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WO1996010565A1 - 6-benzyl-4-oxopyrimidines substituees, leur procede de preparation et compositions pharmaceutiques les contenant - Google Patents

6-benzyl-4-oxopyrimidines substituees, leur procede de preparation et compositions pharmaceutiques les contenant Download PDF

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Publication number
WO1996010565A1
WO1996010565A1 PCT/EP1995/003912 EP9503912W WO9610565A1 WO 1996010565 A1 WO1996010565 A1 WO 1996010565A1 EP 9503912 W EP9503912 W EP 9503912W WO 9610565 A1 WO9610565 A1 WO 9610565A1
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WO
WIPO (PCT)
Prior art keywords
compounds
reacted
preparation
alkyl
hiv
Prior art date
Application number
PCT/EP1995/003912
Other languages
English (en)
Inventor
Marino Artico
Silvio Massa
Antonello Mai
Paolo La Colla
Maria Elena Marongiu
Enzo Tramontano
Original Assignee
Istituto Superiore Di Sanita'
Universita' Degli Studi Di Cagliari
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Istituto Superiore Di Sanita', Universita' Degli Studi Di Cagliari filed Critical Istituto Superiore Di Sanita'
Priority to AU38036/95A priority Critical patent/AU3803695A/en
Publication of WO1996010565A1 publication Critical patent/WO1996010565A1/fr

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D239/00—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings
    • C07D239/02—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings
    • C07D239/24—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings having three or more double bonds between ring members or between ring members and non-ring members
    • C07D239/28—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings having three or more double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, directly attached to ring carbon atoms
    • C07D239/46—Two or more oxygen, sulphur or nitrogen atoms
    • C07D239/52—Two oxygen atoms
    • C07D239/54—Two oxygen atoms as doubly bound oxygen atoms or as unsubstituted hydroxy radicals
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D239/00—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings
    • C07D239/02—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings
    • C07D239/24—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings having three or more double bonds between ring members or between ring members and non-ring members
    • C07D239/28—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings having three or more double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, directly attached to ring carbon atoms
    • C07D239/46—Two or more oxygen, sulphur or nitrogen atoms
    • C07D239/52—Two oxygen atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D239/00—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings
    • C07D239/02—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings
    • C07D239/24—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings having three or more double bonds between ring members or between ring members and non-ring members
    • C07D239/28—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings having three or more double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, directly attached to ring carbon atoms
    • C07D239/46—Two or more oxygen, sulphur or nitrogen atoms
    • C07D239/56—One oxygen atom and one sulfur atom

Definitions

  • the present invention refers to compounds having general formula (I):
  • X is selected from the group consisting of 0 and S;
  • R is selected from the group consisting of C 1-4 alkyl and C 5-6 cycloalkyl
  • AIDS acquired immunodeficiency syndrome
  • the transcription phase of the viral genome (a single RNA filament) in double strand DNA is the most studied one.
  • RT reverse transcriptase
  • DABO 3,4-dihydro-2-alkoxy-6- benzyl-4-oxopyrimidines
  • X is selected from the group consisting of 0 and S;
  • R is selected from the group consisting of C 1 -4 alkyl and C 5-6 cycloalkyl
  • the compounds of the present invention differ from the DABO described in the above reported literature owing to the presence of one S atom in the place of the 0 atom or owing to the presence of substituents on the benzylic ring.
  • Tables 3 and 4 the activity of some compounds according to the invention is reported, while in the Table 5 the data obtained with the above mentioned DABO compounds are reported by comparison.
  • methyl iodide (8 mmol; 1.13 g) is added to a solution containing the suitable 2-thiouracil derivative (4 mmol) dissolved in anhydrous N,N-dimethylformamide (2 ml) and the mixture is left under agitation at room temperature until the starting material disappears by the thin-layer chromatography check (silica gel/n-hexane: ethyl acetate: methanol 12:3:1).
  • the solution is diluted with water (200 ml), the aqueous phase is extracted with ethyl acetate (3 x 50 ml) and the reunited organic extracts are washed with a solution saturated with sodium thiosulfate (100 ml), with a solution saturated with NaCl (100 ml), dried (Na 2 SO 4 ) and deprived of the solvent.
  • anhydrous potassium carbonate (4.2 mmol) and the suitable alkyl halide (4.4 mmol) are added to a solution containing the suitable 2-thiouracil derivative (4 ml) dissolved in anhydrous N,N-dimethylformamide (2 ml) and the resulting mixture is left under agitation at room temperature (method B) or at 80 °C (method C) until the starting material disappears by the thin-layer chromatography check (silica gel/n-hexane: ethyl acetate: methanol 12:3:1).
  • the solution is diluted with water (200 ml), it is acidified to pH 5 with 0.5 N acetic acid and the aqueous phase is extracted with ethyl acetate (3 x 50 ml).
  • the reunited organic extracts are washed with a saturated solution of sodium thiosulfate (100ml), with a saturated solution of NaCl (100 ml), dried (Na 2 SO 4 ) and concentrated at reduced pressure.
  • the agitation has been continued for 1 hour at 0 °C and for a further hour at room temperature, then the mixture has been poured on ice and treated with 2N HCl.
  • the organic layer has been picked up and the aqueous solution washed two times with CH 2 CI 2 .
  • the organic phase and the extracts have been reunited, washed with brine and dried.
  • the mixture has been diluted with water (100 ml) after cooling, acidified to pH 5 with 0.5 N acetic acid and extracted with ethyl acetate (3 x 50 ml).
  • rRT recombinant enzyme
  • the cells used in this study were MT-4 and C8l66, both T4 lymphocytes lines permissive for the HIV replication.
  • the cells were suspended in RPMI 1640 added with fetal calf serum (FCS) at 10%, penicillin 100 U/ml and streptomycin 100 ⁇ g/ml.
  • FCS fetal calf serum
  • the cell cultures were incubated at 37 °C in 5% CO 2 atmosphere and were periodically checked to verify the absence of mycoplasmas contamination.
  • cytotoxicity a colorimetric method has been employed based on the use of a tetrazolium salt, the 3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium-bromide (MTT), which is transformed by the mitocondrial enzyme succinic dehydrogenase into a blue coloured product, the formazane, the amount of which turns out to be directly proportional to the number of viable cells.
  • MTT 3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium-bromide
  • the amount of formazane was then determined at the spectrophotometer by evaluation of the optical density at 570 nm.
  • the values shown in the columns CC 50 represent the compound concentrations required to reduce by 50% the MTT metabolization and, therefore, the cell viability; the mitocondrial metabolic process is, in fact, in a linear relation with the cell viability.
  • the major part of the compounds has low or null cytotoxicity in non infected cells, even at the maximum concentrations tested.
  • the inhibition of the virus-induced cytopathogenicity constituted the estimation criterion of the anti-HIV- 1 activity of the compounds.
  • the virus used in the antiviral tests (HIV-1, strain III B ) has been obtained from the chronically infected H9/III B cells supernatant.
  • the virus stock solutions were titled in C8166 and mantained at -80 °C till the moment of use.
  • MT-4 cells seeded at a density equal to 1 x 10 /ml, were infected with HIV-1 at a multiplicity of infection (m.o.i.) equal to 0.01. After 1 hour of incubation at 20 °C and subsequent removal of the inoculum, the cells were washed three times and then suspended again at a density equal to 1 x 10 ⁇ /ml, in absence or in presence of the test compounds.
  • the cell survival was determined with the above mentioned MTT method, in order to compute the values of EC 50 representing the compound concentration necessary to reduce by 50% the virus-induced cytopathogenicity.
  • test compounds are active in inhibiting the HIV-1 multiplication in MT-4 cells. They, owing to the lack of citotoxicity, have a selectivity index (meant as ratio between cytotoxicity and anti-HIV activity) particularly favourable.
  • RT reverse transcriptase
  • the gene of this enzyme has been formerly cloned in an expression vector, the protein has been expressed in E.coli and subsequently has been purified to obtain a preparation with a high purity degree.
  • the tests with the recombinant RT (rRT) have been carried out at 37 °C for 30 minutes in 50 ⁇ l containing 50 mM tris-HCl (pH 7.8), 1 mM dithiotreitol, 80 mM KCl, 6 mM MgCl 2 .
  • a unit is defined as the amount of enzyme necessary to incorporate 1 nmol of [ 3 H]-dTTP in the "template-primer" Poly(rA)-oligo(dT) 10 in one minute at 37 °C. After incubation, 40 ⁇ l of the reaction have been transferred on Whatman GF/A glass fiber filters and processed for the determination of the acid insoluble radioactivity after treatment with trichloroacetic acid. The values reported in the Tables (IC 50 ) represent the compound concentration required to reduce the enzyme activity by 50%.

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  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Saccharide Compounds (AREA)

Abstract

On décrit des 6-benzyl-4-oxopyrimidines substituées de la formule (I), les sels pharmacologiquement acceptables ainsi que les dérivés solubles de celles-ci. Dans cette formule, X est choisi parmi le groupe comprenant O et S; R est choisi parmi le groupe comprenant alkyle C1-4 et cycloalkyle C5-6; R', R' et Z sont égaux ou différents et représentent H ou alkyle C1-4, étant entendu que lorsque X = O, R et R' ne peuvent représenter tous deux H. On décrit également un des procédés de préparation de ces composés ainsi que l'utilisation de ceux-ci dans la préparation de compositions pharmaceutiques utiles dans le traitement d'infections virales.
PCT/EP1995/003912 1994-10-04 1995-10-04 6-benzyl-4-oxopyrimidines substituees, leur procede de preparation et compositions pharmaceutiques les contenant WO1996010565A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU38036/95A AU3803695A (en) 1994-10-04 1995-10-04 Substituted 6-benzyl-4-oxopyrimidines, process for their preparation and pharmaceutical compositions containing them

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
ITMI94A002023 1994-10-04
ITMI942023A IT1270122B (it) 1994-10-04 1994-10-04 6-benzil-4-ossopirimidine sostituite, processo per la loro preparazione e composizioni farmaceutiche che le contengono

Publications (1)

Publication Number Publication Date
WO1996010565A1 true WO1996010565A1 (fr) 1996-04-11

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Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/EP1995/003912 WO1996010565A1 (fr) 1994-10-04 1995-10-04 6-benzyl-4-oxopyrimidines substituees, leur procede de preparation et compositions pharmaceutiques les contenant

Country Status (3)

Country Link
AU (1) AU3803695A (fr)
IT (1) IT1270122B (fr)
WO (1) WO1996010565A1 (fr)

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2001007027A3 (fr) * 1999-07-22 2001-08-09 Vertex Pharma Inhibiteurs d'helicase virale
CN100439343C (zh) * 2006-08-04 2008-12-03 复旦大学 2-烷基硫-5-烷基-6-(1-氰基芳甲基)取代尿嘧啶类化合物及其制备方法和用途
CN100519540C (zh) * 2007-01-09 2009-07-29 云南大学 S-dabo类化合物、其合成方法和用途

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0123402A2 (fr) * 1983-03-25 1984-10-31 Fujisawa Pharmaceutical Co., Ltd. Dérivés de pyrimidine, leur préparation et leur utilisation
WO1991018887A1 (fr) * 1990-06-06 1991-12-12 Smithkline Beecham Intercredit B.V. Composes de diaminopyrimidine

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0123402A2 (fr) * 1983-03-25 1984-10-31 Fujisawa Pharmaceutical Co., Ltd. Dérivés de pyrimidine, leur préparation et leur utilisation
WO1991018887A1 (fr) * 1990-06-06 1991-12-12 Smithkline Beecham Intercredit B.V. Composes de diaminopyrimidine

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
A.MAI ET AL.: "SYNTHESIS AND ANTI-HIV-1 ACTIVITY OF THIO ANALOGUES OF DIHYDROALKOXYBENZYLOXOPYRIMIDINES.", JOURNAL OF MEDICINAL CHEMISTRY, vol. 38, no. 17, 18 August 1995 (1995-08-18), WASHINGTON US, pages 3258 - 3263 *
ANTIVIRAL CHEM. CHEMOTHER., vol. 6, no. 1, 1995, ENGL., pages 1 - 8 *
CHEMICAL ABSTRACTS, vol. 122, no. 1, 1995, Columbus, Ohio, US; abstract no. 122513c, S.MASSA,A.MAI: "SYNTHESIS AND ANTIVIRAL ACTIVITY OF 3,4-DIHYDRO-2-ALKOXY-6-BENZYL-4-OXOPYRIMIDINES" page 23; *

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2001007027A3 (fr) * 1999-07-22 2001-08-09 Vertex Pharma Inhibiteurs d'helicase virale
CN100439343C (zh) * 2006-08-04 2008-12-03 复旦大学 2-烷基硫-5-烷基-6-(1-氰基芳甲基)取代尿嘧啶类化合物及其制备方法和用途
CN100519540C (zh) * 2007-01-09 2009-07-29 云南大学 S-dabo类化合物、其合成方法和用途

Also Published As

Publication number Publication date
IT1270122B (it) 1997-04-28
ITMI942023A1 (it) 1996-04-04
ITMI942023A0 (it) 1994-10-04
AU3803695A (en) 1996-04-26

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