MC1146A1 - PEPTIDIC DERIVATIVES AND PROCESS FOR THEIR PREPARATION - Google Patents

Description

The invention relates to peptide derivatives, more specifically peptide derivatives of phosphonic and phosphinic acids, and a process for preparing them, as well as pharmaceutical preparations containing them.

5. The peptide derivatives of the invention are compounds of general formula

R-

I

-c:

(has)

H2N—CH—0

R

I

-NH—CH—C0-(b)

R1 0

-NH—GH—P—R

he

(=) i

H

10

15

1 1!

! >

12 3

in which R, R and R individually represent the characteristic group of a cc-amino acid of the usual type in proteins, to con-

3

edition that R does not represent a hydrogen atom 3

and R represents a methyl group when n = lj

4

R represents the hydroxyl or methyl group; n = 1, 2 or 3; the configuration at carbon atom (a) is D; that at carbon atom (b)

2

is l when R is not hydrogen; and the configuration at the carbon atom (c) is (R) when R"*" has a meaning other than a hydrogen atom,

20 and their therapeutically compatible salts,

i?

j: ; In this context, the term "characteristic group of a

; ; ' cc-amino acid of the usual type in „ .

• proteins" means that the R radical of a natural α-amino acid with the general formula

25

h2n-

-CH-

k

-COOH

is of the type found in proteins. Thus, for example, if the amino acid is glycine, the R radical will be hydrogen, and if the amino acid is alanine, the radical

2

!

!

R will represent the methyl group; the radical R will also represent the isopropyl group in valine, the isobutyl group in leucine, the 2-carboxyethyl group in glutamic acid, and the benzyl group in phenylalanine. R- can also represent a radical linked to the nitrogen atom of the amino group (with the loss of one of the hydrogen atoms attached to it), thus forming a nitrogen ring as in proline and pyroglutamic acid.

i i

When R"*" in the general formula I does not represent hydrogen, the configuration at the carbon atom (c) will be 1Q (R), that is, the one that would be obtained by replacing the carboxy group of an L α-amino acid existing in nature with a phosphoric nucleus.

Note that in the general formula I, when n =

2

2 or 3, R will have equal or different values.

15 'The preferred compounds of general formula I are those

' in which R^ represents the hydroxy group, as well as those

■I ?

; in which R = methyl, or those in which R =

^ 3

: methyl and R represents a methyl, isopropyl or isobutyl group.

20. Examples of compounds with the general formula I include:

j acid (1R)-1-(D-alanyl-L-alanylamino)-ethylphosphonic,

: !

; acid (1R)-1-(D-alanyl-L-alanyl-L-alanylamino)-ethyl-

; ; phosphonic,

1 i

25 acid (1R)-1- (D-valyl-L-alanyl-L-alanylamino) -ethyl-

; phosphonic,

(1R)-1-(D-leucyl-L-alanyl-L-alanylamino)-ethylphosphonic acid,

acid (1R)-1-(D-alanyl-L-alanyl-L-alanyl-L-alanylamino)-

30 ethylphosphonic acid.

According to the process of the invention, the above-mentioned peptide derivatives (namely, the compounds of general formula I and their therapeutically compatible salts) are prepared by a process characterized in that

3

. aj splits the protecting group(s) of a compound

general formula

! i B?° _ R20 R10 0

5 i

R —Eïï—CH—CO

(has)

(b)

II

-NH—CH—COH-NH—CH—P—R40 II

Ui n R

in which R, R and R have respectively i 12 3

: 5 the same meanings as R, R and R except that

^ 'any Amino group present can be in the form

I: protected and any other functional group that may

being present is, where applicable, in protected form, R^ represents the methyl group or R^/

41

10R represents a hydroxy group or an alkoxy group

- C

lower protective, R represents a hydrogen atom or a protecting group: the configuration at

the carbon atom (a) is D; that at the atom of

20

carbon (b) is L when R is not hydrogen; 15 and the configuration at the carbon atom (c) is (R)

when R"^ has a meaning other than a hydrogen atom,

or b) separates a diastereomer compound (R,S) of general formula I into its diastereomers and isolates the diastereomer 20 (R), and converts where appropriate a compound of general formula I obtained into a therapeutically compatible salt.

The amino group(s) possibly present in the R, R^® and R^ radicals of the general formula II can be protected by any amino protecting group well known in peptide chemistry. Amino protecting groups particularly suitable for the purposes of the invention include: i. iaralcoxycarbonyl, especially benzyloxycarbonyl, and t.-butoxy-

carbonyl. The amino protecting group can also be a formyl, trityl, or trifluoroacetyl group. Any carboxy or 30-hydroxy group that may be present in the R^, R^, and R^ groups of the general formula II can be protected respectively by a conventional carboxy or hydroxy protecting group. A group

4

carboxyl groups can, for example, be protected by conversion to an ester.

. alcoholic (for example, t-butyl ester) or an ester

; aralcoyl (for example, benzyl ester). Furthermore, a

. a hydroxyl group can for example be protected by means of a

5. Aralkoxy-carbonyl group (e.g., benzyloxy-carbonyl),

• alcanoyl (e.g., acetyl, propionyl, etc.), aroyl (e.g., benzoyl), alkoyl (e.g., t-butyl) or aralcoyl

[ (e.g., benzyl). The protection of other functional groups ' x o *"* n *3 0

The presence of R10, R20, and R30 in radicals can be addressed in a known manner. The protecting group represented by R^ in the general formula II can be any amino protecting group previously mentioned in relation to R10, R20, and R30.

j i i

The splitting of the existing protective group(s)

1

: in a compound of general formula II is carried out according to 15 known methods, that is, methods actually used, described or reported in the literature on the cleavage of protecting groups. Thus, for example, an ar-koxy-carbonyl group (e.g., benzyloxy-carbonyl) or t-butoxycarbonyl group can be cleaved by hydrolysis (i.e., by treatment with a mixture of hydrogen bromide and glacial acetic acid). An ar-koxy-carbonyl group (e.g., benzyloxy-carbonyl) can also be cleaved by hydrogenolysis (e.g., in the presence of palladium carbon or palladium oxide). The t-butoxy-carbonyl group can also be cleaved by means of hydrochloric acid in dioxane. A lower alkoxy group represented by R^ and/or R^ can be linear or branched, preferably containing 1-6 atoms

'j!

j | of carbon (for example methoxy, ethoxy, propoxy, isopropoxy,

■ i !

(Butoxy, etc.), and can be converted to a hydroxyl group by treatment with a mixture of hydrogen bromide in glacial acetic acid or by means of trimethylchlorosilane, followed by aqueous hydrolysis. It should be noted that the cleavage of protecting groups can be carried out in one or more phases depending on the nature of the protecting groups present.

The resolution of a diastereomer compound (R,S) of general formula I into its diastereomers and the isolation of the diastereomer (R) can be carried out according to known methods, for example by crystallization or by pressure liquid chromatography, etc.

5

"The compounds of general formula I are amphoteric and form therapeutically compatible salts with strong acids (e.g. methane- and p-toluenesulfonic acids, hydrochloric, hydrobromic, sulfuric acids etc.) and with 5, bases (e.g. sodium hydroxide).

i i

: j Starting substances of general formula II can

; for example, be prepared by condensation of a compound of ; general formula

1°

15

with an adequately protected a-ammo-aciae, aipeptiae, a rripepriae or tetrapeptide, or one of their reactive derivatives, as appropriate.

Thus, when a compound of general formula III is used in which m = 0, such a compound can be condensed with an adequately protected dipeptide, or one of its reactive derivatives, to give a compound of general formula II, in which n = 1, or with an adequately protected tripeptide, or one of its reactive derivatives, to give a compound of general formula II, in which n = 2, or with an adequately protected tetrapeptide, or one of its reactive derivatives, to give a compound of general formula II, in which n = 3.

30 Furthermore, a compound of general formula III, in which m = 1, can be condensed with a protected α-amino acid

R

20„10

I

-c: (b)

R 0

■ I' 40

H—HlH—CH—C 0-HNH—CH—P—R^u III

'41

m R

in which m = 0, 1, 2 or 3 and R"^, R2^, R^- and R^ have the same meaning as above/ the configuration

20

the configuration at the carbon atom (b) is L when R is not hydrogen and the configuration at the carbon atom (c) is (R) when R has a meaning other than a hydrogen atom,

20

25

6

appropriately, or one of its reactive derivatives, to give a compound of general formula II, in which n = 1, i or with an appropriately protected dipeptide, or one of its reactive derivatives, to give a compound of general formula II, in which n = 2, or with a tripeptide protected by i

■ ; ; appropriate manner, or one of its reactive derivatives, for

! give a compound of general formula II, in which n = 3,

On the other hand, a compound of general formula III, in which m = 2, can be condensed with an adequately protected α-amino acid, or one of its reactive derivatives,

; to give a compound of general formula II, in which n = 2, or with a suitably protected dipeptide, or one of its reactive derivatives, to give a compound of general formula II, in which n = 3.

15 Finally, a compound of general formula III, in which m = 3, can be condensed with an adequately protected α-amino acid, or one of its reactive derivatives, to give a compound of general formula II, in which n = 3.

Furthermore, compounds of general formula II can be prepared by carrying out the above condensation using a compound (R,S) of general formula III and separating the compound (R) from the resulting product (R,S) in a known manner, for example by crystallization, chromatography or fractional crystallization using a suitable base, by example 1'α-methylbenzylamine.

The condensation mentioned above can be carried out according to methods known in peptide chemistry, for example by the mixed anhydride, azide, activated ester or acid chloride method.

3.0 According to one method, a suitable compound of formula ge

General III can be condensed with a suitably protected α-amino acid (di-, tri-, or tetrapeptide, as the case may be), the terminal carboxy group being a mixed anhydride radical formed with an organic or inorganic acid. Advantageously, such a di-, tri-, or tetrapeptide amino acid

7

bearing a free carboxy group is treated with a tertiary base such as a lower trialcoylamine (e.g.

■ a triethylamine) or an N-ethylmorpholine in an inert organic solvent (e.g. tetrahydrofuran, 1,2-dimethoxy-

: 5 ethane, dichloromethane, toluene, petroleum ether or mixtures thereof) and the resulting salt is reacted with a chloroformic acid ester (e.g. butyl or isobutyl ester) at low temperature. The resulting mixed anhydride is then advantageously condensed in situ 10 with the compound of general formula III.

In another method, a suitable compound of general formula III can be condensed with an appropriately protected α-amino acid (di-, tri-, or tetrapeptide, as appropriate), the terminal carboxy group being in the form of a 15-acid azide. This condensation is preferably carried out in an inert organic solvent such as dimethylformamide or ethyl acetate at low temperature.

Another method involves condensing a suitable general formula III compound with an appropriately protected α-amino acid (di-, 20-tri- or tetrapeptide as appropriate), the terminal carboxy function being in the form of an active ester group (e.g. p-nitrophenyl, 2,4,5-trichlorophenyl or N-hydroxysuccinimidic ester group).

This condensation is advantageously carried out in an inert organic solvent such as dimethylformamide or,

■ - in the case where R^ and/or R^"*" represent a lower alkoxy group, in an aqueous alkanol (for example aqueous ethanol).

According to yet another method, a suitable compound of general formula III can be condensed with an appropriately protected α-amino-α30 acid (di-, tri-, or tetrapeptide, as the case may be), the terminal carboxy group being in the form of an acid chloride. This condensation is preferably carried out in the presence of a base and at low temperature.

35 The peptide derivatives of the present invention po-

They stimulate the activity of D-cycloserine. The derivatives

8

The peptides of the invention also possess antibacterial activity against organisms such as Escherichia coli, Klebsiella aerogenes, Streptococcus faecalis and Haemophilus influenzae.

5. The peptide derivatives of the invention can therefore be used as medicinal products, for example in the form of pharmaceutical preparations containing them, in combination with a compatible pharmaceutical vehicle which may be organic or inorganic, suitable for enteral or parenteral administration, for example water, lactose, starch, magnesium stearate, acacia gum, gelatin, polyalkylene glycols, petrolatum, etc. These preparations may be in solid form, for example tablets,

Dragees, suppositories, capsules; or in liquid form, for example, solutions, suspensions, or emulsions. Where appropriate, the preparations may be sterilized or contain adjuvants, for example, preservatives, stabilizers, wetting agents, or emulsifiers. They may also contain osmotic pressure-regulating salts or buffer compounds.

The peptide derivatives of the invention and D-cycloserine can be administered in combination or separately, if necessary by different routes. The amount of peptide, as well as the peptide derivative/D-cycloserine ratio, can vary within wide limits depending on factors such as the derivative chosen, the route of administration.

»

The peptide derivative and D-cycloserine can, for example, be administered in a ratio of approximately 100:1 to 1:100.

9

Example 1

(A) Procedure:

i 4.3 g (7.4 mmol) of benzylamine salt of acid

(1R)-1-[(N-benzyloxycarbonyl-D-alanyl-L-alanyl-L-alanyl)-amino]-5 ethylphosphonic acid is added to 12 mL of a stirred 45% hydrogen bromide solution in glacial acetic acid, then washed with 4 mL of glacial acetic acid. The mixture is stirred at room temperature for 6 hours, after which 100 mL of ether is added while stirring. The supernatant is decanted, and the residue is treated again with 100 mL of ether. The resulting gum is resuspended in 30 mL of methanol, and the solution is treated with a solution of 3 mL of propylene oxide in 5 mL of methanol, giving a white precipitate. After being left to stand overnight at room temperature, the precipitate is filtered, washed successively with methanol and ether, and then dried. Crystallization in a water/ethanol mixture gives 1.96 g of (1R)-1-(D-alanyl-L-alanyl-L-alanylamino)-ethylphosphonic acid melting at 318-320° (dec); = -10^° (c = in 20 sodium hydroxide IN).

(B) Preparation of the starting substance:

2.7 g (10 mmol) of (1R)-1-(L-alanyl-L-alanylamino)-ethylphosphonic acid are stirred in 50 mL of water at 5°C, while 2.0 g (20 mmol) of triethylamine and 50 mL of ethano-25 nol are added. The resulting clear solution is cooled to 0°C, and then 3.8 g (12 mmol) of solid N-benzyloxycarbonyl-D-alanine N-hydroxysuccinic acid ester is added in a single portion. This ester is washed with 25 mL of ethanol. The mixture is stirred for 2 hours at 0°C, then for 16 hours at room temperature (30°C), yielding a clear solution. The solvents are removed by evaporation, and the residue is divided between 150 mL of water and 100 mL of chloroform. The aqueous layer is washed with another 100 ml of chloroform, then the extracts are rinsed with 50 ml of water. The aqueous extracts are combined and evaporated. The residue is resuspended in a mixture of 40 ml of water and 40 ml of methanol, then passed through a resin column.

10

cation exchanger (sulfonated polystyrene, freshly regenerated in the acid cycle). Elution is carried out with the same solvent system. The resulting acidic eluate is evaporated, the residue is resuspended in 250 ml of water and extracted twice with 100 ml of ether each time. The ether extracts are rinsed with 50 ml of water. The combined aqueous extracts are concentrated to approximately 100 ml.

i.e., after which an equal quantity of methanol is added and the resulting solution is titrated to pH 4.5 using 1M aqueous benzylamine. The solution is evaporated to dryness and the resulting solid residue is recrystallized in a mixture of 100 ml of hot water.

400 ml of ethanol and 500 ml of ether. This yields 4.38 g of the benzylamine salt of (1R)-1-[(N-benzyloxycarbonyl-D-alanyl-L-alanyl-L-alanyl)-amino]-ethylphosphonic acid, melting at 240-243° (dec); = ""3®'-'-° (c = 0.53% in the acid

15 acetic acid).

Example 2

(A) Procedure:

In a manner analogous to that described in Example 1, and starting from the benzylamine salt of (1R)-1-[(N-benzyloxy-20 carbonyl-D-valyl-L-alanyl-L-alanyl)-amino]-ethylphosphonic acid, (1R)-1-(D-valyl-L-alanyl-L-alanylamino)-ethylphosphonic acid is obtained, which melts at 301-304° (dec.); [a]20 = -105°

(c = 0.45% in sodium hydroxide IN).

(B) Preparation of the starting substance

25 3.8 g (15 mmol) of N-benzyloxycarbonyl-D-valine are stirred in 200 ml of petroleum ether (boiling point 60-80°C). Then 1.5 g (15 mmol) of triethylamine are added and the mixture

The mixture is cooled to -5°C. 2.1 g (15 mmol) of isobutyl chloroformate is added and the mixture is maintained at -5°C for 30 minutes. While stirring at -5°C, a solution of 2.7 g (10 mmol) of (1R)-1-(L-alanyl-L-alanylamino)-ethylphosphonic acid is added dropwise to a mixture of 2.0 g (20 mmol) of triethylamine and 15 ml of water. This solution is washed with 5 ml of water. Stirring continues between -5°C and 0°C.

11

for another 2 hours, then overnight at room temperature. 150 mL of water is added, and the organic and aqueous phases are separated. The organic phase is evaporated, and the residue is reconstituted in a mixture of 40 mL of water and 40 mL of methanol. The residue is then passed through a column of cation-exchange resin freshly regenerated in the acid cycle. The column is eluted with the same solvent mixture, and the acid fraction is then evaporated and re-evaporated with 50 mL of water. The residue is triturated with 100 mL of ether, and the solid is separated by filtration. This solid is reconstituted in a mixture of 400 mL of methanol and 400 mL of water, after which the solution is titrated to pH 4.5 using 4 M aqueous benzylamine. The solution is evaporated to dryness and the resulting solid is recrystallized in a mixture of 100 ml of methanol and 150 ml of ether to obtain 3.24 g of the benzylamine salt of acid

(1R)-1-[(N-benzyloxycarbonyl-D-valyl-L-alanyl-L-alanyl)-amino]-ethylphosphonic melting at 238-244° (dec.).

Example 3

(A) Procedure:

20. In a manner similar to that described in example 1,

and from benzylamine salt of (1R)-1-[N-benzyloxy-carbonyl-D-alanyl-L-alanyl-L-alanyl-L-alanyl)-amino]-ethylphosphonic acid, we obtain (1R)-1-(D-alanyl-L-alanyl-L-alanyl-L-alanylamino)-ethylphosphonic acid melting at 323-325° 25 (dec);[a]20 = -121° (c = 0.48% in sodium hydroxide IN).

(B) Preparation of the starting substance

In a manner analogous to that described in example 1 (B) and from the N-hydroxysuccinic ester of N-benzyloxycarbonyl-L-alanine and (1R)-1-(L-alanyl-L-alanylamino)-ethyl-30 phosphonic acid, we obtain (1R)-1-[(N-benzyloxycarbonyl-L-alanyl-L-alanyl-L-alanyl) -amino] -ethylphosphonic acid melting at 255-257° (dec.) j [a]2^ = -62.0° (c = 0.4% in glacial acetic acid).

12

In a manner analogous to that described in example 1(A) and starting from (1R)-1-[(N-benzyloxycarbonyl-L-alanyl-L-alanyl-L-alanyl)-amino]-ethylphosphonic acid, we obtain (1R)-1-(L-alanyl-L-alanyl-L-alanylamino)-ethylphosphonic acid 5 melting at 312-313° (dec); = -101° (c = 0.53% in sodium hydroxide IN).

In a manner analogous to that described in Example 1(B), but with acidification to pH 2 and subsequent extraction with ether replacing passage through the resin column 10 followed by extraction with ether, and starting from the ester

N-hydroxysuccinic acid of N-benzyloxycarbonyl-D-alanine and (1R)-1-(L-alanyl-L-alanyl-L-alanylamino)-ethylphosphonic acid yields the benzylamine salt of (1R)-1-[(N-benzyloxy-carbonyl-D-alanyl-L-alanyl-L-alanyl-L-alanyl)-amino]-ethyl-15-phosphonic acid, which melts at 272-277°C (dec.); = (c =

0.5% in acetic acid).

Example 4

(A) Procedure:

10.85 g of dimethyl (1R)-1-[(N-benzyloxycarbonyl-D-leucyl-20 L-alanyl-L-alanyl)-amino]-ethylphosphonate are dissolved in 30 mL of a 35% hydrogen bromide solution in glacial acetic acid, and the mixture is stirred at room temperature for 4 hours. While stirring, 130 mL of ether is added, after which stirring is stopped and the ether is allowed to settle. This process is repeated twice with portions of 80 mL of ether. The residue is then dissolved in 70 mL of methanol and the resulting solution is supplemented with 10 mL of propylene oxide. After cooling overnight, the resulting white precipitate is separated by filtration and then washed with ethanol and ether. The product is dried under vacuum to a constant weight of 7.99 g. p.f. 293-295° (dec.). By recrystallization in 500 ml of cold water and 700 ml of ethanol, 6.67 g of (1R)-1-(D-leucyl-L-alanyl-L-alanylamino)-ethylphosphonic acid is obtained, melting at 300-302° (dec.); = -129.3°

3 5 (c = 1% in water).

13

(B) Preparation of the starting substance:

12.9 g (0.03 mol) of dimethyl (1R)-1-[(N-benzyloxycarbonyl-L-alanyl-L-alanyl)amino]-ethylphosphonate is dissolved in 150 mL of methanol containing 0.032 mol of hydrogen chloride. The solution is hydrogenated at room temperature and pressure in the presence of 1 g of 10% palladium charcoal until hydrogen absorption ceases. The catalyst is separated by filtration, the filtrate is evaporated under vacuum, and the oily hydrochloride is evaporated twice with ethyl acetate.

The product obtained in the preceding paragraph and 10.9 g (0.03 mol) of N-benzyloxycarbonyl-D-leucine N-hydroxysuccinimide ester are stirred in the presence of 100 mL of dry dimethylformamide. While stirring and maintaining a temperature below 15°C, 4.2 mL of dry triethylamine are added dropwise. The mixture is then stirred at room temperature until the following day. The triethylamine hydrochloride is separated by filtration and washed with a small amount of dimethylformamide. The filtrate is evaporated under 20°C vacuum in an oil aspirator and at a bath temperature below 40°C. The residual oil is treated with 50 mL of water, and then the residual mixture is extracted with 4 times 50 mL of chloroform. The combined organic phases are washed with a small volume of 20% potassium carbonate solution, then dried over sodium sulfate. The sodium sulfate is separated by filtration, and the filtrate is evaporated under vacuum using a water aspirator. The residue is evaporated twice again with ethyl acetate. By trituration with dry ether, filtration, and vacuum drying, 15.5 g of dimethyl (1R)-l-r[(N-benzyloxycarbonyl-D-leucyl-L-alanyl-L-alanyl)-amino]ethylphosphonate is obtained, melting at 163-167°C. After recrystallization in acetonitrile, the melting point is 173-176°C; α = -36.6°C (1% in methanol).

14

Example 5

; t (A) Procedure:

In a manner analogous to that described in Example 4(A), 9.92 g of dimethyl (1R)-1-[(N-benzyloxycarbonyl-D-alanyl-L-.5-alanyl)-amino]-ethylphosphonate is treated with 45 mL of a 35% hydrogen bromide in acetic acid solution. Working according to the data of Example 4(A), 6.16 g of crude product melting at 282–285°C (dec.) is obtained, which, after recrystallization in the water/ethanol mixture, gives 5.47 g of pure (1R)-1-(D-alanyl-L-alanylamino)-ethylphosphonic acid at a melting point of 292–293°C (dec.); α = -101.2°C (c = 1% in

1'water).

(B) Preparation of the starting substance:

In a manner analogous to that described in Example 4(B) 15 and starting from 7.82 g (0.03 mole) of (1R)-1- hydrochloride

(L-alanylamino)-ethylphosphonate of dimethyl and 9.6 g (0.03 mol) of the N-hydroxysuccinimidic ester of N-benzyloxycarbonyl-D-alanine, we obtain 11.1 g of (1R)-1-[(N-benzyloxycarbonyl-D-alanyl-L-alanyl)-amino]-ethylphosphonate of dimethyl melting 20 at 115-117°; = "28.4° (c = 0.5% in methanol).

15

The following example illustrates a typical pharmaceutical preparation containing a peptide derivative according to the present invention:

Example 6

5. Prepare 1000 ml of an injection solution containing the following ingredients:

per 1000 ml acid (1R)-1-(D-alanyl-L-alanylamino)-

ethylphosphonic acid 100.0 g chlorocresol 1.0 g

10 glacial acetic acid, 1.2 g sodium hydroxide solution 0.1N q.s. pH 4.5

water for injection ad 1000 ml

(1R)-1-(D-alanyl-L-alanylamino)-ethylphosphonic acid is dissolved in 500 mL of water for injection. Chlorocresol 15 is dissolved in 200 mL of water for injection and added to the first solution. Acetic acid is then added with stirring. A 0.1 N sodium hydroxide solution in water for injection is added with stirring until pH 4.5 is reached. The solution is then made up to 1000 mL with 20 mL of water for injection, filtered through a sterile 0.22-micron membrane, and filled into ampoules. The ampoules are sealed.

then sterilized at 121° for 20 minutes in an autoclave.

16

Claims (1)

DEMANDS 1. Process for the preparation of peptides of general formula R' H2N—CH—C Oj-NH—CH—CO (has) R I (b) R1 0 I "H < -m—CH—P—R (c) n OH 10 15 12 3 in which R, R and R individually represent the characteristic group of an α-amino- acid of the usual type in proteins, at con-3 edition that R does not represent a hydrogen atom 3 and R represents a methyl group when n = 1; 4 R represents the hydroxyl or methyl group; n = 1, 2 or 3; the configuration at carbon atom (a) is D; that at carbon atom (b) 2 is L when R is not hydrogen; and the configuration at the carbon atom (c) is (R) when R"*" has a meaning other than a hydrogen atom, and their therapeutically compatible salts, characterized in that one 20 a) splits the protecting group(s) of a compound with general formula >30 R' 20 R-! 5 i R —m—CH—COH-HH—CH—co- (has) r i t TVTTT Q (b) R10 0 I H -3m—CH—P—R {41 40 II n R in which R, R and R have respectively 12 3 the same meanings as R, R and R except that any amino group present can be in protected form and any other functional group can 17 being present is, where applicable, in protected form; represents the methyl group or R^/ 41 R represents a protecting hydroxyl group or inferior alkoxy group, R~* represents a hydro- atom 5 gene or a protective group; the configuration at the carbon atom (a) is D; that at the atom of 20 . carbon (b) is L when R is not hydrogen; and the configuration at the carbon atom (c) is (R) when R^® has a meaning other than a 1Q hydrogen atom, n = 1, 2 or 3, using known methods, or b) separates a diastereomeric compound (R,S) of general formula I into its diastereomers and isolates the diastereomer (R), and converts where appropriate a compound of general formula I obtained into a therapeutically compatible salt. 2". A process according to claim 1, characterized in that a peptide is prepared, in which R^ represents a hydroxy group. 3. Method according to claim 1 or 2, characterized in pre; methyl group. 20 in that we prepare a peptide, in which R"'" represents a 4. A method according to any one of claims 1, 2 or 3, ^ - 2 characterized in that a peptide is prepared, in which R represents 3 represents a methyl group and R represents a methyl, iso-25 propyl or isobutyl group. 5. Process according to claim 1, characterized in that (1R)-1-(D-alanyl-L-alanylamino)-ethylphosphonic acid is prepared. 6. Process according to claim 1, characterized in that 3Q. is prepared the (1R)-1-(D-alanyl-L-alanyl-L-alanylamino)-ethylphosphonic acid. 7- Process according to claim 1, characterized in that (1R)-1-(D-valyl-L-alanyl-L-alanylamino)-ethylphosphonic acid is prepared. M 18 8- Process according to claim 1, characterized in that the (1R)-1-(D-leucyl-L-alanyl-L-alanylamino)-ethylphosphonic acid is prepared. 9*- Process according to claim 1, characterized in that the (1R)-1-(D-alanyl-L-alanyl-L-alanyl-L-alanylamino)-ethylphosphonic acid is prepared. 10. A process according to claim 1, characterized in that a compound of general formula II is obtained by condensation of a compound of general formula R' 20 H-+NH—CH-(b) R10 0 C0—MH—CH—P—R 40 III (c) m R' 41 in which m = 0, 1, 2 or 3; R^, R2^, R^- and R41 have the same meaning as in claim 1 } the configuration 20 The configuration at the carbon atom (b) is L when R is not hydrogen, and the configuration at the carbon atom (c) is (R) when R has a meaning other than a hydrogen atom. with an adequately protected α-amino acid, dipeptide, tripeptide or tetrapeptide, or one of their reactive derivatives, as appropriate. 11. A process for the manufacture of pharmaceutical preparations, characterized in that a compound of general formula I according to claim 1 is mixed, as an active substance, with solid or liquid, non-toxic, inert and therapeutically compatible carriers, commonly used in such preparations, and/or excipients, and that the resulting mixture is formed into pharmaceutically appropriate dosages. 12. A process for the manufacture of pharmaceutical preparations, characterized in that a compound of general formula I 19 according to claim 1, and D-cycloserine, are mixed, as activating substances, with solid or liquid, non-toxic, inert and therapeutically compatible carriers commonly used in such preparations, and/or excipients, and that the resulting mixture is formulated into pharmaceutically appropriate dosage forms. ORIGINAL in M pages 4 containing xxuCuf\References O? word added -..al. word crossed out null José JURAU Property Consulting tataW* Monte-cahi-o Pu proxy d»