FR2705097A1 - New lupane derivatives, their preparation and pharmaceutical compositions containing them. - Google Patents

Abstract

Lupane derivatives of general formula (I) in which R is: (CF DRAWING IN BOPI) in which X is a bond, optionally substituted carbamoyl or aminocarbonyl, Y is a bond or phenylene, Rdegre and Rdegre degree identical or different are H or methyl and, n is 6 to 12, m is 0 to 3 and p is 0 to 2, with 6 <= m + n + p <= 14, and R1 is methyl, hydroxymethyl, -CH2 OR 'or -CH2 SR' (R 'being alkyl or hydroxyalkyl), the alkyl radicals (1 to 4C) being straight or branched, as well as their salts and their preparation. The products according to the invention are particularly advantageous in the field of AIDS. (CF DRAWING IN BOPI)

Description

leurs sels leur préparation et les compositions pharmaceutiques qui les contiennent.NEW LUPANE DERIVATIVES. PREPARATION AND COMPOSITIONS
PHARMACEUTICLES YES CONTAINING
The present invention relates to lupane derivatives of the general formula

their salts, their preparation and the pharmaceutical compositions containing them.

dans laquelle R1 et R2 sont hydroxy ou acyloxy et R3 est notamment méthyle. Ces dérivés sont utiles dans le domaine des anticancéreux.Japanese Patent Application No. 1,143,832 has disclosed derivatives of betulin of the general formula

wherein R1 and R2 are hydroxy or acyloxy and R3 is especially methyl. These derivatives are useful in the field of anticancer drugs.

It has now been found that lupane derivatives of general formula (I) in which
R represents a radical of general formula

X is a bond or represents a carbamoyl, N-methyl carbamoyl, aminocarbonyl or N-methylaminocarbonyl radical,
Y is a bond or represents a meta or para phenylene radical, R "and ROO, which may be identical or different, are hydrogen atoms or methyl radicals (it being understood that not all the -CR R units are necessarily identical to each other) and, n is an integer from 6 to 12, m is an integer from 0 to 3 and p is an integer from 0 to 2 with the proviso that m + n + p is from 6 to 14, and
R1 is a methyl, hydroxymethyl radical or a radical -CH2OR 'or -CH2SR' for which R1 is alkyl or hydroxyalkyl, and their pharmaceutically acceptable salts, exhibit a cytoprotective effect of cells infected with an HIV virus (Human
Immunodeficiency Virus) as well as inhibitory activity of the reverse transcriptase production of an HIV virus.

In the general formula above, it is understood that the alkyl radicals are straight and branched and contain 1 to 4 carbon atoms.

In cases where RO and ROO are different, the products of general formula (I) have stereoisomeric forms, it is also understood that stereoisomeric forms and mixtures thereof are also within the scope of the present invention.

dans laquelle R1 RO, ROO, X, Y, n, m, et p sont définis comme précédemment.According to the invention, the lupane derivatives of general formula (I) may also be prepared by the action of an azide on a nitrile of general formula

wherein R1 RO, ROO, X, Y, n, m, and p are defined as above.

The azide is advantageously chosen from tributyltin tributyl azide, trimethylsilyl azide or an alkaline azide.

When the n-butyltin azide is used, the reaction is carried out in an organic solvent such as 1,2-dimethoxy ethane, 2-ethoxyethanol or a mixture of such solvents at the reflux temperature of the mixture. reaction. When the trimethylsilyl azide is reacted, the reaction is carried out in the presence of aluminum trichloride, in a chlorinated solvent such as, for example, dichloromethane, at a temperature of between 10 and 400 ° C.

dans laquelle R1 est défini comme précédemment et R" représente un radical de formule générale :
(CH2)n-COOH (IVa)
ou
(CH2) n-NH-R',, (IVb) dans lesquelles n est défini comme précédemment et R"' est un atome d'hydrogène ou un radical méthyle, par action respectivement d'une amine ou d'un acide de formules générales :

dans lesquelles RO, ROO, m et p sont définis comme précédemment et
R"' est défini comme ci-dessus.According to the invention, the lupane derivative of general formula (I) for which X (in the radical R) represents an optionally N-methylated carbamoyl or aminocarbonyl radical and the other radicals are defined as above, can also be obtained from a lupane derivative of general formula

in which R 1 is defined as above and R "represents a radical of general formula:
(CH2) n-COOH (IVa)
or
(CH 2) n -NH-R ', (IVb) in which n is defined as above and R "' is a hydrogen atom or a methyl radical, by action respectively of an amine or an acid of formulas General

in which RO, ROO, m and p are defined as before and
R "'is defined as above.

It is understood that the hydroxyl radicals of the lupane derivative of general formula (IV) are preferably protected by compatible radicals which can be put in place and removed without affecting the rest of the molecule. By way of example, the protective groups may be chosen from the radicals described by TW GREENE,
Protective Groups in Organic Synthesis, J. Wiley-Interscience
Publication (1991) or by Mc OMIE, Protective Groups in Organic
Chemistry, Plenum Press (1973). Radicals that can be removed in a neutral, basic or acidic medium will be selected.

In particular, the hydroxy radicals can be protected in the ester state (formyloxy, acetoxy, i.butoxy, trichloroacetyloxy, phenoxyacetyloxy, benzyloxy); in this case, the elimination is carried out by hydrolysis in a basic medium, especially in the presence of sodium hydroxide at a temperature of between 10 and 50 ° C. The protection may also be carried out in the form of a ketone, in the form of carbonate by a -COORa radical in which Ra is an optionally substituted alkyl or benzyl radical or else a trialkylsilyl radical.

The reaction is carried out according to the usual methods of condensing an amine on an acid. In particular, it operates in the presence of an acid acceptor such as a nitrogenous organic base (trialkylamine, pyridine, N-methyl-morpholine, 1,8-diaza-bicyclo [5.4.0] undecene-7, diaza1,5 bicycloE4 .3.01 nonene-5 for example) in an organic solvent such as a chlorinated solvent (dichloromethane, dichloroethane, chloroform for example) or an amide (dimethylformamide for example). It is also possible to operate in the presence of a condensing agent such as a carbodiimide (dicyclohexylcarbodiimide, hydrochloride of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide for example) and optionally in the presence of a catalyst such as N- hydroxybenzotriazole or N-hydroxysuccinimide, at a temperature between -20 and 400C.

The condensation of the amine of general formula (Va) is preferably carried out from the hydrochloride of the amine.

The nitrile of general formula (III) can be prepared by dehydration of the corresponding amide by any known method which does not alter the rest of the molecule.

It is carried out in particular using trifluoroacetic anhydride in an ether (tetrahydrofuran, for example) at a temperature of between 10 and 660C.

The amide corresponding to the nitrile of general formula (III) may itself be obtained from the acid according to the known methods and / or cited below in the examples.

dans laquelle RO, ROO, X, Y, n, m, et p sont définis comme précédemment et dont la fonction acide est préalablement protégée, sur le chlorure de l'acide de formule générale :

dans laquelle R1 est défini comme précédemment et dont les radicaux hydroxy sont préalablement protégés, suivie de l'élimination du/des radicaux protecteurs.The corresponding acid may be prepared by the action of an amino acid of general formula:

in which RO, ROO, X, Y, n, m, and p are defined as above and whose acid function is protected beforehand, on the chloride of the acid of general formula:

in which R 1 is defined as above and whose hydroxy radicals are protected beforehand, followed by the elimination of the protective radical (s).

The reaction of the amine on the lupen-20 (29) oic acid chloride of general formula (VII) is carried out according to the usual methods which do not affect the rest of the molecule. It is carried out especially in the presence of a nitrogenous organic base such as a trialkylamine (triethylamine for example) in an organic solvent such as a chlorinated solvent (chloroform, 1,2-dichloroethane, dichloromethane) or in tetrahydrofuran or in a mixture of these solvents at a temperature between 15 and 300C.

It is understood that the hydroxy group (s) are preferably protected beforehand. The protection and release of the protective radicals are carried out as described above.

By way of example, the acid-protecting radicals may be chosen from alkyl (methyl, ethyl, tert-butyl), substituted alkyl (trichloroethyl, haloethyl, p-toluenesulfonylethyl, benzyl, benzyl substituted by a nitro radical, benzhydryl radicals). triphenylmethyl, benzyloxymethyl ...), alkyloxyalkyl (methoxymethyl), tetrahydropyranyl or trimethylsilyl. The establishment and elimination of these radicals is carried out according to the usual methods mentioned above.

dans lesquelles R"', RO, Roua, Y, m, et p sont définis comme précédemment, sur un dérivé du lupane de formule générale (IVa) ou (IVb).When the radical X (in R) is an optionally N-methylated carbamoyl or aminocarbonyl radical, the nitrile of general formula (III) may also be prepared by condensation of an amine or an acid of general formula

in which R "', RO, R a, Y, m, and p are defined as above, on a derivative of lupane of general formula (IVa) or (IVb).

The reaction is carried out under conditions identical to those for the preparation of a lupane derivative of general formula (I) starting from an amine or an acid of general formula (Va) or (Vb) and from a lupane derivative of general formula (IV).

The acid derived from lupane, of general formula (IVa), can be prepared by analogy with the reaction of the amino acid of general formula (VI) with the chloride of lupane-derived acid of general formula (VII), by an amino acid of general formula:
H2N- (CH2), - COOH (VIII) in which n is defined as above and whose acid function is previously protected.

The amine derived from lupane, of general formula (IVb), for which
R '''is a hydrogen atom, can be prepared from a lupane derivative of general formula (VII), by action of a diamine of general formula:
H2N- (CH2) n -NH2 (IX) wherein n is defined as above.

The derivatives (IVb) for which R '' is a methyl radical can be obtained by N-methylation according to the method described by
A. KATRITSKY, J. Chem. Soc. Perkin Trans. I, 2539 (1987), from the product of general formula (IVb) for which R "'is a hydrogen atom.

The products of general formula (Va) or (Vb) can be prepared according to or by analogy with the methods mentioned or described hereinafter in the examples.

The lupane derivatives of general formula (IV) or (VII) for which
R1 is a hydroxymethyl radical can be prepared by the action of silver acetate on the corresponding brominated derivative for which R1 is bromomethyl and for which the hydroxy radical is previously protected. The reaction is advantageously carried out in an organic solvent such as, for example, toluene, at a temperature of between 200 ° C. and the reflux temperature of the reaction mixture. The starting brominated derivative can be obtained by the action of a brominating agent such as, for example, tetrabutylammonium tribromide or N-bromosuccinimide on the corresponding derivative of lup-20 (29) en-28-oic acid. The reaction is carried out in a chlorinated solvent such as methylene chloride, chloroform, or carbon tetrachloride at a temperature of 250C.

The lupane derivatives of general formula (VII) for which R 1 is a radical -CH 2 OR 'or -CH 2 SR' can be prepared from the lupane derivative for which R 1 is bromomethyl, by the action of the corresponding alkoxide or thiolate.

The acid chloride of the lupane derivative of general formula (VII) can be prepared according to known methods. By way of example, oxalyl chloride or thionyl chloride is reacted with a chlorinated solvent such as chloroform, dichloroethane or dichloromethane.

The new lupane derivatives of general formula (I) can be purified, if appropriate, by physical methods such as crystallization or chromatography.

When the lupane derivative of general formula (I) has stereoisomeric forms, the preparation is carried out by means of chiral derivatives of general formula (Va), (Vb), (VI), (VIIa) or (VIIb). It is understood that when a chiral intermediate is to be obtained, the separation is carried out according to the usual methods which do not affect the remainder of the molecule, such as, for example, chiral phase chromatography, or by conversion of the racemic mixture. in diastereoisomers and separation by crystallization or chromatography.

The products according to the invention can be converted into metal salts with strong bases according to the methods known per se.

These salts can be obtained by the action of a metal base in a suitable solvent. The salt formed precipitates after optional concentration of the solution; it is separated by filtration.

As examples of pharmaceutically acceptable salts may be mentioned salts with alkali metals (sodium, potassium, lithium).

The novel lupane derivatives according to the present invention are particularly useful for the prophylaxis and treatment of AIDS (Acquired Immune Deficiency Syndrome) and Associated Syndromes (ARC). who have been exposed to HIV viruses, especially asymptomatic seropositive patients who are at risk of developing the disease in the months or years to come after primary infection.

The products according to the invention, which are inhibitors of the cytopathic effect of HIV and inhibitors of the production of reverse transcriptase in cell culture at concentrations devoid of a cytotoxic or cytostatic effect, are particularly interesting.

The activities were highlighted in the following tests: Actlvité vis-à-vis the cytopathogenic effect of the HIV virus
The powdered products were dissolved in a proportion of 2 mg of product per 2 ml (approximately 4 × 10 3 M) in dimethylformamide (DMF) to obtain a stock solution of 100% DMF product. The test is carried out on the lymphoblastoid line CEM M4. In a 96-well microplate, 25 μl / well of a test product solution is deposited in isotonic phosphate buffer (TPI) or TPI alone in the case of the controls. The products are studied at 8 concentrations, at the rate of 6 wells per concentration. 125 μl of a suspension of CEM cells (8 × 10 4 cells per ml) are then added to RPMI medium containing 10% fetal calf serum. IU / ml penicillin, 100 μg / ml streptomycin and 2 pmoles / ml glutamine and the microplates are incubated for 1 hour at 370C under an atmosphere containing 5% carbon dioxide. For each concentration, the assay is divided into two parts: one part (3 wells) on infected cells, for the determination of the antiviral activity and the other part (3 wells) on uninfected cells, to determine the cytotoxicity of the cells. products.On then infects the first series with
HIV-1 (100 μl per well of a suspension of LAV-1-BRU virus containing 200-300 TCID50) while the other set receives 100 μl of RPMI medium as defined above. After 5 days of incubation, 100 μl of cells are taken to measure the cell viability [determined according to a modification of the technique described by R.

Pauwels et al., J. Virol. Meth., 20, 309-321 (1988)]. 10 μl of a solution containing 7 mg of MrT (3- (4,5-dimethyl-2-thiazolyl) -2,5-diphenyltetrazolium bromide) per ml of isotonic phosphate buffer are added to this sample. incubation at 370 ° C. The supernatant is removed The MTT is converted into a formazan salt (blue) only inside the living cells 100 μl of isopropanol (containing 0.04 mol / l hydrochloric acid) is then added and the microplates are shaken until the solubilization of formazan blue.The absorbance at 540 nm is read with an automatic reader of microplate ELISA reactions.This absorbance is proportional to the amount of living cells.

The protection rate (in%) of a given product is determined from the optical densities (OD) by the formula:
OD (cell treated and inf.) - DO (cell untreated and inf.)
OD (cell treated and not inf) - DO (cell untreated and inf.)
If necessary, the 50% inhibitory concentration is determined.

The results demonstrate that for tested product concentrations of between 0.01 and 10 μg / ml, a significant decrease in the cytopathic effect is obtained.

The degree of protection provided by the products according to the invention is between 20 and 100%.

The 50% inhibitory concentration of the products according to the invention is between 0.05 and 50 μg / ml when it can be determined.

Determination of vascular multiplication by assaying the reverse transcriptase of the HIV virus
The activity of the reverse transcriptase is measured in 10 μl of culture supernatant. Using assay kit poly r (A) reverse transcriptase [3H] -SPA enzyme assay (RPQN 0100, Amersham, GB) according to the procedure recommended by the manufacturer. The reaction time is 6 hours and the radioactivity counted using a microbeta counter [TOPCOUNT PACKARD].

Inhibitions of virus-associated reverse transcriptase production are observed between 20 and 100% at the tested concentrations (0.01 to 10 μg / ml).

The following examples given by way of non-limiting example illustrate the present invention.

Example 1
To a solution of 0.5 g of N- (35-acetoxylup-20 (29) -en-28-yl) -8-
3 aminooctanoic in 90 cc of dichloromethane, 125 mg of 5-aminomethyl tetrazole hydrochloride, 141 mg of 1-hydroxybenzotriazole hydrate, 380 mg of 1-β-dimethylaminohydrochloride are added.
3 propyl) -3-ethylcarbodiimide and 0.48 cm 3 of triethylamine. The solution is stirred for 24 hours at 200C then the reaction mixture is
The organic phase is then dried over 100 cm 3 of distilled water. The organic phase is then dried over magnesium sulphate and evaporated to dryness under reduced pressure (2.7 kPa) at a temperature in the region of 400 ° C. The residue obtained (600 mg) is chromatographed on a silica column (0.020.045 mm) eluted first with 2 liters of a mixture of cyclohexane and ethyl acetate (30-70 by volume), then with 2 liters. a mixture of methanol and ethyl acetate (30-70 by volume); the fractions containing the product are combined and concentrated under reduced pressure (2.7 kPa) at a temperature in the region of 400C. The residue is taken up with 10 cm of ethyl ether, filtered, washed twice.
3 with 5 cm 3 of ethyl ether and dried under reduced pressure (13.5 Pa) at 200 ° C.. Thus 290 mg of 5-CN '- [N- (3 P-acetoxylup-20 (29) -en- oyl) -8-aminooctanoyl] -aminomethyl} -tetrazole as a white powder (Rf = 0.26, thin layer chromatography of silica, eluent: chloroform-methanol-20% ammonia (12-3-0 , In volume)).

A solution of 1.07 g of 5- {N '- [N- (3ss-acetoxylup-20 (29) -En-28-oyl) -8-aminooctanoyl] -aminomethyl} -tetrazole obtained as described above. 3 3 above, 5.7 cm of 5N sodium hydroxide, 18.4 cm 3 of tetrahydrofuran and 36.9 cm 3 of methanol are stirred for 20 hours at a temperature in the region of 200 ° C. The mixture is cooled to 50C, then acidified with chlorhy acid
3 g, 5N, and diluted with 100 cm distilled water. After stirring for 1 hour at 50 ° C., the solid obtained is separated by filtration, washed 3 times with 25 cm 3 of distilled water and dried under reduced pressure (15.5 Pa) at a temperature in the region of 200 ° C. Thus 880 mg is obtained 5-tN '- [N- (3-hydroxylup-20 (29) -En-28-oyl) -8-aminooctanoyl] -amino-methyl] -tetrazole, as a white solid melting at about 15 ° C.

N- (3 (3-Acetoxy-20 (29) -En-28-alkyl) -8-aminooctanoic acid can be prepared in the following manner
After refluxing for 3 hours a mixture of 330 mg of 8-aminooctanoic acid and 410 mg of chlorotrimethylsilane in
50 cm 3 of dichloromethane, the solution is cooled to 200 ° C. and 1 g of 35-acetoxylup-20 (29) -en-28-oyl chloride (prepared with
From 313-acetoxylup-20 (29) -en-28-oic acid) followed by 0.81 cm 3 of triethylamine. Stirring is maintained for 15 hours at a temperature in the region of 200 ° C. and the solvent is evaporated to dryness under reduced pressure (2.7 kPa) at a temperature in the region of 350 ° C. The residue is chromatographed on a silica column (0.02 ° C.). -0.045 mm) eluted with a mixture of cyclohexane and ethyl acetate (60-40 by volume). The fractions containing the product are combined and concentrated to dryness under reduced pressure (2.7 kPa) at a temperature of 350C. There is thus obtained 1.1 g of N- (3ss-acetoxylup- (29) -En-28-alkyl) -8-aminooctanolec acid in the form of a white meringue (Rf = 0.4; thin silica eluent: cyclohexane, ethyl acetate (60-40 by volume)).

The 5-aminomethyl-tetrazole hydrochloride can be prepared according to the method described in the patent application DE 3,626,130.

Exemole 2
To a solution of 640 mg of N- (3ss-acetoxylup-20 (29) -en-28-oyl) -
8-aminooctanoic acid in 50 cm 3 of chloroform is added 134 mg of 5-amino-1H-tetrazole, 168 mg of 1-hydroxybenzotriazole hydrate,
3 50 cm 3 of dimethylformamide, 692 mg of 1- (3-dimethyl) hydrochloride
3 laminopropyl) -3-ethylcarbodiimide, and 0.70 cm 3 of triethylamine
3 solution in 5 cm of chloroform.The solution is stirred for 2
3 days at 200 ° C., then the reaction mixture is washed 3 times with 50 cm of 1N hydrochloric acid and then 5 times with 100 cm 3 of distilled water.
The organic phase is dried over magnesium sulphate and evaporated to dryness under reduced pressure (2.7 kPa) at a temperature in the region of 400C. The residue obtained (730 mg) is chromatographed on a column
3 silica (0.02-0.045 mm) eluted with 800 cm 3 of ethyl acetate then
3 with 800 cm 3 of a mixture of dichloromethane and methanol (95-5 by volume). The fractions containing the product are combined and concentrated under reduced pressure (2.7 kPa) at a temperature in the region of 400C. The residue is taken up in 15 cm 3 of isopropyl ether, filtered and dried under reduced pressure (13.5 Pa) at 200 ° C. Thus 265 mg of 5-CN '- CN- (313-acetoxylup-20 (29 ) -En-28-oyl) -8-aminooctanoyl] -amino) -tetrazole as a beige powder (Rf = 0.40; silica thin layer chromatography; eluent: chloroform-methanol-20% ammonia) (12-3-0.5 volumes)).

A solution of 260 mg of 5- {N '- [N- (3ss-acetoxylup-20 (29) -en-28-oyl) -8-aminooctanoyl] -amino} -tetrazole, 0.92 cm3 of sodium hydroxide. 1N lithium, 2.5 cm3 of tetrahydrofuran and 5 cm of methanol is stirred for 20 hours at a temperature of 200C. The mixture is acidified
3 with 1N hydrochloric acid, and diluted with 80 cm distil water
3 lé. The mixture is then extracted with 100 cm 3 of ethyl acetate and then twice with 25 cm 3 of ethyl acetate. The organic phases are combined, washed 4 times with 40 cm 3 of distilled water, dried over magnesium sulphate and evaporated to dryness under reduced pressure (2.7 kPa) at a temperature in the region of 450 ° C. The residue obtained (170 mg ) is chromatographed on a silica column (0.020.045 mm) eluted with a mixture of chloroform, methanol and 20% ammonia (12-3-0.5 by volume). The fractions containing the product are combined and concentrated under reduced pressure (2.7 kPa) at
3 a temperature close to 400C. The residue is taken up by 5 cm
3 isopropyl ether, filtered, washed 2 times with 1 cm of isopropyl ether and dried under reduced pressure (15.5 Pa) at 200C. 80 mg of 5- {N '- [N- (3ss-hydroxylup-20 (29) -En-28-alkyl) -8-aminooctanoyl] -amino} -tetrazole are thus obtained in the form of a white solid. melting at 165-1700C.

Example 3
To a solution of 100 mg of N- (313,30-diacetoxylup-20 (29) -en-28-
3 μl) -8-aminooctanoic in 50 cm 3 of dichloromethane, 22 mg of 5-aminomethyl tetrazole hydrochloride, 25 mg of 1-hydroxybenzotriazole hydrate, 70 mg of 1- (3-dimethylamino) hydrochloride are added.
3 propyl) -3-ethylcarbodiimide and 0.09 cm triethylamine. The solution is stirred for 24 hours at 200 ° C., then the reaction mixture is washed successively 3 times with 50 cm 3 of distilled water, 2 times with 25 cm 3 of 1N hydrochloric acid and then 3 times with 50 cm 3 of distilled water. The organic phase is then dried over magnesium sulphate and evaporated to dryness under reduced pressure (2.7 kPa) at a temperature in the region of 400 ° C. The residue obtained (110 mg) is chromatographed on a silica column (0.02 ° C.). 0.045 mm) eluted with a mixture of chloroform, methanol and 20% ammonia (12-3-0.5 vol. The fractions containing the product are combined and concentrated under reduced pressure (2.7 kPa) at a temperature in the region of 400C. There is thus obtained 65 mg of 5- {N '- [N- (3ss, 30-diacetoxylup-20 (29) -en-28-alkyl) -8-aminooctanoyl] -aminomethyl} -tetrazole in the form of a meringue. white (Rf = 0.40, silica thin layer chromatography, eluent: chloroform-methanol-20% ammonia (12-3-0.5 by volume)).

A solution of 160 mg of 5- {N '- [N- (3ss, 30-diacetoxylup-20 (29) -en-28-
3-yl) -8-aminooctanoyl] -aminomethyl} -tetrazole, 1.3 cm of 5N sodium hydroxide,
36 cm 3 of tetrahydrofuran and 3 cm 3 of methanol are stirred for 20 hours at a temperature in the region of 200 ° C. The mixture is cooled to 50C, then
3 acidified with 5N hydrochloric acid, and diluted with 30 cm of water
Distilled. After stirring for 1 hour at 50 ° C., 150 cm.
3 ml of distilled water and the mixture is extracted 5 times with 50 cm of a mixture of dichloromethane and methanol (9-1 by volume). The organic phases are combined, dried over magnesium sulphate and evaporated to dryness under reduced pressure (2.7 kPa) at a temperature in the region of 400 ° C. The residue obtained (140 mg) is chromatographed on a silica column (0.02 g). -0.045mm) eluted with a mixture of chloroform, methanol and ammonia at 20% (12-3-0.5 volumes). The fractions containing the product are combined and concentrated under reduced pressure (2.7 kPa) at a temperature in the region of 400C. The residue is taken up in 2 cm of acetonitrile, filtered and dried under reduced pressure (15.5 Pa) at 200 ° C. 48 mg of 5- {N '- [N- (3ss, 30-dihydroxylup-20 (29) -En-28-oyl) -8-aminooctanoyl] -aminomethyl) -tetrazole are thus obtained in the form of a white solid melting at 145-1500C.

N- (3 (3,30-Diacetoxylup-20 (29) -En-28-oyl) -8-aminooctanoic acid is synthesized by analogy with N- (3ss-acetoxylup-20 (29) -en 32-amino-octanoic acid from 3β, 30-diacetoxylup- (29) -en-28-alkyl chloride and methyl 8-aminooctanoate (R = 95%, Rf = 0.34; thin layer of silica, eluent: cyclohexane-ethyl acetate (50-50 by volume)).

3β, 30-Diacetoxylup-20 (29) -en-28-oyl chloride is prepared in the following manner:
To a solution of 560 mg of 3ss, 30-diacetoxylup-20 (29) -en-28-
In 25 cm 3 of dichloromethane, 0.22 cm 3 of oxalyl chloride is added. After stirring for 15 hours at a temperature in the region of 20 ° C., the solvent is evaporated under reduced pressure (22 kPa) and at a temperature in the region of 400 ° C. 600 mg of a white meringue is obtained which is used without further purification.

3ss, 30-diacetoxylup-20 (29) -en-28-oic acid is obtained as follows:
A suspension of 1.8 g of 3,3-acetoxy-30-bromolup-20 (29) -en-28-enic acid and 800 mg of silver acetate in 50 cm of toluene is stirred for 48 hours at a temperature of close to 200C then filtered. The solution obtained is then concentrated to dryness under reduced pressure (2 kPa).

3
The residue is dissolved in 50 cm 3 of ethyl acetate. The
The organic phase is decanted and washed with a total of 60 cm 3 of distilled water, dried over anhydrous magnesium sulfate and then concentrated under reduced pressure (2.7 kPa) at a temperature of 400 ° C. The white solid obtained (1.7 g) is chromatographed on a column 1.7 cm in diameter and containing 35 g of silica (0.020.045 mm) eluted with a mixture of cyclohexane and ethyl acetate.
80-20 (by volume), collecting 15 cm fractions. The first 4 fractions obtained are removed, the following 10 are concentrated to dryness under reduced pressure (13.5 Pa) at a temperature in the region of 400 ° C. Thus, 1.1 g of 3β, 30-diacetoxylup- (29) acid is obtained. ) -en-28-oic in the form of a white meringue sufficiently pure for subsequent transformations.

3ss-Acetoxy-30-bromolup-20 (29) -en-28-oic acid is obtained in the following manner:
A solution of 2.5 g of 313-acetoxy-lup-20 (29) -en-28-oic acid and
3.41 g of tetrabutylammonium tribromide in 50 cm 3 of chloroform is stirred for 5 days at a temperature in the region of 25 ° C. The reaction mixture is washed successively twice with 25 cm of distilled water,
2 times per 25 cm of 0.1 N sodium thiosulfate solution and
2 x 25 cm of distilled water and dried over anhydrous magnesium sulfate. The organic phase is concentrated to dryness under reduced pressure (2.7 kPa) and at a temperature in the region of 400 ° C. A yellow pasty solid (4 g) is obtained which is chromatographed on a column 3.2 cm in diameter containing 100 g. of silica (0.02-0.045 mm) and eluted with a mixture of cyclohexane and ethyl acetate 85-15 (in
3 volumes) collecting 20 cm fractions. The first 5 fractions obtained are removed, the following 10 are evaporated under reduced pressure (13.5 Pa) at a temperature of 400C. 2.7 g of 3β-acetoxy-30-bromolup-20 (29) -en-28-oic acid are thus obtained in the form of a white solid melting at a temperature in the region of 1900 ° C.

3ss-Acetoxy-20 (29) -en-28-oic acid can be prepared according to
BRUCKNER, KOVACS and KOCZKA, J. Chem. Soc., 948 (1948).

EXAMPLE 4
To a solution of 400 mg of N- [3ss-acetoxylup-30- (2'-acetoxyethylthio) -20 (29) -en-28-alkyl] -8-aminooctanoic acid, 90 mg of hydrochloride of 5 aminomethyltetrazole, 100 mg of 1-hydroxybenzotriazole hydrate, 250 mg of 1- (3-dimethylaminopropyl) -3-ethyl-carbodiimide hydrochloride in 100 cm 3 of dichloromethane, is added in approximately
10 minutes, 0.32 cm 3 of triethylamine. The solution is agitated during
3 hours at a temperature in the region of 200 ° C., plus 50 cm
3 of distilled water and then acidified to a pH of about 1 per 15 cm of an aqueous solution of N hydrochloric acid. The organic phase is decanted.
3, the aqueous phase is extracted with 3 times 50 cm of dichloromethane. The combined organic phases are then dried over magnesium sulphate and evaporated to dryness under reduced pressure (2.7 kPa) at a temperature in the region of 400C. The residue obtained (400 mg) is chromatographed on a silica column (0.02-0.045 mm), eluted with a mixture of dichloromethane and methanol (90-10 by volume). The fractions containing the product are combined and concentrated under reduced pressure (2.7 kPa) at a temperature in the region of 400C. 300 mg of 5-CN '- [N- (313-acetoxylup-30- (2'-acetoxyethylthio) - (29) -En-28-alkyl) -8-aminooctanoyl] aminomethyl) -tetrazole are thus obtained under the white meringue (Rf = 0.40, thin layer chromatography of silica, eluent: 20% chloroform-methanol-ammonia (12-3-0.5 by volume)).

A solution of 300 mg of 5-CN '- [N- (3P-acetoxylup-30- (2'-acetoxyethylthio) -20 (29) -en-28-alkyl) -8-aminooctanoyl] aminomethyl-tetrazole ,
3.2 cm 3 of N sodium hydroxide, 10 cm 3 of tetrahydrofuran and 20 cm 3 of methanol are stirred for 48 hours at a temperature in the region of 200 ° C. The mixture 3 is acidified to pH 1 by addition of 4 cm of an aqueous solution
3 of 4N hydrochloric acid and then diluted with 70 cm of distilled water.

After stirring for 2 hours at a temperature in the region of 200.degree.
3 solid is filtered and washed 5 times with 10 cm of distilled water. The O
The liquid is dissolved in 20 cm 3 of ethanol. The solution is filtered and the filtrate is evaporated under reduced pressure (2.7 kPa) at a temperature in the region of 40 ° C. The residue obtained (300 mg) is chromatographed on a silica column (0.02-0.045 mm), eluted with a mixture of chloroform, methanol and ammonia at 20% (24-6-1 by volume) The fractions containing the product are combined and concentrated under reduced pressure (2.7 kPa) at a temperature in the region of 400 ° C. The solid obtained 3 (150 mg) is treated with 5 cm 3 of isopropyl ether, the solid is
The mixture is filtered, washed twice with 1 cm 3 of isopropyl ether and then dried over potassium hydroxide. In this way 100 mg of 5- {N '- [N- (3-hydroxylup-30- (2'-hydroxyethylthio) is obtained. -20 (29) -En-28-alkyl) -8-aminooctano-1-aminomethyl-tetrazole as a white solid melting at about 140 ° C.

N- [3ss-acetoxylup-30- (2'-acetoxyethylthio) -20 (29) -en-28-alkyl] -8-amino-octanolic acid can be obtained as follows:
To a solution of 500 mg of N- [3ss-acetoxylup-30- (2'-hydroxyethylthio) -20 (29) -en-28-alkyl] -8-aminooctanoic acid and 600 mg of acetate of 3 Sodium in 35 cm 3 of acetic anhydride is brought to a temperature close to reflux for about 3 hours. The reaction mixture is cooled to 200 ° C. and then concentrated under reduced pressure (2.7 kPa) at a temperature in the region of 40 ° C. The residue is treated with a mixture of
The reaction mixture is concentrated to dryness under reduced pressure (2.7 kPa) at a temperature in the region of 50.degree. C. and the mixture is stirred for 12 hours at a temperature in the region of 200.degree. The residue is treated with 50 cm 3 of distilled water and 10 cm 3 of an aqueous solution of N hydrochloric acid. The organic phase is
3 decanted, the aqueous phase is extracted with 3 times 40 cm of dichloromethane. The combined organic phases are washed 3 times with 20 cm 3 of distilled water, dried over magnesium sulphate and then evaporated to dryness under reduced pressure (2.7 kPa) at a temperature of
3 sine of 400C. The residue obtained (500 mg) is recrystallized in 5 cm of an ethanol-water mixture (75-25 by volume). The solid obtained is
The mixture is filtered, then washed twice with 1 cm of an ethanol-water mixture (75-25 by volume) and dried under reduced pressure (13.5 Pa) over phosphorus pentoxide. 400 mg of N-acid are thus obtained. [3 P-acetoxylup-30- (2'-acetoxyethylthio) -20 (29) -en-28-alkyl] -8-aminooctanoic acid as a white meringue (Rf = 0.25; silica thin layer chromatography) eluent: chloroform-methanol-ammonia 20% (24-6-1 by volume)).

N- [3ss-acetoxylup-30- (2'-hydroxyethylthio) -20 (29) -en-28-alkyl] -8-aminooctanoic acid can be obtained as follows:
To a solution of 1.3 g of N- [3ss-acetoxylup-30-bromo-20 (29) -nn-
28-alkyl-8-amino-octanol in 10 cm 3 of ethanol, a solution is added
0.60 g of sodium 2-hydroxyethanethiolate in 35 cm 3 of ethanol. The reaction mixture is stirred for 48 hours at a temperature in the region of 200 ° C. It is added drop by drop in 10 minutes
Approximately 5 cm 3 of an aqueous solution of hydrochloric acid N and then
3 diluted by 350 cm of water. The reaction mixture is extracted 3 times
3 by 50 cm of dichloromethane.The combined organic phases are
3 washed with 30 cm 3 of distilled water and then dried over magnesium sulphate and evaporated to dryness under reduced pressure (2.7 kPa) at a temperature in the region of 400 ° C. The oil obtained (1.1 g) is chromatographed on a silica column (0.02-0.045 mm), eluted with a mixture of dichloromethane and methanol (98-2 by volume). The fractions containing the product are combined and concentrated under reduced pressure (2.7 kPa) at a temperature in the region of 40 ° C. This gives 500 mg of N- [3ss-acetoxylup-30- (2'-hydroxyethylthio) - (29) -En-28-alkyl] -8-aminooctanoic acid in the form of a white meringue (Rf = 0.40 thin layer silica chromatography, eluent: chloroformmethanol-20% ammonia (12-3-0); , In volume)).

N- [3ss-acetoxylup-30-bromo-20 (29) -en-28-alkyl] -8-aminooctanoic acid can be obtained as follows:
To a mixture of 640 mg of N- [3ss-acetoxylup-20 (29) -en-28-alkyl] -8-aminooctanoic acid and 30 cm 3 of carbon tetrachloride, add at once 350 mg of N bromosuccinimide. The suspension is stirred for 12 hours at a temperature in the region of 2000. The solid is filtered, washed 3 times with 10 cm 3 of carbon tetrachloride. The filtrate is evaporated under reduced pressure (2.7 kPa) at a temperature in the region of 400 ° C. 800 mg of N- [3ss-acetoxylup-30-bromo-20 (29) -En-28-alkyl] -8-aminooctanoic acid are obtained in the form of a white meringue (Rf = 0.25; thin layer of silica, eluent: chloroform-methanol-ammonia 20% (24-6-1 by volume)).

N- [3ss-acetoxylup-30-bromo-20 (29) -en-28-alkyl] -8-aminooctanoic acid is prepared as described in Example 1.

Sodium hydroxyethanethiolate is prepared by the action of sodium ethoxide on 2-hydroxyethanthiol dissolved in methanol
3ss-acetoxylup-20 (29) -en-28-oyl chloride is prepared from J.

PROVITA and A. VYSTRCIL, Collect. Czech. Chem. Common. 41, 1200 (1976).

Exemole 5
To a solution of 1.9 g of N- [[313-acetoxylup-20 (29) -En-28-alkyl] -8-aminooctanoic acid in 55 cm of dichloromethane is added 2.2 cm of thionyl chloride. . The solution is stirred for 24 hours at 200C then the reaction mixture is concentrated to dryness under reduced pressure (2.7 kPa) at a temperature of 350C. The residue obtained is dissolved in 25 cm 3 of dichloromethane and the solution is again evaporated to dryness under reduced pressure (2.7 kPa) at a temperature in the region of 350 ° C. The solid obtained (1.97 g) and 485 mg of 5- (4-aminophenyltetrazole) are dissolved in 60 cm of dichloromethane.
3 ml of ethane and 1.41 cm 3 of triethylamine are added dropwise. The solution obtained is stirred for 24 hours at a temperature in the region of 2000. The solution obtained is concentrated under reduced pressure (2.7 kPa) at a temperature in the region of 350 ° C. To the solid obtained is added 50 cm 3 of dichloromethane and 25 cm 3 of an N hydrochloric acid solution. The organic phase is decanted, washed twice with 10 cm 3 of a solution of hydrochloric acid N and then twice. with 10 cm distilled water. The organic phase is dried over sodium sulphate and then concentrated to dryness under reduced pressure (2.7 kPa) at a temperature in the region of 350 ° C. This gives 2.0 g of a solid which is chromatographed on a silica column (0.degree. 02-0.045 mm) eluted with a mixture of methanol and ethyl acetate (10-90 by volume), the fractions containing the product are combined and concentrated under reduced pressure (2.7 kPa) at a temperature in the region of 4000. The residue (280 mg) is chromatographed on a column of silica (0.020.045 mm) eluted with a mixture of chloroform, methanol and ammonia (24-6-1 by volume), the fractions containing the product are combined and concentrated. under reduced pressure (2.7 kPa) at a temperature in the region of 350 ° C. 310 mg of 5- [4-fN- [N'- (3ss-acetoxylup-20 (29) -en-28-oyl) are thus obtained. 8-aminooctanoyl] amino] phenyl] tetrazole as a white powder (Rf = 0.32, silica thin layer chromatography, eluent: chloroform) rme-methanol-ammonia 20% (24-6-1 by volume)).

A solution of 0.31 g of 5- [4- {N- [N '(3ss-acetoxylup-20 (29) -en-28-
3 3 -aryl-8-aminoctyllamino} -phenyl] -tetrazole, 1 cm 4N sodium hydroxide, 3 cm
3 ml of tetrahydrofuran and 1.5 cm 3 of methanol are stirred for 26 hours at a temperature in the region of 2000. The mixture is evaporated under reduced pressure (2.7 kPa) at a temperature in the region of 350 ° C. and then diluted with 20 cm 3 of distilled water and acidified with 1.2 cm 4N hydrochloric acid solution After stirring for 1 hour, at a temperature in the region of 200 ° C., the solid is separated by filtration, washed 3 times with 5 cm of distilled water and dried under reduced pressure (15.5 Pa) at a temperature in the region of 3000. This gives 250 mg of 5- [4-IN- [N'- (313-hydroxylup-20 (29) -en-28-alkyl ) -8-aminooctanoyl] -aminol-phenyl] -tetrazole, as a white solid melting at 2000C.

5- (4-aminophenyl) tetrazole can be prepared from J.M. McMANUS and R. HERBST, J. Org. Chem., 11, 1044 (1959).

Example 6
To a solution of 1.9 g of N- (313-acetoxylup-20 (29) -en-28-alkyl) -8-aminooctanoic acid in 55 cm 3 of dichloromethane is added thionyl. The solution is stirred for 24 hours at 200C then the reaction mixture is concentrated to dryness under reduced pressure (2.7 kPa) at a temperature of 350C. The resulting residue is put
3 in solution in 25 cm 3 of dichloromethane and the solution is again concentrated to dryness under reduced pressure (2.7 kPa) at a temperature in the region of 3500. The solid obtained (1.97 g) and 485 mg of 5- (3
3 aminophenyltetrazole) are dissolved in 60 cm of dichloromethane
3 ml of ethane and 1.41 cm 3 of triethylamine are added dropwise. The solution obtained is stirred for 24 hours at a temperature in the region of 200 ° C. The solution obtained is concentrated under reduced pressure (2.7 kPa) at a temperature in the region of 35 ° C. C. To the solid obtained is added 50 cm 3 of dichloromethane and 25 cm 3 of a solution of N hydrochloric acid. The organic phase is decanted, washed twice with 10 cm 3 of a solution of hydrochloric acid N and then 2 times with 10 cm of distilled water. The organic phase is dried over sodium sulphate and then concentrated to dryness under reduced pressure (2.7 kPa) at a temperature in the region of 35 ° C. 2.2 g are thus obtained. a solid which is chromatographed on a column of silica (0.02-0.045 mm) eluted with a mixture of chloroform, methanol and ammonia (24-6-1 by volume), the fractions containing the product are combined and concentrated under reduced pressure (2.7 kPa) at a temperature of about 35au. 840 mg of a solid is thus obtained, which is again chromatographed on a column of silica (0.02-0.045 mm) eluted, firstly with ethyl acetate and then with a mixture of chloroform, methanol and dichloromethane. ammonia (24-6-1 by volume), the fractions containing the product are combined and concentrated under reduced pressure (2.7 kPa) at a temperature in the region of 350C. 390 mg of 5- [3-fN- [N '- (3f3-acetoxylup-20 (29) -En-28-alkyl) -8-aminooctanoyl] amino] -phenyl] -tetrazole are thus obtained in the form of a white powder (Rf = 0.36, thin layer chromatography of silica eluent: chloroform-methanol-ammonia 20% (24-6-1 by volume)).

A solution of 380 mg of 5- [3-FN- [N '- (3 (3-acetoxylup-20 (29) -en-28-
3 μl) -8-aminooctanoyl] -amino} -phenyl] -tetrazole, 1.2 cm 4N sodium hydroxide, 3 cm 3 of tetrahydrofuran and 2 cm of methanol is stirred for 22 hours at a temperature in the region of 200 ° C. The mixture is evaporated under reduced pressure (2.7 kPa) at a temperature in the region of 350.degree. C., then diluted with 3.times.25 cm of distilled water and acidified with 1.4 cm of a 4N hydrochloric acid solution.After 1 hour of stirring, at a temperature of 200C, the solid is separated by filtration, washed 3 times with 5 cm of distilled water and dried under reduced pressure (15.5 Pa) at a temperature in the region of 3000. This gives 270 mg 5- [3- {N-tN '- (3ss-hydroxylup-20 (29) -En-28-oyl) -8-aminooctanoyl] amino} -phenyl-tetrazole, as a white solid around 17000.

5- (3-aminophenyl) tetrazole can be prepared from J.M. McMANUS and R. HERBST, J. Org. At., 24, 1044 (1959).

Example 7 670 mg of N- (313-acetoxylup-20 (29) -en-28-alkyl) -11-aminoundecanonitrile
3 g and 1 g of tributyltin azide are dissolved in 5 cm of 1,2-dimethoxyethane. The mixture is refluxed for 48 hours. 0.5 g of tributyltin azide are added and the mixture is again heated under reflux for 48 hours. The reaction mixture
3% is diluted with 50 cm of distilled water and extracted with
3100 cm 3 of ethyl acetate. The organic phase is then washed with
75 cm 3 of distilled water, dried over magnesium sulphate and evaporated to dryness under reduced pressure (2.7 kPa) at a temperature in the region of 400 ° C. The residue obtained (2.2 g) is chromatographed on a silica column (0.02-0.045 mm) eluted with a mixture of chloroform, methanol and 20% ammonia (12-3-0.5 by volume). The fractions containing the product are combined and concentrated under reduced pressure (2.7 kPa) at a temperature in the region of 400C. 700 mg of 5- [N- (3ss-acetoxylup-20 (29) -En-28-oyl) -10-aminodecyl] tetrazole are thus obtained in the form of a white meringue (Rf = 0.32 silica thin layer chromatography, eluent: chloroform-methanolammonia 20% (12-3-0.5 volumes)).

A solution of 290 mg of 5- [N- (3t3-acetoxylup-20 (29) -en-28-oyl) -10-
3 3 aminodecyl] -tetrazole, 0.82 cm 5N sodium hydroxide, 3 cm 3 of tetrahydrofuran and 6 cm 3 of methanol are stirred for 20 hours at a temperature in the region of 2000. The mixture is acidified with 35N hydrochloric acid, and diluted with 40 cm of distilled water. The mixture is extracted by
3. 3 50 cm, then 2 times 25 cm, of ethyl acetate.The organic phases
3 are combined, washed with 100 cm 3 of distilled water, dried over magnesium sulphate and evaporated to dryness under reduced pressure (2.7 kPa) at a temperature in the region of 4000. The residue obtained (280 mg) is chromatographed on a silica column (0.020.045 mm) eluted with a mixture of chloroform, methanol and 20% ammonia (12-3-0.5 by volume). The fractions containing the product are combined and concentrated under reduced pressure (2.7 kPa) at a temperature in the region of 4000. The residue is taken up in 30 cm 3 of isopropyl ether, filtered, washed with 10 cm 3 of isopropyl oxide and dried under reduced pressure (15.5 Pa) at 2000. 5- [N- (3ss-hydroxylup-20 (29) -he-28-alkyl) -10-aminodecyl] tetrazole, as a white solid melting at about 200 ° C.

Tributyltin azide may be prepared from H.R. KRICHELDORF and E LEPPERT, Synthesis, 329 (1976).

N- (313-acetoxylup-20 (29) -en-28-alkyl) -11-aminoundecanonitrile can be obtained as follows
To a solution of 2.5 g N- (3f3-acetoxylup-20 (29) -en-28-oyl) -11-ami-
3 noundecanamide in 300 cm dry tetrahydrofuran, add drop
3 drops 1.92 cm 3 of trifluoroacetic anhydride. The solution is acted
After 15 hours at room temperature, an additional 1.9 cm 3 of trifluoroacetic anhydride is added and the solution is stirred for 48 hours at room temperature. The solution is neutralized by addition of a saturated solution of sodium bicarbonate and then concentrated under reduced pressure (2.7 kPa). The residue is taken up in 3 100 cm of distilled water and the mixture is extracted twice with 150 cm. of ethyl acetate. The organic phases are collected, washed 3
3 times with 100 cm 3 of distilled water, dried over magnesium sulphate and evaporated to dryness under reduced pressure (2.7 kPa) at a temperature in the region of 4000. The residue obtained (2.5 g) is chromatographed on a silica column (0.02-0.045 mm) eluted with dichloromethane. The fractions containing the product are combined and concentrated under reduced pressure (2.7 kPa) at a temperature in the region of 4000. Thus 1.6 g of N- (3ss-acetoxylup-20 (29) -en-28-alkyl are obtained. ) Aminoundecanenitrile in the form of a white meringue (Rf = 0.6 silica thin layer chromatography, eluent: ethyl cyclohexaneacetate (60-40 by volume)).

N- (3B-acetoxylup-20 (29) -en-28-alkyl) -11-aminoundecanamide can be obtained as follows:
To a solution of 2.6 g of N- [3ss-acetoxylup-20 (29) -en-28-oyl] -
3 11-aminoundecanoic in 240 cm of dichloromethane is added
3.5 cc of 20% aqueous ammonia, 580 mg of 1-hydroxybenzotriazole hydrate, and 1.45 g of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride. The solution is stirred for 2 days at 200 ° C. then the organic phase is decanted, washed successively with 50 cm 3 of distilled water, 3 times 25 cm of chloric acid
3 ml of water, 2 times 40 cm 3 of distilled water, then dried over magnesium sulphate, filtered and evaporated to dryness under reduced pressure (2.7 kPa) at a temperature in the region of 400 ° C. 2.7 g of N- (313-acetoxylup-20 (29) -En-28-alkyl) -11-aminoundecanamide are thus obtained in the form of a cream-colored meringue (Rf = 0.15, thin layer chromatography). silica, eluent: cyclohexane-ethyl acetate (30-70 by volume)).

N- (3ss-Acetoxy-20 (29) -En-28-oyl) -11-aminoundecanoic acid is synthesized by analogy with N- (3ss-acetoxylup-20 (29) -en-28-alkyl ) -8-aminooctanoic acid from 3ss-acetoxylup-20 (29) -en-28-oyl chloride and methyl 11-aminoundecanoate (R = 38%, Rf = 0.57, silica thin layer chromatography; dichloromethane-methanol eluent (90-10 by volume)).

Example 8 1 g of N- (3 -3-acetoxylup-20 (29) -en-28-alkyl) -8-aminooctanenitrile and
1.6 g of tributyltin azide are dissolved in 25 cm of 1,2-dimethoxyethane. The mixture is refluxed for 24 hours. 35 cm 2 of 2-ethoxyethanol are added, 1,2-dimethoxyethane is distilled off and the mixture is heated again under reflux for 48 hours. The reaction mixture is cooled to room temperature, poured into a solution of 100 cm 1N hydrochloric acid. After stirring for 15 minutes, the mixture is extracted with 3 times 50 cm 3 of ethyl acetate. The organic phases are combined,
3 washed with 7 times 30 cm of distilled water, dried over magnesium sulphate and concentrated to dryness under reduced pressure (2.7 kPa) at a temperature in the region of 400C. The residue obtained (2.7 g) is chromatographed on a silica column (0.02-0.045 mm) eluted with a mixture of chloroform, methanol and 20% ammonia (12-3-0.5 by volume). The fractions containing the product are combined and concentrated under reduced pressure (2.7 kPa) at a temperature in the region of 400 ° C. The residue obtained is washed with 5 times 75 cm 3 of pentane, filtered and dried under reduced pressure (2.7 kPa). ) at 400C. 920 mg of 5- [N- (3ss-acetoxylup-20 (29) -en-28-alkyl) -7-aminoheptyl] tetrazole are thus obtained in the form of a cream-white powder (Rf = 0.28 chromatography). thin layer of silica, eluent: chloroformmethanol-ammonia 20% (12-3-0.5 volumes)).

A solution of 900 mg of 5- [N- (3ss-acetoxylup-20 (29) -en-28-alkyl) -7-aminoheptyl] -tetrazole, 2.7 cm 5N sodium hydroxide, 10 cm 3 of tetrahydrofu
3 rane and 20 cm of methanol is stirred for 15 hours at a temperature in the region of 2000. The mixture is acidified with 1N hydrochloric acid and diluted with 175 cm 3 of distilled water. The mixture is ex
3 times with 3 times 100 cm of ethyl acetate. Organic phases
3 are combined, washed with 250 cm of distilled water, dried over magnesium sulphate and concentrated to dryness under reduced pressure (2.7 kPa) at a temperature in the region of 4000. The residue obtained (865 mg) is chromatographed on a silica column (0.020.045 mm) eluted with a mixture of chloroform, methanol and 20% ammonia (12-3-0.5 by volume). The fractions containing the product are combined and concentrated under reduced pressure (2.7 kPa) at
3 a temperature close to 400C. The residue is taken up by 10 cm
3 isopropyl ether, filtered, washed with 6 cm of isopropyl ether and dried under reduced pressure (15.5 Pa) at 200 ° C., The powder obtained (510 mg) is again chromatographed on a silica column (0.020.045 mm) eluted with a mixture of dichloromethane and methanol (95-5 by volume). The fractions containing the product are combined and concentrated under reduced pressure (2.7 kPa) at a similar temperature.
3 'of 4000. The residue is taken up in 5 cm of isopropyl ether, filtered,
3 was washed with 2 cm 3 of isopropyl ether and dried under reduced pressure (15.5 Pa) at 2000. Thus 224 mg of 5- [N- (3e-hydroxylup-20 (29) -en-28- oyl) -7-aminoheptyl] -tetrazole, in the form of a white solid melting at about 150-1550C.

N- (3 P-acetoxylup-20 (29) -en-28-alkyl) -8-aminooctanenitrile can be obtained as follows
To a solution of 2.4 g of N- (3t3-acetoxylup-20 (29) -en-28-oyl) -8-
3 aminooctanamide in 200 cm dry tetrahydrofuran, add
Dropwise 1.97 cm 3 of trifluoroacetic anhydride. The solution
3 is stirred for 15 hours at room temperature then an additional 5.0 cm 3 of trifluoroacetic anhydride is added and the solution is stirred for 48 hours at room temperature. The solution is neutralized by adding a saturated solution of sodium bicarbonate
3 'sodium, then diluted with 100 cm of distilled water.Tetrahydrofuran is evaporated under reduced pressure (2.7 kPa) to 4000. The mixture is
3 then taken up with 200 cm 3 of ethyl acetate. The aqueous phase is
3 extracted with 50 cm of ethyl acetate. The organic phases are
3 pooled, washed twice with 100 cm 3 of distilled water, dried over magnesium sulfate, filtered and evaporated to dryness under reduced pressure (2.7 kPa) at a temperature in the region of 400C. 2.2 g of N- (3ss-acetoxylup-20 (29) -En-28-oyl) -8-aminooctanenitrile are thus obtained in the form of a cream-white meringue (Rf = 0.23, thin layer chromatography). silica, eluent: cyclohexane-ethyl acetate (80-20 by volume)).

N- (3-acetoxylup-20 (29) -en-28-alkyl) -8-aminooctanamide can be obtained as follows
To a solution of 2.4 g of N- (3ss-acetoxylup-20 (29) -en-28-oyl) -8-
In a 240 cm 3 dichloromethane solution, 3.8 cm 3 of 20% ammonia, 580 mg of 1-hydroxybenzotriazole hydrate and 1.45 g of 1- (3-dimethylaminopropyl) hydrochloride were added. The solution is stirred for 24 hours at room temperature, then the organic phase is decanted, washed with 5 times 250 cm 3 of distilled water, dried over magnesium sulphate, filtered and evaporated to dryness under reduced pressure (2 ° C.). 7 kPa) at a temperature in the region of 400C. There is thus obtained 2.4 g of N- (3ss-acetoxylup-20 (29) -En-28-alkyl) -8-amino-octanamide in the form of a white meringue (Rf = 0.1; thin silica, eluent: cyclohexane-ethyl acetate (50-50 by volume)).

Example 9 1.3 g of N- (313-acetoxylup-20 (29) -en-28-alkyl) -12-aminododecanenitrile
3 and 1.27 g of tributyltin azide are dissolved in 50 cm 2 of 2-ethoxyethanol. The mixture is refluxed during
3 48 hours. The reaction mixture is poured into 300 cm ice-cold distilled water. After stirring for 5 minutes, the mixture is acidified with a 2N hydrochloric acid solution and then extracted with 3 times.
The organic phases are combined, washed until neutral with distilled water, dried over magnesium sulphate and evaporated to dryness under reduced pressure (2.7 kPa) at a temperature in the region of 400.degree. The residue obtained (1.4 g) is chromatographed on a silica column (0.02-0.045 mm) eluted with a mixture of chloroform, methanol and 20% ammonia (12-3-0.5 vol). The fractions containing the product are combined and concentrated under reduced pressure (2.7 kPa) at a temperature in the region of 400C. 1.2 g of 5- [N- (3ss-acetoxylup-20 (29) -en-28-yl) -11-aminoundecyl] -tetrazole are thus obtained in the form of a white meringue (Rf = 0.45 thin layer chromatography of silica, eluent chloroform-methanol-ammonia 20% (12-3-0.5 volumes)).

A solution of 1.2 g of 5- [N- (3ss-acetoxylup-20 (29) -en-28-oyl) -11-
3 3 aminoundecyll-tetrazole, 3.3 cm 5N sodium hydroxide, 12 cm tetrahydrofu
3 rane and 24 cm of methanol is stirred for 20 hours at a temperature of about 2000. The mixture is acidified with 5N hydrochloric acid, and diluted with 150 cm of distilled water. The mixture is ex
The organic phases are combined, washed until neutral with distilled water, dried over magnesium sulphate and evaporated to dryness under reduced pressure (2.7 kPa) at a temperature of 3 × 60 ml of dichloromethane. close to 400C. The residue obtained (1.1 g) is chromatographed on a silica column (0.02-0.045 mm) eluted with a mixture of dichloromethane, methanol and 20% ammonia (12-3-0.5 by volume). The fractions containing the product are combined and concentrated under reduced pressure (2.7 kPa) at a temperature of
The residue is taken up with 15 cm 3 of ethyl ether.
3 ml, filtered, washed with 12 cm 3 of ethyl ether and dried under reduced pressure (15.5 Pa) to 2000. Thus 340 mg of 5- [N- (3ss-hydroxylup-20 (29) - η-28-oyl) -11-aminoundecyl] -tetrazole, in the form of a white solid melting at about 200-2050 ° C.

N- (3ss-acetoxylup-20 (29) -en-28-alkyl) -12-aminododecanenitrile can be obtained as follows:
To a solution of 1.5 g of N- (3 (3-acetoxylup-20 (29) -en-28-alkyl) -12-ami-
3 nododecanamide in 150 cm dry tetrahydrofuran, add drop
1.9 cm 3 of trifluoroacetic anhydride. The solution is acted
3 60 hours at room temperature, then diluted with 30 cm of distilled water. Tetrahydrofuran is evaporated to dryness under pressure.
The mixture is taken up in 100 cm 3 of distilled water, neutralized by addition of a saturated solution of sodium bicarbonate 3 and then extracted with 3 times 50 cm 3 of dichloromethane. The
3 organic phases are combined, washed with 3 times 40 cm of distilled water, dried over magnesium sulfate and evaporated to dryness under reduced pressure (2.7 kPa) at a temperature in the region of 400 ° C., thus obtaining 1.36 g. N- (3ss-acetoxylup-20 (29) -En-28-alkyl) -12-aminododecanonitrile as a white meringue (Rf = 0.35 silica thin layer chromatography, eluent: cyclohexaneacetate; ethyl (80-20 by volume)).

N- (3-acetoxylup-20 (29) -en-28-alkyl) -1,2-aminododecanamide can be obtained as follows
To a solution of 1.5 g of N- (3ss-acetoxylup-20 (29) -en-28-oyl) -
3 12-aminododecanoic in 250 cm of dichloromethane is added
2.15 cc of 20% ammonia, 330 mg of 1-hydroxybenzotriazole hydrate, and 826 mg of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride. The solution is stirred for 15 hours at room temperature then the organic phase is decanted, washed successively.
3 × with 5 times 80 cm of distilled water, 3 times 80 cm of 1 N hydrochloric acid, 5 × 80 cm of distilled water, then dried over magnesium sulfate, filtered and concentrated to dryness under reduced pressure ( 2.7 kPa) at a temperature in the region of 400C. 1.5 g of N- (3 -3-acetoxylup-20 (29) -En-28-alkyl) -1,2-aminododecanamide are thus obtained in the form of a cream-colored meringue (Rf = 0.47, layer chromatography). thin silica, eluent: dichloromethane-methanol (90-10 by volume)).

N- (3ss-acetoxylup-20 (29) -En-28-alkyl) -12-aminododecanoic acid is synthesized by analogy with N- (3ss-acetoxylup-20 (29) -en-28-alkyl ) -8-aminooctanoic acid from 3ss-acetoxylup-20 (29) -en-28-oyl chloride and methyl 12-aminododecanoate (R = 75%, Rf = 0.45, silica thin layer chromatography; eluent: dichloromethane-methanol (90-10 by volume)).

EXAMPLE 10 490 mg of N '- [N- (3 (3-acetoxylup-20 (29) -En-28-alkyl) -8-aminooctanoyl] -3-aminopropanenitrile and 700 mg of tributyltin azide are used.
3 solution in 10 cm of 1,2-dimethoxyethane. The mixture is refluxed for 6 days. The reaction mixture is cooled to room temperature, diluted with 100 cm 3 of dichloromethane and acidified.
3 with 25 cm 2N hydrochloric acid After 5 minutes of stirring, the
The organic phase is decanted, washed with 5 times 40 cm of distilled water, dried over magnesium sulfate, filtered and concentrated to dryness under reduced pressure (2.7 kPa) at a temperature of
The residue obtained is washed with 5 times 10 cm 3 of pentane, chromatographed on a silica column (0.02-0.045 mm) eluted with a mixture of ethyl acetate and methanol (80-20%). volumes). The fractions containing the product are combined and concentrated under reduced pressure (2.7 kPa) at a temperature in the region of 400C. 435 mg of 5-CN'- [N- (3 P -acetoxy-20 (29) -En-28-alkyl) -8-aminooctanoyl] aminoethyl-tetrazole are thus obtained in the form of a white meringue.

(Rf = 0.39, silica thin layer chromatography, eluent chloroform-methanol-ammonia 20% (12-3-0.5 volumes)).

A 600 mg solution of 5- {N '- [N- (3ss-acetoxylup-20 (29) -en-28-oyl) -
3 3 8-aminooctanoyl] aminoethyl) -tetrazole, 1.6 cm 5N sodium hydroxide, 10 cm
3 tetrahydrofuran and 20 cm of methanol is stirred for 15 hours at a temperature in the region of 2000. The mixture is acidified with acid
1N hydrochloric acid, and diluted with 120 cm of distilled water. The mixture is stirred for 1 hour at 200 ° C., filtered and washed to neutrality with
The resulting powder (553 mg) is chromatographed on a silica column (0.02 g), distilled water and 5 times 5 cm of pentane, and dried under reduced pressure (15.5 Pa). 0.045mm) eluted with a mixture of chloroform, methanol and 20% ammonia (12-3-0.5 by volume). The fractions containing the product are combined and concentrated under reduced pressure (2.7 kPa) at a similar temperature. of
3 400C. The residue is taken up in 15 cm 3 of isopropyl ether, filtered and dried under reduced pressure (15.5 Pa) at 2000. This gives 430 mg of 5-fN '- [N- (3-hydroxylup-20 (29) -En-28-oyl) -8-aminooctamoyl] aminoethyl) -tetrazole as a white solid melting at about 155-1600C.

N '- [N- (3ss-acetoxylup-20 (29) -En-28-oyl) -8-aminooctanoyl] -3-aminopropanenitrile can be obtained as follows:
To a solution of 960 mg N '- [N- (3ss-acetoxylup-20 (29) -en-28-oyl) -8-
3 aminooctanoyl] -3-aminopropanamide in 70 cm 3 of tetrahydrofuran
After 3 sec, 0.94 cm 3 of trifluoroacetic anhydride is added dropwise. The solution is stirred for 15 hours at room temperature, and then the tetrahydrofuran is evaporated under reduced pressure (2.7 kPa) at
3 400C. The mixture is then taken up in 70 cm 3 of dichloromethane. The organic phase phase is decanted, washed with 3 times 50 cm of distilled water, dried over magnesium sulfate and evaporated to dryness under reduced pressure (2.7 kPa) at a temperature in the region of 4000. The residue obtained ( 887 mg) is chromatographed on a silica column (0.020.045 mm) eluted with a mixture of ethyl acetate and cyclohexane (70-30 by volume). The fractions containing the product are combined and concentrated under reduced pressure (2.7 kPa) at a temperature in the region of 4000. 692 mg of N '- [N- (3ss-acetoxylup-20 (29) -en-28 are thus obtained. -oyl) -8-aminooctanoyl] -3-aminopropanenitrile as a white meringue (Rf = 0.27, silica thin layer chromatography, eluent: cyclohexane-ethyl acetate (30-70 by volume) ).

N '- [N- (3ss-acetoxylup-20 (29) -En-28-oyl) -8-aminooctanoyl] -3-aminopropanamide can be obtained as follows:
To a solution of 967 mg of N '- [N- (3ss-acetoxylup-20 (29) -en-28-
3 oyl) -8-aminooctanoyl] -3-aminopropanoic in 150 cm of dichloro
3 ml of 20% ammonia, 208 mg of 1-hydroxybenzotriazole hydrate, and 521 mg of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride are added. The solution is stirred for 50 hours at room temperature and then organic phase is
3 decanted, washed with 60 cm of 1N hydrochloric acid, 5 times 50 cm of distilled water, dried over magnesium sulphate, filtered and evaporated to dryness under reduced pressure (2.7 kPa) at a temperature in the region of 4000 960 mg of N '- [N- (3ss-acetoxylup- (29) -En-28-oyl) -8-aminooctanoyl] -3-aminopropanamide are thus obtained in the form of a cream-white meringue (Rf = 0.68, thin layer chromatography of silica, eluent: 20% chloroform-methanol-ammonia (12-3-0.5 by volume)).

N '- [N- (3ss-Acetoxy-20 (29) -En-28-oyl) -8-aminooctanoyl] -3-aminopropanoic acid can be obtained as follows
3
To an anhydrous solution of 2.3 g of lithium iodide and 100 cm 3 of collidine was added 1.3 g of N - [(3ss-acetoxylup-20 (29) -en-28-alkyl) -8-aminooctanoyl. The solution is refluxed for 3 hours and then concentrated under reduced pressure (2.7 kPa) at a temperature in the region of 400C. The residue is taken up by
3 '3 150 cm of distilled water and 150 cm of ethyl acetate, then cooled
3 to OOC. The organic phase is extracted, washed with 3 times 50 cm of 1N hydrochloric acid, then with distilled water until neutral, dried over magnesium sulphate and evaporated to dryness under reduced pressure (2.7 kPa) at room temperature. a temperature close to 400C. The residue
3 is taken up in 20 cm 3 of ethyl acetate and the mixture is concentrated under reduced pressure (2.7 kPa) at a temperature in the region of 4000. Thus, 1.07 g of N '- [N- ( 313-acetoxylup-20 (29) -en-28-oyl) -8-aminooctanoyl] -3-aminopropanol in the form of a beige meringue (Rf = 0.22, silica thin-layer chromatography eluent: chloroform-methanol 20% ammonia (12-3-0.5)).

Methyl N '- [N- (3ss-acetoxylup-20 (29) -en-28-oyl) -8-aminooctanoyl] -3-aminopropionate can be prepared in the following manner:
To a solution of 2 g of N- (3ss-acetoxylup-20 (29) -en-28-oyl) -8-
3 Aminooctanoic in 260 cm 3 of dichloromethane, add 524 mg of methyl 3-amino-propionate hydrochloride, 480 mg of 1-hydroxybenzotriazole hydrate, 1.2 g of 13-dimethylaminopropyl hydrochloride-3 ethylcarbodiimide, and 1.1 g of triethylamine in solution
The solution is stirred for 50 hours at room temperature, then the organic phase is decanted, washed with
3 times 150 cm 3 of distilled water, dried over magnesium sulphate, filtered and evaporated to dryness under reduced pressure (2.7 kPa) at a temperature in the region of 400 ° C. The residue obtained (2.4 g) is chromatographed on a silica column (0.02-0.045 mm) eluted with a mixture of ethyl acetate and cyclohexane (50/50 by volume). The fractions containing the product are combined and concentrated under reduced pressure (2.7 kPa) at a temperature in the region of 4000. This gives 2 g of N '- [N- (3ss-acetoxylup-20 (29) -en-28-oyl) -8 Methylaminooyl] -3-amino propionate as a white meringue (Rf = 0.62, silica thin layer chromatography, eluent: ethyl acetate) Example 11 720 mg of [N- (3ß- acetoxylup-20 (29) -En-28-alkyl-8-aminooctanoyl-methylamino] -acetonitrile and 1.1 g of tributyltin azide are dissolved in 15 cm of 1,2-dimethoxyethane. The reaction mixture is diluted with 40 3 3 cm of distilled water and 15 cm of 2N hydrochloric acid, and then extracted.
3 with 3 times 80 cm of dichloromethane. The organic phase is dried over magnesium sulfate, filtered and evaporated to dryness under reduced pressure (2.7 kPa) at a temperature in the region of 4000. The residue obtained (1.8 g) is chromatographed on a silica column (0.degree. , 02-0.045 mm) eluted with a mixture of chloroform, methanol and 20% ammonia (12-3-0.5 volumes). The fractions containing the product are combined and concentrated under reduced pressure (2.7 kPa) at a
The residue is washed 5 times with 20 cm 3 of pentane and then dried under reduced pressure (15.5 Pa) at 2000. 820 mg of 5- [N- (3-acetoxylup-20 29) -En-28-oyl) -8-aminooctanoyl-methyl-aminomethyl] -tetrazole as a white meringue (Rf = 0.33, thin-layer silica chromatography, eluent: chloroform-methanol-aqueous ammonia; 20% (12-3-0.5 in volumes)).

To a solution of 800 mg of 5- [N- (3ss-acetoxylup-20 (29) -en-28-oyl) -8-
3 aminooctanoyl-methyl-aminomethyl] -tetrazole, 2.7 cm 5N sodium hydroxide, 10
3 cm 3 of tetrahydrofuran and 20 cm 3 of methanol are stirred for 48 hours at a temperature in the region of 20 ° C. The mixture is acidified with 1N hydrochloric acid and diluted with 100 cm 3 of distilled water. The mixture is stirred for 1 hour at 200 ° C., filtered, washed to neutrality with 3 of the distilled water and then with 5 times 5 cm 3 of pentane, and dried under reduced pressure (15.5 Pa) at 2000. The product obtained (740 mg) is chromatographed on a column of silica (0.02-0.045 mm) eluted with a mixture of chloroform, methanol and 20% ammonia (12-3-0.5 by volume). The fractions containing the product are pooled and concentrated under reduced pressure (2.7 kPa) at a temperature in the region of 400C. The residue is taken up in 15 cm 3 of isopropyl ether, filtered, and dried under reduced pressure (15.5 Pa) in 2000. 360 mg of 5- [N- (3ss-hydroxylup-20 (29) is thus obtained. 1-en-28-yl) -8-aminooctanoyl-methyl-aminomethyl] -tetrazole, as a white solid melting at about 15100.

[N- (3-acetoxylup-20 (29) -En-28-oyl) -8-aminooctanoyl-methyl-amino] -acetonitrile can be obtained as follows:
To a solution of 1.2 g of [N- (3ss-acetoxylup-20 (29) -en-28-oyl) -8-
3 aminooctanoyl-methyl-amino] -acetamide in 70 cm 3 of tetrahydrofuran
3 sec, 1 cm 3 of trifluoroacetic anhydride is added dropwise.

The solution is stirred for 15 hours at room temperature and then neutralized with a saturated solution of sodium bicarbonate. Tetrahydrofuran is evaporated under reduced pressure (2.7 kPa) at 400C.

3
The mixture is then taken up in 200 cm of distilled water and extracted
3 by 3 times 80 cm of dichloromethane. The organic phases are re
3, washed with 3 times 50 cm of distilled water, dried over magnesium sulphate and evaporated to dryness under reduced pressure (2.7 kPa) at a temperature in the region of 400C. The residue obtained (1.0 g) is chromatographed on a silica column (0.02-0.045 mm) eluted with a mixture of ethyl acetate and cyclohexane (30-70 by volume). The fractions containing the product are combined and concentrated under reduced pressure (2.7 kPa) at a temperature in the region of 400C. 720 mg of [N- (3 P-acetoxylup-20 (29) -En-28-alkyl) -8-aminooctanoyl-methyl-amino] -acetonitrile are thus obtained in the form of a white meringue (Rf = 0.32 silica thin layer chromatography, cyclohexane-ethyl acetate eluent (50-50 by volume)).

[N- (3 -3-acetoxylup-20 (29β-en-28-oyl) -8-aminooctanoyl-methyl-amino] -acetamide can be obtained as follows:
To a solution of 700 mg of [N- (313-acetoxylup-20 (29) -en-28-oyl) -
8-aminooctanoyl-methyl-amino] -acetic acid in 100 cm of
3 mmol, 0.72 cm 3 of 20% ammonia, 150 mg of 1-hydroxybenzotriazole hydrate, and 380 mg of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride are added. The solution is stirred for 15 hours at room temperature. ambient then the organic phase is
3 decanted, washed with 30 cm hydrochloric acid 1N and then with 5 times
40 cm 3 of distilled water, dried over magnesium sulphate, filtered and concentrated to dryness under reduced pressure (2.7 kPa) at a temperature in the region of 4000. The residue obtained (630 mg) is chromatographed on a silica column. (0.02-0.045 mm) eluted with a mixture of dichloromethane and methanol (97-3 by volume). The fractions containing the product are combined and concentrated under reduced pressure (2.7 kPa) at a temperature in the region of 4000. 500 mg of [N- (313-acetoxylup-20 (29) -En-28-alkyl) -8-aminooctanoyl-methyl-amino] -acetamide are thus obtained in the form of a cream-white meringue (Rf = 0, Thin layer chromatography of silica, eluent: dichloromethane-methanol (95-5 by volume).

[N- (3-Acetoxy-20 (29) -En-28-oyl) -8-aminooctanoyl-methylamino] -acetic acid can be obtained as follows:
A solution of 167 mg of sarcosine, 406 mg of chlorotrimethylsilane 3 and 75 cm 3 of dichloromethane is refluxed for 15 hours. This solution is then added to a solution containing 1 g of N- (313-acetoxylup-20 (29) -en-28-alkyl) -8-aminooctanoic acid, 239 mg of 1-hydroxybenzotriazole hydrate, 600 g. mg of chlorine
3 - (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrate, and 75 cm -1
3 dichloromethane. 1.5 cm 3 of triethylamine are then added
3% solution in 10 cm 3 of dichloromethane.The solution is stirred for 50 hours at room temperature and the reaction mixture is washed.
3 with 70 cm 2N hydrochloric acid, then with 3 times 80 cm 3 of distilled water, dried over magnesium sulfate, filtered and concentrated to dryness under reduced pressure (2.7 kPa) at a temperature in the region of 4000. The residue obtained (2.4 g) is chromatographed on a silica column (0.02-0.045 mm) eluted with a mixture of dichloromethane, methanol and 20% ammonia (12-3-0.5 by volume). Fractions containing the product are combined and concentrated under reduced pressure (2.7 kPa) at a temperature in the region of 400C. 712 mg of [N- (313-acetoxylup-20 (29) -En-28-alkyl) -8-aminooctanoylmethylamino] -acetic acid are thus obtained in the form of a white meringue (Rf = 0.32 thin layer chromatography of silica, eluent chloroform-methanol-ammonia 20% (12-3-0.5 volumes)).

Example 12
To a solution of 1.4 g N- (3ss-acetoxylup-20 (29) -en-28-oyl) -8-
3 aminooctylcarbamoylacetonitrile in 20 cm 3 of dichloromethane,
3 adds 0.8 cm of trimethylsilyl azide and 270 mg of aluminum trichloride. The reaction mixture is stirred for 72 hours at 200 ° C.
3 and then poured dropwise over 30 minutes over 50 cm of a 10% aqueous solution of sodium hydrogencarbonate. The organic phase is
3, the aqueous phase is extracted with twice 50 cm of dichloromethane. The combined organic phases are washed 3 times with 50 cm 3 of distilled water, dried over magnesium sulphate and then evaporated to dryness under reduced pressure (2.7 kPa). ) at a temperature close to 300C. The residue obtained (1.9 g) is chromatographed on a silica column (0.02-0.045 mm), eluted with a mixture of chloroform, methanol and 20% ammonia (24-6-1 by volume). The fractions containing the product are combined and concentrated under reduced pressure (2.7 kPa) at a temperature in the region of 400C. There is thus obtained 1 g of 5- [N- (3ss-acetoxylup-20 (29) -En-28-alkyl) -8-aminooctylcarbamoylmethyl] -tetrazole in the form of a white meringue (Rf = 0.25). thin layer chromatography of silica, eluent: chloroform-methanol-ammonia 20% (24-6-1 by volume)).

A solution of 1 g of 5- [N- (3-t-acetoxylup-20 (29) -en-28-alkyl)
3 3 Noctylcarbamoylmethyl] -tetrazole, 5.4 cm sodium hydroxide N, 20 cm
3 tetrahydrofuran and 40 cm of methanol is stirred for 72 hours at a
3 temperature close to 2000. The mixture is poured into 10 cm of a
3 N hydrochloric acid solution and then diluted with 250 cm of distilled water. After stirring for 1 hour at a temperature close to
3 200C, the solid is filtered and washed 4 times with 10 cm of distilled water
3 then dissolved in 30 cm of ethanol. The solution is filtered, the filtrate is evaporated under reduced pressure (2.7 kPa) at a temperature
The residue obtained (140 mg) is taken up in 15 cm 3 of isopropyl ether. The solid obtained is filtered, washed with 2 times 35 cm 3 of isopropyl ether and then dried under vacuum (13.degree. 5 Pa) at a temperature in the region of 4000. Thus 700 mg of 5- [N- (3H-hydroxylup-20 (29) -En-28-alkyl) -8-aminooctylcarbamoylmethyl] -tetrazole is obtained under the form of a white solid melting around 17500.

N- (3ss-acetoxylup-20 (29) -En-28-oyl) -8-aminooctylcarbamoylacetonitrile can be obtained as follows:
To a solution of 3.6 g of N- [313-acetoxylup-20 (29) -En-28-oyl] -1,8-diaminooctane, 500 mg of cyanoacetic acid, 1.06 g of hydrate of 1 2-hydroxybenzotriazole, 2.76 g of 1- (3-dimethylamino-3-propyl) -3-ethylcarbodiimide hydrochloride in 75 cm 3 of dichloromethane is
3 ml of triethylamine was added in about 10 minutes. The solution is stirred for 15 hours at a temperature in the region of 200.degree.
The reaction mixture is washed 3 times with 40 cm 3 of distilled water.

The organic phase is then dried over magnesium sulphate and evaporated to dryness under reduced pressure (2.7 kPa) at a temperature in the region of 4000. The residue obtained (4.8 g) is chromatographed on a silica column (0.02). -0.045 mm), eluted with a mixture of dichloromethane and methanol (99-1 by volume). The fractions containing the product are combined and concentrated under reduced pressure (2.7 kPa) at a temperature in the region of 400C. 3.7 g of N- (3f3-acetoxylup-20 (29) -En-28-alkyl) -8-aminooctylcarbamoylacetonitrile are thus obtained in the form of a yellow meringue (Rf = 0.40; silica, eluent: 20% chloroform-methanol-ammonia (12-3-0.5 by volume)).

N- (313-acetoxylup-20 (29) -en-28-alkyl) -1,8-diaminooctane can be obtained as follows:
50 cm 3 of a solution in dichloromethane of 3β-acetoxyl-20 (29) -en-28-oyl chloride (prepared from 5 g of 3β-acetoxyl-20-acid (29)). een-28-oic) are added dropwise in about 3 hours to a solution of 8.6 g of 1,8-diaminooctane in 20 cm 3 of dichloromethane. The reaction mixture is stirred for 2 hours at a temperature in the region of 200 ° C. and then concentrated to dryness under reduced pressure (2.7 kPa) at a temperature in the region of 400 ° C. The residue obtained is chromatographed on a silica column (0.02-0.045 mm), eluted with a mixture of chloroform, methanol and 20% ammonia (24-6-1 by volume). The fractions containing the product are combined and concentrated under reduced pressure (2.7 kPa) at a temperature in the region of 400C. There is thus obtained 5.9 g of N- [3ss-acetoxylup-20 (29) -En-28-alkyl] 1,8-diaminooctane in the form of a yellow meringue (Rf = 0.25 thin layer chromatography). silica: eluent: chloroformmethanol-ammonia 20% (24-6-1 by volume)).

Example 13
To a solution of 2.1 g N- (3ss-acetoxylup-20 (29) -en-28-oyl) -7-amine
3 Noheptycarbamoylacetonitrile in 25 cm of dichloromethane,
3 add 1.07 cm of trimethylsilyl azide and 410 mg of aluminum trichloride. The reaction mixture is stirred for 72 hours at 200 ° C.
3 and then poured dropwise over 30 minutes over 50 cm of a 10% aqueous solution of sodium hydrogencarbonate. The organic phase is
3, the aqueous phase is extracted with twice 60 cm 3 of dichloromethane. The combined organic phases are washed 3 times with 50 cm 3 of distilled water, dried over magnesium sulphate and then concentrated to dryness under reduced pressure (2.7 kPa). ) at a temperature in the region of 4000. The residue obtained (2.5 g) is chromatographed on a silica column (0.02-0.045 mm), eluted with a mixture of chloroform, methanol and 20% ammonia (24-6%). 1 in volume). The fractions containing the product are combined and concentrated under reduced pressure (2.7 kPa) at a temperature in the region of 400C. 1.4 g of 5- [N- (3ss-acetoxylup-20 (29) -En-28-alkyl) -7-aminoheptylcarbamoylmethyl] -tetrazole are thus obtained in the form of a white meringue (Rf 0, Thin layer chromatography of silica, eluent: chloroform-methanol-20% ammonia (24-6-1 by volume)).

A solution of 1 g of 5- [N- (3B-acetoxylup-20 (29) -en-28-oyl) -7-
3 3 aminoheptylcarbamoylmethyl] -tetrazole, 8 cm sodium hydroxide N, 25 cm
3 tetrahydrofuran and 50 cm of methanol is stirred for 48 hours at a temperature of 20 OC. The reaction mixture is partially evaporated under reduced pressure (2.7 kPa) at a temperature in the region of 400C until the appearance of a haze. The mixture is then acidified to a pH of between 1 and 2 by adding an aqueous solution of N hydrochloric acid. After stirring for 2 hours at a temperature in the region of 20 ° C., the solid is filtered and washed 5 times with 3 times. The solution is filtered and the filtrate is evaporated under reduced pressure (2.7 kPa) at a temperature in the region of 400 ° C. The residue obtained is taken up in the presence of 50 ml of distilled water and then dissolved in 50 cm 3 of ethanol.
3 cm 3 of isopropyl oxide. The solid obtained is filtered, washed by 2
3 times 5 cm 3 of isopropyl ether and then dried under reduced pressure (13.5 Pa) at a temperature in the region of 4000. The residue obtained is chromatographed on a silica column (0.02-0.045 mm), eluted with a mixture of of chloroform, methanol and ammonia at 20% (24-6-1 by volume). The fractions containing the product are combined and concentrated under reduced pressure (2.7 kPa) at a temperature in the region of 4000. The solid obtained is dissolved in 20 cm 3 of chloroform, the solution is filtered and the filtrate is evaporated under pressure. reduced (2.7 kPa) at a temperature close to 400C, the
The residue obtained is taken up in 20 cm 3 of isopropyl ether. The solid
3 is obtained, washed with 2 times 5 cm 3 of isopropyl ether and then dried under vacuum (13.5 Pa) at a temperature in the region of 4000. 900 mg of 5- [N- (3ss-hydroxylup-) (29) -En-28-alkyl) -7-aminoheptylcarbamoylmethyl] -tetrazole as a white solid melting at about 1600 ° C.

N- (3ss-acetoxylup-20 (29) -en-28-yl) -7-aminoheptylcarbamoyl-acetonitrile can be obtained as follows:
To a solution of 2.45 g of N- (3f3-acetoxylup-20 (29) -en-28-oyl) -1,7-diaminoheptane, 340 mg of cyanoacetic acid, 730 g of 1-hydroxybenzotriazole hydrate 1.9 g of 1- (3-dimethylamino) hydrochloride
3-propyl) -3-ethylcarbodiimide in 120 cm of dichloromethane, is
Added in about 10 minutes, 2.5 cm 3 of triethylamine. The solution is stirred for 15 hours at a temperature in the region of 200.degree.
The reaction mixture is washed 3 times with 50 cm 3 of distilled water.

The organic phase is then dried over magnesium sulphate and evaporated to dryness under reduced pressure (2.7 kPa) at a temperature in the region of 4000. The residue obtained (2.7 g) is chromatographed on a silica column (0.02). -0.045 mm), eluted with a mixture of dichloromethane and methanol (99-1 by volume). The fractions containing the product are combined and concentrated under reduced pressure (2.7 kPa) at a temperature in the region of 4000. This gives 3.7 g of N- (3ss-acetoxylup-20 (29) -en-28-ol ) -7-aminoheptylcarbamoylacetonitrile as a white meringue (Rf = 0.40, silica thin layer chromatography, eluent: chloroform-methanol-20% ammonia (12-3-0.5 by volume)).

N- [3ss-acetoxylup-20 (29) -En-28-oyl] -1,7-diaminOheptane can be obtained as follows:
1000 cm 3 of a solution in dichloromethane of 3β-acetoxylup-20 (29) -en-28-oyl chloride (prepared from 30 g of 3-acetoxylup-20 (29) -en-28-oic acid) ) are added dropwise over a period of about 10 hours to a solution of 46.8 g of 1,7-diaminoheptane in 3120 cm 3 of dichloromethane. The reaction mixture is stirred for 15 hours at a temperature in the region of 200 ° C. and then concentrated to dryness under reduced pressure (2.7 kPa) at a temperature in the region of 400 ° C. The re
3 sidu is taken up in 2000 cm of distilled water.The solid is filtered,
3 washed with 3 times 200 cm of distilled water, then dissolved in
3,500 cm of ethanol. The solution is filtered, the solid is washed twice
3 with 50 cm 3 of ethanol and the filtrates are evaporated to dryness under reduced pressure (2.7 kPa) at a temperature in the region of 5000. The residue obtained is chromatographed on a silica column (0.02-0.045 mm), eluted with a mixture of chloroform, methanol and ammonia at 20% (24-6-1 by volume). The fractions containing the product are combined and concentrated under reduced pressure (2.7 kPa) at a temperature in the region of 4000. thus 28 g of N- (3ss-acetoxylup-20 (29) -en-28-oyl) -1,7-diaminoheptane as a yellow meringue (Rf = 0.25, silica thin layer chromatography; eluent: chloroform-methanol-ammonia 20% (24-6-1 by volume)).

Example 14
To a solution of 2.3 g of N- (3 (3-acetoxylup-20 (29) -en-28-alkyl) -7-amine
3-Noheptylcarbamoylpropionitrile in 25 cm 3 of dichloromethane,
Add 1.30 cm 3 of trimethylsilyl azide and 450 mg of aluminum trichloride. The reaction mixture is stirred for 48 hours at 200C then 32 hours at a temperature close to reflux. We add to
1.30 cm 3 of trimethylsilyl azide and 450 mg of aluminum trichloride are added and the mixture is heated for 24 hours at a temperature close to reflux. The reaction mixture is cooled to 200 ° C. and then poured dropwise over 30 minutes over 40 cm of a 10% aqueous solution of sodium hydrogen carbonate. The organic phase is decanted, the phase
The aqueous solution is extracted 3 times with 50 cm 3 of dichloromethane. The combined organic phases are washed 3 times with 50 cm 3 of distilled water, dried over magnesium sulphate and then evaporated to dryness under reduced pressure (2.7 kPa) at a temperature in the region of 400 ° C. The residue obtained 3
(2.5 g) is treated with 25 cm 3 of isopropyl ether and washed 3 times
3 by 5 cm of isopropyl ether. The solid obtained (1.8 g) is put into
3 solution in 25 cm of dichloromethane. The solution is filtered on celite. The filtrate is concentrated to dryness under reduced pressure
(2.7 kPa) at a temperature in the region of 400C. The solid is treated with 3 cm 3 of isopropyl ether and then dried under reduced pressure
(2.7 kPa) at a temperature in the region of 400C. 1.5 g of 5- [N- (3ss-acetoxylup-20 (29) -En-28-alkyl-7-aminoheptylcarbamoylethyl] -tetrazole are thus obtained in the form of a white meringue (Rf = 0). Silica thin layer chromatography, eluent: chloroformmethanol-ammonia 20% (24-6-1 by volume)).

A solution of 1 g of 5- [N- (3 (3-acetoxylup-20 (29) -en-28-oyl) -7-
3 3 aminoheptylcarbamoylethyl] -tetrazole, 20 cm sodium hydroxide N, 25 cm
3 tetrahydrofuran and 50 cm 3 of methanol are stirred for 72 hours at a temperature close to 2000. The mixture is diluted with 250 cm 3 of distilled water and acidified to pH 1 by addition of an aqueous solution of N hydrochloric acid. 1 hour with stirring at a temperature of 20 ° C., the solid is filtered and washed 5 times with 10 cm of water.
The combined organic phases are evaporated under reduced pressure (2.7 kPa) at a temperature in the region of 4000. The residue obtained (700 mg) is chromatographed on a column of dichloromethane which is then distilled and then treated with 5 times 40 cm 3 of ethyl acetate. silica (0.02-0.045 mm), eluted with a mixture of chloroform, methanol and 20% ammonia (24-6-1 by volume). The fractions containing the product are combined and concentrated under reduced pressure (2.7 kPa) at a temperature close to 4000. The solid obtained (600 mg) is treated with 15 cm 3 of isopropyl ether, the solid is filtered off. washed twice with 5 cm 3 of isopropyl ether and then dried over potassium hydroxide. 140 mg of 5- [N- (3ss-hydroxylup-20 (29) -en-28-oyl) are thus obtained. aminoheptylcarbamoylmethyl] -tetrazole in the form of a white solid melting at 1100C,
N- (3ss-acetoxylup-20 (29) -en-28-alkyl) -7-aminoheptylcarbamoylproponitrile can be obtained as follows:
To a solution of 2.45 g of N- (3f3-acetoxylup-20 (29) -en-28-oyl) -1,7-diaminoheptane, 400 mg of 2-cyanopropionic acid, 730 g of hydrate of 1 2-hydroxybenzotriazole, 1.9 g of 1- (3-dimethylamino-3-propyl) -3-ethylcarbodiimide hydrochloride in 120 cm 3 of dichloromethane, is
The solution is stirred for 15 hours at a temperature in the region of 2000.
The reaction mixture is washed 3 times with 50 cm 3 of distilled water. The organic phase is then dried over magnesium sulphate and evaporated to dryness under reduced pressure (2.7 kPa) at a temperature in the region of 400C. The residue obtained (3.5 g) is chromatographed on a silica column (0.02-0.045 mm), eluted with a mixture of dichloromethane and methanol (99-1 by volume). The fractions containing the product are combined and concentrated. under reduced pressure (2.7 kPa) at a temperature in the region of 4000. 2.3 g of N- (3e acetoxylup-20 (29) -En-28-alkyl) -7-aminoheptylcarbamoylpropionitrile are thus obtained in the form of a white meringue (Rf = 0.40, thin layer chromatography of silica, eluent: chloroform-methanol-ammonia 20% (12-3-0.5 volumes)).

N- (3ss-acetoxylup-20 (29) -En-28-oyl) -1,7-diaminOheptane can be obtained as follows:
1000 cm 3 of a solution in dichloromethane of 3β-acetoxylup-20 (29) -en-28-oyl chloride (prepared from 30 g of 3β-acetoxylup-20 (29) -n-28- oique) are added dropwise in about 10 hours to a solution of 46.8 g of 1,7-diaminoheptane in
3,120 cm 3 of dichloromethane. The reaction mixture is stirred for 15 hours at a temperature in the region of 200 ° C. and then concentrated to dryness under reduced pressure (2.7 kPa) at a temperature in the region of 400 ° C. The re
3 'sidu is taken up in 2000 cm of distilled water.The solid is filtered,
3 washed with 3 times 200 cm of distilled water, then dissolved in
3,500 cm of ethanol. The solution is filtered, the solid is washed twice
3 with 50 cm 3 of ethanol and the filtrates are evaporated to dryness under reduced pressure (2.7 kPa) at a temperature in the region of 5000. The residue obtained is chromatographed on a silica column (0.02-0.045 mm), eluted with a mixture of chloroform, methanol and ammonia at 20% (24-6-1 by volume). The fractions containing the product are combined and concentrated under reduced pressure (2.7 kPa) at a temperature in the region of 4000. thus 28 g of N- (3ss-acetoxylup-20 (29) -en-28-oyl) -1,7-diaminoheptane as a yellow meringue (Rf = 0.25, silica thin layer chromatography; eluent: chloroform-methanol-ammonia 20% (24-6-1 by volume)).

The present invention also relates to pharmaceutical compositions containing at least one product of general formula (I) optionally in salt form, in the pure form or in the form of an association with one or more compatible and pharmaceutically acceptable diluents or adjuvants, or with another agent for the treatment of AIDS, an antiviral, immunomodulatory or antimicrobial agent.

The composition according to the invention is capable of keeping cells infected with an HIV virus alive and thus of reducing progression to AIDS or of decreasing its severity in already infected subjects by reducing the mortality of infected cells. The compositions can be used orally, parenterally or rectally.

The compositions may be used for curative or preventive use in subjects with immunodeficiency and / or infected with an HIV virus. Of course, the constitution of these compositions will be adapted to the particular case of the digestive tract of immunodepressed.

As solid compositions for oral administration may be used tablets, pills, capsules, powders or granules. In these compositions, the active product according to the invention is mixed with one or more inert diluents or adjuvants, such as sucrose, lactose or starch.

These compositions may comprise substances other than diluents, for example a lubricant such as magnesium stearate or a coating intended for controlled release.

As liquid compositions for oral administration, pharmaceutically acceptable solutions, suspensions, emulsions, syrups and elixirs containing inert diluents such as water or paraffin oil can be used. These compositions may also comprise substances other than diluents, for example wetting, sweetening or flavoring products.

The compositions for parenteral administration may be sterile solutions or emulsions. As the solvent or vehicle, propylene glycol, a polyethylene glycol, vegetable oils, in particular olive oil, injectable organic esters, for example ethyl oleate, may be used.

These compositions may also contain adjuvants, in particular wetting agents, isotonic agents, emulsifiers, dispersants and stabilizers.

Sterilization can be done in several ways, for example using a bacteriological filter, irradiation or heating.

They can also be prepared as sterile solid compositions which can be dissolved at the time of use in sterile water or any other sterile injectable medium.

The compositions by rectal administration are suppositories or rectal capsules, which contain in addition to the active ingredient, excipients such as cocoa butter, semisynthetic glycerides or polyethylene glycols.

In human therapeutics, the doctor will determine the dosage he considers the most appropriate based on a preventive or curative treatment, depending on the age, weight, degree of infection and other factors specific to the subject to treat. Generally, the doses are between 10 and 100 mg / kg orally for an adult.

The present invention also relates to the combinations consisting of one or more lupane derivatives of general formula (I), and / or where appropriate their salts, and of another active ingredient known for its anti-retrovirus activity, possibly in the presence pharmaceutically acceptable excipients.

The anti-retrovirus agents that can be combined are chosen from agents that are compatible and inert with respect to the lupane derivative of general formula (I). As a non-limiting example, these agents are chosen from inhibitors of the reverse transcriptase tzidovudine (AZT), didanosine (DDI), dideoxycytidine (DDC), TIBO, nevirapine, PMEA, D4T, pyridones, a-APA, HEPT ...], among protease inhibitors tomme for example RO 31-8959 or A 77003], or among inhibitors of tat protein [such as RO 24-7429].

The following example illustrates a composition according to the invention.

Example
Ampoules containing a solution of 10 mg of 5 tN '- [N- (3ss-hydroxylup-20 (29) -en-28-alkyl) -8-aminooctanoyl] -aminomethyl] -tetrazole are prepared by adding 2 % of glycine and 0.1% of cysteine at 3500 cm of a 0.2% solution of 5- [N '- [N- (3f3-hydroxylup-20 (29) -en-28-oyl) - 8-aminooctanoyl] -aminomethyl] -tetrazole in isotonic phosphate buffer. The solution is divided and distributed aseptically into ampoules. Each ampoule contains 10 mg of active product.

Claims (4)

1 - A lupane derivative of general formula:

 in which : R represents a radical of general formula:

R1 is a methyl, hydroxymethyl radical or a radical -CH2OR 'or -CH2SR' for which R 'is alkyl or hydroxyalkyl, it being understood that the alkyl radicals are straight or branched and contain 1 to 4 carbon atoms, as well as their salts and where appropriate their stereoisomeric forms and mixtures thereof. Y is a bond or represents a meta or para phenylene radical, RO and ROO, which are identical or different, are hydrogen atoms or methyl radicals (it being understood that not all the -CR R motifs are necessarily identical to each other) and, n is an integer from 6 to 12, m is an integer from 0 to 3 and p is an integer from 0 to 2 with the proviso that m + n + p is from 6 to 14, and X is a bond or represents a carbamoyl, N-methyl carbamoyl, aminocarbonyl or N-methylaminocarbonyl radical, 2 - Process for the preparation of a lupane derivative according to claim 1, characterized in that an azide is activated on a nitrile of general formula:

 wherein R1 RO, ROO, X, Y, n, m, and p are defined as in claim 1, then optionally eliminates protective groups and optionally converts the resulting product into a salt. 3 - Process for the preparation of a lupane derivative according to claim 1, for which X (in the radical R) represents an optionally N-methylated carbamoyl or aminocarbonyl radical and the other radicals are defined as in claim 1, characterized in that that an amine or an acid of general formulas is made to act:

 in which R ", RO", m and p are defined as in claim 1 and R "'is a hydrogen atom or a methyl radical, on a lupane derivative of the general formula:

 wherein R 1 is defined as in claim 1 and R "is a radical of the general formula: ## STR2 ##  or (CH 2) n-NH-R "'in which n is defined as in claim 1 and R"' is defined as above, then optionally eliminates protecting groups and optionally converts the resulting product to a salt. 4 - Pharmaceutical composition characterized in that it contains at least one product according to claim 1, in pure form or in combination with any compatible diluent or adjuvant and pharmaceutically acceptable and / or in combination with another antiviral active ingredient, immunomodulator or antimicrobial.