CA1090785A - Peptide derivatives and process for the preparation thereof - Google Patents
Peptide derivatives and process for the preparation thereofInfo
- Publication number
- CA1090785A CA1090785A CA278,276A CA278276A CA1090785A CA 1090785 A CA1090785 A CA 1090785A CA 278276 A CA278276 A CA 278276A CA 1090785 A CA1090785 A CA 1090785A
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-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
- C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic Table
- C07F9/02—Phosphorus compounds
- C07F9/28—Phosphorus compounds with one or more P—C bonds
- C07F9/30—Phosphinic acids [R2P(=O)(OH)]; Thiophosphinic acids ; [R2P(=X1)(X2H) (X1, X2 are each independently O, S or Se)]
- C07F9/301—Acyclic saturated acids which can have further substituents on alkyl
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
- C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic Table
- C07F9/02—Phosphorus compounds
- C07F9/28—Phosphorus compounds with one or more P—C bonds
- C07F9/38—Phosphonic acids [RP(=O)(OH)2]; Thiophosphonic acids ; [RP(=X1)(X2H)2(X1, X2 are each independently O, S or Se)]
- C07F9/3804—Phosphonic acids [RP(=O)(OH)2]; Thiophosphonic acids ; [RP(=X1)(X2H)2(X1, X2 are each independently O, S or Se)] not used, see subgroups
- C07F9/3808—Acyclic saturated acids which can have further substituents on alkyl
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
- C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
- C07K5/06—Dipeptides
- C07K5/06191—Dipeptides containing heteroatoms different from O, S, or N
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
- C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
- C07K5/08—Tripeptides
- C07K5/0827—Tripeptides containing heteroatoms different from O, S, or N
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
- C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
- C07K5/10—Tetrapeptides
- C07K5/1027—Tetrapeptides containing heteroatoms different from O, S, or N
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/55—Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups
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- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Biochemistry (AREA)
- Molecular Biology (AREA)
- Medicinal Chemistry (AREA)
- Biophysics (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Genetics & Genomics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Oncology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Communicable Diseases (AREA)
- Animal Behavior & Ethology (AREA)
- Pharmacology & Pharmacy (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Peptides Or Proteins (AREA)
Abstract
ABSTRACT OF THE DISCLOSURE
This invention relates to a process for the manufacture of peptide derivatives of the general formula (I) wherein n stands for 1, 2 or 3; R1 is methyl and R2 and R3 are methyl, isopropyl or isobutyl with the proviso that R3 is methyl when n is 1; R4 represents a hydroxy or methyl group; the con-figuration at the carbon atom designated as (a) is D; the con-figuration at the carbon atom designated as (b) is L; and the configuration at the carbon atom denoted as (c) is (R), and of pharmaceutically acceptable salts thereof, which process comprises (a) cleaving off the protecting group(s) present in a compound of the general formula (II) wherein n, R1, R and R3 have the significances given above;
R40 represents a methyl group or R41; R41 represents a hydroxy group or lower alkoxy protecting group; R5 represents a hydrogen atom or a protecting group; and the configurations at the car-bon atoms designated as (a), (b) and (c) are as defined above, or (b) separating an (R,S)-diastereomeric compound corresponding to formula I into its diastereomers and isolating the (R)-diastereomer, and, if desired, converting an obtained compound of formula I into a pharmaceutically acceptable salt. me peptide derivatives of formula I potentiate the activity of D-cycloserine. In addition, they possess an antibacterial activity against organisms such as Escherichia coli, Klebsiella aerogenes, Streptococcus faecalisand Haemophilus influenzae.
This invention relates to a process for the manufacture of peptide derivatives of the general formula (I) wherein n stands for 1, 2 or 3; R1 is methyl and R2 and R3 are methyl, isopropyl or isobutyl with the proviso that R3 is methyl when n is 1; R4 represents a hydroxy or methyl group; the con-figuration at the carbon atom designated as (a) is D; the con-figuration at the carbon atom designated as (b) is L; and the configuration at the carbon atom denoted as (c) is (R), and of pharmaceutically acceptable salts thereof, which process comprises (a) cleaving off the protecting group(s) present in a compound of the general formula (II) wherein n, R1, R and R3 have the significances given above;
R40 represents a methyl group or R41; R41 represents a hydroxy group or lower alkoxy protecting group; R5 represents a hydrogen atom or a protecting group; and the configurations at the car-bon atoms designated as (a), (b) and (c) are as defined above, or (b) separating an (R,S)-diastereomeric compound corresponding to formula I into its diastereomers and isolating the (R)-diastereomer, and, if desired, converting an obtained compound of formula I into a pharmaceutically acceptable salt. me peptide derivatives of formula I potentiate the activity of D-cycloserine. In addition, they possess an antibacterial activity against organisms such as Escherichia coli, Klebsiella aerogenes, Streptococcus faecalisand Haemophilus influenzae.
Description
10~078S
The present invention relates to peptide derivatives.
More particularly, the invention is concerned with peptide derivatives of phosphonic and phosphinic acids, a process for the manufacture thereof and pharmaceutical preparations containing same.
The peptide derivatives provided by the present inven-tion are compounds of the general formula :, R3 R2 Rl O
H2N - CH - CO- -NH - CH - CO ~ NH - CH - P - R4 (I) (a) (b) (c) _ _ n OH
wherein n stands for 1, 2 or 3; R1 is methyl and R2 and R3 are methyl, isopropyl or isobutyl with the proviso that R3 is methyl when n is l; R4 represents a hydroxy or methyl group;
.` the configuration at the carbon atom designated as (a) is D;
the configuration at the carbon atom designated as (b) is L;
and the configuration at- the carbon atom denoted as (c) is (R), and pharmaceutically acceptable salts thereof.
.; As used in this specification, the expression "the . characterising group of an ~-amino acid of the type normally found in proteins" means the residue R in a natural ~-amino ~ acid of the general formula : H2N - CH - COOH
. 20 R
... .
;~.
. .
~ which is of the type normally occurring in proteins. Thus, .-: :
for example, if the amino acid is glycine then R represents a ;; hydrogen atom and if the amino acid is alanine then R represents . the methyl group. In valine R represents the isopropyl group, ....
, in leucine R represents the isobutyl group and in glutamic
The present invention relates to peptide derivatives.
More particularly, the invention is concerned with peptide derivatives of phosphonic and phosphinic acids, a process for the manufacture thereof and pharmaceutical preparations containing same.
The peptide derivatives provided by the present inven-tion are compounds of the general formula :, R3 R2 Rl O
H2N - CH - CO- -NH - CH - CO ~ NH - CH - P - R4 (I) (a) (b) (c) _ _ n OH
wherein n stands for 1, 2 or 3; R1 is methyl and R2 and R3 are methyl, isopropyl or isobutyl with the proviso that R3 is methyl when n is l; R4 represents a hydroxy or methyl group;
.` the configuration at the carbon atom designated as (a) is D;
the configuration at the carbon atom designated as (b) is L;
and the configuration at- the carbon atom denoted as (c) is (R), and pharmaceutically acceptable salts thereof.
.; As used in this specification, the expression "the . characterising group of an ~-amino acid of the type normally found in proteins" means the residue R in a natural ~-amino ~ acid of the general formula : H2N - CH - COOH
. 20 R
... .
;~.
. .
~ which is of the type normally occurring in proteins. Thus, .-: :
for example, if the amino acid is glycine then R represents a ;; hydrogen atom and if the amino acid is alanine then R represents . the methyl group. In valine R represents the isopropyl group, ....
, in leucine R represents the isobutyl group and in glutamic
- 2 -~ .
. : .
1(J~078S
acid R represents the 2-carboxyethyl group and in phenyl-alanine R represents the benzyl group. R can also represent a residue which is linked with the amino nitrogen (with the loss of one of the hydrogen atoms attached thereto), thus ; forming a nitrogen-containing ring such as in proline and pyroglutamic acid.
The configuration at the carbon atom denoted as (c) is (R); that is to sayl the configuration which would be ob-tained by replacing the carboxyl group of a naturally occurr-ing L ~-amino acid by a phosphorus moiety.
It will be appreciated that when n in formula I
he~einbefore stands for 2 or 3, the value of R2 can be the same or different.
Preferred compounds of formula I hereinbefore are those in which R4 represents a hydroxy group. Also preferred ~-are those compounds of formula I in which R represents a methyl group.
~, Examples of compounds of formula I are the following:
i~ (lR)-l-(D-alanyl-L-alanylamino)-ethylphosphonic acid, ,;, ~lR)-l-(D-alanyl-L-alanyl-L-alanylamino)-ethylphos-phonic acid, i (lR)-l-(D-valyl-L-alanyl-L-alanylamino)-ethylphos-:``
phonic acid, (lR)-l-(D-leucyl-L-alanyl-L-alanylamino)-ethylphos-phonic acid and (lR)-l-(D-alanyl-L-alanyl-L-alanyl-L-alanylamino)-~,' ethylphosphonic acid.
According to the process provided by the present invention, the peptide derivatives aforesaid ~i.e. the compounds of formula I and their pharmaceutically acceptable salts) are manufactured by ~,.i -~ (a) cleaving off by methods known per se the protecting group(s) present in a compound of the general formula ~ A -3_ !-, . .
, , .
ogoq85 r- _ R3 R2 Rl (a) (b) n R41 (II) ::`
', wherein n, R1, R2 and R3 have the significances given earlier;
R40 represents a methyl group or R41; R41 represents a `- hydroxy group or lower alkoxy protecting group; R5 repre-sents a hydrogen atom or a protecting group; the configur-` ation at the carbon atom designated as (a) is D; the con-. i ;;~ figuration at the carbon atom designated as (b) is L;
.~
hydrogen atom; and the configuration at the carbon atom designated as (c) is (R), (b) separating an (R,S)-diastereomeric compound .., corresponding to formula I into its diastereomers and iso-lating the (R)-diastereomer, and, if desired, converting an obtained compound of formula I into a pharmaceutically acceptable salt.
The protecting group denoted by R5 in formula II can be any amino-protecting group which is well-known in peptide chemistry. Especially suitable amino-protecting groups for ! the purpose of the present invention are aralkoxycarbonyl groups, particularly the benzyloxycarbonyl group, and the tertbutoxycarbonyl group. The amino-protecting group may also be a formyl, trityl or trifluoroacetyl group.
. ;.~ . .
., ;~. '~
4 ~
' 7~' ,~ :
,.. : . . . : .
.,.. . ~ . , .
:'... ~ ' ' ' . .
109078~i .
- The cleavage of the protecting group or protecting groups present in a compound of formula II is carried out in accordance with methods known per se; that is to say, ~i1 methods in actual use for or described in the literature on the cleavage of protecting groups. Thus, for example, an aralkoxycarbonyl group (e.g. benzyloxycarbonyl) or a tertbutoxycarbonyl group may be cleaved off by hydrolysis (e.g. treatment with a mixture of hydrogen bromide and glacial acetic acid). An aralkoxycarbonyl group (e.g.
benzyloxycarbonyl) can also be cleaved off by hydrogenolysis (e.g. in the presence of palladium-on-charcoal or palladium .
oxide). The tertbutoxycarbonyl group may also be cleaved ; off by means of hydrogen chloride in dioxan. A lower alkoxy group denoted by R40 and/or R41 can be a straight-chain or branched-chain alkoxy group preferably containing from 1 to 6 carbon atoms (e.g. methoxy, ethoxy, propoxy, isopropoxy, butoxy etc.) and may be converted into a hydroxy group by treatment with a mixture of hydrogen ?:
bromide in glacial acetic acid or by means of trimethyl-,~ 20 chlorosilane followed by aqueous hydrolysis. It will be , ~ .
' appreciated that the cleavage of the protecting groups can .:, be carried out in a single step or in more than one step i depending on the nature of the protecting groups present.
The separation of an (R,S) diastereomeric compound corresponding to formula I into its diastereomers and iso-lation of the (R)-diastereomer can be carried out according i~ to known methods~ for example, by fractional crystallisation or by high pressure liquid chromatography.
.i,;,. ..
.....
_ 5 _ .
''' '' ' " ' .
1090~8S
Compounds of formula I are amphoteric in nature and form pharmaceutically acceptable salts with strong acids (e.g. methanesulphonic acid, paratoluenesulphonic acid, hydrochloric acid, hydrobromic acid, sulphuric acid etc.) and with bases (e.g. sodium hydroxide etc.).
The starting materials of formula II hereinbefore ` may be prepared, for example, by condensing a compound of the general formula R2 Rl O
H- -NH - CH - C0- - NH - CH - P - R40 (III) (b) m R 1 ~' ' wherein m stands for zero, 1, 2 or 3; Rl, R , R and 41 have the significance given earlier; the configuration at the carbon atom designated as (b) is L; and the con-figuration at the carbon atom designated as (c) is (R), with an appropriately protected -amino acid, an appropriately protected dipeptide, an appropriately protected tripeptide, an appropriately protected tetrapeptide or a reactive de-rivative thereof as the case may require.
Thus, a compound of formula III in which m stands for zero can be condensed with an appropriately protected -dipeptide or a reactive derivative thereof to give a com-.i ~ pound of formula II in which n stands for 1, or with an ,~ ~
f, ~ ' ' ~(~9078S
,.
appropriately protected tripeptide or a reactive derivative thereof to give a compound of formula II in which n stands for 2 or with an appropriately protected tetrapeptide or a reactive derivative thereof to give a compound of formula II in which n stands for 3.
Again, a compound of formula III in which m stands for 1 can be condensed with an appropriately protected amino acid or a reactive derivative thereof to give a --compound of formula II in which n stands for 1, or with '.!
10 an appropriately protected dipeptide or a reactive deriva-. tive thereof to give a compound of formula II in which n j stands for 2 or with an appropriately protected tripeptide or a reactive derivative thereof to give a compound of formula II in which n stands for 3.
Yet again, a compound of formula III in which m ~ stands for 2 can be condensed with an appropriately pro- ~:
;.,~ tected ~-amino ;
..~ i ~s . .
, '~, .
...
., . . :
;
. . .
::~, , .
;, ;. ~ : .
~a, ~~' . . . .
acid or a reactive derivative thereof to give a compound of formula II in which n stands for 2 or with an appropriately protected dipeptide or a reactive derivative thereof to give a f compound of formula II in which n stands for 3.
i~
. .
'. 5 Finally, a compound of formula III in which m stands for
. : .
1(J~078S
acid R represents the 2-carboxyethyl group and in phenyl-alanine R represents the benzyl group. R can also represent a residue which is linked with the amino nitrogen (with the loss of one of the hydrogen atoms attached thereto), thus ; forming a nitrogen-containing ring such as in proline and pyroglutamic acid.
The configuration at the carbon atom denoted as (c) is (R); that is to sayl the configuration which would be ob-tained by replacing the carboxyl group of a naturally occurr-ing L ~-amino acid by a phosphorus moiety.
It will be appreciated that when n in formula I
he~einbefore stands for 2 or 3, the value of R2 can be the same or different.
Preferred compounds of formula I hereinbefore are those in which R4 represents a hydroxy group. Also preferred ~-are those compounds of formula I in which R represents a methyl group.
~, Examples of compounds of formula I are the following:
i~ (lR)-l-(D-alanyl-L-alanylamino)-ethylphosphonic acid, ,;, ~lR)-l-(D-alanyl-L-alanyl-L-alanylamino)-ethylphos-phonic acid, i (lR)-l-(D-valyl-L-alanyl-L-alanylamino)-ethylphos-:``
phonic acid, (lR)-l-(D-leucyl-L-alanyl-L-alanylamino)-ethylphos-phonic acid and (lR)-l-(D-alanyl-L-alanyl-L-alanyl-L-alanylamino)-~,' ethylphosphonic acid.
According to the process provided by the present invention, the peptide derivatives aforesaid ~i.e. the compounds of formula I and their pharmaceutically acceptable salts) are manufactured by ~,.i -~ (a) cleaving off by methods known per se the protecting group(s) present in a compound of the general formula ~ A -3_ !-, . .
, , .
ogoq85 r- _ R3 R2 Rl (a) (b) n R41 (II) ::`
', wherein n, R1, R2 and R3 have the significances given earlier;
R40 represents a methyl group or R41; R41 represents a `- hydroxy group or lower alkoxy protecting group; R5 repre-sents a hydrogen atom or a protecting group; the configur-` ation at the carbon atom designated as (a) is D; the con-. i ;;~ figuration at the carbon atom designated as (b) is L;
.~
hydrogen atom; and the configuration at the carbon atom designated as (c) is (R), (b) separating an (R,S)-diastereomeric compound .., corresponding to formula I into its diastereomers and iso-lating the (R)-diastereomer, and, if desired, converting an obtained compound of formula I into a pharmaceutically acceptable salt.
The protecting group denoted by R5 in formula II can be any amino-protecting group which is well-known in peptide chemistry. Especially suitable amino-protecting groups for ! the purpose of the present invention are aralkoxycarbonyl groups, particularly the benzyloxycarbonyl group, and the tertbutoxycarbonyl group. The amino-protecting group may also be a formyl, trityl or trifluoroacetyl group.
. ;.~ . .
., ;~. '~
4 ~
' 7~' ,~ :
,.. : . . . : .
.,.. . ~ . , .
:'... ~ ' ' ' . .
109078~i .
- The cleavage of the protecting group or protecting groups present in a compound of formula II is carried out in accordance with methods known per se; that is to say, ~i1 methods in actual use for or described in the literature on the cleavage of protecting groups. Thus, for example, an aralkoxycarbonyl group (e.g. benzyloxycarbonyl) or a tertbutoxycarbonyl group may be cleaved off by hydrolysis (e.g. treatment with a mixture of hydrogen bromide and glacial acetic acid). An aralkoxycarbonyl group (e.g.
benzyloxycarbonyl) can also be cleaved off by hydrogenolysis (e.g. in the presence of palladium-on-charcoal or palladium .
oxide). The tertbutoxycarbonyl group may also be cleaved ; off by means of hydrogen chloride in dioxan. A lower alkoxy group denoted by R40 and/or R41 can be a straight-chain or branched-chain alkoxy group preferably containing from 1 to 6 carbon atoms (e.g. methoxy, ethoxy, propoxy, isopropoxy, butoxy etc.) and may be converted into a hydroxy group by treatment with a mixture of hydrogen ?:
bromide in glacial acetic acid or by means of trimethyl-,~ 20 chlorosilane followed by aqueous hydrolysis. It will be , ~ .
' appreciated that the cleavage of the protecting groups can .:, be carried out in a single step or in more than one step i depending on the nature of the protecting groups present.
The separation of an (R,S) diastereomeric compound corresponding to formula I into its diastereomers and iso-lation of the (R)-diastereomer can be carried out according i~ to known methods~ for example, by fractional crystallisation or by high pressure liquid chromatography.
.i,;,. ..
.....
_ 5 _ .
''' '' ' " ' .
1090~8S
Compounds of formula I are amphoteric in nature and form pharmaceutically acceptable salts with strong acids (e.g. methanesulphonic acid, paratoluenesulphonic acid, hydrochloric acid, hydrobromic acid, sulphuric acid etc.) and with bases (e.g. sodium hydroxide etc.).
The starting materials of formula II hereinbefore ` may be prepared, for example, by condensing a compound of the general formula R2 Rl O
H- -NH - CH - C0- - NH - CH - P - R40 (III) (b) m R 1 ~' ' wherein m stands for zero, 1, 2 or 3; Rl, R , R and 41 have the significance given earlier; the configuration at the carbon atom designated as (b) is L; and the con-figuration at the carbon atom designated as (c) is (R), with an appropriately protected -amino acid, an appropriately protected dipeptide, an appropriately protected tripeptide, an appropriately protected tetrapeptide or a reactive de-rivative thereof as the case may require.
Thus, a compound of formula III in which m stands for zero can be condensed with an appropriately protected -dipeptide or a reactive derivative thereof to give a com-.i ~ pound of formula II in which n stands for 1, or with an ,~ ~
f, ~ ' ' ~(~9078S
,.
appropriately protected tripeptide or a reactive derivative thereof to give a compound of formula II in which n stands for 2 or with an appropriately protected tetrapeptide or a reactive derivative thereof to give a compound of formula II in which n stands for 3.
Again, a compound of formula III in which m stands for 1 can be condensed with an appropriately protected amino acid or a reactive derivative thereof to give a --compound of formula II in which n stands for 1, or with '.!
10 an appropriately protected dipeptide or a reactive deriva-. tive thereof to give a compound of formula II in which n j stands for 2 or with an appropriately protected tripeptide or a reactive derivative thereof to give a compound of formula II in which n stands for 3.
Yet again, a compound of formula III in which m ~ stands for 2 can be condensed with an appropriately pro- ~:
;.,~ tected ~-amino ;
..~ i ~s . .
, '~, .
...
., . . :
;
. . .
::~, , .
;, ;. ~ : .
~a, ~~' . . . .
acid or a reactive derivative thereof to give a compound of formula II in which n stands for 2 or with an appropriately protected dipeptide or a reactive derivative thereof to give a f compound of formula II in which n stands for 3.
i~
. .
'. 5 Finally, a compound of formula III in which m stands for
3 can be condensed with an appropriately protected a-amino acid or a reactive derivative thereof to give a compound of , formula II in which n stands for 3.
,, , . Alternatively, the compounds of formula II can be prepared by carrying out the foregoing condensation using an (R,S) compound corresponding to formula III and separating the ~;~' (R) compound from the resulting (R,S) product in a manner ~'3 known per se; for example, by crystallisation, chromatography . ,,1 ~j or fractional crystallisation using a suitable base such as ;~
a-methylkenzylamine.
The aforementioned condensation can be carried out in accordance with methods which are known per se in peptide chemistry; for example, by the mixed anhydride, azide, ~, activated ester or acid chloride method.
....
'ii 20 In one method, an appropriate compound of formula III
. can be condensed with an appropriately protected ~-amino acid, i~l di-, tri- or tetrapeptide as the case may require in which the terminal carboxy function is a mixed anhydride residue formed .,:;
::l with an organic or inorganic acid. Suitably, such an amino ~ 25 acid, di-, tri- or tetrapeptide carrying a free carboxy ,., ~' function is treated with a tertiary base such as a tri-(lower ,~ .
;: ~
~ ~ 8 , .. .
;, '': .
_.
~0907fl~i alkyl)-amine (e.y. triethylamine) or ~-ethyImorplloline in an inert organic solvent (e.g. tetrahydrofuran, 1,2-dimethoxy-ethane, dichloromethane, toluene, petroleum ether or mixtures thereof) and the resulting salt is reacted with a chloroformic acid ester (e.g. the ethyl or isobutyl ester) at a low temperature. The mixed anhydride obtained is then suitably condensed in situ with the compound of formula III.
.~ .
In another method, an appropriate compound of formula III
can be condensed with an appropriately protected a-amino acid, di-, tri- or tetrapeptide as the case may require in which the terminal carboxy group is in the form of an acid azide. This condensation is preferably carried out in an inert organic soivent such as dimethylformamide or ethyl acetate at a low temperature.
In yet another method, an appropriate compound of formula ~; III can be condensed with an appropriately protected a-amino acid, di-, tri- or tetrapeptide as the case may require in which the terminal carboxy function is in the form of an active ester group (e.g. the p-nitrophenyl, 2,4,5-trichlorophenyl or N-hydroxysuccinimide ester group). This condensation is suitably carried out either in an inert organic solvent such as dimethylformamide or, in the case where R40 and/or R4l ~r,,~ represents a lower alkoxy group, in an aqueous alkanol (e.g.
l aqueous ethanol).
,$ 25 In a further method, an appropriate compound of formula III can be condensed with an appropriately protected -amino acid, di-, tri- or tetrapeptide as the case may require in '. ' ~, 9 ', which the terminal carboxy function is in the form of an acid chloride, This condensation is preferably carried out in the presence of a base and at a low temperature.
- .
The peptide derivatives provided by the present invention potentiate the activity of D-cycloserine. In addition, they possess an antibacterial activity against organisms such as Escherichia coli, Klebsiella aerogenes, Streptococcus faecalis and Haemophilus influenzae.
,, ,, The present peptide derivatives may accordingly be used . 10 as medicaments; for example, in the form of pharmaceutical . . .
preparations which contain them in association with a compatible pharmaceutical carrier material. This carrier ~ material can be an organic or inorganic carrier material 'lr suitable for enteral (e.g. oral) or parenteral administration.
Examples of such carrier materials are water, lactose, starch, magnesium stearate, gum arabic, gelatin, polyalkyleneglycols, petroleum jelly and the like. The pharmaceutical preparations can be made up in a solid form (e.g. as tablets, dragées, .,:.
~' suppositories or capsules) or in a liquid form (e.g. as ,,.:., solutions, suspensions or emulsions). The pharmaceutical preparations may be subjected to conventional pharmaceutical ~, .
`l operations such as sterilisation and may contain adjuvants such as preservatives, stabilisers, wetting agents or salts for ~ altering the osmotic pressure.
`: .
~, 25 The peptide derivatives provided by the present invention can be administered in combination with D-cycloserine.
', Alternatively, the peptide derivative and D-cycloserine can be . : , :,' -1()907S5 administered separately, if necessary by different routes.
The amount of the peptide derivatives to be administered as well as the ratio in which the peptide derivative and D-cyclo-serine can be administered can vary within wide limits depending on such factors as the particular derivative chosen, the route of administration and the organism to be combatted.
For example, the peptide derivative and D-cycloserine may be administered in a ratio of from ca 100:1 to 1:100.
' '', ,;
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. ,~ .
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-` 109078~
t ;
The following Examples illustrate the process provided by the present invention :
, ` Example 1 . ~
(A) The process:
- 5 4.3 g (7.4 mmol) of the benzylamine salt of (lR)-l-[(N--benzyloxycarbonyl-D-alanyl-L-alanyl-L-alanyl)-amino]-ethyl-phosphonic acid were added to 12 ml of a stirred 45% solution `~ of hydrogen bromide in glacial acetic acid and washed in with ; 4 ml of glacial acetic acid. The mixture was stirred at room temperature for 6 hours and then 100 ml of ether were added while stirring. The supernatant was decanted and the residue was again treated with 100 ml of ether. The resulting gum was taken up in 30 ml of methanol and the solution treated with a solution of 3 ml of propylene oxide in 5 ml of methanol to give a white precipitate. After standing overnight at room temperature, the precipitate was filtered off, washed succes-';, sively sith methanol and ether and then dried. Crystallisation ,~ fron water/ethanol gave 1.96 g of (lR)-l-(D-alanyl-L-alanyl-L--alanylamino)-ethylphosphonic acid of melting point 318-320C
(decomposition); [a]D = -107 (c = 0.5% in 1 N sodium hydroxide).
(B) The preparation of the starting material:
2.7 g (10 mmol) of (lR)-l-(L-alanyl-L-alanylamino)-z -ethylphosphonic acid were stirred in 50 ml of water at 5C
, 25 while 2,0 g (20 mmol) of triethylamine followed by 50 ml of ethanol were added. The resulting clear solution was cooled to 0C and there were added thereto 3.8 g (12 mmol) of solid N-hydroxysuccinimide ester of N-benzyloxycarbonyl-D-alanine in , .
~ 12 , .
` `` i(J9078S
a single portion. This N-hydroxysuccinimide ester was washed in with 25 ml of ethanol. The mixture was stirred for 2 hours at 0C and then for 16 hours at room temperature when a clear solution was obtained. The solvents were removed by evaporation and the residue was partitioned between 150 ml of water and 100 ml of chloroform. The aqueous layer was washed with a further 100 ml of chloroform and the solvent extracts were back-washed with 50 ml of water. The aqueous extracts were combined and evaporated. The residue was taken up in a mixture of 40 ml of water and 40 ml of methanol and passed down a column of cation exchange resin (sulfonated polystyrene, freshly regenerated in the acid cycle).
Elution was carried out with the same solvent system. The acid eluate obtained was evaporated, the residue taken up in 250 ml of water and extracted twice with 100 ml of ether each time. The ether extracts were back-washed with 50 ml of water.
The combined aqueous extracts were concentrated to ca 100 ml.
An equal amount of methanol was added and the resulting solution was titrated to pH 4.5 with l-M aqueous benzylamine. The solution ' ,' 20 was evaporated to dryness and the solid obtained was recrystal-;' lised from a mixture of 100 ml of hot water to which was added - 400 ml of ethanol followed by 500 ml of ether. There were obtained 4.38 g of the benzylamine salt of (lR)-l-[(~-benzyloxy-carbonyl-D-alanyl-L-alanyl-L-alanyl~amino]-ethylphosphonic acid of melting point 240-243C (decomposition); []D = -38.1 (c = 0.53% in acetic acid).
' :
' :' l-.. 13 ~, , , Example 2 (A) The process:
;; In a manner analogous to that described in Example 1, from the benzylamine salt of (lR)-l-[(N-benzyloxycarbonyl-D-valyl-L-alanyl-L-alanyl)-amino]-ethylphosphonic acid there was ~ obtained (lR)-l-(D-valyl-L-alanyl-L-alanylamino)-ethylphosphonic ,'!, acid of melting point 301-304C (decomposition); [~]D0 =
-105 (c = 0.45% in 1 N sodium hydroxide).
(B) The preparation of the starting material:
3.8 g (15 mmol) of N-benzyloxycarbonyl-D-valine were ,1 stirred in 200 ml of petroleum ether (boiling point 60-30 C), 1.5 g (15 mmol) of triethylamine were added and the mixture was cooled to -5C. 2.1 g (15 mmol) of isobutyl chloroformate were added and the mixture was maintained at -5C for 30 3~ 15 minutes. While stirring this mixture at -5C there was added , dropwise a solution of 2.7 g (10 mmol) of (lR)-l-(L-alanyl-L-;1 ¦ alanylamino)-ethylphosphonic acid in a mixture of 2.0 g (20 mmol) of triethylamine and 15 ml of water. This solution was washed in with 5 ml of water. The stirring was continued at -5C to 0C for a further 2 hours and then overnight at room ..
:l temperature. 150 ml of water were added and the aqueous and Si organic phases were separated. The aqueous phase was evaporated, the residue taken up in a mixture of 40 ml of water and 40 ml of methanol and the residue passed down a column of cation exchange resin freshly regenerated in the acid cycle. The :.
column was eluted with the same solvent mixture and the acid fraction was evaporated and re-evaporated with five 50 ml . ., -' portions of water. The residue was triturated with 100 ml of ether and the solid filtered off. The solid was taken up in a ,.~
'' ' ` 109078S
mixture of 400 ml of methanol and 400 ml of water and the solution was titrated to pH 4.5 with 4 M aqueous benzylamine.
The solution was evaporated to dryness and the resulting solid was crystallised from a mixture of 100 ml of methanol and 150 ml of ether to give 3.24 g of the benzylamine salt of (lR)-l-[(N-benzyloxycarbonyl-D-valyl-L-alanyl-L-alanyl)-amino]--ethylphosphonic acid of melting point 238-244 (decomposi-tion).
Example 3 ., , .... .
: lo (A) The process:
In a manner analogous to that described in Example 1, :~ from the benzylamine salt of (lR)-l-[(N-benzyloxycarbonyl- .
, D-alanyl-L-alanyl-L-alanyl-L-alanyl)-amino]-ethylphosphonic acid there was obtained (lR)-l-(D-alanyl-L-alanyl--L-alanyl-L-alanylamino)-ethylphosphonic acid of melting point 323-325C (decomposition); [a]D = -121 (c = 0.48~ in 1 N
sodium hydroxide).
(B) The preparation of the starting material:
- .
In a manner analogous to that described in part (B) of . 20 Example 1, from the N-hydroxysuccinimide ester of N-benzyloxy-- carbonyl-L-alanine and (lP;)-l-(L-alanyl-L-alanylamino)-ethyl-;l phosphonic acid there was obtained (lR)-l-[(N-benzyloxv-carbonyl-L-alanyl-L-alanyl-L-alanyl)-amino]-ethylphosphonic acid of melting point 255-257C (decomposition); []DO = -62.0 . 25 (c = 0.4% in glacial acetic acid).
. ~ . , .
. .
. In a manner analogous to that described in part (A) of .~ .
~ Example 1, from (lR)-l-[(N-benzyloxycarbonyl-L-alanyl-L-alanyl-;::
` ~ 15 ,"~
. . .
, , .~. . ..
.... .
~1090785 -L-alanyl)-amino]-ethylphosphonic acid there was obtained (lR)-l-(L-alanyl-L-alanyl-L-alanylamino)-ethylphosphonic acid of melting point 312-313C (decomposition); [a]D =
~; -101 (c = 0.53% in 1 N sodium hydroxide).
, In a manner analogous to that described in part (B) - of Example 1, but with acidification to pH 2 and further :, ~ solvent extraction with ether replacing passage through the ,.:
resin and subsequent solvent extraction with ether, from the N-hydroxysuccinimide ester of N-benzyloxycarbonyl-D-alanine and (lR)-l-(L-alanyl-L-alanyl-L-alanylamino)-ethylphosphonic .j .
acid there was obtained the benzylamine salt of (lR)-l-[(N-benzyloxycarbonyl-D-alanyl-L-alanyl-L-alanyl-L-alanyl)-amino]-~ ethylphosphonic acid of melting point 272-277C (decomposition);
!,~., [a]20 = _47.oo (c = 0,5% in acetic acid).
~J ~
~ 15 Example 4 . ~
" ~ .
(A) The process:
~, 10,85 g of dimethyl (lR)-l-[(N-benzyloxycarbonyl-D-. .
~`~, -leucyl-L-alanyl-L-alanyl)-amino]-ethylphosphonate were ; dissolved in 30 ml of a 35~ solution of hydrogen bromide ,;:
in glacial acetic acid and the mixture was stirred at room ', temperature for 4 hours. 130 ml of ether were then added - while stirring, the stirring was discontinued and the ether : decanted off. This procedure was repeated twice using 80 ml -, of ether each time. The residue was dissolved in 70 ml of methanol and tc the resulting solution were added 10 ml of , propylene oxide. After refrigerating overnight, the resulting , - white precipitate was filtered off and washed with ethanol and ether. The product was dried in vacuo to a constant weight of '' ', ~ 16 ,1 . ~ ~
:
~L090785 7.99 g; melting point 293-295C (decomposition). Recrystal-lisation from 500 ml of cold water and from 700 ml of ethanol gave 6.67 g of (lR)-l-(D-leucyl-L-alanyl-L-alanylamino)--ethylphosphonic acid of melting point 300-302C (decomposi-tion); [a]D = -129,3 (c = 1% in water).
., .
(B) The preparation of the starting material:
12.9 g (0.03 mol) of dimethyl (lR)-l-[(N-benzyloxy-carbonyl-L-alanyl-L-alanyl)-amino]-ethylphosphonate were dissolved in 150 ml of methanol containing 0.032 mol of hydrogen chloride. The solution was hydrogenated at room temperature and pressure in the presence of 1 g of 10~
palladium-on-charcoal until the hydrogen uptake ceased.
The catalyst was filtered off, the filtrate evaporated in vacuo and the oily hydrochloride evaporated twice with , 15 ethyl acetate.
,, .
; The product obtained according to the preceding paragraph and 10.9 g (0.03 mol) of the N-hydroxysuccinimide ester of i N-benzyloxycarbonyl-D-leucine were stirred in the presence ; of 100 ml of dry dimethylformamide. While stirring and ' 20 maintaining the temperature below 15C, 4.2 ml of dry . !
triethylamine were added dropwise. The mixture was then stirred overnight at room temperature. The triethylamine hydrochloride was filtered off and washed with a small amount of dimethylformamide. The filtrate was evaporated under an oil-pump vacuum and at a bath temperature of below - 40C. The residual oil was treated with 50 ml of water and the resulting mixture extracted with four 50 ml portions of chloroform. The combined or~anic phases were washed with a small vclume of 20% potassium carbonate solution and then .~.
, .
,,.
, .. .
~L090785 .
dried over sodium sulphate. The sodium sulphate was filtered . off and the filtrate evaporated under a water-pump vacuum.
The residue was re-evaporated twice with ethyl acetate.
Trituration with dry ether. filtration and vacuum drying yielded 15.5 g of dimethyl (lR)-l-[(N-benzyloxycarbonyl-D-leucyl-L-alanyl-L-alanyl)-amino]-ethylphosphonate of melting . point 163-167C. After recrvstallisation from a_etonitrile, . the melting point was 173-176C; [~]D0 = -36.6 (c = 1% in ; methanol).
! .
lo Example 5 ~' ` (A) The process:
In a manner analogous to that described in Example
,, , . Alternatively, the compounds of formula II can be prepared by carrying out the foregoing condensation using an (R,S) compound corresponding to formula III and separating the ~;~' (R) compound from the resulting (R,S) product in a manner ~'3 known per se; for example, by crystallisation, chromatography . ,,1 ~j or fractional crystallisation using a suitable base such as ;~
a-methylkenzylamine.
The aforementioned condensation can be carried out in accordance with methods which are known per se in peptide chemistry; for example, by the mixed anhydride, azide, ~, activated ester or acid chloride method.
....
'ii 20 In one method, an appropriate compound of formula III
. can be condensed with an appropriately protected ~-amino acid, i~l di-, tri- or tetrapeptide as the case may require in which the terminal carboxy function is a mixed anhydride residue formed .,:;
::l with an organic or inorganic acid. Suitably, such an amino ~ 25 acid, di-, tri- or tetrapeptide carrying a free carboxy ,., ~' function is treated with a tertiary base such as a tri-(lower ,~ .
;: ~
~ ~ 8 , .. .
;, '': .
_.
~0907fl~i alkyl)-amine (e.y. triethylamine) or ~-ethyImorplloline in an inert organic solvent (e.g. tetrahydrofuran, 1,2-dimethoxy-ethane, dichloromethane, toluene, petroleum ether or mixtures thereof) and the resulting salt is reacted with a chloroformic acid ester (e.g. the ethyl or isobutyl ester) at a low temperature. The mixed anhydride obtained is then suitably condensed in situ with the compound of formula III.
.~ .
In another method, an appropriate compound of formula III
can be condensed with an appropriately protected a-amino acid, di-, tri- or tetrapeptide as the case may require in which the terminal carboxy group is in the form of an acid azide. This condensation is preferably carried out in an inert organic soivent such as dimethylformamide or ethyl acetate at a low temperature.
In yet another method, an appropriate compound of formula ~; III can be condensed with an appropriately protected a-amino acid, di-, tri- or tetrapeptide as the case may require in which the terminal carboxy function is in the form of an active ester group (e.g. the p-nitrophenyl, 2,4,5-trichlorophenyl or N-hydroxysuccinimide ester group). This condensation is suitably carried out either in an inert organic solvent such as dimethylformamide or, in the case where R40 and/or R4l ~r,,~ represents a lower alkoxy group, in an aqueous alkanol (e.g.
l aqueous ethanol).
,$ 25 In a further method, an appropriate compound of formula III can be condensed with an appropriately protected -amino acid, di-, tri- or tetrapeptide as the case may require in '. ' ~, 9 ', which the terminal carboxy function is in the form of an acid chloride, This condensation is preferably carried out in the presence of a base and at a low temperature.
- .
The peptide derivatives provided by the present invention potentiate the activity of D-cycloserine. In addition, they possess an antibacterial activity against organisms such as Escherichia coli, Klebsiella aerogenes, Streptococcus faecalis and Haemophilus influenzae.
,, ,, The present peptide derivatives may accordingly be used . 10 as medicaments; for example, in the form of pharmaceutical . . .
preparations which contain them in association with a compatible pharmaceutical carrier material. This carrier ~ material can be an organic or inorganic carrier material 'lr suitable for enteral (e.g. oral) or parenteral administration.
Examples of such carrier materials are water, lactose, starch, magnesium stearate, gum arabic, gelatin, polyalkyleneglycols, petroleum jelly and the like. The pharmaceutical preparations can be made up in a solid form (e.g. as tablets, dragées, .,:.
~' suppositories or capsules) or in a liquid form (e.g. as ,,.:., solutions, suspensions or emulsions). The pharmaceutical preparations may be subjected to conventional pharmaceutical ~, .
`l operations such as sterilisation and may contain adjuvants such as preservatives, stabilisers, wetting agents or salts for ~ altering the osmotic pressure.
`: .
~, 25 The peptide derivatives provided by the present invention can be administered in combination with D-cycloserine.
', Alternatively, the peptide derivative and D-cycloserine can be . : , :,' -1()907S5 administered separately, if necessary by different routes.
The amount of the peptide derivatives to be administered as well as the ratio in which the peptide derivative and D-cyclo-serine can be administered can vary within wide limits depending on such factors as the particular derivative chosen, the route of administration and the organism to be combatted.
For example, the peptide derivative and D-cycloserine may be administered in a ratio of from ca 100:1 to 1:100.
' '', ,;
., f, '',~`;li ' , ~
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. ,~ .
:!
' ~1 ' . .
,..
. ~
. .
: A 11 ... .
. .....
-` 109078~
t ;
The following Examples illustrate the process provided by the present invention :
, ` Example 1 . ~
(A) The process:
- 5 4.3 g (7.4 mmol) of the benzylamine salt of (lR)-l-[(N--benzyloxycarbonyl-D-alanyl-L-alanyl-L-alanyl)-amino]-ethyl-phosphonic acid were added to 12 ml of a stirred 45% solution `~ of hydrogen bromide in glacial acetic acid and washed in with ; 4 ml of glacial acetic acid. The mixture was stirred at room temperature for 6 hours and then 100 ml of ether were added while stirring. The supernatant was decanted and the residue was again treated with 100 ml of ether. The resulting gum was taken up in 30 ml of methanol and the solution treated with a solution of 3 ml of propylene oxide in 5 ml of methanol to give a white precipitate. After standing overnight at room temperature, the precipitate was filtered off, washed succes-';, sively sith methanol and ether and then dried. Crystallisation ,~ fron water/ethanol gave 1.96 g of (lR)-l-(D-alanyl-L-alanyl-L--alanylamino)-ethylphosphonic acid of melting point 318-320C
(decomposition); [a]D = -107 (c = 0.5% in 1 N sodium hydroxide).
(B) The preparation of the starting material:
2.7 g (10 mmol) of (lR)-l-(L-alanyl-L-alanylamino)-z -ethylphosphonic acid were stirred in 50 ml of water at 5C
, 25 while 2,0 g (20 mmol) of triethylamine followed by 50 ml of ethanol were added. The resulting clear solution was cooled to 0C and there were added thereto 3.8 g (12 mmol) of solid N-hydroxysuccinimide ester of N-benzyloxycarbonyl-D-alanine in , .
~ 12 , .
` `` i(J9078S
a single portion. This N-hydroxysuccinimide ester was washed in with 25 ml of ethanol. The mixture was stirred for 2 hours at 0C and then for 16 hours at room temperature when a clear solution was obtained. The solvents were removed by evaporation and the residue was partitioned between 150 ml of water and 100 ml of chloroform. The aqueous layer was washed with a further 100 ml of chloroform and the solvent extracts were back-washed with 50 ml of water. The aqueous extracts were combined and evaporated. The residue was taken up in a mixture of 40 ml of water and 40 ml of methanol and passed down a column of cation exchange resin (sulfonated polystyrene, freshly regenerated in the acid cycle).
Elution was carried out with the same solvent system. The acid eluate obtained was evaporated, the residue taken up in 250 ml of water and extracted twice with 100 ml of ether each time. The ether extracts were back-washed with 50 ml of water.
The combined aqueous extracts were concentrated to ca 100 ml.
An equal amount of methanol was added and the resulting solution was titrated to pH 4.5 with l-M aqueous benzylamine. The solution ' ,' 20 was evaporated to dryness and the solid obtained was recrystal-;' lised from a mixture of 100 ml of hot water to which was added - 400 ml of ethanol followed by 500 ml of ether. There were obtained 4.38 g of the benzylamine salt of (lR)-l-[(~-benzyloxy-carbonyl-D-alanyl-L-alanyl-L-alanyl~amino]-ethylphosphonic acid of melting point 240-243C (decomposition); []D = -38.1 (c = 0.53% in acetic acid).
' :
' :' l-.. 13 ~, , , Example 2 (A) The process:
;; In a manner analogous to that described in Example 1, from the benzylamine salt of (lR)-l-[(N-benzyloxycarbonyl-D-valyl-L-alanyl-L-alanyl)-amino]-ethylphosphonic acid there was ~ obtained (lR)-l-(D-valyl-L-alanyl-L-alanylamino)-ethylphosphonic ,'!, acid of melting point 301-304C (decomposition); [~]D0 =
-105 (c = 0.45% in 1 N sodium hydroxide).
(B) The preparation of the starting material:
3.8 g (15 mmol) of N-benzyloxycarbonyl-D-valine were ,1 stirred in 200 ml of petroleum ether (boiling point 60-30 C), 1.5 g (15 mmol) of triethylamine were added and the mixture was cooled to -5C. 2.1 g (15 mmol) of isobutyl chloroformate were added and the mixture was maintained at -5C for 30 3~ 15 minutes. While stirring this mixture at -5C there was added , dropwise a solution of 2.7 g (10 mmol) of (lR)-l-(L-alanyl-L-;1 ¦ alanylamino)-ethylphosphonic acid in a mixture of 2.0 g (20 mmol) of triethylamine and 15 ml of water. This solution was washed in with 5 ml of water. The stirring was continued at -5C to 0C for a further 2 hours and then overnight at room ..
:l temperature. 150 ml of water were added and the aqueous and Si organic phases were separated. The aqueous phase was evaporated, the residue taken up in a mixture of 40 ml of water and 40 ml of methanol and the residue passed down a column of cation exchange resin freshly regenerated in the acid cycle. The :.
column was eluted with the same solvent mixture and the acid fraction was evaporated and re-evaporated with five 50 ml . ., -' portions of water. The residue was triturated with 100 ml of ether and the solid filtered off. The solid was taken up in a ,.~
'' ' ` 109078S
mixture of 400 ml of methanol and 400 ml of water and the solution was titrated to pH 4.5 with 4 M aqueous benzylamine.
The solution was evaporated to dryness and the resulting solid was crystallised from a mixture of 100 ml of methanol and 150 ml of ether to give 3.24 g of the benzylamine salt of (lR)-l-[(N-benzyloxycarbonyl-D-valyl-L-alanyl-L-alanyl)-amino]--ethylphosphonic acid of melting point 238-244 (decomposi-tion).
Example 3 ., , .... .
: lo (A) The process:
In a manner analogous to that described in Example 1, :~ from the benzylamine salt of (lR)-l-[(N-benzyloxycarbonyl- .
, D-alanyl-L-alanyl-L-alanyl-L-alanyl)-amino]-ethylphosphonic acid there was obtained (lR)-l-(D-alanyl-L-alanyl--L-alanyl-L-alanylamino)-ethylphosphonic acid of melting point 323-325C (decomposition); [a]D = -121 (c = 0.48~ in 1 N
sodium hydroxide).
(B) The preparation of the starting material:
- .
In a manner analogous to that described in part (B) of . 20 Example 1, from the N-hydroxysuccinimide ester of N-benzyloxy-- carbonyl-L-alanine and (lP;)-l-(L-alanyl-L-alanylamino)-ethyl-;l phosphonic acid there was obtained (lR)-l-[(N-benzyloxv-carbonyl-L-alanyl-L-alanyl-L-alanyl)-amino]-ethylphosphonic acid of melting point 255-257C (decomposition); []DO = -62.0 . 25 (c = 0.4% in glacial acetic acid).
. ~ . , .
. .
. In a manner analogous to that described in part (A) of .~ .
~ Example 1, from (lR)-l-[(N-benzyloxycarbonyl-L-alanyl-L-alanyl-;::
` ~ 15 ,"~
. . .
, , .~. . ..
.... .
~1090785 -L-alanyl)-amino]-ethylphosphonic acid there was obtained (lR)-l-(L-alanyl-L-alanyl-L-alanylamino)-ethylphosphonic acid of melting point 312-313C (decomposition); [a]D =
~; -101 (c = 0.53% in 1 N sodium hydroxide).
, In a manner analogous to that described in part (B) - of Example 1, but with acidification to pH 2 and further :, ~ solvent extraction with ether replacing passage through the ,.:
resin and subsequent solvent extraction with ether, from the N-hydroxysuccinimide ester of N-benzyloxycarbonyl-D-alanine and (lR)-l-(L-alanyl-L-alanyl-L-alanylamino)-ethylphosphonic .j .
acid there was obtained the benzylamine salt of (lR)-l-[(N-benzyloxycarbonyl-D-alanyl-L-alanyl-L-alanyl-L-alanyl)-amino]-~ ethylphosphonic acid of melting point 272-277C (decomposition);
!,~., [a]20 = _47.oo (c = 0,5% in acetic acid).
~J ~
~ 15 Example 4 . ~
" ~ .
(A) The process:
~, 10,85 g of dimethyl (lR)-l-[(N-benzyloxycarbonyl-D-. .
~`~, -leucyl-L-alanyl-L-alanyl)-amino]-ethylphosphonate were ; dissolved in 30 ml of a 35~ solution of hydrogen bromide ,;:
in glacial acetic acid and the mixture was stirred at room ', temperature for 4 hours. 130 ml of ether were then added - while stirring, the stirring was discontinued and the ether : decanted off. This procedure was repeated twice using 80 ml -, of ether each time. The residue was dissolved in 70 ml of methanol and tc the resulting solution were added 10 ml of , propylene oxide. After refrigerating overnight, the resulting , - white precipitate was filtered off and washed with ethanol and ether. The product was dried in vacuo to a constant weight of '' ', ~ 16 ,1 . ~ ~
:
~L090785 7.99 g; melting point 293-295C (decomposition). Recrystal-lisation from 500 ml of cold water and from 700 ml of ethanol gave 6.67 g of (lR)-l-(D-leucyl-L-alanyl-L-alanylamino)--ethylphosphonic acid of melting point 300-302C (decomposi-tion); [a]D = -129,3 (c = 1% in water).
., .
(B) The preparation of the starting material:
12.9 g (0.03 mol) of dimethyl (lR)-l-[(N-benzyloxy-carbonyl-L-alanyl-L-alanyl)-amino]-ethylphosphonate were dissolved in 150 ml of methanol containing 0.032 mol of hydrogen chloride. The solution was hydrogenated at room temperature and pressure in the presence of 1 g of 10~
palladium-on-charcoal until the hydrogen uptake ceased.
The catalyst was filtered off, the filtrate evaporated in vacuo and the oily hydrochloride evaporated twice with , 15 ethyl acetate.
,, .
; The product obtained according to the preceding paragraph and 10.9 g (0.03 mol) of the N-hydroxysuccinimide ester of i N-benzyloxycarbonyl-D-leucine were stirred in the presence ; of 100 ml of dry dimethylformamide. While stirring and ' 20 maintaining the temperature below 15C, 4.2 ml of dry . !
triethylamine were added dropwise. The mixture was then stirred overnight at room temperature. The triethylamine hydrochloride was filtered off and washed with a small amount of dimethylformamide. The filtrate was evaporated under an oil-pump vacuum and at a bath temperature of below - 40C. The residual oil was treated with 50 ml of water and the resulting mixture extracted with four 50 ml portions of chloroform. The combined or~anic phases were washed with a small vclume of 20% potassium carbonate solution and then .~.
, .
,,.
, .. .
~L090785 .
dried over sodium sulphate. The sodium sulphate was filtered . off and the filtrate evaporated under a water-pump vacuum.
The residue was re-evaporated twice with ethyl acetate.
Trituration with dry ether. filtration and vacuum drying yielded 15.5 g of dimethyl (lR)-l-[(N-benzyloxycarbonyl-D-leucyl-L-alanyl-L-alanyl)-amino]-ethylphosphonate of melting . point 163-167C. After recrvstallisation from a_etonitrile, . the melting point was 173-176C; [~]D0 = -36.6 (c = 1% in ; methanol).
! .
lo Example 5 ~' ` (A) The process:
In a manner analogous to that described in Example
4(A), 9.92 g of dimethyl (lR)-l-[(N-benzyloxycarbonyl-D-alanyl-L-alanyl)-amino]-ethylphosphonate were treated with 45 ml of a 35~ solution of hydrogen bromide in acetic acid.
,~
Working-up in the same manner as described in Example 4(A) : gave 6.16 g of crude product of melting point 282-285C
(decomposition) which, after recrystallisation from water/ethanol, yielded 5.47 g of pure (lR)-l-(D-alanyl-L-alanylamino)-ethylphosphonic acid of melting point 292-293C (decomposition);
]D0 = -101.2 (c = 1% in water).
ji; ~
.: (B) The preparation of the starting material:
;
~l In a manner analogous to that described in Example 4(B) . from 7.82 g (0,03 mol) of dimethyl (lR)-l-(L-alanylamino)-, -: 25 -e~hylphosphonate hydrochloride and 9.6 g (0,03 mol) of the - N-hydroxysuccinimide ester of N-benzyloxycarbonyl-D-alanine -. there were obtained 11.1 g of dimethyl (lR)-l-[(N-benzyloxy-carbonyl-D-alanyl-L-alanyl)-amino]-ethylphosphonate of melting ,....
~ ~ 18 - .
:10~0785 point 115-117C; [a]D = -28.4 (c = 0.5~ in methanol).
The following Example illustrates a typical pharmaceutical preparation containing a peptide derivative provided by the , present invention:
Example 6 , . .
A 1000 ml injection solution containing the following ingredients was prepared:
Per lO00 ml (lR)-l-(D-Alanyl-L-alanylamino)- ---ethylphosphonic acid 100.0 g ; Chlorocresol 1.0 g Acetic acid (glacial) 1.2 g Sodium hydroxide solution (0.1 N) ~;, q.s. pH 4.5 ~ 15 Water for injectionad 1000 ml `~ The (lR)-l-(D-alanyl-L-alanylamino)-ethylphosphonic acid was suspended in 500 ml of water for injection. The ~,1 chlorocresol was dissolved in 200 ml of water for injection "
and added to the first solution. The acetic acid was then added while stirring. A 0.1 N solution of sodium hydroxide in water for injection was added while stirring until a pH
value of 4.5 was obtained. The solution was then made up to 1000 ml with water for injection, filtered through a sterile .... ~! 0.22 micron membrane filter and filled into ampoules. The , . ..
- 25 ampoules were sealed and then sterilised by autoclaving at '~ 121C for 20 minutes.
- s.
~,, i ~
;
,,-.
,,."~ `, 1 1 9 ~ ~ i `''~;
. ," .
,~
Working-up in the same manner as described in Example 4(A) : gave 6.16 g of crude product of melting point 282-285C
(decomposition) which, after recrystallisation from water/ethanol, yielded 5.47 g of pure (lR)-l-(D-alanyl-L-alanylamino)-ethylphosphonic acid of melting point 292-293C (decomposition);
]D0 = -101.2 (c = 1% in water).
ji; ~
.: (B) The preparation of the starting material:
;
~l In a manner analogous to that described in Example 4(B) . from 7.82 g (0,03 mol) of dimethyl (lR)-l-(L-alanylamino)-, -: 25 -e~hylphosphonate hydrochloride and 9.6 g (0,03 mol) of the - N-hydroxysuccinimide ester of N-benzyloxycarbonyl-D-alanine -. there were obtained 11.1 g of dimethyl (lR)-l-[(N-benzyloxy-carbonyl-D-alanyl-L-alanyl)-amino]-ethylphosphonate of melting ,....
~ ~ 18 - .
:10~0785 point 115-117C; [a]D = -28.4 (c = 0.5~ in methanol).
The following Example illustrates a typical pharmaceutical preparation containing a peptide derivative provided by the , present invention:
Example 6 , . .
A 1000 ml injection solution containing the following ingredients was prepared:
Per lO00 ml (lR)-l-(D-Alanyl-L-alanylamino)- ---ethylphosphonic acid 100.0 g ; Chlorocresol 1.0 g Acetic acid (glacial) 1.2 g Sodium hydroxide solution (0.1 N) ~;, q.s. pH 4.5 ~ 15 Water for injectionad 1000 ml `~ The (lR)-l-(D-alanyl-L-alanylamino)-ethylphosphonic acid was suspended in 500 ml of water for injection. The ~,1 chlorocresol was dissolved in 200 ml of water for injection "
and added to the first solution. The acetic acid was then added while stirring. A 0.1 N solution of sodium hydroxide in water for injection was added while stirring until a pH
value of 4.5 was obtained. The solution was then made up to 1000 ml with water for injection, filtered through a sterile .... ~! 0.22 micron membrane filter and filled into ampoules. The , . ..
- 25 ampoules were sealed and then sterilised by autoclaving at '~ 121C for 20 minutes.
- s.
~,, i ~
;
,,-.
,,."~ `, 1 1 9 ~ ~ i `''~;
. ," .
Claims (10)
PROPERTY OR PRIVILEGE IS CLAIMED ARE DEFINED AS FOLLOWS:
1. A process for the manufacture of peptide derivatives of the general formula (I) wherein n stands for 1, 2 or 3; R1 is methyl and R2 and R3 are methyl, isopropyl or isobutyl with the proviso that R3 is methyl when n is 1;
R4 represents a hydroxy or methyl group; the configuration at the carbon atom designated as (a) is D; the configuration at the carbon atom desig-nated as (b) is L; and the configuration at the carbon atom denoted as (c) is (R), and of pharmaceutically acceptable salts thereof, which process comprises (a) cleaving off the protecting group(s) present in a compound of the general formula (II) wherein n, R1, R2 and R3 have the significances given above; R40 represents a methyl group or R41; R41 represents a hydroxy group or lower alkoxy protecting group; R5 represents a hydrogen atom or a protecting group; and the configurations at the carbon atoms designated as (a), (b) and (c) are as defined above, or (b) separating an (R,S)-diastereomeric compound corresponding to formula I into its diastereomers and isolating the (R)-diastereomer, and, if desired, converting an obtained compound of formula I into a pharmaceutically acceptable salt.
R4 represents a hydroxy or methyl group; the configuration at the carbon atom designated as (a) is D; the configuration at the carbon atom desig-nated as (b) is L; and the configuration at the carbon atom denoted as (c) is (R), and of pharmaceutically acceptable salts thereof, which process comprises (a) cleaving off the protecting group(s) present in a compound of the general formula (II) wherein n, R1, R2 and R3 have the significances given above; R40 represents a methyl group or R41; R41 represents a hydroxy group or lower alkoxy protecting group; R5 represents a hydrogen atom or a protecting group; and the configurations at the carbon atoms designated as (a), (b) and (c) are as defined above, or (b) separating an (R,S)-diastereomeric compound corresponding to formula I into its diastereomers and isolating the (R)-diastereomer, and, if desired, converting an obtained compound of formula I into a pharmaceutically acceptable salt.
2. A process as claimed in claim 1, wherein there is used a starting material of formula II in which R40 is hydroxy or a lower alkoxy group.
3. A process as claimed in claim 1 or claim 2, wherein there is used a starting material of formula II in which R2 is methyl.
4. A process as claimed in claim 2, wherein there is used a starting material of formula II in which R2 and R3 are methyl and n is 1.
5. A process as claimed in claim 2, wherein there is used a starting material of formula II in which R2 and R3 are methyl and n is 2.
6. A process as claimed in claim 2, wherein there is used a starting material of formula II in which R2 and R3 are methyl and n is 3.
7. A process as claimed in claim 2, wherein there is used a starting material of formula II in which R2 is methyl, R3 is isopropyl and n is 2.
8. A process as claimed in claim 2, wherein there is used a starting material of formula II in which R2 is methyl, R3 is isobutyl and n is 2.
9. A process according to claim 1, wherein a compound of formula II
is obtained by condensing a compound of the general formula (III) wherein m stands for zero, 1, 2 or 3; R1, R2, R40 and R41 have the significance given in claim 1; the configuration at the carbon atom desig-nated as (b) is L; and the configuration at the carbon atom designated as (c) is (R), with an appropriately protected .alpha.-amino acid, an appropriately protected dipeptide, an appropriately protected tripeptide, an appropriately protected tetrapeptide or a reactive derivative thereof.
is obtained by condensing a compound of the general formula (III) wherein m stands for zero, 1, 2 or 3; R1, R2, R40 and R41 have the significance given in claim 1; the configuration at the carbon atom desig-nated as (b) is L; and the configuration at the carbon atom designated as (c) is (R), with an appropriately protected .alpha.-amino acid, an appropriately protected dipeptide, an appropriately protected tripeptide, an appropriately protected tetrapeptide or a reactive derivative thereof.
10. A peptide derivative of formula I set forth in claim 1, when manufactured by the process claimed in claim 1 or 9, or by an obvious chemical equivalent thereof.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| GB20014/76A GB1577232A (en) | 1976-05-14 | 1976-05-14 | Peptide derivatives of phosphonic and phosphinic acids |
| GB20014/76 | 1976-05-14 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA1090785A true CA1090785A (en) | 1980-12-02 |
Family
ID=10138919
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA278,276A Expired CA1090785A (en) | 1976-05-14 | 1977-05-12 | Peptide derivatives and process for the preparation thereof |
Country Status (23)
| Country | Link |
|---|---|
| JP (1) | JPS52139023A (en) |
| AT (1) | AT355740B (en) |
| AU (1) | AU511452B2 (en) |
| BE (1) | BE854591A (en) |
| CA (1) | CA1090785A (en) |
| DE (1) | DE2721760A1 (en) |
| DK (1) | DK209877A (en) |
| ES (1) | ES458748A1 (en) |
| FI (1) | FI771518A7 (en) |
| FR (1) | FR2351124A1 (en) |
| GB (1) | GB1577232A (en) |
| GR (1) | GR73010B (en) |
| IL (1) | IL52040A (en) |
| IT (1) | IT1075484B (en) |
| LU (1) | LU77319A1 (en) |
| MC (1) | MC1146A1 (en) |
| NL (1) | NL7704370A (en) |
| NO (1) | NO771717L (en) |
| NZ (1) | NZ184047A (en) |
| PH (1) | PH14715A (en) |
| PT (1) | PT66544B (en) |
| SE (1) | SE7705628L (en) |
| ZA (1) | ZA772732B (en) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0002039A1 (en) * | 1977-11-19 | 1979-05-30 | Ciba-Geigy Ag | Phosphonous acid derivatives, processes for their preparation and their use in combating microorganisms |
| EP0010067B1 (en) * | 1978-10-05 | 1983-08-31 | Ciba-Geigy Ag | Process for influencing plant growth |
| EP0103867B1 (en) * | 1982-09-17 | 1986-12-17 | Kyowa Hakko Kogyo Co., Ltd. | Phosphorus-containing peptide derivative |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS51639B1 (en) * | 1971-07-28 | 1976-01-09 | ||
| IL48835A (en) * | 1975-01-27 | 1979-05-31 | Sparamedica Ag | Amino acyl and peptidyl derivatives of phophonic acids, their preparation and pharmaceutical compositions containingthem |
-
1976
- 1976-05-14 GB GB20014/76A patent/GB1577232A/en not_active Expired
-
1977
- 1977-04-21 NL NL7704370A patent/NL7704370A/en not_active Application Discontinuation
- 1977-04-29 IT IT23054/77A patent/IT1075484B/en active
- 1977-05-06 ZA ZA00772732A patent/ZA772732B/en unknown
- 1977-05-06 IL IL52040A patent/IL52040A/en unknown
- 1977-05-09 NZ NZ184047A patent/NZ184047A/en unknown
- 1977-05-09 AU AU25008/77A patent/AU511452B2/en not_active Expired
- 1977-05-10 PH PH19754A patent/PH14715A/en unknown
- 1977-05-12 CA CA278,276A patent/CA1090785A/en not_active Expired
- 1977-05-12 FI FI771518A patent/FI771518A7/fi not_active Application Discontinuation
- 1977-05-12 FR FR7714549A patent/FR2351124A1/en active Granted
- 1977-05-12 LU LU77319A patent/LU77319A1/xx unknown
- 1977-05-13 DE DE19772721760 patent/DE2721760A1/en not_active Withdrawn
- 1977-05-13 NO NO771717A patent/NO771717L/en unknown
- 1977-05-13 SE SE7705628A patent/SE7705628L/en unknown
- 1977-05-13 PT PT66544A patent/PT66544B/en unknown
- 1977-05-13 MC MC771242A patent/MC1146A1/en unknown
- 1977-05-13 JP JP5447577A patent/JPS52139023A/en active Pending
- 1977-05-13 AT AT345877A patent/AT355740B/en not_active IP Right Cessation
- 1977-05-13 DK DK209877A patent/DK209877A/en unknown
- 1977-05-13 ES ES458748A patent/ES458748A1/en not_active Expired
- 1977-05-13 BE BE177536A patent/BE854591A/en unknown
- 1977-05-13 GR GR53459A patent/GR73010B/el unknown
Also Published As
| Publication number | Publication date |
|---|---|
| ES458748A1 (en) | 1978-03-01 |
| IT1075484B (en) | 1985-04-22 |
| SE7705628L (en) | 1977-11-15 |
| AT355740B (en) | 1980-03-25 |
| PT66544B (en) | 1979-04-12 |
| FR2351124A1 (en) | 1977-12-09 |
| AU2500877A (en) | 1978-11-16 |
| ATA345877A (en) | 1979-08-15 |
| DK209877A (en) | 1977-11-15 |
| PH14715A (en) | 1981-11-13 |
| JPS52139023A (en) | 1977-11-19 |
| PT66544A (en) | 1977-06-01 |
| NZ184047A (en) | 1980-04-28 |
| IL52040A0 (en) | 1977-07-31 |
| NO771717L (en) | 1977-11-15 |
| NL7704370A (en) | 1977-11-16 |
| GR73010B (en) | 1984-01-24 |
| LU77319A1 (en) | 1978-06-26 |
| FI771518A7 (en) | 1977-11-15 |
| MC1146A1 (en) | 1978-01-30 |
| FR2351124B1 (en) | 1981-02-13 |
| GB1577232A (en) | 1980-10-22 |
| BE854591A (en) | 1977-11-14 |
| ZA772732B (en) | 1978-04-26 |
| DE2721760A1 (en) | 1977-12-01 |
| AU511452B2 (en) | 1980-08-21 |
| IL52040A (en) | 1979-11-30 |
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