IE893717L - A process for the preparation of tripeptides - Google Patents
A process for the preparation of tripeptidesInfo
- Publication number
- IE893717L IE893717L IE893717A IE371789A IE893717L IE 893717 L IE893717 L IE 893717L IE 893717 A IE893717 A IE 893717A IE 371789 A IE371789 A IE 371789A IE 893717 L IE893717 L IE 893717L
- Authority
- IE
- Ireland
- Prior art keywords
- denotes
- compound
- general formula
- formula
- protective group
- Prior art date
Links
- 238000000034 method Methods 0.000 title claims abstract description 29
- 238000002360 preparation method Methods 0.000 title claims description 7
- 150000001875 compounds Chemical class 0.000 claims abstract description 13
- 229910052739 hydrogen Inorganic materials 0.000 claims abstract description 5
- 239000001257 hydrogen Substances 0.000 claims abstract description 5
- 238000007327 hydrogenolysis reaction Methods 0.000 claims abstract description 4
- 229910014033 C-OH Inorganic materials 0.000 claims abstract description 3
- 229910014570 C—OH Inorganic materials 0.000 claims abstract description 3
- 125000000217 alkyl group Chemical group 0.000 claims abstract description 3
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 24
- PAQZWJGSJMLPMG-UHFFFAOYSA-N 2,4,6-tripropyl-1,3,5,2$l^{5},4$l^{5},6$l^{5}-trioxatriphosphinane 2,4,6-trioxide Chemical compound CCCP1(=O)OP(=O)(CCC)OP(=O)(CCC)O1 PAQZWJGSJMLPMG-UHFFFAOYSA-N 0.000 claims description 12
- 238000006243 chemical reaction Methods 0.000 claims description 11
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 11
- 102000004142 Trypsin Human genes 0.000 claims description 6
- 108090000631 Trypsin Proteins 0.000 claims description 6
- 125000001584 benzyloxycarbonyl group Chemical group C(=O)(OCC1=CC=CC=C1)* 0.000 claims description 6
- 239000012588 trypsin Substances 0.000 claims description 6
- 239000000203 mixture Substances 0.000 claims description 5
- UYWQUFXKFGHYNT-UHFFFAOYSA-N phenylmethyl ester of formic acid Natural products O=COCC1=CC=CC=C1 UYWQUFXKFGHYNT-UHFFFAOYSA-N 0.000 claims description 5
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 claims description 4
- 150000001412 amines Chemical class 0.000 claims description 4
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 claims description 4
- 125000003118 aryl group Chemical group 0.000 claims description 3
- 230000002255 enzymatic effect Effects 0.000 claims description 3
- 125000006239 protecting group Chemical group 0.000 claims description 3
- CFYIUBWVKZQDOG-UHFFFAOYSA-N 4-[[2-[[2-[[1-(4-nitroanilino)-1-oxo-3-phenylpropan-2-yl]amino]-2-oxoethyl]amino]-2-oxoethyl]amino]-4-oxobutanoic acid Chemical compound C=1C=C([N+]([O-])=O)C=CC=1NC(=O)C(NC(=O)CNC(=O)CNC(=O)CCC(=O)O)CC1=CC=CC=C1 CFYIUBWVKZQDOG-UHFFFAOYSA-N 0.000 claims description 2
- 125000004432 carbon atom Chemical group C* 0.000 claims description 2
- 239000003795 chemical substances by application Substances 0.000 claims 1
- 150000001413 amino acids Chemical class 0.000 abstract 1
- 125000004435 hydrogen atom Chemical group [H]* 0.000 abstract 1
- 238000004519 manufacturing process Methods 0.000 abstract 1
- 239000012071 phase Substances 0.000 description 12
- 239000008346 aqueous phase Substances 0.000 description 10
- 150000002148 esters Chemical class 0.000 description 10
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 9
- 239000000243 solution Substances 0.000 description 9
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 description 6
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 5
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 5
- KDLHZDBZIXYQEI-UHFFFAOYSA-N Palladium Chemical compound [Pd] KDLHZDBZIXYQEI-UHFFFAOYSA-N 0.000 description 5
- 108090000765 processed proteins & peptides Proteins 0.000 description 5
- HVCNXQOWACZAFN-UHFFFAOYSA-N 4-ethylmorpholine Chemical compound CCN1CCOCC1 HVCNXQOWACZAFN-UHFFFAOYSA-N 0.000 description 4
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 4
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 4
- 229910000343 potassium bisulfate Inorganic materials 0.000 description 4
- 239000002244 precipitate Substances 0.000 description 4
- 239000000047 product Substances 0.000 description 4
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 4
- 102000004190 Enzymes Human genes 0.000 description 3
- 108090000790 Enzymes Proteins 0.000 description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 description 3
- 230000015572 biosynthetic process Effects 0.000 description 3
- 229940088598 enzyme Drugs 0.000 description 3
- 239000000543 intermediate Substances 0.000 description 3
- 239000011541 reaction mixture Substances 0.000 description 3
- 238000003786 synthesis reaction Methods 0.000 description 3
- AHYFYYVVAXRMKB-KRWDZBQOSA-N (2s)-3-(1h-indol-3-yl)-2-(phenylmethoxycarbonylamino)propanoic acid Chemical compound N([C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)O)C(=O)OCC1=CC=CC=C1 AHYFYYVVAXRMKB-KRWDZBQOSA-N 0.000 description 2
- JTNCEQNHURODLX-UHFFFAOYSA-N 2-phenylethanimidamide Chemical compound NC(=N)CC1=CC=CC=C1 JTNCEQNHURODLX-UHFFFAOYSA-N 0.000 description 2
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 2
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 2
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 2
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 239000000872 buffer Substances 0.000 description 2
- 230000008878 coupling Effects 0.000 description 2
- 238000010168 coupling process Methods 0.000 description 2
- 238000005859 coupling reaction Methods 0.000 description 2
- 238000004128 high performance liquid chromatography Methods 0.000 description 2
- 235000017557 sodium bicarbonate Nutrition 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- 239000002904 solvent Substances 0.000 description 2
- 239000007858 starting material Substances 0.000 description 2
- VPNIQGRFZCTBEZ-SPTGULJVSA-N 3-n-[(2s,3r)-4-(cyclopropylamino)-3-hydroxy-1-phenylbutan-2-yl]-5-[methyl(methylsulfonyl)amino]-1-n-[(1r)-1-phenylethyl]benzene-1,3-dicarboxamide Chemical compound C([C@H](NC(=O)C=1C=C(C=C(C=1)C(=O)N[C@H](C)C=1C=CC=CC=1)N(C)S(C)(=O)=O)[C@H](O)CNC1CC1)C1=CC=CC=C1 VPNIQGRFZCTBEZ-SPTGULJVSA-N 0.000 description 1
- QTBSBXVTEAMEQO-UHFFFAOYSA-M Acetate Chemical compound CC([O-])=O QTBSBXVTEAMEQO-UHFFFAOYSA-M 0.000 description 1
- DKPFZGUDAPQIHT-UHFFFAOYSA-N Butyl acetate Natural products CCCCOC(C)=O DKPFZGUDAPQIHT-UHFFFAOYSA-N 0.000 description 1
- 102400000932 Gonadoliberin-1 Human genes 0.000 description 1
- 101500026183 Homo sapiens Gonadoliberin-1 Proteins 0.000 description 1
- FXHOOIRPVKKKFG-UHFFFAOYSA-N N,N-Dimethylacetamide Chemical compound CN(C)C(C)=O FXHOOIRPVKKKFG-UHFFFAOYSA-N 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 229960000583 acetic acid Drugs 0.000 description 1
- 230000000975 bioactive effect Effects 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- 239000003054 catalyst Substances 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- 230000009089 cytolysis Effects 0.000 description 1
- 125000000118 dimethyl group Chemical group [H]C([H])([H])* 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 230000008030 elimination Effects 0.000 description 1
- 238000003379 elimination reaction Methods 0.000 description 1
- 125000001495 ethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 description 1
- 239000012362 glacial acetic acid Substances 0.000 description 1
- XLXSAKCOAKORKW-AQJXLSMYSA-N gonadorelin Chemical compound C([C@@H](C(=O)NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N1[C@@H](CCC1)C(=O)NCC(N)=O)NC(=O)[C@H](CO)NC(=O)[C@H](CC=1C2=CC=CC=C2NC=1)NC(=O)[C@H](CC=1N=CNC=1)NC(=O)[C@H]1NC(=O)CC1)C1=CC=C(O)C=C1 XLXSAKCOAKORKW-AQJXLSMYSA-N 0.000 description 1
- 229960001442 gonadorelin Drugs 0.000 description 1
- GNOIPBMMFNIUFM-UHFFFAOYSA-N hexamethylphosphoric triamide Chemical compound CN(C)P(=O)(N(C)C)N(C)C GNOIPBMMFNIUFM-UHFFFAOYSA-N 0.000 description 1
- FUZZWVXGSFPDMH-UHFFFAOYSA-N hexanoic acid Chemical compound CCCCCC(O)=O FUZZWVXGSFPDMH-UHFFFAOYSA-N 0.000 description 1
- 239000000601 hypothalamic hormone Substances 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 1
- 235000019796 monopotassium phosphate Nutrition 0.000 description 1
- 238000005580 one pot reaction Methods 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 description 1
- 239000002798 polar solvent Substances 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 150000003462 sulfoxides Chemical class 0.000 description 1
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
- C07K5/0821—Tripeptides with the first amino acid being heterocyclic, e.g. His, Pro, Trp
-
- 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
-
- 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
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Analytical Chemistry (AREA)
- Peptides Or Proteins (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
Process for preparing tripeptides of the general formula I U-A-B-C-OH I in which U is hydrogen or a urethane protective group and A, B and C are amino acids, by reacting a compound of the general formula II U'-B-OH II in which U' is a urethane protective group which can be eliminated by hydrogenolysis, with a compound of the general formula III H-C-OR III in which R is alkyl, by the PPA method, eliminating U', reacting the compound obtained in this way with a compound of the general formula IV H-B-C-OR IV by the PPA method and subsequently eliminating R enzymatically.
Description
(rt- HOECHST AKTIENGESELLSCHAFT HOE 88/F 327 67293 Dr. MY/je Description A process for the preparation of tripeptides Tripeptides are important intermediates in the synthesis 5 of bioactive peptides such as, for example, the hypothalamus hormone gonadorelin and its analogs. For this purpose the tripeptides must be available in the most straightforward manner possible, on the one hand in good yields and, on the other hand, in high purity. The 10 processes hitherto disclosed for the preparation of tripeptides do not meet these requirements in an optimal manner and are associated with disadvantages, some of which are serious. Thus, for example, even the products prepared by the process described in EP-A 156,280 are 15 contaminated with byproducts which become disadvantageous ly evident in the subsequent synthetic steps. Thus the object of the present invention is to provide a process for the preparation of tripeptides which does not have the said disadvantages and provides, in a straight-20 forward manner, products of high purity in good yields.
Accordingly, the invention relates to a process for the preparation of tripeptides of the general formula I U-A-B-C-OH 1 in which 25 U denotes hydrogen or a urethane protective group A denotes a natural a-amino acid or derivatives thereof B denotes a natural a-amino acid or derivatives thereof and 30 C denotes an aromatic a-amino acid, which comprises reacting a compound of the general formula II V - B - OH II - 2 - in which D' is a urethane protective group which can be eliminated by hydrogenolys is, and B has the above-mentioned meaning, with a compound of the general formula III H - C - OR 5 III in which R represents alkyl having 1 to 4 carbon atoms, and C has the abovementioned meaning, in the presence of propylphosphonic anhydride, eliminating the protective group U' by hydrogenolysis, reacting the resulting 10 compound of the general formula IV H - B - C - OR IV with a compound of the general formula V U - A - OH V in the presence of propylphosphonic anhydride, and 15 finally eliminating R enzymatically.
The urethane protective groups representing U are preferably the urethane protective groups customary in peptide chemistry, as are described, for example, in Kontakte Merck 3/79, page 14. 20 The benzyloxycarbonyl and the tert.-butyloxycarbonyl groups are particularly preferred.
A urethane protective group U' which can be eliminated by hydrogenolys is is preferably the benzyloxycarbonyl group.
Natural a-amino acids or their derivatives representing 25 A and/or B are preferably Gly, Ala, Ser, Thr, Val, Leu, lie, Glu, Gin, p-Glu, Tyr, Phe, Trp and His. Ser, Thr, Trp and Phe are particularly preferred.
An aromatic a-amino acid representing C is preferably Tyr or Phe. - 3 - R in the general formula IV preferably denotes methyl.
A process in which U and V denote benzyloxycarbonyl, A denotes Trp, B denotes Ser, C denotes Tyr and R denotes methyl is very particularly preferred. 5 The formation of a peptide linkage in the presence of propylphosphonic anhydride is known as the PPA method (Angew. Chem. Int. Ed. 1&, 133 (1980)). This reaction is preferably carried out in polar solvents such as, for example, dimethylacetamide, dimethylformamide, dimethyl 10 sulfoxide, phosphoric tris(dimethylamide), N-methyl-pyrro1idone or water. However, chloroform, methylene chloride or ethyl acetate are also employed.
It is also possible in an advantageous manner to use mixtures of the said solvents with water. An ethyl 15 acetate/water mixture is particularly preferred. The synthesis can be carried out between -10°C and room temperature. It is preferable to start at about 0°C and subsequently to raise to room temperature.
The elimination of the U' protective group by hydrogeno-20 lysis is advantageously carried out in a known manner with hydrogen on a Pd/C catalyst.
The enzymatic esterolysis in the last reaction step is preferably carried out with trypsin and/or a-chymotrypsin (Hoppe-Seylers Zeitschrift f. physiol. Chemie, 336. 248 25 (1964)). Trypsin is particularly preferred. Where appropriate, enzymes which are immobilized by known methods on a support are also used, such as described, for example, in EP-A 178,553. In this case, the enzymes are advantageously employed in amounts of 0.01 to 20 % by 30 weight relative to the amount of substrate. An amount of 2 % by weight of enzyme is particularly preferred.
Examples of solvents which can be employed are water, dimethylformamide, methanol, ethanol, isopropanol, butanol, ethyl acetate, butyl acetate, toluene or - 4 - methylene chloride.
An ethyl acetate/water mixture is preferred. The temperatures are advantageously between 0 and 60°C. A temperature range from 20 to 35°C is preferred. The pH of the reaction 5 medium is preferably in the range between 4 and 10, particularly preferably between 4 and 8.
The process according to the invention can be carried out in such a way that each intermediate is isolated. However, it is preferably carried out in a one-pot process, 10 that is to say without isolation of the intermediates.
The starting compounds of the general formulae II, III and V are known and can be obtained by the customary methods.
The process according to the invention surprisingly 15 provides products of high chemical and optical purity, which can be employed without difficulty in further syntheses. The yields are likewise excellent and are between 40 and 50 % based on the amount of the compound of the general formula III employed. 20 It has to be regarded as particularly surprising that the process according to the invention is distinctly superior, in terms both of purity and of yield, to the process of BP-A 156,280, which has only three stages.
Example 25 Z-Trp-Ser-Tyr-OH a) 350 ml of water are placed in a 2 1 stirred apparatus, and 47.8 g (0.200 mol) of Z-Ser-QH, 46.4 g (0.200 mol) of H-Tyr-QMexHCl and 150 g of sodium chloride are introduced. Also added are 700 ml of 30 ethyl acetate and, after everything has dissolved, the pH of the mixture is adjusted to 5.0 by addition of about 25 ml of N-ethylmorpholine. During the addition of about 220 ml (0.42 mol) of PPA solution (w(PPA) in % = 50) in about 30 minutes at a maximum of 30°C (cool somewhat at the end), about 110 ml (0.86 mol) of N-ethylmorpholine are added via a pH-stat pump at pH 5.0. The PPA addition is terminated when a precipitate forms in the reaction mixture. The precipitate is redissolved by subsequent addition of 350 ml of water. The aqueous phase is separated off in a separating funnel and then the ester phase is washed with 700 ml of potassium bisulfate solution (w(KHS04) in % ■ 10) and 700 ml of sodium bicarbonate solution (w(NaHC03) in % = 5). The aqueous phase from the reaction and the wash phases are discarded.
About 700 ml of ester phase from the 1st coupling, 200 ml of water and 3.3 g of palladium on carbon w(Pd) in % = 2.5 are placed in a 2 1 stirred apparatus and a stream of hydrogen is passed in at 25-30°C. During the reaction the pH is maintained at 4.0 with a pH-stat pump and addition of about 160 ml (0.16 mol) of hydrochloric acid c(HCl) = 1 mol/1. After the reaction is complete, when no more hydrochloric acid is consumed, (about 30 minutes) the reaction mixture is filtered through a suction funnel, and the aqueous phase is separated from the ester phase in a separating funnel. The ester phase is discarded.
About 430 ml of aqueous phase from the hydrogeno-lysis and 700 ml of ethyl acetate are placed in a 2 1 stirred apparatus and 50.7 g (0.15 mol) of Z-Trp-OH and 125 g of sodium chloride are added. After everything has dissolved, the pH is adjusted to 5.0 with about 19 ml of N-ethylmorpholine. During the addition of about 220 ml (0.42 mol) of PPA solution (w(PPA) in % - 50) in about 30 minutes at a maximum of 30°C (cool somewhat at the end), about 110 ml (0.86 mol) of N-ethylmorpholine are added via a pH-stat pump at pH 5.0. The PPA addition is terminated when a precipitate forms in the reaction mixture. The precipitate is redissolved by subsequent addition of 350 ml of water. The aqueous phase is separated off in a separating funnel, and then the ester phase is washed with 700 ml of potassium bisulfate solution (w(KHS04) in % = 10) and several times with 700 ml portions of sodium bicarbonate solution (w(NaHC03) in % = 5) until Z-Trp-OH has been completely removed (according to TLC analysis). The aqueous phase from the reaction and the wash phases are discarded.
About 700 ml of ester phase from the 2nd coupling and 700 ml of water are placed in a 2 1 stirred apparatus and heated to 35-40°C, and 1 g of trypsin is initially added. The reaction starts immediately and, during it, the pH is maintained constant at pH 7.0 with about 110 ml (0.11 mol) of sodium hydroxide solution (c(NaOH) = 1 mol/1). The reaction lasts about 7 hours and, during this, the rate is increased now and again by further addition of 0.5 g of trypsin. It is complete when trypsin addition now brings about only a slight increase in the rate of absorption of sodium hydroxide solution, or TLC analysis shows hardly any starting material remaining. The reaction solution is clarified through a suction funnel, and the ester phase is separated from the aqueous phase in a separating funnel. The ester phase is discarded.
The aqueous phase is initially extracted by shaking twice at pH 5.8-6.0, by addition and dissolution of 4.0 g of potassium dihydrogen phosphate, with 700 ml of ethyl acetate each time. The ester phases are discarded. The aqueous phase is then extracted by shaking three times at pH 5.0, adjusted by addition of about 5 ml of glacial acetic acid, with 700 ml of ethyl acetate each time. The aqueous phase is discarded. The ester phases contain the tripeptide - 7 - which, on evaparation to dryness in vacuo, remains in the form of loosely packed crystals. The product is dried in a vacuum oven at 40°C.
Weights 51.2 g 5 Yields 42.0 % based on H-Tyr-OMexHCl Puritys 98.2 % (determined with HPLC LiChrosorb Si 60/peptide buffer) Comparison Example Z-Trp-Ser-Tyr-OH was prepared by the process specified in 10 EP-A 156,280.
Yields 30 % Puritys 78.8 % (determined with HPLC LiChrosorb Si 60/peptide buffer)
Claims (12)
1. A process for the preparation of tripeptides of the general formula I U-A-B-C-OH 1 in which U denotes hydrogen or a urethane protective group A denotes a natural a-amino acid or derivatives thereof B denotes a natural a-amino acid or derivatives thereof and C denotes an aromatic a-amino acid, which comprises reacting a compound of the formula II U' - B - OH 11 in which U' is a urethane protective group which can be eliminated by hydrogenolys is, and B has the above-mentioned meaning, with a compound of the formula III H - C - OR in in which R represents alkyl having 1 to 4 carbon atoms, and C has the abovementioned meaning, in the presence of propylphosphonic anhydride, eliminating the protective group U' by hydrogenolysis, reacting the resulting compound of the formula IV H - B - C - OR IV with a compound of the formula V U - A - OH V in the presence of propylphosphonic anhydride, and finally eliminating R enzymatic ally.
2. The process as claimed in claim 1, wherein U denotes benzyloxycarbonyl or tert. -butyloxycarbonyl.
3. The process as claimed in claim 1 and/or 2, wherein A and/or B denote Gly, Ala, Ser, Thr, Val, Leu, lie, Glu, Gin, p-Glu, Tyr, Phe, Trp or His.
4. The process as claimed in one or more of claims 1 to 3, wherein A and/or B denote Ser, Thr, Trp or Phe.
5. The process as claimed in one or more of claims 1 to 4, wherein C denotes Tyr or Phe.
6. The process as claimed in one or more of claims 1 to 5, wherein U' denotes benzyloxycarbonyl.
7. The process as claimed in one or more of claims 1 to 6, wherein R denotes methyl.
8. The process as claimed in one or more of claims 1 to 7, wherein the reactions are carried out in the presence of propylphosphonic anhydride in an ethyl acetate/water mixture.
9. The process as claimed in one or more of claims 1 to 8, wherein the enzymatic esterolysis is carried out with trypsin and/or a-chymotrypsin.
10. The process as claimed in one or more of claims 1 to. 9, wherein U and U' denote benzyloxycarbonyl, A denotes Trp, B denotes Ser, C denotes Tyr and R denotes methyl.
11. A process according to claim 1 for the preparation of a tripeptide of the general formula I given and defined therein, substantially as hereinbefore described and exemplified.
12. A tripeptide of the general formula I given and defined in claim 1, whenever prepared by a process claimed in a preceding claim. - 10 - Dated this the 21st day of November, 198? F. R. KELLY & CO. pv-M|f t| — 4p EXECUTIVE ClVde 27 Clyde Road, pa-nTsbridge f Dublin 4. AGENTS FOR THE APPLICANTS.
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| DE3839379A DE3839379A1 (en) | 1988-11-22 | 1988-11-22 | METHOD FOR PRODUCING TRIPEPTIDES |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| IE893717L true IE893717L (en) | 1990-05-22 |
| IE67293B1 IE67293B1 (en) | 1996-03-20 |
Family
ID=6367619
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| IE371789A IE67293B1 (en) | 1988-11-22 | 1989-11-21 | A process for the preparation of tripeptides |
Country Status (10)
| Country | Link |
|---|---|
| EP (1) | EP0370399B1 (en) |
| JP (1) | JP2843618B2 (en) |
| AT (1) | ATE124053T1 (en) |
| AU (1) | AU626608B2 (en) |
| CA (1) | CA1335493C (en) |
| DE (2) | DE3839379A1 (en) |
| DK (1) | DK173690B1 (en) |
| ES (1) | ES2075028T3 (en) |
| IE (1) | IE67293B1 (en) |
| IL (1) | IL92368A0 (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE102004029812A1 (en) | 2004-06-19 | 2006-05-24 | Clariant Gmbh | Process for the preparation of nitriles from aldehyde oximes by reaction with alkylphosphonic anhydrides |
| DE102004029811A1 (en) * | 2004-06-19 | 2006-01-12 | Clariant Gmbh | Process for the preparation of alkenes by elimination of water from alcohols with alkylphosphonic anhydrides |
Family Cites Families (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| SE7801373L (en) * | 1978-02-07 | 1979-08-08 | Kabi Ab | EASY SPLABLE SUBSTRATE FOR QUANTIFIATION OF PROTEASES |
| US4293648A (en) * | 1979-12-12 | 1981-10-06 | G. D. Searle & Co. | Process for esterification of α-L-aspartyl-L-phenylalanine |
| DE3411224A1 (en) * | 1984-03-27 | 1985-10-10 | Hoechst Ag, 6230 Frankfurt | METHOD FOR THE RACEMATAR PRODUCTION OF INTERMEDIATE PEPTIDE PRODUCTS OF THE GONADORELIN AND GONADORELINANALOGA SYNTHESIS AND NEW INTERMEDIATE PRODUCTS IN THIS METHOD |
| DE3438189A1 (en) * | 1984-10-18 | 1986-04-24 | Hoechst Ag, 6230 Frankfurt | METHOD FOR PRODUCING AROMATICALLY SUBSTITUTED L-AMINO ACIDS |
| US4888385A (en) * | 1987-04-30 | 1989-12-19 | Millipore Corporation | BOP reagent for solid phase peptide synthesis |
| US4946942A (en) * | 1988-03-11 | 1990-08-07 | Bioresearch, Inc. | Urethane-protected amino acid-N-carboxyanhydrides |
-
1988
- 1988-11-22 DE DE3839379A patent/DE3839379A1/en not_active Withdrawn
-
1989
- 1989-09-29 CA CA000614545A patent/CA1335493C/en not_active Expired - Lifetime
- 1989-11-17 ES ES89121277T patent/ES2075028T3/en not_active Expired - Lifetime
- 1989-11-17 AT AT89121277T patent/ATE124053T1/en not_active IP Right Cessation
- 1989-11-17 DE DE58909308T patent/DE58909308D1/en not_active Expired - Lifetime
- 1989-11-17 EP EP89121277A patent/EP0370399B1/en not_active Expired - Lifetime
- 1989-11-20 IL IL92368A patent/IL92368A0/en not_active IP Right Cessation
- 1989-11-21 AU AU45328/89A patent/AU626608B2/en not_active Expired
- 1989-11-21 IE IE371789A patent/IE67293B1/en not_active IP Right Cessation
- 1989-11-21 DK DK198905844A patent/DK173690B1/en not_active IP Right Cessation
- 1989-11-21 JP JP1300960A patent/JP2843618B2/en not_active Expired - Lifetime
Also Published As
| Publication number | Publication date |
|---|---|
| AU626608B2 (en) | 1992-08-06 |
| IL92368A0 (en) | 1990-07-26 |
| DK173690B1 (en) | 2001-06-25 |
| ES2075028T3 (en) | 1995-10-01 |
| ATE124053T1 (en) | 1995-07-15 |
| IE67293B1 (en) | 1996-03-20 |
| JPH02219587A (en) | 1990-09-03 |
| DK584489A (en) | 1990-05-23 |
| CA1335493C (en) | 1995-05-09 |
| EP0370399B1 (en) | 1995-06-21 |
| DK584489D0 (en) | 1989-11-21 |
| DE58909308D1 (en) | 1995-07-27 |
| AU4532889A (en) | 1990-05-31 |
| EP0370399A2 (en) | 1990-05-30 |
| DE3839379A1 (en) | 1990-05-23 |
| JP2843618B2 (en) | 1999-01-06 |
| EP0370399A3 (en) | 1991-09-18 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US6251625B1 (en) | Process for preparing peptides and N-carbamoyl-protected peptides | |
| US5015728A (en) | Process for the preparation of insulin derivatives, the B chain of which is lengthened c-terminally | |
| EP0575844B1 (en) | Hydroxamic acid derivatives and pharmaceutical compositions thereof | |
| HU205091B (en) | Process for producing amino acid-n-carboxyanhydrides protected with urethane group | |
| FI79860C (en) | Process for the preparation of human insulin, Des-PheB1 human insulin or derivatives thereof from porcine insulin, Des-PheB1 porcine insulin or derivatives thereof | |
| HU185668B (en) | Process for the enzymatic substitution of aminoacid in position b-30 of | |
| CA1174973A (en) | Process for preparing polypeptide derivatives | |
| EP0149594A2 (en) | Enzymatic coupling of n-formyl amino acids and/or peptide residues | |
| FI78119C (en) | FOERFARANDE FOER FRAMSTAELLNING AV MAENSKLIGT INSULIN ELLER B-30-ESTRAR DAERAV. | |
| US5191065A (en) | Process for the preparation of tripeptides | |
| AU626608B2 (en) | A process for the preparation of tripeptides | |
| US3435046A (en) | Trialkyl-silyl ethers of n-carboxy anhydrides of monohydroxy alpha-amino acids and processes for preparing them | |
| US5157145A (en) | Process for the preparation of dipeptides with c-terminal non-proteinogenous amino acids | |
| US4426325A (en) | Process for the preparation of compounds containing carboxylic acid amide groups, in particular or peptides | |
| US5138071A (en) | Process for preparing peptides with n-terminal non-proteinogenous amino acids | |
| CA2223911C (en) | The preparation of active peptides | |
| Hemmi et al. | STUDIES ON A NEW IMMUNOACTIVE PEPTIDE, FK-156 IV. SYNTHESIS OF FK-156 AND ITS GEOMETRIC ISOMER | |
| Ohno et al. | Partial enzymic deprotection in the synthesis of a protected octapeptide bearing a free terminal carboxyl group | |
| HU221619B1 (en) | Process for synthetise peptides and intermediates thereof | |
| CA1304388C (en) | Process of preparing carboxyalkyl dipeptides | |
| CA1285698C (en) | Process for the preparation of compounds containing carboxamide groups,in particular of peptides | |
| CS255616B1 (en) | Process for the preparation of (2-O-methyltyrosine) deamino-1-carbaoxytocin | |
| US20140213758A1 (en) | Versatile native chemical ligation technologies | |
| EP0375047A2 (en) | New process for the synthesis of N alpha-fluorenylmethoxycarbonyl-Ng-trityl-arginine | |
| Kalashnikov et al. | Protection of the carboxy group in the form of the 2-cyanoethyl ester in synthesis of peptides |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| MK9A | Patent expired |