CA1285698C - Process for the preparation of compounds containing carboxamide groups,in particular of peptides - Google Patents
Process for the preparation of compounds containing carboxamide groups,in particular of peptidesInfo
- Publication number
- CA1285698C CA1285698C CA000463073A CA463073A CA1285698C CA 1285698 C CA1285698 C CA 1285698C CA 000463073 A CA000463073 A CA 000463073A CA 463073 A CA463073 A CA 463073A CA 1285698 C CA1285698 C CA 1285698C
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Classifications
-
- 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
- C07K5/0802—Tripeptides with the first amino acid being neutral
- C07K5/0804—Tripeptides with the first amino acid being neutral and aliphatic
- C07K5/0806—Tripeptides with the first amino acid being neutral and aliphatic the side chain containing 0 or 1 carbon atoms, i.e. Gly, Ala
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/06—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length using protecting groups or activating agents
- C07K1/08—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length using protecting groups or activating agents using activating agents
- C07K1/082—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length using protecting groups or activating agents using activating agents containing phosphorus
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- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Biophysics (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Medicinal Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Analytical Chemistry (AREA)
- Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
- Peptides Or Proteins (AREA)
- Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
Abstract of the disclosure:
The invention relates to a process for the pre-paration of compounds containing carboxamide groups, in particular peptides, by reaction of compounds which con-tain a carboxyl group, in the presence of dialkylphos-phinic anhydrides, with compounds which contain a free amino group, which comprises maintaining the hydrogen ion concentration of the reaction mixture within a defined range during the reaction by metering in a base and, after the reaction is complete, eliminating, where appropriate, radicals introduced to protect other functional groups.
The invention relates to a process for the pre-paration of compounds containing carboxamide groups, in particular peptides, by reaction of compounds which con-tain a carboxyl group, in the presence of dialkylphos-phinic anhydrides, with compounds which contain a free amino group, which comprises maintaining the hydrogen ion concentration of the reaction mixture within a defined range during the reaction by metering in a base and, after the reaction is complete, eliminating, where appropriate, radicals introduced to protect other functional groups.
Description
A large number of processes is known for the pre-paration of carboxamide and peptide bonds tsee, for example, Houben-Weyl, Methoden der organ;schen Chemie (Methods of Organic Chemistry~, Vol. XV, Part II, pages 1-364. Also Angew~ Chemie _ , 129 ~1980)). All these processes aim, w;th vary;ng success, at ensuring the cri-ter;a, wh;ch are necessary for the synthesis of pepticles, of freedom from racemization, and of straightforward pro-cedure under mild conditions w;th high yields and ~lith 10 readily accessible starting ma.erials.
A process for the preparation of carboxamides is known from European Patent A1~56,618, in which compounds containing carboxyl groups are reacted with compounds ~Ihich contain a free amino group in the presence of 15 dialkylphosph;nic anhydrides. The dialkylphosphinic acid which is liberated during the reaction is bouncl by an excess of an organic base or a basic buffer which is added to the reaction mixture at the start.
The present process represents a new way OT opti-20 mi~ing tlle said conditions for an economic syntheo;s of pept;des and amides~
It has been found that compounds containin~ car-boxamide groups, in particular oligopep~ides, can be pre-pared under mild conditions and in ~ood yield by reacting ,~ 25 compounds which contain a ~ree amino group~ in particular ? aminocarboxylic acid derivatives or pept;des whose carboxyl ., .
,`~ " ' $~
;!;
/, ` ~
~ 5~
group is protected, in the presence of the anhydride of a dialkylphosphinic acid, with compounds which contain a free carboxyl group, in particular aminocarboxyl;c acids or peptides whose am;no ~roup is acylated. The ne~ pro-cess comprises mainta;ning the hydrogen ion concentrationof the react;on m;xture within a defined range during the reaction by metering in a base.
The radicals introduced to protect the functiona groups can, in the case of synthesis of peptides~ subse-1û quently be el;minated ;n a customary manner.
Anhydr;des of dialkylphosph;n;c acids are to be understood to be compounds of the formula I
R R
- O - P ~ O -- P. =. O ~ I
t t R R
.
in wh;ch R denotes alkyl. The substituents R shown in the formula can be identical or different. Anhydrides in ~Jh1ch both P atoms have identical substituents are preferred~
~ Jith;n the scope of the ;nvention, anhydrides o-f the formula I in which R ;s in each case a lower alkyl, prefcrably one havin~ 1 to l~ carbon atolns, are particu 2n larly suitable~
The dialkylphosph;nic anhydridès use~ accordin0 to the invent;on are colorless liquids. They are stable at room temperature ancl can be d;stilled under reduced pressure witllout decompos-ition. They are readily soluble in most ~ ~5~
non-aqueous solvents, especially in lipid solvents, such as chloroform or rnethylene chloride, but also in polar solvents, such as DMF and DMA.
Examples of anhydrides of dialkylphosphinic acid ~hich may be mentioned are: methylethylphosphinic anhy-dride, methylpropylphosphinic anhydride, methylbutylphos-phinic anhydride, diethylphosphinic anhydride, di-n-propyl-phosphinic anhydride and di-n-butylphosphinic anhydride.
The preparation of the dialkylphosphinic anhydrides can be carried out in a manner known per se by, for exam ple, rcact;on of the d;alkyLphosph;noyl chlorides with alkyl dialkylphosphinates at 150-160C ~Houben-Weylr Methoden der Organischen ~hemie, G. Thieme Verl., Stuttgart ~963r Vol. XII, pages 266 et seq.). Processes in which 1S dialkylphosphinic acids, their salts or esters are reacted with phosgene are particularly preferred (German Patent 2,129,583, German Offenlegungsschrift 2~225,545). ~' The process according to the invention is pre-ferably carr;ed out in a mixed aqueous, single- or 2-phase Z0 system~ within a narrow pH range, preferably at approxi-mately constan~ pH. It is possible for the pH of the reac~
tion mixtures to be 5-10 and, advantageously, it should be ;n the neutral or weakly acid range; pH values between '~
5.0 and 7.0 are particularly preferred. However, it ;s Z5 also possible to carry out the syntheses ;n the ~leakly ~
alkaline range. The p~l ;s preferably controlled by metered ~', addi.ion o-f concentrated aqueous solutions of alkali metal ~;~
hydrox;des~ but it i5 also po~sible to use organic bases, 5j~
such as N-ethylmorpholirle~ triethylamine or trialkylamine ';~
i~:
- ~ . - : .
' .~ : .
:
:~ 2~
having up to 6 carbon atorns.
For the preparation of oligopeptides by the pro-cess 2ccording to the ;nvent;on, the starting mater;als used are, on the one hand~ an am;noacid or a pept;de hav;ng a protected carboxy( group and, on the other-hand, an am;noacid or a peptide having a protected amino group.
It ;s poss;ble to use for the protection of the carboxyl groups all the protective ~roups customary in peptide synthesis. Esters of straight-chain or branched aliphatic alcohols, such as methanol, ethanol and tert~-butanol, are particularly suitable. Esters of aral;phatic alcohols, s~ch as benzyl alcohol or diphenylmethylcarb;nol, can also be used~
~ikew;se, it ;s possible to use for the protection of the amino groups all the protective groups custom3ry in peptide synthesis~ Examples of particularly suitable groups ~Ihich may be mentioned are the carboben~oxy radical and the carbo~tert.-butyloxy radical.
It is poss;ble to use as the solvent all the anhydrous ;nert solvents customary in pept;de synthesis~
for example methylene chloride, chloroform, d~methyl,or~
mamide, dimethylacetamide~ d;oxane or tetrahydrofurarl.
Solvents wh;ch can be used in the sin~le phase mixed a~lueous procedure for the reaction are mixtures of water and an organic solvent which is misc;ble with water~
such as, for example, d;oxane, tetrahydrofuran, dimethyl-formamide or dime~hylacetamide. The use of systems of this type ;s especially advantageous when linking pep~i~es ~hich ~re ma;nly solubLe in wrter.
.
For the two-phase mixed aqueous procedure for the react,on, it is possible to use systems such as~ for example, ethyl acetate, propyl acetate, n-butyl acetate, methylene chloride, glycol dimethyl ether, 3-methyltetra-S hydrofuran and chlorof~rm, each ;n a heterogeneous mix-ture with water.
As a rule, the reaction takes place suff;c;ently rap;dly at room temperature. Gentle warm;ng is not injur;ous. Higher tcmperatures~ say above 30Cr are not 10 adv;sable, espec;ally ;n pept;de synthes;s, because of the dar1ger of racemization. A reaction temperature between 0 and 3~C is preferred, and one between 5 and ?5C is particularly preferred.
The process accord;ng to the ;nvent;on makes it possible for the first time to follo~ the course of the reaction by the consumption of the base which is used when the pH is approx;mately constant. For this purposer the reaction is preferably carried out in an automatic record~
ing pi~-stat, advarlta~eously with v;gorous mixin~ of the m;xed aqueous system conta;n;ng the reactants, the carboxyl component, thc am;rle component and the d;alkylphosphin1c anhydride. The graph provided by the recorder of the con~
sumptiol1 of base as a function of t;me sho~s a curvc which becoll1es asymptotic toward the end of the reaction ~cf. the curves repl~oduced ;n the Fig.). Thus, tl)e process accord-ing to the invention makes it possible to detect the end-point of the synthet;c react;on ;n a s~raightforward manner~
It ought to be noted that the "apparent" pH va~ues in these m;xed aqueous systems measured us;ng the rnethod .
~ "
, ' : `
~2~
descr;bed can differ from the true pH values in these systems.
When buf-fer solutions are used in a known nanner to trap the d;all;ylphosphinic acid, then it is unavoidable 5 that excess salts, composed of inorganic salts, occur ;n the mother l;quors of the batches. When carrying out the process industrially, they make it difficult to recover the dialkylphosphinic acid produced from the condensing agent, and they can give rise to effluent problems.
Durins isolatiorl of the reaction procluGt by extrac tion from alkali solution with a lipoid phase which is mis-cible l~ith water to only a limited extent, the alkali ~etal salts wll;ch are forrned dur;ng the course of the re-action when alkali metal hydroxides are used as the bases 1S in this variant of the process according to the ;nvention are, because of their relat;ve~y h;gh hyclroph;licity and in contr~st to the salts resulting when some tertiary organic bases are used, relatively easy to remove.
Furthcrmore~ contam;nation of the lipo;d phase by er~tracted 20 tert;ary base ;s avoided in this procedure.
The process accord;ng to the invent;on ;s stra;~ht-forward ~o carry out and provides pep~ides of high op~ical pur;ty and in hi~h yield~ In addition, ;t is economical ar1d env;ror1lnentally acceptable.
Z5 The dialkylphosphirl;c anhydr;des have low Molecular weights, are readily obtained and pur;fied and have a high proportion o~ reac~ive groups per unit wei~ht and have h;gh lipophil;c;tyu The dialkylphosphin;c anhydr;des and the corresponding dialkylphosphinic acids are lipicl~
, soluble. Th`i 5 nlakes it possible to work up water-soluble peptide derivat;ves via a first precipitation scep using suitable lipid solvents7 The dialkylphosphinic acid obtained from the d;alkylphosphin;c anhydr;de dur;ng the course of the pep t;de synthesis can be recovered fro~n the solut;ons remain-in3 after the synthet;c react;on~ Recovery of the ~ialkyl-phosph;nic acids from a relatively large amount of aqueous mother liquors from the synthesis can be carried out by extract;on w;th solvents such as chloroform and isobutanol followed by work-up by distillation. In this context, it ;s of particular industrial interest that the dialkyl~
phosphinic ac;ds can be d;stilled in vacuo without decom-pos;t;on~ The d;a~kylphosph;n;c acids thus recovered from the ~lork-up can then readily be converted ;nto the corres-pondin~ dialkylphos~hinic anhydrides by the process of German Offenlegungsschrift 2,225,545.
In the mixed aqueous procedure described, it is possible to replace the organic base by alkali metal hydroxide solutions, and this considerably simplifies the recovery of the dialkylphosphinic acids during the work~up after the synthes;s as clescri~ed above. In th;s case, it is pos3;ble to d1spense w;th the extract10n step. After liberationr the dialkylpho3phinic ac;d can then be dis-t;lled directly out of the evaporated mother liquor fromthe synthesis or, ;n the two-phase procedure, it js recovered From the aqueous phase, after removal of the l;poid phase, by acidification and extraction. The neutral inorgarlîc salts then relnain in the aqueous phase.
~ . ~
: . '' '' ~, , ':
,, . ' ~2~5~
g Example 1:
Ethy~ ester of carbobenzoxyglycine:
__ ____~
7.0 g (~.OS mole) of H-Gly-OCH3.HCl are added, with stirring at -5C, to a solution of 10.5 9 (0.05 Inole~ oF
5 carbobenzoxyglyc;ne in 120 ml of ethyl acetate. 20 9 of methylethyLphosphinic anhydride are added dropwise to the resulting suspension~ The pH of the reaction mixture is then adjusted to 7.0 at a temperature of 0 to ~10C by dropuise adcJition of 4 N NaOH using an autotitrator, and 10 the batch~ ~Ihlch is thoroughly mixed by h;gh-speed stirr ing, is maintailled a~ this pH un~il the recorder connected to the autotitrator shows that the curve of consumption of sodium hydroxide SOtUtiOIl has become asymp.otic with the time axis. This is the case after about 60 r,linutes.
15 The ethyl acetate phase is now removed, extracted twice w;th 50 rnl of saturated sodium bicarbonate sotution, dried with sodium sulfate, and, a~ter removal o-f the solvent in vacuo at room temperature, 1~.25 g (84% of theory) of Z~
dipeptide ester having a meltin~ point of 81C are 20 obta;ned.
Example 2:
Carboben,oxyphenyla~anine Gyclohexylam;de:
A process for the preparation of carboxamides is known from European Patent A1~56,618, in which compounds containing carboxyl groups are reacted with compounds ~Ihich contain a free amino group in the presence of 15 dialkylphosph;nic anhydrides. The dialkylphosphinic acid which is liberated during the reaction is bouncl by an excess of an organic base or a basic buffer which is added to the reaction mixture at the start.
The present process represents a new way OT opti-20 mi~ing tlle said conditions for an economic syntheo;s of pept;des and amides~
It has been found that compounds containin~ car-boxamide groups, in particular oligopep~ides, can be pre-pared under mild conditions and in ~ood yield by reacting ,~ 25 compounds which contain a ~ree amino group~ in particular ? aminocarboxylic acid derivatives or pept;des whose carboxyl ., .
,`~ " ' $~
;!;
/, ` ~
~ 5~
group is protected, in the presence of the anhydride of a dialkylphosphinic acid, with compounds which contain a free carboxyl group, in particular aminocarboxyl;c acids or peptides whose am;no ~roup is acylated. The ne~ pro-cess comprises mainta;ning the hydrogen ion concentrationof the react;on m;xture within a defined range during the reaction by metering in a base.
The radicals introduced to protect the functiona groups can, in the case of synthesis of peptides~ subse-1û quently be el;minated ;n a customary manner.
Anhydr;des of dialkylphosph;n;c acids are to be understood to be compounds of the formula I
R R
- O - P ~ O -- P. =. O ~ I
t t R R
.
in wh;ch R denotes alkyl. The substituents R shown in the formula can be identical or different. Anhydrides in ~Jh1ch both P atoms have identical substituents are preferred~
~ Jith;n the scope of the ;nvention, anhydrides o-f the formula I in which R ;s in each case a lower alkyl, prefcrably one havin~ 1 to l~ carbon atolns, are particu 2n larly suitable~
The dialkylphosph;nic anhydridès use~ accordin0 to the invent;on are colorless liquids. They are stable at room temperature ancl can be d;stilled under reduced pressure witllout decompos-ition. They are readily soluble in most ~ ~5~
non-aqueous solvents, especially in lipid solvents, such as chloroform or rnethylene chloride, but also in polar solvents, such as DMF and DMA.
Examples of anhydrides of dialkylphosphinic acid ~hich may be mentioned are: methylethylphosphinic anhy-dride, methylpropylphosphinic anhydride, methylbutylphos-phinic anhydride, diethylphosphinic anhydride, di-n-propyl-phosphinic anhydride and di-n-butylphosphinic anhydride.
The preparation of the dialkylphosphinic anhydrides can be carried out in a manner known per se by, for exam ple, rcact;on of the d;alkyLphosph;noyl chlorides with alkyl dialkylphosphinates at 150-160C ~Houben-Weylr Methoden der Organischen ~hemie, G. Thieme Verl., Stuttgart ~963r Vol. XII, pages 266 et seq.). Processes in which 1S dialkylphosphinic acids, their salts or esters are reacted with phosgene are particularly preferred (German Patent 2,129,583, German Offenlegungsschrift 2~225,545). ~' The process according to the invention is pre-ferably carr;ed out in a mixed aqueous, single- or 2-phase Z0 system~ within a narrow pH range, preferably at approxi-mately constan~ pH. It is possible for the pH of the reac~
tion mixtures to be 5-10 and, advantageously, it should be ;n the neutral or weakly acid range; pH values between '~
5.0 and 7.0 are particularly preferred. However, it ;s Z5 also possible to carry out the syntheses ;n the ~leakly ~
alkaline range. The p~l ;s preferably controlled by metered ~', addi.ion o-f concentrated aqueous solutions of alkali metal ~;~
hydrox;des~ but it i5 also po~sible to use organic bases, 5j~
such as N-ethylmorpholirle~ triethylamine or trialkylamine ';~
i~:
- ~ . - : .
' .~ : .
:
:~ 2~
having up to 6 carbon atorns.
For the preparation of oligopeptides by the pro-cess 2ccording to the ;nvent;on, the starting mater;als used are, on the one hand~ an am;noacid or a pept;de hav;ng a protected carboxy( group and, on the other-hand, an am;noacid or a peptide having a protected amino group.
It ;s poss;ble to use for the protection of the carboxyl groups all the protective ~roups customary in peptide synthesis. Esters of straight-chain or branched aliphatic alcohols, such as methanol, ethanol and tert~-butanol, are particularly suitable. Esters of aral;phatic alcohols, s~ch as benzyl alcohol or diphenylmethylcarb;nol, can also be used~
~ikew;se, it ;s possible to use for the protection of the amino groups all the protective groups custom3ry in peptide synthesis~ Examples of particularly suitable groups ~Ihich may be mentioned are the carboben~oxy radical and the carbo~tert.-butyloxy radical.
It is poss;ble to use as the solvent all the anhydrous ;nert solvents customary in pept;de synthesis~
for example methylene chloride, chloroform, d~methyl,or~
mamide, dimethylacetamide~ d;oxane or tetrahydrofurarl.
Solvents wh;ch can be used in the sin~le phase mixed a~lueous procedure for the reaction are mixtures of water and an organic solvent which is misc;ble with water~
such as, for example, d;oxane, tetrahydrofuran, dimethyl-formamide or dime~hylacetamide. The use of systems of this type ;s especially advantageous when linking pep~i~es ~hich ~re ma;nly solubLe in wrter.
.
For the two-phase mixed aqueous procedure for the react,on, it is possible to use systems such as~ for example, ethyl acetate, propyl acetate, n-butyl acetate, methylene chloride, glycol dimethyl ether, 3-methyltetra-S hydrofuran and chlorof~rm, each ;n a heterogeneous mix-ture with water.
As a rule, the reaction takes place suff;c;ently rap;dly at room temperature. Gentle warm;ng is not injur;ous. Higher tcmperatures~ say above 30Cr are not 10 adv;sable, espec;ally ;n pept;de synthes;s, because of the dar1ger of racemization. A reaction temperature between 0 and 3~C is preferred, and one between 5 and ?5C is particularly preferred.
The process accord;ng to the ;nvent;on makes it possible for the first time to follo~ the course of the reaction by the consumption of the base which is used when the pH is approx;mately constant. For this purposer the reaction is preferably carried out in an automatic record~
ing pi~-stat, advarlta~eously with v;gorous mixin~ of the m;xed aqueous system conta;n;ng the reactants, the carboxyl component, thc am;rle component and the d;alkylphosphin1c anhydride. The graph provided by the recorder of the con~
sumptiol1 of base as a function of t;me sho~s a curvc which becoll1es asymptotic toward the end of the reaction ~cf. the curves repl~oduced ;n the Fig.). Thus, tl)e process accord-ing to the invention makes it possible to detect the end-point of the synthet;c react;on ;n a s~raightforward manner~
It ought to be noted that the "apparent" pH va~ues in these m;xed aqueous systems measured us;ng the rnethod .
~ "
, ' : `
~2~
descr;bed can differ from the true pH values in these systems.
When buf-fer solutions are used in a known nanner to trap the d;all;ylphosphinic acid, then it is unavoidable 5 that excess salts, composed of inorganic salts, occur ;n the mother l;quors of the batches. When carrying out the process industrially, they make it difficult to recover the dialkylphosphinic acid produced from the condensing agent, and they can give rise to effluent problems.
Durins isolatiorl of the reaction procluGt by extrac tion from alkali solution with a lipoid phase which is mis-cible l~ith water to only a limited extent, the alkali ~etal salts wll;ch are forrned dur;ng the course of the re-action when alkali metal hydroxides are used as the bases 1S in this variant of the process according to the ;nvention are, because of their relat;ve~y h;gh hyclroph;licity and in contr~st to the salts resulting when some tertiary organic bases are used, relatively easy to remove.
Furthcrmore~ contam;nation of the lipo;d phase by er~tracted 20 tert;ary base ;s avoided in this procedure.
The process accord;ng to the invent;on ;s stra;~ht-forward ~o carry out and provides pep~ides of high op~ical pur;ty and in hi~h yield~ In addition, ;t is economical ar1d env;ror1lnentally acceptable.
Z5 The dialkylphosphirl;c anhydr;des have low Molecular weights, are readily obtained and pur;fied and have a high proportion o~ reac~ive groups per unit wei~ht and have h;gh lipophil;c;tyu The dialkylphosphin;c anhydr;des and the corresponding dialkylphosphinic acids are lipicl~
, soluble. Th`i 5 nlakes it possible to work up water-soluble peptide derivat;ves via a first precipitation scep using suitable lipid solvents7 The dialkylphosphinic acid obtained from the d;alkylphosphin;c anhydr;de dur;ng the course of the pep t;de synthesis can be recovered fro~n the solut;ons remain-in3 after the synthet;c react;on~ Recovery of the ~ialkyl-phosph;nic acids from a relatively large amount of aqueous mother liquors from the synthesis can be carried out by extract;on w;th solvents such as chloroform and isobutanol followed by work-up by distillation. In this context, it ;s of particular industrial interest that the dialkyl~
phosphinic ac;ds can be d;stilled in vacuo without decom-pos;t;on~ The d;a~kylphosph;n;c acids thus recovered from the ~lork-up can then readily be converted ;nto the corres-pondin~ dialkylphos~hinic anhydrides by the process of German Offenlegungsschrift 2,225,545.
In the mixed aqueous procedure described, it is possible to replace the organic base by alkali metal hydroxide solutions, and this considerably simplifies the recovery of the dialkylphosphinic acids during the work~up after the synthes;s as clescri~ed above. In th;s case, it is pos3;ble to d1spense w;th the extract10n step. After liberationr the dialkylpho3phinic ac;d can then be dis-t;lled directly out of the evaporated mother liquor fromthe synthesis or, ;n the two-phase procedure, it js recovered From the aqueous phase, after removal of the l;poid phase, by acidification and extraction. The neutral inorgarlîc salts then relnain in the aqueous phase.
~ . ~
: . '' '' ~, , ':
,, . ' ~2~5~
g Example 1:
Ethy~ ester of carbobenzoxyglycine:
__ ____~
7.0 g (~.OS mole) of H-Gly-OCH3.HCl are added, with stirring at -5C, to a solution of 10.5 9 (0.05 Inole~ oF
5 carbobenzoxyglyc;ne in 120 ml of ethyl acetate. 20 9 of methylethyLphosphinic anhydride are added dropwise to the resulting suspension~ The pH of the reaction mixture is then adjusted to 7.0 at a temperature of 0 to ~10C by dropuise adcJition of 4 N NaOH using an autotitrator, and 10 the batch~ ~Ihlch is thoroughly mixed by h;gh-speed stirr ing, is maintailled a~ this pH un~il the recorder connected to the autotitrator shows that the curve of consumption of sodium hydroxide SOtUtiOIl has become asymp.otic with the time axis. This is the case after about 60 r,linutes.
15 The ethyl acetate phase is now removed, extracted twice w;th 50 rnl of saturated sodium bicarbonate sotution, dried with sodium sulfate, and, a~ter removal o-f the solvent in vacuo at room temperature, 1~.25 g (84% of theory) of Z~
dipeptide ester having a meltin~ point of 81C are 20 obta;ned.
Example 2:
Carboben,oxyphenyla~anine Gyclohexylam;de:
3.0 ~t ~0~01 mole~ of Z-Phe-0!t and 1nO ~ ~0~01 nnole, 1~2 nnl) of cyclohexylalnine are dissolve(l in a nlixture of 25 2n ml of tetrahydrofuran and 5 Ml oF water~ After cool;ng the reac~ion rnixture to ~-5C~ 4 y of methylethylphos phinic anhydride are added, with stirting, and the p~l of ~he reaction solution is acljusted to 6.0 with ~ N NaOH~
and is maintained constant throughout the react;on time by : , .. ' . ' , metered addition of sod;um hydroxide solution as described in Example 1~ lhe reaction is virtually complete after 60 minutes, as can be seen from the consumption of alkali metal hydroxide. The reaction sqlution is evaporated in S vacuo at room temperature, the residue is taken up with ethyl acetate, and the ethyl acetate solution is washed with 5% strength potassium bisulfate soLut;on, saturated sodium b;carbona-te solution and water, dried over sod;um sulfate and, after rernoval of the solven~ in vacuo at room telnperature and dr~ of the product in vacuo over P20S~
3.0 g of final product of melting point 167C are obtained, Ca~g = -3~0 ~c - 1, DM~).
Example 3:
Z-Trp-Gly-OCH
3r35 B (0~01 mole) of Z Trp-~ll are dissolved in 20 ml of isopropyl acetate, 1.25 9 ~0.01 mole) of H~Gly-OCH~
is added, ~he vigorously stirred suspension is cooled to -5C, and 4.n ml of methylethylphosphinic anhydride are added at this temperature, at the same time adjustin~ the pH to 5.7 by metered addition of 4 N NaOH using an auto-titrator as described in Exa0ple 1. ~Ihen the phases are thoroughly mixed~ the react;on is finished within ~0 min-utes. The ~thyl ace~ate phase is rernoved and ~lorked up as described in ~xampLe 2. ~ield: 3.53 g ~86% of theory) ~a~D-13.5 ~c ~ 0~1, glacial acet1c acid).
Exam~_e 4:
Z-Phe-Arq-Trp-Gly-OCH7 1.55 g (0.005 mole) of H Trp-Gly-OCH~ cl is added to a solution o~ 2~Z6 ~ ~0~005 rnole) o~ Z Phe-Ar~-O~ in ~-:'-.' ' , ~ - " ' ' - ' -,: .
. . .
25 ml of isopropyl acetate, and the v;gorously st;rred suspens;on ls cooled to -5C. Then ~ ml of nlethylethyl-phosphinic ar,hydride are added dropwise, maintaining the pH constant at 5.2 using 4 N NaOH as describeà in Example 1. After 15 minutes, the temperature of the reaction m;xture is allowed to reach room temperaturen The reaction ;s virtually complete after ~0 m;nutes as is shown by the graph of the consumption of NaOH with reaction time. The ethyl acetate phase is no~l removed, washed with water and saturated sodium bicarbonate solution, and the reaction product is isolated from the dried solution by evaporation in vacuo at room temperature and di0estion of the residue with absolute diethyl ether~
Yield after recrystallizatiorl -From ethanol/ether:
; 15 3.0 9 (84.5% of theol~y~, Cu~2DO~ -25.7 (c ~- n.1~ DMF)~ ;
Example 5 _ Z-Lys(~oc~-va~-T~ 3 -1~91 ~ tO.OOS mole~ of Z-Lys(Boc)-OH are dissolved ;n 2S ml of butyl acetate ~Ihich is saturated with ~later~
and 1.65 ~ ~0~005 mole) of H~Val-Tyr~OCH~.HCl are added, the ~igorously stirred reaction mixture is cooled to -5C~
and the pH of the rapidly stirred mixture is brought to 7.0 ith ~) N NaOH as descr;L)ed in Example 1, and it is main~
ta;ned at this value throu~llout the reaction time ~70 mirl-utes). The temperature during the first 10 minutes of ther~act;on t;me ;s maintained at 5C ~o 0C, and is th~n ~llowed to ~arm to room temperature. The worl;ing up of the but~l acetate phase containing the final product is carried out as indicated in Example 2~
-.
' ~L2'~
~- 12 -Yield after recrystallization from ethanol/ether:
3.0 9 (91% of theory), Ca~ = ~25 (c = 1~ ethanol)r Example 6 Z-Gly-Leu-Arg-OCH
-- ~ 3 1.3 ml of methylethylphosph;nic anhydride is added to a so!.ution of 642 mg of Z-Gly-Leu-OH and 449 mg of H-Arg-OMe.HCl in 3 ml of dimethylacetamide and 0.5 ml of water, ancl th~ pll dur;ng the reaction which no~ takes place is maintained at 7.2 with a mixture of N ethylmor-phol;ne and wa~er t1~1, vollvol) us;ng a pH-statO The reaction is co~plete after 40 minutes accord-in~ to the graph on the recorder~ The work;ng up is carried out as described in Example 2~ but without the extraction of the ethyl acetate phase with 5% potassium bisulfate solution mentioned there~
Yield: 700 mg (71X of theory~
C~X]2DO = -2404 (C = 1, CH30H)
and is maintained constant throughout the react;on time by : , .. ' . ' , metered addition of sod;um hydroxide solution as described in Example 1~ lhe reaction is virtually complete after 60 minutes, as can be seen from the consumption of alkali metal hydroxide. The reaction sqlution is evaporated in S vacuo at room temperature, the residue is taken up with ethyl acetate, and the ethyl acetate solution is washed with 5% strength potassium bisulfate soLut;on, saturated sodium b;carbona-te solution and water, dried over sod;um sulfate and, after rernoval of the solven~ in vacuo at room telnperature and dr~ of the product in vacuo over P20S~
3.0 g of final product of melting point 167C are obtained, Ca~g = -3~0 ~c - 1, DM~).
Example 3:
Z-Trp-Gly-OCH
3r35 B (0~01 mole) of Z Trp-~ll are dissolved in 20 ml of isopropyl acetate, 1.25 9 ~0.01 mole) of H~Gly-OCH~
is added, ~he vigorously stirred suspension is cooled to -5C, and 4.n ml of methylethylphosphinic anhydride are added at this temperature, at the same time adjustin~ the pH to 5.7 by metered addition of 4 N NaOH using an auto-titrator as described in Exa0ple 1. ~Ihen the phases are thoroughly mixed~ the react;on is finished within ~0 min-utes. The ~thyl ace~ate phase is rernoved and ~lorked up as described in ~xampLe 2. ~ield: 3.53 g ~86% of theory) ~a~D-13.5 ~c ~ 0~1, glacial acet1c acid).
Exam~_e 4:
Z-Phe-Arq-Trp-Gly-OCH7 1.55 g (0.005 mole) of H Trp-Gly-OCH~ cl is added to a solution o~ 2~Z6 ~ ~0~005 rnole) o~ Z Phe-Ar~-O~ in ~-:'-.' ' , ~ - " ' ' - ' -,: .
. . .
25 ml of isopropyl acetate, and the v;gorously st;rred suspens;on ls cooled to -5C. Then ~ ml of nlethylethyl-phosphinic ar,hydride are added dropwise, maintaining the pH constant at 5.2 using 4 N NaOH as describeà in Example 1. After 15 minutes, the temperature of the reaction m;xture is allowed to reach room temperaturen The reaction ;s virtually complete after ~0 m;nutes as is shown by the graph of the consumption of NaOH with reaction time. The ethyl acetate phase is no~l removed, washed with water and saturated sodium bicarbonate solution, and the reaction product is isolated from the dried solution by evaporation in vacuo at room temperature and di0estion of the residue with absolute diethyl ether~
Yield after recrystallizatiorl -From ethanol/ether:
; 15 3.0 9 (84.5% of theol~y~, Cu~2DO~ -25.7 (c ~- n.1~ DMF)~ ;
Example 5 _ Z-Lys(~oc~-va~-T~ 3 -1~91 ~ tO.OOS mole~ of Z-Lys(Boc)-OH are dissolved ;n 2S ml of butyl acetate ~Ihich is saturated with ~later~
and 1.65 ~ ~0~005 mole) of H~Val-Tyr~OCH~.HCl are added, the ~igorously stirred reaction mixture is cooled to -5C~
and the pH of the rapidly stirred mixture is brought to 7.0 ith ~) N NaOH as descr;L)ed in Example 1, and it is main~
ta;ned at this value throu~llout the reaction time ~70 mirl-utes). The temperature during the first 10 minutes of ther~act;on t;me ;s maintained at 5C ~o 0C, and is th~n ~llowed to ~arm to room temperature. The worl;ing up of the but~l acetate phase containing the final product is carried out as indicated in Example 2~
-.
' ~L2'~
~- 12 -Yield after recrystallization from ethanol/ether:
3.0 9 (91% of theory), Ca~ = ~25 (c = 1~ ethanol)r Example 6 Z-Gly-Leu-Arg-OCH
-- ~ 3 1.3 ml of methylethylphosph;nic anhydride is added to a so!.ution of 642 mg of Z-Gly-Leu-OH and 449 mg of H-Arg-OMe.HCl in 3 ml of dimethylacetamide and 0.5 ml of water, ancl th~ pll dur;ng the reaction which no~ takes place is maintained at 7.2 with a mixture of N ethylmor-phol;ne and wa~er t1~1, vollvol) us;ng a pH-statO The reaction is co~plete after 40 minutes accord-in~ to the graph on the recorder~ The work;ng up is carried out as described in Example 2~ but without the extraction of the ethyl acetate phase with 5% potassium bisulfate solution mentioned there~
Yield: 700 mg (71X of theory~
C~X]2DO = -2404 (C = 1, CH30H)
Claims (11)
1. A process for the preparation of a compound containing carboxamide groups by reaction of a compound which contains a carboxyl group, in the presence of dialkylphosphinic anhydrides, with a compound which contains a free amino group, which comprises maintaining the measured pH value in the reaction mixture approximately constant within the pH range of 5-10 during the reaction by metering in a base.
2. The process as claimed in claim 1 for the preparation of peptides.
3. The process as claimed in claim 1, wherein the reaction is carried out in a homogeneous or heterogeneous mixed aqueous medium.
4. The process as claimed in claim 3, wherein the pH
during the reaction is maintained at 7-10 by metered addition of bases.
during the reaction is maintained at 7-10 by metered addition of bases.
5. The process as claimed in claim 1, wherein the bases used are aqueous solutions of alkali metal hydroxides.
6. The process as claimed in claim 1, wherein trialkylamines are used as the bases.
7. The process as claimed in claim 1, wherein the end-point of the synthetic reaction is established with the aid of the graph recorded by a pH-stat (consumption of base versus reaction time).
8. The process as claimed in claim 2 or 3, wherein the reaction is carried out between 0 and 30°C.
9. The process as claimed in claim 4, 5 or 6, wherein the reaction is carried out between 0 and 30°C.
10. The process as claimed in claim 7, wherein the reaction is carried out between 0 and 30°C.
11. The process as claimed in claim 1 further comprising the step of eliminating, after the reaction is complete, radicals which have been introduced to protect other functional groups.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| DEP3333456.0 | 1983-09-16 | ||
| DE19833333456 DE3333456A1 (en) | 1983-09-16 | 1983-09-16 | METHOD FOR PRODUCING COMPOUNDS CONTAINING CARBONIC ACID AMIDE, IN PARTICULAR PEPTIDES |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA1285698C true CA1285698C (en) | 1991-07-02 |
Family
ID=6209237
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA000463073A Expired - Lifetime CA1285698C (en) | 1983-09-16 | 1984-09-13 | Process for the preparation of compounds containing carboxamide groups,in particular of peptides |
Country Status (13)
| Country | Link |
|---|---|
| EP (1) | EP0135183B1 (en) |
| JP (1) | JPS6084249A (en) |
| AT (1) | ATE33251T1 (en) |
| AU (1) | AU570724B2 (en) |
| CA (1) | CA1285698C (en) |
| DE (2) | DE3333456A1 (en) |
| DK (1) | DK160041C (en) |
| ES (1) | ES535919A0 (en) |
| GR (1) | GR80370B (en) |
| HU (1) | HU189936B (en) |
| IE (1) | IE58082B1 (en) |
| IL (1) | IL72943A (en) |
| PT (1) | PT79195B (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE3333455A1 (en) * | 1983-09-16 | 1985-04-11 | Hoechst Ag, 6230 Frankfurt | METHOD FOR PRODUCING N-ALKYLATED DIPEPTIDES AND THEIR ESTERS |
| DE3333454A1 (en) * | 1983-09-16 | 1985-04-11 | Hoechst Ag, 6230 Frankfurt | METHOD FOR PRODUCING N-ALKYLATED DIPEPTIDES AND THEIR ESTERS |
Family Cites Families (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE2901843A1 (en) * | 1979-01-18 | 1980-07-31 | Hoechst Ag | METHOD FOR PRODUCING CARBONIC ACID AMIDES AND PEPTIDES |
| DE3101427A1 (en) * | 1981-01-17 | 1982-09-02 | Hoechst Ag, 6000 Frankfurt | "METHOD FOR PRODUCING COMPOUNDS CONTAINING CARBONIC ACID AMIDE, IN PARTICULAR PEPTIDES" |
| DE3333455A1 (en) * | 1983-09-16 | 1985-04-11 | Hoechst Ag, 6230 Frankfurt | METHOD FOR PRODUCING N-ALKYLATED DIPEPTIDES AND THEIR ESTERS |
| DE3333454A1 (en) * | 1983-09-16 | 1985-04-11 | Hoechst Ag, 6230 Frankfurt | METHOD FOR PRODUCING N-ALKYLATED DIPEPTIDES AND THEIR ESTERS |
-
1983
- 1983-09-16 DE DE19833333456 patent/DE3333456A1/en not_active Withdrawn
-
1984
- 1984-09-07 AT AT84110679T patent/ATE33251T1/en not_active IP Right Cessation
- 1984-09-07 EP EP84110679A patent/EP0135183B1/en not_active Expired
- 1984-09-07 DE DE8484110679T patent/DE3470162D1/en not_active Expired
- 1984-09-10 HU HU843418A patent/HU189936B/en unknown
- 1984-09-13 PT PT79195A patent/PT79195B/en active IP Right Revival
- 1984-09-13 CA CA000463073A patent/CA1285698C/en not_active Expired - Lifetime
- 1984-09-14 DK DK440184A patent/DK160041C/en not_active IP Right Cessation
- 1984-09-14 AU AU33073/84A patent/AU570724B2/en not_active Expired
- 1984-09-14 IE IE235284A patent/IE58082B1/en not_active IP Right Cessation
- 1984-09-14 JP JP59191870A patent/JPS6084249A/en active Granted
- 1984-09-14 GR GR80370A patent/GR80370B/en unknown
- 1984-09-14 ES ES535919A patent/ES535919A0/en active Granted
- 1984-09-14 IL IL72943A patent/IL72943A/en not_active IP Right Cessation
Also Published As
| Publication number | Publication date |
|---|---|
| DE3470162D1 (en) | 1988-05-05 |
| DK160041C (en) | 1991-06-10 |
| ES8505939A1 (en) | 1985-06-16 |
| GR80370B (en) | 1985-01-11 |
| EP0135183A2 (en) | 1985-03-27 |
| EP0135183B1 (en) | 1988-03-30 |
| ES535919A0 (en) | 1985-06-16 |
| PT79195A (en) | 1984-10-01 |
| HU189936B (en) | 1986-08-28 |
| AU3307384A (en) | 1985-03-21 |
| DK160041B (en) | 1991-01-21 |
| ATE33251T1 (en) | 1988-04-15 |
| AU570724B2 (en) | 1988-03-24 |
| EP0135183A3 (en) | 1986-03-05 |
| IL72943A (en) | 1988-07-31 |
| HUT36083A (en) | 1985-08-28 |
| PT79195B (en) | 1986-09-10 |
| DE3333456A1 (en) | 1985-04-11 |
| JPH0417177B2 (en) | 1992-03-25 |
| JPS6084249A (en) | 1985-05-13 |
| IE842352L (en) | 1985-03-16 |
| IE58082B1 (en) | 1993-06-30 |
| DK440184D0 (en) | 1984-09-14 |
| IL72943A0 (en) | 1984-12-31 |
| DK440184A (en) | 1985-03-17 |
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