl (1 ml), 0.1 mmol). After 4 h of stirring at 0 C, the reaction medium was concentrated and the pyridine was removed by co-distillation with toluene. The residue was extracted with ether (3 x 20 ml) and the organic phase was dried (magnesium sulfate), concentrated, and subjected to flash column chromatography on silica gel (petroleum ether / ethyl acetate. 3: 2) to provide 4-hydroxybutyl benzoate (9.9 g, 55%) as a colorless oil.

RF 0.36 (petroleum ether / ethyl acetate 1: 1).

1H NMR (CDCl s, 200 MHz): ô 1.79 (m, 4H, CHCHCHCH.

3.60 (t, 2H, J = 7.0Hz, CHOH). 4.34 (t, 2 H, J = 8. 0 Hz, PhCOOCHs),
7.45-8.02 (m, 5H, Ph).

B. 1- (4-Benzoyloxybutyloxymethyl) -6- (3,5-dimethylbenzyl) -5-ethyluracil To a solution of 4-hydroxybutyl benzoate (1 g, 5.2 mmol) in dichloromethane (18 ml), it Paraformaldehyde (100 mg, 5.2 mmol) was added.

In the mixture, brought to 0 ° C., hydrochloric acid was bubbled for 2 h. After this time, the container was stoppered and stirred at 4 ° C for 16 h. The reaction mixture was diluted with dichloromethane (10 mL), dried (magnesium sulfate), then concentrated. This solution was added to a suspension of 6- (3,5-dimethylbenzyl) 5-ethyluracil (obtained as described in

<Desc / Clms Page number 23>

l'Exemple 3) (400 mg, 1, 55 mmoles) et de N, O-bis (triméthylsilyl) acétamide (1, 1 ml, 4,4 mmoles) dans du dichlorométhane (10 ml) préalablement agitée 30 mn à 20 C puis additionnée d'iodure de tétrabutylammonium (20 mg, 0, 05 mmole). Le mélange a été chauffé à reflux pendant 16 h puis ramené à 20 Il a ensuite été versé dans une solution aqueuse saturée d'hydrogénocarbonate de sodium maintenue à 0 C et le mélange a été agité pendant 30 mn. La phase organique, décantée, a été lavée avec une solution aqueuse saturée de chlorure de sodium (15 ml), séchée (sulfate de magnésium), concentrée et soumise à une chromatographie sur colonne flash de gel de silice (éther de pétrole/acétate d'éthyle 7 : 3) qui fournissait le 1- (4-benzoyloxy- butyloxyméthyl)-6-(3,5-diméthylbenzyl)-5-éthyluracile (460 mg, 60 %) sous forme d'une huile.

Example 3) (400 mg, 1.55 mmol) and N, O-bis (trimethylsilyl) acetamide (1.1 ml, 4.4 mmol) in dichloromethane (10 ml) previously stirred for 30 min at 20 C then added with tetrabutylammonium iodide (20 mg, 0.05 mmol). The mixture was heated at reflux for 16 h then brought back to 20 It was then poured into a saturated aqueous solution of sodium hydrogencarbonate maintained at 0 C and the mixture was stirred for 30 min. The organic phase, decanted, was washed with a saturated aqueous solution of sodium chloride (15 ml), dried (magnesium sulfate), concentrated and subjected to flash column chromatography on silica gel (petroleum ether / acetate d. ethyl 7: 3) which provided 1- (4-benzoyloxy-butyloxymethyl) -6- (3,5-dimethylbenzyl) -5-ethyluracil (460 mg, 60%) as an oil.

RF 0.71 (dichtoromethane / methano) 19: 1).

1H NMR (CDC) 3.200 MHz): 8.08 (t, 3H J = 8.0Hz, CHsCHg), 1.78 (m, 4H, CH? CH2CH2CH2), 2.29 (s , 6H, Me2Ph), 2.48 (q, 2H, CH2CH3), 3.61 (t, 2H, J = 5.5Hz, (CH2hCH20CH2), 4.09 (s, 2H, CH2Ph) , 4.34 (t, 2H, J = 6.0Hz, PhCOOCH2), 5.11 (s, 2H, OCH2N), 6.60 (tel s, 2H, Hoo-benzyl), 6, 91 (s, 1H, Hpara-benzyl), 7.40-8.09 (m, 5H, PhCO), 8.38 (sg., 1H, NH).

IR (NaCl): 3050 (vc-H arom.), 2924 (VC-H aliph), 1702 and 706 (vc = o) cm-1.

UV (EtOH): vmax (v) 202 (51500), 220 (33300), 267 (15150) nm.

MS (m / z (%)) 464 (1, M- "), 270 (58, M. + - PhCO2 (CH2) 40), 177 (15, PhCC (CH),
105 (100, PhCO +).

Elemental analysis for C27H32N2O5 (464, 57):
Calculated: C, 69, 81; H, 6.94; N, 6.03.

Found: C, 69.66; H. 7.02; N, 5, 92.

C. 6- (3, 5-Dimethytbenzyi) -5-ethyt-1- (4-hydroxybuty! Oxymethy)) uraci) e.

1- (4-Benzoyloxybutyloxymethyl) -6- (3.5-dimethylbenzyl) -5-ethyluracil (500 mg, 1.08 mmol) was added to a solution of sodium hydroxide (360 mg, 9 mmol) in methanol (50 mL) and the mixture was stirred at 20 C for 3 h. After neutralization (concentrated hydrochloric acid) and concentration, the residue was subjected to flash column chromatography (dichloromethane / methanol 19: 1)

<Desc / Clms Page number 24>

which provided 6- (3,5-dimethylbenzyl) -5-ethyl-1- (4-hydroxybutyloxvmethyl) uracil (190 mg, 48%) as a white solid.

Melting point: 101.4-102.8 C; RF 0.29 (dichloromethane / methanoI19: 1).

1H NMR (CD30D, 200 MHz): 8 1.00 (t, 3H, J = 7.5Hz, CHs), 1.54 (m, 4H, CH2CH2CH2CH2), 2.26 (s, 6H, Me2Ph). 2,41 (q, 2H, CHCHs), 3,52 (m, 4H, CH2CH2CH2CH2), 4, 11 (s, 2H, CHPh), 5, 10 (s, 2H, OCH2N), 6, 75 (s, 2H, Hortho-benzyl), 6.90 (s, 1H, Hpara-benzyl).

IR (KBr) 3395 (vo-H), 1694 and 1637 (vc = o) cm '.

UV (methanol): Vmax (v) 270 nm (11174).

MS (m / z (%)): 360 (1, M. +), 271 (7, M. + - O (CH2) 4OH),
258 (100, M '+ - HO (CH2) 40CH2).

Elemental analysis for C20H2sH204 (360, 45):
Calculated: C, 66.64; H, 7.83; N, 7.77.

Found: C, 66.39; H, 7.87; N, 7.63.

D. 1- (4-N1-hydroxyureidobutyloxymethyl) -6- (3,5-dimethylbenzyl) -5-ethyluracil (Compound Comp. 2) To a solution of 6- (3,5-dimethylbenzyl) -5-ethyl-1 - (4-hydroxybutyloxymethyl) uracil (180 mg, 0.5 mmol), N, O-bis (phenoxycarbonyl) hydroxylamine (150 mg, 0.55 mmol) and triphenylphosphine (262 mg, 1 mmol) in tetrahydrofuran ( 5 ml), a solution of DIAD (diisopropylazodicarboxylate) (202 mg, 1 mmol) in tetrahydrofuran (81 ml) was added dropwise at 0 C.

conduire à 260 mg du composé intermédiaire 6- (3, 5-diméthylbenzyl) -1-[4- (Nphénoxycarbonyl-N-phénoxycarbonyloxyamino) butyloxyméthyl]-5-éthyluracile Ce composé intermédiaire a été immédiatement dissous dans une solution méthanolique saturée d'ammoniac (5 ml) et le milieu réactionnel agité à 20 C pendant 24 h. A ce moment, le composé intermédiaire avait intégralement réagi (chromatographie sur couche mince) et le milieu réactionnel, concentré, a été soumis The reaction mixture was then brought to 20 ° C., concentrated and subjected to column chromatography on silica gel (ethyl acetate / ethyl ether 1: 9), then to flash column chromatography (methanol / chloroform 3: 97). ) for

lead to 260 mg of the intermediate compound 6- (3, 5-dimethylbenzyl) -1- [4- (Nphenoxycarbonyl-N-phenoxycarbonyloxyamino) butyloxymethyl] -5-ethyluracil This intermediate compound was immediately dissolved in a methanolic solution saturated with ammonia (5 ml) and the reaction medium stirred at 20 ° C. for 24 h. At this time, the intermediate compound had fully reacted (thin layer chromatography) and the reaction medium, concentrated, was subjected to

<Desc / Clms Page number 25>

to flash column chromatography on silica gel (methanol / ethyl acetate 1: 9) to provide 1- (4-N1-hydroxyureidobutyloxymethyl) -6- (3, 5-dimethylbenzyl) -5-ethyluracil (45 mg , 23%) as a solid.

Melting point: 148-150 C, RF 0.40 (methanol / ethyl acetate 1: 9).

1H NMR (CD3OD, 200 MHz): # 1.00 (t, 3H, J = 7.5Hz, CHCH).

3, 42 et 3, 51 (2 t, 2x2 H J = 6, 25 Hz, NCH2 (CH2) 2CH20 et NCH2 (CH2) 2CH20), 4, 11 (s, 2 H, CH2Ph), 5, 10 (s, 2 H, OCH2N), 6, 72 (s, 2 H, Ho-benzyle), 6, 90 (s, 1 H, Hpara-benzyle). 1.56 (m, 4H, CH2CH2CH2CH2), 2.25 (s, 6H, MePh), 2.41 (q, 2H, CH2CH3),

3, 42 and 3, 51 (2 t, 2x2 HJ = 6, 25 Hz, NCH2 (CH2) 2CH20 and NCH2 (CH2) 2CH20), 4, 11 (s, 2 H, CH2Ph), 5, 10 (s, 2H, OCH2N), 6.72 (s, 2H, Ho-benzyl), 6.60 (s, 1H, Hpara-benzyl).

IR (KBr): 3485, 3356 and 3158 (VNH and VOH), 1682 and 1652 cm-1 (vc = o). MS (m / z (%)): 373 (1, 5, M. + - NH2COH), 355 (20, M. + - NH2COH - H2O),
271 (40, M '+ - NH2CONOH (CH2) 40), 258 (55, BH +), 257 (29, B +).

Elemental analysis for C21H30N4Os. H2O (436, 51):
Calculated: C, 59.00; H, 7.30; N, 13.10.

Found: C, 59.40; H, 7.28; N, 13.09.

Synthèse du 1- (2-N-hydroxyuréidoéthoxyméthy !)-6-benzyt-5-éthyturaci ! e (Composé Comp. 3) (Composé de Formule 1 comparatif dans lequel n = 2, R1 = éTHYLE, R2 = H, R3 = OH, R4 = H).

Example 4

Synthesis of 1- (2-N-hydroxyureidoethoxymethy!) - 6-benzyt-5-ethyturaci! e (Compound Comp. 3) (A compound of Comparative Formula 1 wherein n = 2, R1 = ETHYL, R2 = H, R3 = OH, R4 = H).

A. 1- (2-Acetoxyethoxymethyl) -6-benzyl-5-ethyluracil A a suspension of 6-benzyl-5-ethyluracil (690 mg, 3 mmol), prepared as described in point A of Example 1 by replacing l '2-bromo-3-methylbutanoic acid by 2-bromobutanoic acid and 3, 5-dimethylphenylethanenitrile by phenylethanenitrile, in HMDS (hexamethyldisilazane) (15 ml, 75 mmol), added, under cover nitrogen, ammonium sulfate (15 mg). The reaction medium was kept at a boil under reflux for 14 h, cooled, then concentrated in vacuo. The residue was taken up, under cover of nitrogen, in anhydrous dichloromethane (12 ml). To the solution obtained was added in

<Desc / Clms Page number 26>

strictly anhydrous conditions, 1-acetoxy-2-acetoxymethoxyethane [A.

Rosowski, SH Kim. M. Wick, J. Med. Chem. (1981) 24, 1177-81] (1.06 g, 6 mmol) and trimethylsilyl triflate (0.57 ml, 3.3 mmol). After 4 h of stirring at 20 ° C, dichloromethane (12 ml) was added. The organic phase, washed with saturated sodium hydrogencarbonate solution (10 ml), then with saturated sodium chloride solution (10 ml), dried (magnesium sulfate) and concentrated, was successively subjected to chromatography on flash column of silica gel (chloroform / ethanol 9: 1) followed by a second flash column chromatography (petroleum ether / ethyl acetate 1: 1) to provide 1- (2-acetoxyethoxymethyl) -6- benzyl- 5-ethyluracil (380 mg, 69%) which was immediately subjected to the next step.

B. 6-Benzyi-S-ethyM- (2-hydroxyethoxymethyl!) Uraci) e.

To a solution of 1- (2-acetoxyethoxymethyl) -6-benzyl-5-ethyluracil (1.74 g, 5.02 mmol in methanol (30 ml), a 1 M methanolic solution of sodium methanolate (7 ml) was added. After stirring for 14 h at 20 ° C., the solution was brought to pH 4 with a 1 M aqueous solution of hydrochloric acid. The reaction medium concentrated, subjected to chromatography on a column of silica gel (chloroform / ethanol 19: 1) provided 6-benzyl-5-ethyl-1- (2hydroxyethoxymethyl) uracil (1 g, 66%).

2, 28 (q, 1 H, CHsCHs), 3, 42 (m, 4 H, CHCH), 4, 10 (s eel., 2 H, CHzPh),
4,65 (t, 1 H, JCH2, OH = 4,5 Hz, CH2OH), 5,01 (s, 2 H, OCH2N),
7,25 (s él., 5 H, Ph), 11,45 (s él, 1 H, NH). Melting point: 120-122 C, RF 0.25 (chloroform / ethanol 19: 1), 1H NMR (200 MHz, CDOs): 8 0.88 (t, 3H, J = 7.25 Hz, CH2CHO,

2, 28 (q, 1H, CHsCHs), 3, 42 (m, 4H, CHCH), 4, 10 (s eel., 2H, CHzPh),
4.65 (t, 1H, JCH2, OH = 4.5Hz, CH2OH), 5.01 (s, 2H, OCH2N),
7.25 (s el, 5H, Ph), 11.45 (s el, 1H, NH).

IR (KBr) 3396 (VOH), 3028 (VC-H arom.), 2832 (# C-H aliph.) 1706 cm-1 (# C = O).

C. 6-Benzyl-5-ethyl-1- (2-N1-hydroxyureidoethoxymethyl) uracil (Compound Comp. 3) To a solution of 6-benzyl-5-ethyl-1- (2-hydroxyethoxymethyl) uracil (150 mg,

<Desc / Clms Page number 27>

du composé intermédiaire 6-benzyl-1-[2- (N-phénoxycarbonyl-Nphénoxycarbonyloxyamino) éthyloxyméthyl) -5-éthyluracile, qui a été immédiatement dissous dans une solution méthanolique saturée d'ammoniac (5 ml). Le milieu réactionnel a été agité 4 h à 20 oC, concentré, puis soumis à une chromatographie sur colonne flash de gel de silice (dichlorométhane/éthanoI9 : 1) pour fournir, après recristallisation (acétate d'éthyle/méthanol) le 1-(2-N1-hydroxyréidoéthoxyméthyl)-6- benzyl-5-éthyluracile (80 mg, 45 %). 0.4 mmol) in tetrahydrofuran (3 ml), triphenylphosphine (126 mg, 0.48 mmol and N, O-bis (phenoxycarbonyl) hydroxylamine (120 mg, 0.44 mmol) were added at 20 ° C. ), then, dropwise, a solution of DIAD (diisopropylazodicarboxylate) (97 mg, 0.48 mmol) in tetrahydrofuran (0.5 ml). The reaction medium, concentrated, was subjected to chromatography on a flash column of silica gel (chloroform / ethanol 19: 1) to yield 300 mg

the intermediate compound 6-benzyl-1- [2- (N-phenoxycarbonyl-Nphenoxycarbonyloxyamino) ethyloxymethyl) -5-ethyluracil, which was immediately dissolved in methanolic solution saturated with ammonia (5 ml). The reaction medium was stirred for 4 h at 20 ° C., concentrated, then subjected to chromatography on a flash column of silica gel (dichloromethane / ethanoI9: 1) to provide, after recrystallization (ethyl acetate / methanol) the 1- ( 2-N1-hydroxyreidoethoxymethyl) -6-benzyl-5-ethyluracil (80 mg, 45%).

Melting point: 276-277 C, RF 0.25 (dichloromethane / methane 9: 1).

1H NMR (200 MHz, (CD3) 2SO, 80 OC): 0.89 (t, 3H, J = 7.3Hz, CHzCHs), 2.30 (q, 2H, CHzCHg), 3.47 ( t, 2H, J = 6.0Hz, NCH2CH2), 3.59 (t, 2H, CHsCHzO), 4.10 (s eel., 2H, CHsPh), 5.03 (s, 2H, OCH2N),
6.00 (s, 2H, NH2), 7.22 (s el, 5H, Ph), 9.13 (s, 1H, NOH),
11.16 (s el, 1H, NH).

IR (KBr): 3475 and 3341 (VNH and voH), 1706 cm-1 (vc = o).

MS (m / z (%)) 317 (3, M. + - NH2COH), 299 (6, M. + - NH2COH-H2O),
243 (94, M. + - NH2CONOH (CH2) 2O), 230 (100, M. + -NH2CONOH (CH2) 2OCH).

UV [MeOH, vmax (v)]: 205 (21210), 269 (11200) nm.

Elemental analysis for C12H22N4Os (362, 39):
Calculated: C, 56.35; H, 6.12; N, 15.46.

Found: C, 55.89; H, 6.07; N, 15.20.

Claims (1)

CLAIMS 1. Compound having the following general formula 1: in which: - n is equal to 3; - R1 represents an isopropyl group; - Each of the R 2 groups represents, independently of one another, a methyl group; - the group R3 represents a group -OR, where R is a pivaloyl group and R4 is a hydrogen atom; or a pharmaceutically acceptable salt thereof. 2. A compound according to claim 1 for use as a medicament. 3. A compound according to claim 1 for use as an antiviral agent. 4. A compound according to claim 3, for use as an anti-HIV-1 agent. <Desc / Clms Page number 29> 5. Process for preparing a compound of the following general formula 1:

in which: - n is equal to 3; - R1 represents an isopropyl group; - Each of the R 2 groups represents, independently of one another, a methyl group; - the group R3 represents a group -ORs, where R5 is a pivaloyl group and R4 is a hydrogen atom; which comprises the steps of: a) reacting a compound of formula II below:

wherein R2 is as defined above, with a compound of the following Formula III:

R1CHBrCOOEt (III) in which R1 is as defined above, in the presence of Zn, to obtain a compound of the following Formula IV: <Desc / Clms Page number 30>

wherein R1 and R2 are as defined above; b) reacting the compound of Formula IV obtained in step a) above a) of thiourea in a basic alcoholic medium to obtain a thiouracil of the following Formula V:

wherein R 1 and R2 are as defined above; c) reacting the thiouracil of Formula V obtained in step b) above with organic acid to obtain a compound of the following Formula VI:

wherein R1 and R2 are as defined above; d) carrying out a glycosidation reaction of the compound of Formula VI obtaining step c) above with a compound of Formula VII below

in which n is equal to 3, to obtain an ester, then solvolysing the ester for furnace the alcohol of Formula IX following: <Desc / Clms Page number 31>

wherein n, R1 and R2 are as defined above; and e) subjecting the alcohol of Formula IX obtained in step d) above to a Mitsunobu reaction using N, O-bis (phenoxycarbonyl) hydroxylamine as a nucleophile to provide the following compound of Formula X:

wherein n, R1 and R2 are as defined above; f) subjecting the compound of Formula X obtained in step e) above to aminolysis, to provide the compound of Formula as follows:

wherein n, R1 and R2 are as defined above; and <Desc / Clms Page number 32> g) protecting the OH group of the compound of Formula Ia obtained in step f) above with a group RI by an O-pivaloylation reaction to obtain the following compound of Formula lb:

where n, R \ R and R are as defined above. 6. A pharmaceutical composition containing as an active ingredient at least the compound according to claim 1 or a pharmaceutically acceptable salt thereof. 7. A pharmaceutical composition containing an antiviral effective amount of said compound according to claim 1 or a pharmaceutically acceptable salt thereof. 8. A pharmaceutical composition according to claim 6 or 7, for use as an anti-HIV-1 drug. 9. Use of a compound according to claim 1, or a pharmaceutically acceptable salt thereof, for the manufacture of an anti-HIV-1 medicament.