PROCESS FOR THE PREPARATION OF C-TERMINALLY AMIDATED PEPTIDES

Abstract: Preparation of C-terminally amidated peptides P-NH2 by reacting a substrate component selected from a) C-terminally unblocked peptides P-OH, b) peptides or peptide derivatives P-X-Y, where X is an amino acid or peptide residue and Y is OH, OMe or C-terminal modification, and c) C-terminally esterified peptides P-OR', where R' is alkyl, aryl, heteroaryl, aralkyl or an alpha -des amino fragment of an amino acid, with a nucleophile component in the presence of an enzyme using as nuclophile a compound NH2-R capable of cleaving the reaction product P-NH-R to P-NH2, and undertaking such cleavage. Preferred nucleophiles are selected from (I), wherein A-F and A'-E' are carbon atoms or up to two hetero atoms, Y is H, alkyl, aryl, aralkyl, oxo or carboxy, or a functional derivative thereof, X<1>-X<5> are H or various substituents. The cleavage may be induced by photolysis, solvolysis, reduction, rearrangement elimination or oxidation. The process is very flexible in that it may be adapted to any type of enzymatic synthesis and lend itself to C-terminal amidation of all types of peptides.

Description: Process for the preparation of C-terminally amidated peptides The present invention relates to a process for the preparation of C-terminally amidated peptides P-NH-, wherein P is a peptide residue which may be N-protected. The use of biologically active peptides for pharmaceutical purposes and in agriculture has increased the importance of being able to synthesize such compounds in bulk-scale. In principle, three methods are available: (a) chemical synthesis based on solid-phase or solution-phase techniques, (b) enzymatic synthesis, and (c) fermentation with genetically manipulated microorganisms. While the methods (a) and (b), or a combination of them, are the preferred for short peptides they are not suitable for bulk-scale production, they are rather expensive and the yields are often low. It therefore becomes more and more apparent that long peptides in the future will be produced through exploitation of the advances in the methods dealing with recombinant DNA. However, these methods do not permit a number of modifications such as incorporation of D-amino acids or C-terminal amide groups, etc., which may be of importance for the biological activity. A subsequent enzymatic modification is therefore highly desirable and such reactions have been studied to a limited extent. An enzyme has been described which catalyses the hydroxyl- ation of C-terminal glycyl residues which subsequently decomposes leaving the penultimate residue amidated (US Patent No. 4.708.934, EP 308067A, DK Application No. 4489/88). This glycine oxidase enzyme which is dependent on Cu + , 0- and ascorbate as cofactors is considered to be the enzyme responsible for in vivo formation of peptide amides- It has been utilised for amidation of peptides in small scale but as it exhibits low activity its applicab¬ ility for large scale work is still questionable. The enzyme, as isolated from natural sources like rat medul- lary thyroid carcinoma, is very costly. Amidation may also by achieved by protease-catalysed condensation reactions (reversal of hydrolysis) using an amino acid amide or peptide amide as nucleophile. The yields of condensation reactions are generally low even in the presence of organic solvents unless the product precipitates in the reaction mixture and this is often not the case with long peptides. In addition, the precursor peptide may exhibit poor solubility in such media. However, serine or thiol-protease catalysed trans- peptidation reactions may be carried out in high yield but it is a prerequisite that the enzyme exhibits specificity for a peptide bond close to the C-terminus. Endopeptidases are not generally suitable since they usually will cleave at other positions in the peptide chain as well. Serine carboxypeptidases, on the other hand, exhibit strict specificity for the C-terminal peptide bond and are able to catalyse the exchange of the C-terminal amino acid with an amino acid amide, added to the " reaction medium to compete with water as nucleophile. This property of serine carboxypeptidases was realized by a group of researchers at Carlsberg Research Center and lead to a family of patents based on DK application No. 1443/79 represented by EP Patent No. 17485, US Patent No. 4.806.534 and its parent US patent No. 4.339.534 and WO 80/02151, which lead to i.a. DK Patent No. 155613. These patents were based on the, at that time, surprising finding that exopeptidases were suitable as catalysts for enzymatic peptide synthesis, while the prior art dealt exclusively with endopeptidases. Dependent on the nature of the reactants (substrate and nucleophile components), and the reaction conditions, particularly the pH, serine and thiol carboxypeptidases may catalyze peptide synthesis by chain elongation or by transpeptidation. The preferred enzyme is carboxypeptidase Y (CPD-Y) from yeast. This enzyme reacts with the substrate component under formation of an acyl enzyme intermediate from which the acyl group is transferred to the nucleophile during a transacylation which according to the reaction conditions . may be a transpeptidation in which the C-terminal amino acid is exchanged. The underlying and subsequent research has been further described in a number of articles (Refs. 14-18), which together with the above-mentioned patents are all incorporated by reference. The general principle of enzymatic peptide synthesis by transpeptidation in the presence of serine or thiol carboxypeptidases is disclosed in US Patent No. 4.806.473 and its parallel Danish Patent No. 155613. With particular reference to the production of peptide amides these patents generally disclose and claim the production of peptide amides A-B-NH- where A represent an N-terminal protected amino acid residue or a peptide residue which may be N-terminal protected, and B represents an L-amino acid residue, by reacting as substrate component . an optionally N-terminal protected peptide A-X-OH, where A is as defined above and X represents an amino acid residue, with a nucleophile (amine) component H-B-NH„ in the presence of an L-specific serine or thiol carboxypeptidase enzyme from yeast, or of animal, vegetable or microbial origin in an aqueous solution or dispersion of a pH from 5 to 10.5. As further explained in Ref. 14 the preferred pH is about neutral if the formation of a peptide amide is desired. As a representative example from US Patent No. 4,806,473 may be mentioned the reaction of Bz-Phe-Gly with Leu-NH„ in the presence of CPD-Y at pH 7.6 and 25°C leading to the formation of Bz-Phe-Leu-NH„ in a 90% yield. Further examples are incorporated in DK Patent No. 155613 e.g. the reaction of Ac(Ala). with Leu-NH- in the presence of CPD-Y leading to Ac(Ala)-.-Leu-NH„ in a yield of 70%. Similarly, the general principle of enzymatic peptide synthesis by chain elongation is disclosed in US patent no. 4,339,534 and the above-mentioned DK patent no. 155,613. With particular reference to the production of peptide amides these patents generally disclose and claim the production of peptide amides A-B-NH„, where A and B are as defined above by reacting a substrate component selected from amino acid and peptide esters, depsi- peptides, optionally N-substituted amino acid amides or peptide amides of the formulae A-0R 1 or A-NR2R3, wherein R represent alkyl, aryl, heteroaryl, aralkyl or an -des- amino fragment of an amino acid residue, and R 2 and R3 are independently selected from H, alkyl, aryl, heteroaryl and aralkyl with a nucleophile (amine) component H-B-NH-. The preferred reaction mechanism may be termed "aminolysis of esters" i.e. a peptide ester or amino acid ester is used as the substrate component. This reaction is exempli¬ fied for a great number of amino acid amide nucleophiles in the above patents. The use of HU * . as nucleophile in the aminolysis under peptide amide formation is shown e.g. in Ref. 15. Further experiments are disclosed in Ref. 14-18 which support the pioneer character of these early patents and the general applicability of serine carboxypetidases as catalysts for C-terminal modification of peptides. In order to provide a better understanding of the present invention which is described in more detail below a general discussion of the transpeptidation principle and the competing reactions is deemed proper. C-terminal amidation of a peptide by means of a serine carboxypeptidase catalysed transpeptidation reaction is dependent on: a) solubility of the peptide in a medium in which the enzyme is active and relatively stable, b) accessibility of the C-terminus to enzymatic cleavage and c) availability of a serine carboxypeptidase of suitable substrate preference. The types of reactions which may take place are outlined in Scheme 1 with the substrate R-NH-A-B-C-OH and the nucleophile H-D-NH^. The enzyme attacks the C-terminal peptide bond producing an acyl-enzyme intermediate which subsequently may be deacylated by the nucleophile, producing an amidated transpeptidation product (Tl) , in competition with water, producing a hydrolysis product (HI ) . The ratio of Tl to HI may be increased by increasing the concentration of the reactive, deprotonated form of the nucleophile, i.e. by increasing either pH or the added amount of nucleophile. The nature of the leaving group, H- C-OH, has, in the case of CPD-Y, been shown to significantly influence this ratio as well (14, 15) albeit with no obvious trend. Since this amino acid residue does not constitute part of the desired amidated product (Tl) it may in principle be chosen freely. A serine carboxypeptidase may also catalyse the formation of an elongated condensation product (Cl ) in competition with HI and Tl. However, at the optimal pH-values for transpeptidation such reactions are energically unfavourable in aqueous solution and consequently, at equilibrium, the amount of Cl which may be formed is limited to a few per cent, even at nucleophile concentra¬ tions exceeding 1 M. Furthermore, the rates of such reactions are normally very low compared to the competing attack on the C-terminal peptide bond, and thus, condensation products are rarely a problem. However, this relation may be reversed in cases where the C-terminal sequence of the peptide does not match the substrate preference of the enzyme with the consequence that small amounts of condensation products are formed in the initial phases of the reaction and then, with time, disappears as the substrate is consumed (16) . The hydrolysis product (HI) may be transformed via the same type of reactions into the products T2, H2 and Tl, followed by similar reactions with H2, etc. However, new products may also arise when the enzyme acts on the C- terminal amide bond of Tl, i.e. ammonia functions as leaving group, producing another transpeptidation product (TT1) or hydrolysis product (HT1). For these products to appear in significant amounts it is required that the amidase activity towards Tl is significant compared to the peptidase activity towards the substrate and this is normally only the case at pH>9 where the peptidase activity is low and the amidase activity is high or in cases where the peptidase activity is low due to lack of preference for the C-terminal peptide bond. It is apparent that the action of a serine carboxy¬ peptidase on the C-terminus of a peptide in the presence of an amino acid amide may lead to numerous products of which only Tl is the desired. ω ro ro CO o Oi o ϋi n R-NH-fl-8-C-D-NHg ( CD R-NH-fl-B-D-OH (HID R-NH-fl-B-C-OH ( S ) R-NH-fl-B-D-NH s (Tl) R-NH-fl-B-D-O-HHg (TT1) R-NH-fl-B~0H (HI) R-NH-fl-D-NH s (T2) R-NH-fl-D-D-NH, <TTE> I R-NH-R-OH <H2> R-NH-fl-D-OH (HT2 ) i: T anspeptidation reac tion c : ' Condensation reac lion Nuc leophi le : H-DηNHg h: Hydrolysis reac lion While reactions with N-blocked dipeptides of a suitable composition relative to the substrate preference of the enzyme in some case lead to yields of Tl approaching 100% (14, 15, 17, 18) the situation is different with larger molecules. Thus in the patent family represented by EP patent no. 45189, US patent no. 4,645,740 and Danish patent no. 148714 incorporated herein by reference a process is described for enzymatic replacement of the B-30 amino acid in insulins, in particular conversion of porcine insulin into human insulin by replacement of the B-30 amino acid alanine with threonine using a serine carboxypeptidase, preferably carboxypeptidase Y from yeast (CPD-Y) or carboxypeptidase P from Penicillium janthinellum (CPD-P) as catalyst and threonine or preferably its amides or esters as the nucleophile component. While the reaction is only illustrated with unsubstituted threonine amide, the use of N-substituted amino acid amides H-B-NR 1R2 is claimed, wherein B is an amino acid and either of R 1 and R2 are independently selected from hydrogen, amino, hydroxy, alkyl, cycloalkyl, aryl, hheetteerrooaarryyll oorr NNRR 1 1RR22 iiss aa hheeterocyclic group which may contain a further hetero atom. Thus, depending on the reaction conditions during the transpeptidation step and optionally N-substituted insulin amide Ins-B-NR 1R2 may be formed. This intermediate may be isolated and if desired subsequently deamidated, preferably by enzymatic cleavage, to form the desired free insulin Ins-B-OH or this deamidation may take place in situ by cleavage with the same enzyme as was used during the transpeptidation, preferably CPD-Y. When porcine insulin was reacted with threonine amide in the presence of CPD-Y at pH 7.5 a complicated mixture of reaction products was obtained (Example 4 of the patents) . The mixture was analysed by ion exchange chromatography and three peaks were detected. The peaks were further analyzed by enzymatic digestion and amino acid analysis leading to the following composition (Ins-Pro-Lys-Ala-OH being used to denominate the porcine insulin starting material), and the symbols from Scheme I are used as illustration. Peak 1; 21% of which 65% Ins-Pro-Lys-Thr-Thr-NH (TT1) 35% Ins-Pro-Lys-Thr-NH 2 (Tl) Peak 2: 61% of which 52% Ins-Pro-Lys-Ala-OH (unreacted porcine insulin) (S) 26% Ins-Pro-Lys-Thr-OH (human insulin) (HT1) 22% Ins-Pro-Thr-NH„ (T2) Peak 3: 18% Breakdown products Thus in the known process only 21% of a mixture of Tl and TT1 was obtained. The predominant fraction consisted of a mixture of unreacted S, HT1 and T2. Also a significant amount of breakdown products was obtained. In example 2 of EP 45187 which was carried out at pH 9.5 80% of the porcine insulin was reacted with threonine amide under formation of what appeared to be human insulin amide (Tl) which, however, was further hydrolyzed in situ to human insulin (HT1). A significant amount (20%) had reacted to form H2. The above-mentioned insulin modification is further analyzed in Refs. 23 and 16. While therefore the formation of human insulin amide by means of the general principle of transpeptidation described above (S -> Tl) is certainly possible, it is not easily monitored. Evidently reaction conditions may be selected which on the face of it seem favourable to the formation of Tl, but such attempts have not been reported. Also the separation of the various reactants may be more or less difficult. As Tl lacks the negative charge at the C-terminus it may easily be separated from HI in pre- parative scale by ion exchange chromatography. However, provided that the side-chain of the C-terminal amino acid residue is uncharged, T2, T3 etc. as well as Cl and TT1 exhibit the same charge and therefore cannot be separated from Tl by ion exchange chromatography. In such cases preparative HPLC with reverse-phase columns appears to be the only, and a much less attractive, alternative. It is thus more desirable to attempt to suppress the formation of such products by selecting the optimal pH, concentra¬ tion of nucleophile and most important, the serine carboxypeptidase of the most suitable substrate preference which rapidly and in the highest possible yield converts the substrate in Tl and produces the lowest possible yield, if any, of the undesirable side-products. To recapitulate the essence of the above observations, incorporation of C-terminal amide groups in peptides by transpeptidation in the presence of a serine carboxy¬ peptidase using the proper amino acid amide as the nucleophile as broadly described and claimed in US patent no. 4,806,473 and the other family members is a very appropriate method virtually applicable for any peptide. However, the process is not always sufficiently selective and necessitates purification procedures in order to remove products of various side reactions in particular when longer peptides are used, in which case the optimal reaction conditions for suppressing the side reactions are difficult to establish. In Ref. 23 some of the experiments underlying EP patent no. 45187 and its family members were discussed. Here porcine insulin Ins-Pro-Lys-Ala was reacted with i.a. Thr- NH^ and it was concluded that Ins-Pro-Lys-OH was a better substrate that Ins-Pro-Lys-Ala-OH, since Ins-Pro-Thr-NH^ was formed in higher yields than Ins-Pro-Lys-Thr-NH». By inference Lys in this reaction was a better leaving group than Ala. Also a significant oligomerization under formation of Ins- Pro-Lys-Thr-Thr-NH„ occurred. These results were further confirmed in Ref. 16 using Bz- Lys-Ala-OH as a model peptide along with porcine insulin. The conclusive message was that for the future use of CPD- Y (the serine carboxypeptidase used in the experiments) in transpeptidation reactions it is important to be aware of the possibility that side products may be formed. For the sake of completeness the enzymatic synthesis of peptide amides by the transacylation mechanism referred to above as "aminolysis of esters" is briefly discussed using the same general symbols as in the above transpeptidation scheme . The reaction is performed according to the following general reaction scheme R-NH-A-B-OMe + H-D-NH 2 -> R-NH-A-B-D-NH 2 + MeOH The yield of aminolysis is 85-95% for most amino acid amides. Besides the above investigations of the applicability of serine carboxypeptidases in C-terminal modifications of insulin, a further experiment with amidation of longer peptides using CPD-Y as a catalyst has been reported. Thus in EP-B2-197794 and the parallel US patent no. 4,709,014 (Tamaoki) human calcitonin-Leu peptide was reacted with ammonia as the nucleophile using CPD-Y as the catalyst under conditions otherwise similar to those used by Breddam et al in Ref. 15. (The discussion of Ref. 15 above has been limited to the use of amino acid amides as nucleophiles, while in fact the main purpose of the article was to compare various nucleophiles also including ammonia) . Tamaoki obtained human calcitonin amide in a yield of 24.7%, leaving 57% unreacted substrate and 17.2% non- amidated side products (including human calcitonin). In its more general aspects the Tamaoki patents disclose a process for the preparation of a peptide having a C- terminal proline amide, which comprises reacting in aqueous solution a peptide substrate having C-terminal Pro-Leu, Pro-lie, Pro-Val or Pro-Phe with carboxypeptidase Y in the presence of ammonia. Without in any way wanting to endorse the statements made by Tamaoki, it should be mentioned that he claims that contrary to the findings of Breddam et al in Ref. 15, where a preference for hydrophilic C-terminal amino acids as leaving groups is expressed, the use of hydrophobic amino acids (Leu, lie, Val and Phe) gives better yields than Gly, when Pro is the penultimate amino acid. Nevertheless the yields of the amidation products in Tamaoki's examples using Cbz-Ala-Pro-X-OH as the sub¬ strate, where X is Leu, Val, Phe and lie, respectively, were only 35,1%, 43%, 15,4%, 13,4%, 22,6%, respectively. The remainder was - to the extent reported - unreacted starting material and non-amidated side-products Cbz-Ala- Pro-OH. The present invention is based on the surprising finding that by using a particular type of nucleophile components in the enzyme catalyzed condensation or transacylation processes generally described above, viz. amino group containing compounds susceptible to cleavage leaving an a- amido group on the C-terminal of the formed peptide, and subjecting the peptide to such cleavage, peptide amides may be formed in high yields and excellent purity. The process therefore lends itself perfectly to bulk scale production of peptide amides by modification of peptides, which have been made by genetic engineering. Consequently, the invention relates to a process for the preparation of C-terminally amidated peptides P-NH 2 wherein P is a peptide residue which may be N-protected by reacting a substrate component selected from a) C-terminally unblocked peptides of the formula P-OH where P is as defined above b) peptides or peptide derivatives of the formula P-X-Y where P is as defined above, X is an amino acid residue or a peptide residue and Y is OH, OMe or any other suitable C-terminal modification, and c) C-terminally esterified peptides of the formula P-OR' where P is as defined above and R'represents alkyl, aryl, heteroaryl, aralkyl or an α-des amino fragment of an amino acid with a nucleophile component in the presence of an enzyme capable of catalyzing the incorporation of the nucleophile, in an aqueous reaction medium, in an organic solvent or a mixture thereof, at a pH sufficient to maintain the enzyme activity, and the process is characterized by using as nucleophile component an amino group containing compound NH„-R which is cleavable so that the reaction product P-NH-R can be converted into P-NH 2 and subsequently subjecting the reaction product to such cleavage. Consequently, the nucleophile NH -R may be incorporated in three types of enzymatically catalyzed reactions: (a) condensation reactions P-OH + NH 2 -R =-=* P-NH-R + H 2 0 P = peptide residue, which may be N-protected, NH„-R = cleavable nucleophile leaving a C-terminal amide group on the preceeding amino acid residue, i.e. forming P-NH 2 in a subsequent reaction. Such reactions are catalysed by all types of proteases, i.e. aspartic-proteases, metallo-proteases, serine- proteases and thiol proteases, both exopeptidases, e.g. CPD-Y, and endopeptidases, e.g. trypsin and thermolysin (as generally described in US patent No. 4,339,534 and the corresponding DK patent No. 155613, i.e. with reference to EP 17938 (Morihara) which reacted des B-30 insulin with an excess amount of a threonine derivative in the presence of trypsin under formation of human insulin.). The equilibrium of such a condensation reaction is normally shifted to the left in aqueous solution unless very high concentrations of the reactants are used. However, addition of solvents may change the equilibrium constant such that a higher yield of product is obtained. Precipitation of the product in the reaction medium can also shift the reaction in favour of the product. (b) transpeptidation reactions P-X-Y + NH 2 -R -> P-NH-R + H-X-Y P and NH^R are defined as in (a) . X = amino acid residue or peptidyl, Y = OH, 0-alkyl, e.g. OMe or any other suitable C-terminal modification of X. Such reactions are catalysed by thiol and serine proteases, both exopeptidases, e.g. CPD-Y as explained above, and endopeptidases, e.g. trypsin. In case a carboxypeptidase is used, X = amino acid residue and Y = OH. In case an endopeptidase is used X and Y are as defined above. (c) aminolysis of peptide and amino acid esters P-OR 1 + NH 2 -R -> P-NH-R + HO-X P and NH ~R are defined as in (a) R = alkyl, aryl, heteroaryl, aralkyl or an α-desamino fragment of an amino acid. Such reactions are catalysed by thiol and serine proteases, both exopeptidases, e.g. CPD-Y (as explained above), and endopeptidases, e.g. trypsin. The selection of the most appropriate of the above three reaction types can be made by the person skilled in the art based especially on the amino acid sequence of the peptide and the specificity of the available enzymes. If the peptide can easily be converted to an ester, the "aminolysis of esters" is normally preferable. This esterification may create difficulties, particularly if the peptide contains acidic amino acids, e.g. Glu or Asp, since their free carboxy groups would then also be esterified. In these cases a transpeptidation type reaction might therefore be the best choice. Generally speaking the person skilled in the art will be able to select the proper reaction type based on experiments with model peptides. The same goes for the selection of the nucleophile. Once a proper enzyme and reaction type has been chosen, one would preferably select a nucleophile with a suitable pKa such that the nucleophilic amino group is partially or completely deprotonated within the optimal pH-range for the enzyme in the given reaction. Again a proper selection may be made by experiments. By the same token, if the use of a particular nucleophile is desired, e.g. because it is easy to cleave from the reaction product, an enzyme and a reaction type may be chosen where the pH-conditions are adapted to the pKa of the nucleophile. A preferred group of such nucleophiles is disclosed in claim 2, and a preferred segment thereof in claim 3. In case the cleavage is to be induced following rearrangement as further described below, an allyl amine, e.g. as defined in claim 10, may be used as nucleophile. All these compounds may be reacted with the above- mentioned substrate component in a condensation or transacylation reaction under formation of a C-terminal modified peptide which may be transformed to a peptide amide. In Ref. 7 carboxypeptidase Y catalyzed reactions between a common acyl component (Bz-Ala-OMe) and a variety of amine compounds are described. The a-amino acid amides tested were - with the exception of isoglutamine - incorporated in high yields (70-95%). Glycinonitril, glycine-N- methylamide and threonine-N-methylamide reacted in low yields (<20%) only. None of the secondary amines tested (sarcosine, sarcosine methyl ester, N-methyl-alanine, proline and proline amide) were incorporated to form an amide bond. However, several structurally interesting primary amines (e.g. L-alaninol, cyclopropylamine, >- alanine amide, taurine, etc.) reacted in good yields (40- 60%). Benzyl amine reacted in 65% yield, at a concentration of 0.1 M. At higher concentrations (0.5 M) there was no turnover, since apparently the formation of an acyl-enzyme intermediate was inhibited. While it has been shown that some amines were applicable as nμcleophile components in enzymatic peptide synthesis, no reference is made to a subsequent cleavage of the aminolysis product leaving an amido group on the C-terminal. The cleavage of the reaction product may be induced inter alia in the following manners: (a) by photolysis (b) by solvolysis (c) by reduction (d) by rearrangement (e) by oxidation (a) PHOTOLYSIS The peptide to be photolyzed is placed in solution in water, possibly mixed with other solvents like dimethyl- formamide, hexamethylphosphortriamide, dimethylsulfoxide, lower alcohols, ethylene glycol or diethylene glycol ethers, etc. , and irradiated with electromagnetic radiation. In a preferred embodiment, the light has wavelengths longer than 300 nm and shorter than 500 nm, but the useful range of wavelengths will be dependent on the substituents of the cleavable group. In a preferred embodiment of the invention, a mildly reducing agent (e.g. sodium hydrogen sulfite) is added before irradiation in order to reduce strongly colored by-products from the photolysis which may react with the substrates or the products or act as internal filter. Photolysis has earlier been used in connection with solid- phase and liquid-phase peptide synthesis for the removal of C-terminal peptide amides from the resins used as supports in the solid-phase synthesis or from the polyethylene glycol (PEG) used as support in liquid-phase synthesis. Rich, D.H. and Gurwara, S.K. (1975), Tetrahedron Letters 301-304 (Ref. 4) synthesized polystyrene resin having a 3- nitro-4-amino-methylbenzoylamide anchoring group: After removal of the Boc-group with 50% TFA/CH 2 C1 2 followed by neutralization with TEA in CH-C1.-,, Boc-Gly and Boc-Val was coupled with the resin. The protected amino acid amides Boc-Gly-NH._ and Boc-Val- NH 2 were liberated from the anchoring group upon photo¬ lysis of the resin. The resin was suspended in methanol and irradiated at 350 nm in the absence of oxygen. The protected amides were isolated in quantitative yields, which establish that hindered amino acid derivatives can be removed efficiently from the resin under mild conditions. In another experiment the decapeptide LH-RH was prepared according to the classical Merrifield-solid phase synthesis using the above resin as support. The protected decapeptide resin was photolyzed as described above liberating the protected decapeptide amide. These results establish that the particular resin can be used in solid-phase synthesis of protected peptides as the C-terminal amides. Hindered C-terminal amino acids are removed under normal photolysis condition. The authors therefore conclude that the resin is useful for the synthesis of C-terminal peptide amides which are difficult to remove from the classical Merrifield resins under mild conditions. In a very recent article Ajayagosh, A. and Pillai, V.N.R. (1990), J. Org. Chem. 55, 2826-2829 (Ref. 1) reports the use of similar photolabile o-nitrobenzyl anchoring groups for solid-phase synthesis of peptide-N-alkylamide. The rationale for this work is that peptide N-alkylamides in a number of cases have been observed to be more active than the parent peptide amide. In other experiments reported by Pillai, V.N.R. , Mutter, M. and Bayer, E. (1979), Tetrahedron Letters 3409-3412 (Ref. 2) and Pillai, V.N.R., Mutter, M. (1980), J. Org. Chem. 45, 5364-5370 (Ref. 3) the 3-nitro-4-aminomethyl benzoyl anchoring group was attached to polyethylene glycol and used as support in liquid-phase peptide synthesis. Also in this method it was possible to release peptide amides from the photolabile anchoring group by photolysis and the authors conclude that several methodological improvements to liquid-phase synthesis on PEG supports are provided. In particular a new soluble polymeric support which permits synthesis and final photolytic removal of peptide amides under neutral conditions was developed. Furthermore, as described by Wang, S.-S. (1976) J. Org. Chem., 41, 3258-3261 (Ref. 5) other photolabile groups may be used in solid or liquid phase peptide chemistry, e.g. CH, Z-Lys(Z Z-Lys(Z)-Phe-Phe-Gly-OH Based on the above articles it can be concluded that it is known to use photolabile anchoring groups for peptide amides in the classical solid-phase and liquid-phase peptide synthesis. However, it has not earlier been suggested to use photolabile amine compounds as the nucleophile in enzymatic peptide synthesis with a view to obtain peptide amides by C-terminal modification following a photolytic cleavage of the reaction product. Also it could not be expected with any certainty that the claimed compounds would act as nucleophiles in the enzymatic peptide synthesis and that both the incorporation of the nucleophile and the subsequent cleavage would proceed with high yields. (b) SOLVOLYSIS The peptide to be solvolyzed is suspended or preferably dissolved in an acid, or an acid medium, e.g. trifluoro- acetic acid (TFA) or TFA containing various amounts of water and left until all or a sufficiently large amount of the starting material has disappeared. The reaction mixture is subsequently treated in such a way as to remove the cleaved off by-product, e.g. by filtration, and the generated peptide amide is purified if necessary by standard methods. Solvolysis is a well known method for cleavage of peptides generated by solid or liquid phase peptide synthesis from the resins used as supports in the solid phase synthesis or from the polyethylene glycol used as support in liquid phase synthesis. The use of 2,4-dimethoxybenzhydrylamine resins in this connection is described by Penke, B, and Rivier, J. (1987), J. Org. Chem., 52, 1197-1200 (Ref. 6). (c) REDUCTION The peptide to be reduced is suspended or dissolved in a suitable solvent, e.g. acetic acid or aqueous acetic acid, methanol or aqueous methanol, a catalyst is added followed by subjection of the reaction mixture to hydrogen at a pressure between 1 Atm and 200 Atm, or another hydrogen donor may be employed, e.g. formic acid, possibly at elevated temperatures. After uptake of the required amount of hydrogen, the catalyst is removed and the generated peptide amide is purified by standard methods. Furthermore, electrochemical or metal/acid mediated reduction may be employed. (d) REARRANGEMENT By using an allyl amine, preferably as defined in claim 10, as the nucleophile a reaction product is obtained which may be transformed to a peptide amide by rearrangement and subsequent hydrolysis. The peptide to be rearranged is suspended or preferably dissolved in water or a mixture of water and an organic solvent, or an organic solvent, e.g. an alkanol, dimethylformamide, hexamethylphosphortriamide, dimethylsulfoxide dioxane or pyridine, etc. Subsequently a catalyst is added, like e.g. triethylamine, lithium oxide, lithium sulfide, etc. The mixture is reacted for 1-48 hours, after which the resulting peptide amide is purified by standard methods. (e) ELIMINATION By using an amine as nucleophile a reaction product is obtained which may be transformed to a peptide amide by elimination. Nucleophiles in this category are preferably compounds that after insertion in the C-terminal of the peptide can be removed in a 1,2-, 1,4- or 1, 6-elimination . reaction, e.g. allylamine derivatives as described in claim 10. In a preferred form of experiment highly conjugated 0- amino acid derivatives are enzymatically inserted in the peptide which is subsequently subjected to an acid/ or base catalyzed 1, 2-elimination which produces a peptide amide. In another preferred form of experiment hydroxy benzylamine derivatives inserted in the peptide can be subjected to a base catalyzed 1, 6-elimination resulting in formation of the peptide amide. (f) OXIDATION The peptide to be oxidized is suspended or preferably dissolved in water or a mixture of water and an organic solvent. Subsequently a relatively mild oxidizer is added, like e.g. hydrogen peroxide, chloramine-T or immobilized chloramine ( Iodobeads) . As soon as the oxidation is finished, excess oxidant is destroyed or removed, and the resulting peptide amide is purified by standard methods. The leaving group on the substrate component should be selected in accordance with the enzymatic properties of the enzyme used. Preferred enzymes at present are serine carboxypeptidases, but also serine- or thiol-endoproteases or other proteases may be used, if the necessary measures are taken to avoid internal cleavage of the reaction products. As mentioned above serine carboxypeptidases are capable of catalyzing both aminolysis of esters, in which case the leaving group should be an alcohol group and transpeptida¬ tion in which case the leaving group is a free amino acid. The preferred carboxypeptidases in the process of the invention are L-specific serine or thiol carboxy¬ peptidases. Such enzymes can be produced by yeast fungi, or they may be of animal, vegetable or other microbial origin. A particularly expedient enzyme is carboxypeptidase Y from yeast fungi (CPD-Y). This enzyme is described in the earlier patents i.a. with reference to Johansen et al (Ref. 28) who developed a particularly expedient purifica¬ tion method by affinity chromatography on an affinity resin comprising a polymeric resin matrix with coupled benzylsuccinyl grups. CPD-Y, which is a serine enzyme is available in large amounts and displays a high stability. Further details are given in Ref. 14. In addition to CPD-Y, which is the preferred enzyme at present, the process of the invention is feasible with other carboxypeptidases, such as those listed in the following survey: Enzyme Origin Fungi Carboxypeptidase S-l Penicillium janthinellum Carboxypeptidase S-2 Penicillium janthinellum Carboxypeptidase(s) from Aspergillus saitoi Carboxypeptidase(s) from Aspergillus oryzae Plants Carboxypeptidase(s) C Orange leaves Orange peels Carboxypeptidase C N Citrus natsudaidai Hayata Phaseolain French bean leaves Carboxypeptidases M-I and M-II Germinating berlay Carboxypeptidase W-II Wheat bran Carboxypeptidases from Germinating cotton plants Tomatoes Watermelons Bromelain(pineapple)powder The close relationship between a number of the above carboxypeptidases is discussed by Kubota et al (Ref. 33). Another type of applicable enzymes are dipeptidyl peptidases which cleave a dipeptide from the C-terminal. In connection with the above discussion of reactions (a), (b) and (c) a number of applicable enzymes has been suggested. Generally speaking, the person skilled in the art will be able to select a suitable enzyme based on the known specificities for particular amino acids or substrates, the structure of the desired peptide amide, etc. The process of the invention may be carried out at pH 3-11 depending on the enzyme used. The preferred pH-value, which is often within a very narrow range, depends upon the enzyme used and of the type of reaction aimed at, viz. (a), (b) or (c) as discussed above. The selected pH-value should preferably be maintained throughout the reaction. The pH-control may be provided for by incorporating a suitable buffer for the selected pH-range in the reaction medium. The pH-value may also be maintained by adding an acid, such as HCl, or a base, such as NaOH, during the reaction. This may conveniently be done by using a pH-stat. However, the conditions may also be influenced upon by varying the enzyme concentration, reaction time, etc. Depending particularly on the enzyme and the reactants, the reaction is carried out in an aqueous reaction medium or a mixture of water and an organic solvent, or in an organic solvent. Preferred organic solvents are alkanols, e.g. methanol and ethanol, glycols, e.g. ethylene glycol or polyethylene glycols, glycerol, alkanoic acids, e.g. acetic acid, dimethyl formamide, dimethyl sulfoxide, tetrahydrofurane, dioxane and dimethoxyethane. The selection of the composition of the reaction medium depends particularly upon the solubility, temperature and pH of the reaction components and the reaction products involved and upon the stability of the enzyme. The reaction medium may also comprise a component that renders the enzyme insoluble, but retains a considerable part of the enzyme activity, such as an ion exchanger resin. Alternatively, the enzyme may be immobilized in known manner, e.g. by bonding to a matrix, such as a cross-linked dextran or agarose, or to a silica, polyamide or cellulose, or by encapsulating in polyacrylamide, alginates or fibres. Besides, the enzyme may be modified by chemical means or site directed mutagenesis or other genetic means to improve its stability or enzymatic properties. Additives like chelating agents, e.g. EDTA, metal ions and salts are included in the reaction medium depending on the requirements of the enzyme salts. The concentration of the two participants in the reaction may vary within wide limits, as explained below. A pre¬ ferred starting concentration for the substrate component is 0,1-20 mM, preferably 1-10 mM, and for the nucleophile component 0.001 to 2 M, preferably 0.025 to 1.5 M, in particular 0.1 - 1.0 M. The enzyme concentration may vary as well, but the concentration is preferably 10 -8 to 10-4 M. The most advantageous activity depends i.a. on the substrate chain and concentration, the nucleophile concentration, the reaction time, the reaction temperature, the pH, and the presence of organic solvents and/or salts. According to the invention the reaction temperature is -30° to 80°C, preferably 20° to 40°C. The most appropriate reaction temperature for a given synthesis can be determined by experiments. An appropriate temperature will usually be about 20° to 37°C, preferably about 25°C, taking into account due consideration for enzyme activity and stability. Similar variations occur for the reaction time which depends very much upon the other reaction parameters, especially the enzyme concentration. The standard reaction time in the process of the invention is about 1 - 3 hours. The abbreviations of amino acids, amino acid derivatives and peptides are according to Guidelines of the IUPAC-IUB Commission on Biochemical Nomenclature and the amino acids are on L-form unless otherwise stipulated. The following additional abbreviations are used: AcOH, acetic acid; MeOH, methanol; Bz, N-benzoyl; Boc, tert.butoxycarbonyl; DMF, N,N-dimethy1formamide; EDTA, ethylene diamine-N,N,N' ,N' -tetraacetic acid; HPLC, High Performance Liquid Chromatography; TLC, Thin Layer, Chromatography; TFA, trifluoro acetic acid; THF, tetra¬ hydrofuran, TEA, triethylamine; TEAP, triethylammonium phosphate. Before the process of the invention will be illustrated by examples, starting materials, apparatus methods of measurement, etc. will be explained in general terms. In a number of examples N-substituted benzamides were used as models to illustrate the invention, where benzamide was the expected product. These were prepared by standard procedures. Furthermore, the peptides used to demonstrate the invention were often labelled with a benzoyl group at the N-terminus, in order to make their detection by the HPLC and UV detector easier. CHEMICALS CPD-Y courtesy of Carlsberg Ltd. (Klaus Breddam). Z-Gly-Ala-Pro-Ala-OH, Z-Gly-Ala-Pro-NH 2 , Tyr-Gly-Trp-Met- Asp-Phe-Ala-OH, Tyr-Gly-Trp-Met-Asp-Phe-NH 2 , Pro-Gln-Thr- Ala-Ile-Gly-Val-Gly-Ala-Pro-Ala-OH and Pro-Gln-Thr-Ala- Ile-Gly-Val-Gly-Ala-Pro-NH 2 were prepared by standard solid-phase peptide synthesis. 2-Nitrophenylglycine amide and 3-(2,4-dimethoxyphenyl)-3- propionic acid amide were prepared as outlined in example 1 and 10, respectively, Acetonitrile (HPLC-grade, RH 1015) from RATHBURN. All other chemicals are from Aldrich. Other organic solvents for HPLC were distilled before use. APPARATUS pH measurements: Radiometer (Copenhagen) pH Meter 28 Electrode GK 2421C (Radiometer, Glass electrode) HPLC system: Waters 994 Programmable Photodiode Array Detector Waters 600E Multisolvent Delivery System Waters 5200 Printer Plotter Photolysis: Bausch & Lomb Mercury Light Source SP200 Bausch & Lomb Mercury Power Aupply SP200 HPLC SOLVENT SYSTEM System 1: Buffer A: 50 mM TEAP , pH~3.0, containing 10% (v/v) CHgC Buffer B: CH.-,CN containing 20% (v/v) buffer A. Gradient: t (min) %A %B 0 60 40 linear 10 0 100 linear 15 60 40 System 2: Buffer A: 50 mN TEAP, pH~3.0 containing 10% (v/v) CH 3 CN Buffer B: CH.-,CN containing 20% (v/v) buffer A Gradient: t (mi ) %A %B 0 100 0 concave 60 40 linear 8 100 0 System 3: Buffer A: 50 mM TEAP, pH~3.0 containing 10% (v/v) CH--CN Buffer B: CHgCN containing 20% (v/v) buffer A Isocratic. %A %B 60 40 Flow: 1.5 ml/min. System 4: Buffer A: H^O containing 0.1% (v/v) trifluoro- acetic aid (TFA) Buffer B: CHgCN containing 10% (v/v) H 2 0 and 0.2% (v/v) TFA Isocratic: %A %B 80 20 Flow: 1.5 ml/min. System 5: Buffer A: H_0 containing 0.1% (v/v) trifluoro- acetic acid (TFA) Buffer B: CHgCN containing 10% (v/v) H 2 0 and 0.1% (v/v) (TFA) Isocratic: %A %B 70 30 Flow: 2 ml/min System 6: Buffer A: H-O containing 0.1 (v/v) trifluoro- acetic acid (TFA) Buffer B: CHgCN containing 10% (v/v) H 2 0 and 0.1% (v/v) (TFA) Gradient: t(min) 0 5 10 12 Flow: 2 ml/min 0.5 M Triethylammonium phosphate buffer: 900 ml H^O containing 33.7 ml orthophosphoric acid (85%). Titrated with triethylamino to pH ~ 3.0. Add H 2 0 to 1000 ml. EXAMPLE 1 Preparation of N-benzoyl-2-nitrophenylglycine amide (model compound) and photolytic cleavage A) Preparation of 2-nitrophenylglycine amide 2-Nitrophenylacetic acid (36.2 g, 0.2 mol) was suspended in thionyl chloride (30 ml, 0.4 mol) and stirred for 16 h at 20°C. The reaction mixture was evaporated to dryness and suspended in dry chloroform (100 ml) . To this solution was added N-bromosuccinimide (50 g, 0.28 mol), dibenzoyl peroxide (10 mg) as catalyst and it was refluxed for 48 h. The reaction mixture was evaporated to dryness, suspended in dry ether, and the precipitated (N-hydroxysuccinimide) was filtered off. The solution was evaporated to dryness and added to cold concentrated ammonium hydroxide (200 ml). The β-bromo-(2-nitrophenyl)acetamide was collected by filtration after 1 h and recrystallized from water/ethanol (4:1 v/v). Yield: 20.0 g (38%); mp 126-129°C, identified by IR and H NMR spectroscopy. α-Bromo-(2-nitrophenyl)acetamide (4.0 g, 0.02 mol) was suspended in DMSO (4 ml) and treated with NH„; after 2 h the reaction mixture was freeze dried and the product was recrystallized from absolute ethanol. Yield: 2.6 g (68%); mp 116-120°C, identified by H NMR spectroscopy. B) Preparation of N-Benzoyl-2-nitrophenylglycine amide 2-Nitrophenylglycine amide, 1 DMSO (500 mg, 1.83 mmol) was suspended in chloroform (20 ml) and triethylamine (390 μl, 2.80 mmol) was added. Subsequently benzoyl chloride (330 μl, 2.8 mmol) was added with stirring. The reaction was followed by TLC using as eluent: CH 2 Cl 2 /MeOH/AcOH (85:10:5) and after 1 h at 20°C the reaction mixture was treated with a saturated solution of NaHCO.-,, dried over anhydrous MgSO. and evaporated to dryness. The product was recrystallized from absolute ethanol. Yield: 600 mg (77%); mp 178-180°C, identified by H NMR spectroscopy. Reaction scheme: C ) Photolysis of N-benzoyl-2-nitrophenylglycine amide N-Benzoyl-2-nitrophenylglycine amide (15 mg, 50 μmol) was dissolved in methanol:water (1:1) (25 ml), and NaHSO,, (52 mg, 500 μmol) was added. The reaction mixture was purged with nitrogen for 15 min and subsequently photolyzed under nitrogen cover at a distance of 20 cm from the SP 200 lamp through Pyrex. The photolysis was followed by taking out samples at 0, 10, 30 and 60 min. Each sample was analyzed using HPLC system-1 and the results were compared with the 0' test as well as with the chromatogram of a 10 mM benzamide solution in methanol. The generated benzamide was determined quantitatively using benzamide standard chromatograms. EXAMPLE 2 Preparation of Bz-Ala-NH 2 by "aminolysis of esters" and photolytic cleavage A) Transacylation of Bz-Ala-OMe with 2-nitrophenylglycine amide as nucleophile using enzyme CPD-Y EDTA (2 ml, 5 mM), pH ~ 7.9 was added to 2-nitrophenyl- glycine amide, 1 DMSO (25 mg), pH was adjusted to ~8.0 with 5 N NaOH, and Bz-Ala-OMe (10 μl) of a solution of 207 mg/ml (1 M) MeOH was added. A 0' sample was prepared by adding CH 3 C (240 ml) to 10 μl reaction mixture. Subsequently CPD-Y (15 μl, 22 mg/ml H„0) was added to the reaction mixture, and the reaction was followed by extracting samples at 15, 30 and 60 min. Each sample was analyzed using HPLC system-3 and the chromatograms compared to those of standard solutions of Bz-Ala-OMe and Bz-Ala-OH, whereby the product distribution was determined. At t = 75 min, 0.1 N HCl was added until pH corresponded to 2.0. The reaction mixture was extracted with CHCl (3 x 2 ml) and the chloroform phase was dried over MgSO * and evaporated. The residue was dissolved in methanol and analyzed using HPLC system-3, which showed that the residue contained ~100% transacylation product which was identified by mass spectrometry as Bz-Ala-2- nitrophenylglycine amide, M = 370. B) Photolysis of Bz-Ala-2-nitrophenylglycine amide Bz-Ala-2-nitrophenylglycine amide (95 mg) purified by semipreparative HPLC as described above was dissolved in MeOH/H 2 0 (25 ml, 1:1), and NaHS0 3 (52 mg, 0.62 mmol) was added to quench the generated 2-nitrosophenylglycolamide which would otherwise form a powerful inner filter, which could reduce the peptide amide yield. The reaction mixture was purged with nitrogen for 15 min and subsequently photolysed with nitrogen cover a distance of 20 cm from the SP 200 lamp through Pyrex. After 30, 60 and 90 min samples were taken out and analyzed using HPLC system-1. The results were compared with that from the starting material and the chromatogram of 1 mM Bz-Ala-NH 2 prepared from Bz-Ala-NHS and 25% aqueous ammonia. The photolysis mixture was extracted with chloroform (2 x 10 ml). 0.5 N NaOH was added to the aqueous phase until pH ~ 9.0 and it was subsequently extracted with chloroform (2 x 20 ml) . The chloroform phase was dried over MgSO. and evaporated. The residue was dissolved in MeOH/H_0 (1:1) and analyzed using HPLC system-1. The residue was found to contain ~ 100% Bz-Ala-NH . The reaction product was identified using UV spectroscopy, HPLC chromatograms and mass spectrometry. EXAMPLE 3 Preparation of Bz-Ala-NH 2 by "aminolysis of as nucleophile ester" and photolytic cleavage A) Transacylation of Bz-Ala-OMe with 2-nitrobenzylamine as nucleophile using CPD-Y 2-Nitrobenzylamine, HCl (94 mg) was added to 5 mM EDTA (2 ml, pH ~ 8.0), pH was adjsuted to 8.0 with 5N NaOH, and Bz-Ala-OMe (20 μl, 207 mg/(ml MeOH)) was added. A 0 sample was prepared by adding CH.,CN (240 μi) to 10 μl of the reaction mixture. Subsequently CPD-Y (15 μl, 22 mg/ml H ^ O) was added, and the reaction was followed by extracting and analyzing samples at 5, 60, 120 and 300 min. Each sample was analyzed using HPLC system-6 and the chromatograms compared to standard solutions of Bz-Ala-OMe and Bz-Ala-OH whereby the product distribution was determined, (the transacylation product yield was >90%). The reaction product was identified using mass spectrometry as Bz-Ala- 2-nitrobenzylamide, M = 305. B) Photolysis of Bz-Ala-2-nitrobenzylamide Bz-Ala-2-nitrobenzylamide (1.5 mg) in MeOH (1.0 ml) was added to NaHS0„ (6.2 mg) ( issolved in 1.0 ml water) and pH was adjusted to 9.5 with 5N NaOH. The reaction mixture was purged with nitrogen for 15 min and subsequently photolysed with nitrogen cover at a distance of 20 cm from the SP 200 lamp through Pyrex. The reaction was followed by extracting and analyzing samples at 0, 1, 5, . 10 and 30 min. Each sample was analyzed using HPLC system-6 and the chromatograms compared with those of a 10 mM standard solution of Bz-Ala-NH ^ . The reaction product (yield >95%) was further identified using mass spectrometry. EXAMPLE 4 Preparation of Bz-Gly-Ala-Pro-2-nitrophenyl- glycine amide by transpeptidation A) Hydrolysis of Bz-Gly-Ala-Pro-Ala-OH with CPD-Y EDTA (95 μl, 5 mM), pH ~ 6.5 (prepared as follows: 146 mg EDTA was dissolved in 2.0 ml 2 N NaOH, pH adjusted to 6.5 with 4 N HCl and R-,0 added to 100 ml) was added to 100 mM Bz-Gly-Ala-Pro-Ala-OH (5 μl, 41 mg/ml DMF) and pH was adjusted to 6.5 with 5 N NaOH. As a 0' test 10 μl of the reaction mixture was added to 200 μl of CH^-CN. After this CPD-Y (10 μl, 22 mg/ml H 2 0) was added to the reaction mixture, and the reaction was followed by extracting 10 μl samples at 1, 5, 10 and 20 min. CPD-Y was denatured by adding CH-CN (200 μl) to each sample. Each sample was analyzed using HPLC system-2 and the chromatograms compared with those of 10 M standard solutions of Bz-Gly- OH and Bz-Gl-Ala-Pro-Ala-OH, whereby retention times for Bz-Gly-Ala-Pro-OH and Bz-Gly-Ala-OH could be determined. B) Transpeptidation of Bz-Gly-Ala-Pro-Ala-OH with 2- nitrophenylglycine amide as nucleophile using CPD-Y 70 mM 2-nitrophenylglycine amide (95 μl), pH ~ 6.5 (prepared as follows: 70 mM 2-nitrophenylglycine amide (1 M DMSO/mol, 19.5 mg) was added to 5 mM EDTA (1 ml, pH ~ 6.5) and pH adjusted to 6.5 with 5 N NaOH) was mixed with 100 mM Bz-Gly-Ala-Pro-Ala-OH (5 μl, 41 mg/ml DMF). 0 min test: 10 μl reaction mixture was added to 200 . μl CH„CN. Subsequently CPD-Y (15 μl, 22 mg/ml H ? 0) was added to the reaction mixture. The reaction was followed by extraction of samples at 1, 5, 10, 20, 40, 60, 120 and 180 min. The enzymatic reaction was stopped by adding CH^CN (200 μl) to each sample. The samples were analyzed using HPLC system- 2, and the chromatograms were compared with the chromatogram corresponding to 0 min and the results from CPD-Y hydrolysis of Bz-Gly-Ala-Pro-Ala-OH. The trans¬ peptidation product was purified using HPLC system-4 and identification performed by determination of retention time, UV, H NMR and mass spectrometry. EXAMPLE 5 Preparation of Z-Gly-Ala-Pro-NH,-, by trans- peptidation and photolytic cleavage A) Transpeptidation of Z-Gly-Ala-Pro-Ala-OH with 2- nitrophenylglycine amide as nucleophile using CPD-Y 300 mM 2-nitrophenylglycine amide (95μl), pH ~ 6.0 (prepared as follows: 2-nitrophenylglycine amide, DMSO (81.9 mg) was added to 5 mM EDTA (1 ml, pH ~ 6.5) and pH was adjusted to 6.0 with 5N NaOH) was mixed with 10 mM Z- Gly-Ala-Pro-Ala-OH (5 μl, 4.1 mg/ml DMF). 0' test: 10 μl reaction mixture was added to 200 μl CH...CN. Subsequently CPD-Y (15 μl, 22 mg/ml H 2 0) was added to the reaction mixture. The reaction was followed by extraction of samples for analysis at 10, 60, 120, 180 min. The enzymatic reaction was stopped by adding CH^CN (200 μl) to each sample. The samples were analyzed using HPLC system- 5, and the chromatograms were compared with the chromatogram corresponding to 0 min and the results from CPD-Y hydrolysis of Z-Gly-Ala-Pro-Ala-OH. The yield of transpeptidation product was ~ 100% and it was purified using HPLC system-4. Identification was performed by determination of retention time, UV spectroscopy, H NMR spectroscopy and by mass spectrometry. B) Photolysis of Z-Gly-Ala-Pro-2-nitrophenylglycine amide Z-Gly-Ala-Pro-2-nitrophenylglycine amide (0.025 mmol) in MeOH (12,5 ml) was added to 80 mM NaHSO.-, (12.5 ml) and pH was adjsuted to 9.5 with 5N NaOH. The reaction mixture was purged with N 2 for 15 min and subsequently photolyzed under nitrogen cover at a distance of 20 cm from the SP 200 lamp through Pyrex. The photolysis was followed by extraction of samples at 0, 30, 60 and 120 min. Each sample was analyzed using HPLC system-5 and the results compared with those from the 0 min sample as well as with the results from the CPD-Y hydrolysis and the trans¬ peptidation mixture. Purification and identification were performed as previously described. EXAMPLE 6 Preparation of Tyr-Gly-Trp-Met-Asp-Phe-NH ? by transpeptidation and photolytic cleavage A) Transpeptidation of Tyr-Gly-Trp-Met-Asp-Phe-Ala-OH with 2-nitrophenylglycine amide as nucleophile using CPD-Y 300 mM 2-nitrophenylglycine amide (95 μl), pH ~ 5.5, (prepared as follows: 2-nitrophenylglycine amide, 1 DMSO (81.9 mg) was added to 5 mM EDTA (1 ml, pH ~ 6.5) and pH was adjusted to 5-5 with 5N NaOH), was mixed with 50 mM Tyr-Gly-Trp-Met-Asp-Phe-Ala-OH (1 μl, 44.4 mg/ml DMF). 0 test: 10 μl reaction mixture was added to CH^CN (200 μl) . Subsequently CPD-Y (2 μl, 22 mg/ml water) was added to the reaction mixture. The reaction was followed by extraction of samples for analysis at 10, 60, 120, 180 min. The enzymatic reaction was stopped by adding CH^CN (200 μl) to each sample. The samples were analyzed using HPLC system- 5, and the chromatograms were compared with the chromato¬ gram corresponding to 0 min and the results from CPD-Y hydrolysis of Tyr-Gly-Trp-Met-Asp-Phe-Ala-OH. The yield of transpeptidation product was ~ 100% and it was purified using HPLC system-4. Identification was performed by determination of retention time, UV spectroscopy, amino acid analysis, and mass spectrometry. B) Photolysis of Tyr-Gly-Trp-Met-Asp-Phe-2-nitrophenyl- glycine amide Tyr-Gly-Trp-Met-Asp-Phe-2-nitrophenylglycine amide (0.025 mmol) in 12.5 ml MeOH was added to 80 mM NaHS0 3 (12.5 ml) and pH was adjusted to 9.5 with 5N NaOH. The reaction mixture was purged with N„ for 15 min and subsequently photolyzed under nitrogen cover at a distance of 20 cm from the SP 200 lamp through Pyrex. The photolysis was followed by extraction of samples for analysis at 0, 30, 60 and 120 min. Each sample was analyzed using HPLC system-5 and the results compared with the 0 min sample as well as with the results from the CPD-Y hydrolysis and from the transpeptidation mixture. Purification and identification were performed as previously described. EXAMPLE 7 Preparation of Pro-Gln-Thr-Ala-Ile-Gly-Val-Gly- Ala-Pro-NH-, by transpeptidation and photolytic cleavage A) Transpeptidation of Pro-Gln-Thr-Ile-Gly-Val-Gly-Ala- Pro-Ala-OH with 2-nitrophenylglycine amide as nucleophile using CPD-Y 220 mM 2-nitrophenylglycine amide (400 μl), .pH ~ 6.0 (prepared as follows: 2-nitrophenylglycine amide, 1 DMSO (60 mg) was added to 5 mM EDTA (1 ml, pH ~ 6.5) and pH was adjusted to 6.0 with 5N NaOH) was mixed with 50 mM Pro- Gln-Thr-Ala-Ile-Gly-Val-Gly-Ala-Pro-Ala-OH (4 μl, 49 mg/ml DMF). 0 min test: 10 μl reaction mixture was added to CH 3 CN (200 μl). Subsequently CPD-Y (2 μl, 22 mg/ml water) was added to the reaction mixture. The reaction was followed by extraction of samples for analysis of 10, 60, 120, 180 min. The enzymatic reaction was stopped by adding CH-CN (200 μl) to each sample. The samples were analyzed using HPLC system-5, and the chromatograms were compared with the chromatogram corresponding to 0 min and the results from CPD-Y hydrolysis of Pro-Gln-Thr-Ala-Ile-Gly- Val-Gly-Ala-Pro-Ala-OH. The yield of transpeptidation product was >90% and it was purified using HPLC. Identification was performed by determination of retention time, UV spectroscopy, amino acid analysis, and mass spectrometry. B) Photolysis of Pro-Gln-Thr-Ala-Ile-Gly-Val-Gly-Ala-Pro- 2-nitrophenylglycine amide Pro-Gln-Thr-Ala-Ile-Gly-Val-Gly-Ala-Pro-2-nitrophenyl- glycine amide (0.025 mmol) in 12,5 ml MeOH was added to 80 mM NaHS0 3 (12.5 ml) and pH was adjusted to 9.5 with 5 N NaOH. The reaction mixture was purged with N„ for 15 min and subsequently photolyzed under nitrogen cover at a distance of 20 cm from the SP 200 lamp through Pyrex. The photolysis was followed by extraction of samples for analysis at 0, 30, 60 and 120 min. Each sample was analyzed using HPLC system-5 and the results compared with the 0 sample as well as with the results from the CPD-Y hydrolysis and from the transpeptidation mixture. The yield of photolysis product was >70% (HPLC). Purification and identification were performed as previously described. EXAMPLE 8 Preparation of Bz-Ala-phenacylamide by "aminolysis of ester" Transacylation of Bz-Ala-OMe with 2-aminoacetophenone as nucleophile using CPD-Y 2-Aminoacetophenone,HCl (17.2 mg) was added to 5 mM EDTA (1 ml, pH ~ 8.0). pH was adjusted to 8.0 with 5 N NaOH, and Bz-Ala-OMe (10 μl, 207 mg/(ml MeOH)) was added. A 0 sample was prepared by adding CH 3 CN (240 μl) to 10 μl of the reaction mixture. Subsequently CPD-Y (5 μl, 18 mg/ml H-0) was added, and the reaction was followed by extracting and analyzing samples at 5, 30, 60 and 120 min. Each sample was analyzed using HPLC system-6 and the chromatograms compared to those of standard solutions of Bz-Ala-OMe and Bz-Ala-OH whereby the product distribution was determined (the transacylation product yield was ~80%) . The reaction product was identified using mass spectrometry as Bz-Ala-phenacylamide, M = 310. EXAMPLE 9 Preparation and solvolysis of benzoylbenzyl- amine model compounds for solvolytic cleavage A) N-benzoyl-2,4-dimethoxybenzylamine 2,4-dimethoxybenzylamine, HCl (1.02 g, 5.0 mmol) was suspended in pyridine (20 ml) and cooled in an ice bath. Subsequently benzoyl chloride (0.52 ml, 6.5 mol) was added with stirring. The ice bath was removed and the reaction mixture was stirred for 3 h at 20°C. The reaction mixture was extracted with 2 x 20 ml methylene chloride and the organic layer was treated with 2 x 10 ml 0.1 N NaOH, 3 x 20 ml 0.5 N HCl, 2 x 20 ml saturated solution of NaCl, dried over anhydrous MgSO- and evaporated to dryness- The product was recrystallized from absolute ethanol. Yield: 760 mg (56%); mp 101-103°C, identified by IN NMR spectroscopy. B) Solvolysis of N-benzoyl-2,4-dimethoxybenzylamine N-benzoyl-2,4-dimethoxybenzylamine (100 mg, 0.37 mmol) was dissolved in 1 ml 95% (v/v) trifluoroacetic acid (TFA) and stirred for 1 h at 20°C. TFA was removed by blowing with N„ and the residue suspended in methanol (1 ml) . The reaction mixture was analyzed by TLC using CH 2 Cl 2 /MeOH/AcOH (85:10:5) as eluent and the generated benzamide was determined quantitatively by HPLC using benzamide standard chromatograms. N-Benzoyl-4-methoxybenzylamine, N-benzoyl-2-methoxybenzyl- amine and N-benzoyl-3,4-dimethoxybenzylamine were prepared and solvolyzed as described above. EXAMPLE 10 Preparation and solvolysis of a propionic amide model compound for solvolytic cleavage A) Preparation of 3-amino-3-(2,4-dimethoxy- phenyl)propionic amide A mixture of 2,4-dimethoxybenzaldehyde (21 g, 0.126 mol), malonic acid (20 g, 0.192 mol), ammonium acetate (30 g, 0.39 mol) and ethanol (40 ml) was stirred and heated on a water bath. After refluxing for 4 h the mixture was cooled to 40°C and the precipitated salt was filtered off and washed with cold ethanol (40 ml) . By additional cooling on ice a further crystalline material precipitated, which was isolated and washed with cold ethanol (40 ml) and ether (40 ml). Recrystallisation from ethanol/water 60:40 gave 3-amino-3-(2,4-dimethoxyphenyl)propanoic acid (10.3 g, 38%). Benzyl chloroformate (2.5 ml, 17.5 mmol) was added dropwise to a solution of 3-amino-3-(2,4-dimethoxy- phenyl)propanoic acid (3 g, 14,1 mmol) in 0.2 N NaOH (20 ml) . The solution was stirred for 2 h and kept basic with additional cone. NaOH. The solution was acidified with cone- HCl whereby Z-3-amiho-3-(2,4-dimethoxy- phenyl)propanoic acid precipitated. The precipitate was isolated and washed with ether to give (3.9 g, 11.29 mmol, 80%). A solution of Z-3-amino-3-(2,4-dimethoxyphenyl)propanoic acid (2.3 g, 6.66 mmol) in methylene chloride (20 ml) was cooled on ice and thionyl chloride (0.5 ml, 6.85 mmol) was added. The solution was stirred for 1 h at room temperature, filtrated and evaporated to dryness under reduced pressure. The residue was dissolved in methylene chloride (20 ml) and stirred vigorously while cone, ammonium hydroxide was added. After 1 h Z-3-amino-3-(2,4- dimethoxyphenyl)propanoic amide precipitated. The precipitate was isolated and washed with ether (2.1 g, 6.1 mmol, 91%). A mixture of Z-3-amino-3-(2,4-dimethoxyphenyl)propanoic amide (0.7 g, 2.03 mmol) and 10% Pd/C (0.5 g) and 4.4% formic acid in water (20 ml) was stirred for 1 h at room temperature. The catalyst was removed by filtration and the clear solution was evaporated under reduced pressure whereby 3-amino-3-(2,4-dimethoxyphenyl)propanoic amide (0.43 g, 1.92 mmol, 94%) was isolated. Mp: 148-149°C, and the compound was identified by H-NMR spectroscopy. B) Preparation of N-benzoyl-3-amino-3-(2,4-dimethoxy- phenyl)propionic amide Prepared analogously with N-benzoyl-2-nitrophenylglycine amide described in example IB. C) Solvolysis of N-benzoyl-3-amino-3-(2,4-dimethoxy- phenyl) ropionic amide N-benzoyl-3-amino-3-(2,4-dimethoxyphenyl)propionic amide (6.09 mmol) was dissolved in TFA/thioanisole/pentamethyl- benzene (2 ml/0.587 ml, 5 mmol/29.7 mg, 0.2 mmol). After 1 h the generated benzamide (4.99 mmol 82%) was determined quantitatively by HPLC using benzamide standard chromatograms. EXAMPLE 11 Transacylation of Bz-Ala-OMe with 2,4- dimethoxybenzylamine as nucleophile using enzyme CPD-Y 2,4-dimethoxybenzylamine, HCl (41 mg) was added to 5 mM EDTA (2 ml, pH ~ 8.0), pH was adjusted to 9.5 with 5 N NaOH, and Bz-Ala-OMe (20 μl, 207 mg/ml MeOH) was added. A 0 sample was prepared by adding CH„CN (240 μl) to 10 μl reaction mixture. Subsequently CPD-Y (10 μl, 22 mg/ml H ^ O) was added to the reaction mixture, and the reaction was followed by extracting and analyzing samples at 5, 30, 60 and 120 min. Each sample was analyzed using HPLC system-6 and the chromatograms compared to standard solutions of Bz-Ala-OMe and Bz-Ala-OH whereby the product distribution was determined, (the transacylation product yield was >60%). The reaction product was identified using mass spectrometry as Bz-Ala-2,4-dimethoxybenzylamide, M 320. The transacylation product may be transformed to Bz-Ala- NH„ by solvolytic cleavage. EXAMPLE 12 Preparation of N-protected Phe-NH.-, by trans¬ peptidation and solvolytic cleavage A) Transpeptidation of Z-Phe-Ala-OH with 2,4-dimethoxy- benzylamine using CPD-Y 2,4-Dimethoxybenzylamine,HCl (166 mg, 0.8 mmol) was added to 5 mM EDTA (2 ml, pH ~ 8.0), pH was adjusted to 8.5 with 5N NaOH, and Z-Phe-Ala-OH (10 μl of a solution of 68 mg/ml DMF) was added. A 0 sample was prepared by adding CH-.CN (240 μl) to 10 μl reaction mixture. Subsequently CPD-Y (15 μl, 18 mg/ml H„0) was added to the reaction mixture, and the reaction mixture was analyzed after 5, 60, 120 and 300 min. Each sample was analyzed using HPLC system-7 and the chromatograms compared to standard solutions of Z-Phe-Ala- OH and Z-Phe-OH, whereby the product distribution was determined; the transpeptidation product yield was ~ 60%. The reaction product was identified using mass spectro- metry as Z-Phe-2,4-dimethoxybenzylamide, M = 448.5, and purified using HPLC system-7. B) Solvolysis of Bz-Phe-2,4-dimethoxybenzylamide Bz-Phe-2,4-dimethoxybenzylamide (4.2 mg) was dissolved in trifluoroacetic acid (1.0 ml) containing 10% (v/v) tri- fluoromethane sulfonic acid (TFMSA) and the reaction mixture was kept in the dark for 1 h at 20°C. TFA/TFMSA was removed by blowing with N and the residue was suspended in 95% (v/v) CHgCN (1.0 ml). The reaction mixture was analyzed by HPLC system-7 and the generated Bz-Phe-NH 2 (>95%) was determined quantitatively using a Bz-Phe-NH standard solution (Bz-Phe-NH 2 was purchased from Bache , Switzerland). EXAMPLE 13 Reductive cleavage of N-benzoylphenacylamine as model compound Preparation of N-benzoylphenacylamine Prepared as N-benzoyl-2-nitrophenylglycine amide described in example IB. Cleavage of N-benzoylphenacylamine N-benzoylphenacylamine (10 mg, 41,8 mmol) was dissolved in 60% aqueous acetic acid (1 ml) and Zn-dust (20 mg) was added. Samples were taken every 15 min and N- benzoylphenacylamine, the generated benzamide and acetophenone were quantified by HPLC using standard samples of the respective compounds. EDTA (2 ml, 5 mM), pH ~ 8.0, was added to allylamine (11 mg). pH was adjusted to ~ 8.5 with 5N NaOH, and Bz-Ala-OMe (20 μl) of a solution of 207 mg/ml methanol (1M) was added- A 0 min sample was prepared by adding 200 μl methanol to 10 μl reaction mixture. Subsequently, CPD-Y (10 μl, 22 mg/ml H 2 0 was added to the reaction mixture, and the reaction was followed by extracting samples at 10, 30 and 60 min. Each sample was analyzed using HPLC and the Bz-Ala-OH, whereby the product distribution was determined. The transacylation product may be transformed to Bz-Ala-NH by rearrangement and hydrolysis. B) Rearrangement and hydrolysis of N-benzoylallylamine (model compound) with LiOH N-Benzoylallylamine (161 mg, 1 mmol) was dissolved in 5 ml acetone and 5 ml 0.5 N LiOH was added. The solution was stirred to 1 h at 20°C. Acetone was removed by blowing with N„ and the aqueous solution, pH was adjusted to ~ 1.0 with 6N HCl. The reaction mixture was qualitatively analyzed by TLC to determine generated benzamide. EXAMPLE 15 Preparation of Bz-Phe-NH^ by oxidative cleavage Oxidation of Bz-Phe-2,4-dimethoxybenzylamide Bz-Phe-2,4-dimethoxybenzylamide prepared analogously with example 12 (8,4 mg) was dissolved in CH 3 CN/H 2 0 (100:1) (2.0 ml). To this solution (NH 4 )Ce(N0 3 ) g (CAN) (15.4 mg) was added and the reaction mixture was kept in the dark for 2 h at 20°C. The reaction mixture was analyzed by HPLC system-7 and the generated Bz-Phe-NH 2 (>65%) was determined quantitatively using a Bz-Phe-NH„ standard solution (Bz-Phe-NH 2 purchased from Bachem, Switzerland). EXAMPLE 16 Preparation of Bz-Ala-Ala-Arg-2-nitrophenyl- glycine amide by transpeptidation A) Transpeptidation of Bz-Ala-Ala-Arg-Ala-Phe-Ala-OH with 2-nitrophenylglycine amide as nucleophile using trypsin To a solution of 400 mM 2-nitrophenylglycine amide and 2 mM CaCl 2 in DMF/H 2 0 (60/40 v/v, 120 μl pH ~ 6-5) was added 80 mM Bz-Ala-Ala-Arg-Ala-Phe-Ala-OH in DMF (3 μl). Trypsin (3 μl, 0.5 mg/ml in Hepes, 2 mM CaCl 2 , pH 7.5) was added to the reaction mixture. The reaction was followed by extracting a sample at 93 min. The sample was analyzed using HPLC, and the chromatogram was compared with the chromatogram corresponding to the results from trypsin hydrolysis of Bz-Ala-Ala-Arg-Ala-Phe-Ala-OH and amino acid analysis of reaction products. The reaction mixture composition was as follows: Bz-Ala-Ala-Arg-Ala-Phe-Ala-OH: 21% Bz-Ala-Ala-Arg-OH: 25% Bz-Ala-Ala-Arg-2-nitrophenylglycine amide: 53% The fraction of aminolysis was ~ 68% and the product was purified by HPLC and identification was performed by determination of retention time, amino acid analysis and by mass spectrometry. Same experiment performed with 800 mM 2-nitrophenylglycine amide and 1 μl trypsin (5 mg/ml). The reaction mixture was analyzed after 55. min and composition was as follows: Bz-Ala-Ala-Arg-Ala-Phe-Ala-OH: 11% Bz-Ala-Ala-Arg-OH: 19% Bz-Ala-Ala-Arg-2-nitrophenylglycine amide: 70% The ' fraction of aminolysis was ~ 79% and the product was purified by HPLC and identification was performed by determination of retention time, amino acid analysis and by mass spectrometry. REFERENCES: 1. Ajayaghosh, A. and Pillai, V.N.R. (1990), J. Org. Chem. 55, 2826-2829 2. Pillai, V.N.R., Mutter, M. and Bayer, F. (1979), Tetrahedron Letters 3409-3412 3- Pillai, V.N.R. and Mutter, M. (1980), J. Org. Chem. 45, 5364-5370 4. Rich, D.H. and Gurwara, S.K. (1975), Tetrahedron Letters 301-304 5. Wang, S.-S. (1976), J. Org. Chem., 41, 3258-3261 6. Penke B. and Rivier, J. (1987), J. Org. Chem., 52, 1197-1200 7. Widmer, F. , Breddam, K. & Johansen, J.T. (1981), Carlsberg Res. Comm. 46, p. 97-106 14. Breddam, K. , Widmer, F. & Johansen, J.T. (1980) Carlsberg Res. Commun. 45, 237-247 15. Breddam, K. , Widmer, F. & Johansen, J.T. (1981) Carlsberg Res. Commun. 46, 121-128 16. Breddam, K. , Johansen, J.T. & Ottesen, M. (1984) Carlsberg Res. Commun. 49, 457-462 17. Breddam, K. (1985) Carlsberg Res. Commun. 50, 309-323 " 18. Breddam, K. (1988) Carlsberg Res. Commun. 53, 309-320 21. Breddam, K. Sørensen, S.B. & Svendsen, I. (1987) Carlsberg Res. Commun. 52, 297-311 23. Breddam, K. , Widmer, F. & Johansen, J.T. (1981) Carlsberg Res. Commun. 46, 361-372 25. Breddam, K. & Ottesen, M. (1984) Carlsberg Res. Commun. 49, 473-481 26. Breddam, K. , Sørensen, S.B. & Ottesen, M. (1985) Carlsberg Res. Commun. 50, 199-209 28. Johansen, J.T., Breddam, K. & Ottesen, M. (1976) Carlsberg Res. Commun. 41, 1-14 30. Breddam, K. , Sørensen, S.B. & Ottesen, M. (1983) Carlsberg Res. Commun. 48, 217-230 33. Kubota et al. Carboxypeptidase C N (1973), J. Biochem. 74, no. 4, 757-770

Claims: C L A I S 1. Process for the preparation of C-terminally amidated peptides P-NH 2 wherein P is a peptide residue which may be N-protected by reacting a substrate component selected from a) C-terminally unblocked peptides of the formula P-OH where P is as defined above b) peptides or peptide derivatives of the formula P-X-Y where P is as defined above, X is an amino acid residue or a peptide residue and Y is OH, OMe or any other suitable C-terminal modification, and c) C-terminally esterified peptides of the formula P-OR' where P is as defined above and R'represents alkyl, aryl, heteroaryl, aralkyl or an α-des amino fragment of an amino acid with a nucleophile component in the presence of an enzyme capable of catalyzing the incorporation of the nucleophile, in an aqueous reaction medium, in an organic solvent or a mixture thereof, at a pH sufficient to maintain the enzyme activity, CHARACTERIZED BY using as nucleophile component an amino group containing compound NH 2 ~R which is cleavable so that the reaction product P-NH-R can be converted into P-NH- and subsequently subjecting the reaction product to such cleavage. 2. Process according to claim 1, wherein the nucleophilic component R-NH„ is selected from derivatives with the general formula wherein A-F represent carbon atoms or up to two nitrogen atoms, Y represents H, straight chain or branched (C.-C fi ) alkyl, aryl (R 2 ) or aralkyl (CH,R 2 or CHR 2 R 3 ), wherein R 2 and R 3 are aryl groups, which may be substituted with halogen, hydroxy, alkoxy, nitro, amino or alkyl, or Y represents an oxo group, a carboxy group, or a functional derivative of a carboxy group, e.g. an ester, a nitrile, an amide which may be substituted on N with (C..-C, ) alkyl, aryl (R 2 ) or aralkyl (CH„R 2 or CHR 2 R 3 ), where R and R 3 are as defined above, Xl-X5 independently represent H, (C--C-.) alkyl, halogen, cyano, methoxy, hydroxy, sulfo, amino or nitro, or any two of them may join to form a fused aliphatic or aromatic ring and when any of A-E is nitrogen, the corresponding X is void or is oxygen, Z represents amino or an amine derivative of the general formula (n={0, 1, ..-, 5}) 2 wherein Z is as defined for Y except for an oxo group, R' and R" are as defined for Y and derivatives of the general formula wherein A'-E' represent carbon atoms, or one of A'-E' is an oxygen or sulphur atom or up to two of A'-E' are 1 1 4 nitrogen atoms, Y, Z and X -X are as defined above, and when any of A'-E' are nitrogen, the corresponding X is void or is oxygen and when any of A'-E' is oxygen or sulphur, the corresponding X is void. 3. Process according to claim 1 or 2, wherein the nucleophile component NH„-R is selected from 2- nitroarylmethylamine derivatives of the general formula wherein R represents H, straight chain or branched (C-- C. ) alkyl, aryl (R ) or aralkyl (CH_R 2 or CHR 2 R 3 ), wherein 2 3 R and R are aryl groups, which may be substituted with halogen, hydroxy, alkoxy, nitro, amino or alkyl, or R represents a carboxy group, or a functional derivative of a carboxy group, e.g. an ester, a nitrile, an amide which 2 may be substituted on N with (C--C fi ) alkyl, aryl (R ) or 2 2 3 2 3 aralkyl (CH 2 R or CHR R ), where R and R are as defined above, X -X independently represent H, (C..-C.-.) alkyl, halogen, cyano, methoxy, hydroxy, sulfo or nitro, or any two of them may join to form a fused aliphatic or aromatic ring, and benzylamine derivatives of the general formula wherein R 1 is as defined above and X1-X5 independently represent H, (C.-C.-,) alkyl, halogen, hydroxy, (C..-C.-,) alkoxy, nitro, sulfo, cyano or carboxy. 4. Process according to any of claims 1 to 3, wherein said cleavage is induced by photolysis. 5. Process according to claim 4, wherein the photolysis is performed on an aqueous solution of the peptide optionally containing an organic solvent or an organic solution optionally containing a reducing agent, preferably at a wave length of 300 - 800 nm, in particular about 300 - 500 nm. 6. Process according to any of claims 1 to 3, wherein said cleavage is performed by solvolysis. 7. Process according to claim 6, wherein the solvolysis is performed in an acid medium, preferably in trifluoro- acetic acid or aqueous trifluoroacetic acid. 8. Process according to any of claims 1 to 3, wherein said cleavage is performed by reduction. 9. Process according to claim 8, wherein the reduction is performed as a catalytic reduction, preferably a catalytic hydrogenation. 10. Process according to claim 1, wherein the nucleophile component R-NH„ is selected from derivatives of allylamine with the general formula wherein R 4-R7 represents H, straight chain or branched (C..-C, ) alkyl, aryl (R 2 ) or aralkyl (CH.-R 2 or CHR 2 R 3 ), 2 3 wherein R or R are aryl groups, which may be substituted with halogen, hydroxy, alkoxy, nitro, amino or alkyl, or R 4-R7 represents a carboxy group, or a functional derivative or a carboxy group, e.g. an ester, a nitrile, an amide which may be substituted on N with (C.-C,-) alkyl, aryl (R ) or aralkyl (CH 2 R 2 or CHR 2 R 3 ), where R and R 3 are as defined above. 11. Process according to claim 10, wherein the reaction product is cleaved by rearrangement and hydrolysis or by elimination. 12. Process according to claim 1, wherein the cleavage is induced by oxidation. 13. The process according to claim 1, wherein the enzyme is a protease, i.e. an endo- or exopeptidase, preferably a serine or thiol exopeptidase or serine or thiol endopeptidase. 14. The process according to claim 13, wherein a serine protease enzyme is used selected from L-specific serine carboxypeptidases from yeast or of animal, vegetable or other microbial origin, preferably selected from the group consisting of carboxypeptidase Y from yeast, carboxy¬ peptidase S-l and S-2 from Penicillium janthinellum, carboxypeptidases from Aspergillus saitoi or Aspergillus oryzae, carboxypeptidases C from orange leaves or organge peels, carboxypeptidase C N from Citrus natsudaidai Hayata, phaseolain from bean leaves, carboxypeptidases M-I and M- II from germinating barley, carboxypeptidase .W-II from wheat bran, carboxypeptidases from germinating cotton plants, tomatoes, watermelons and Bromelein (pineapple) powder. 15. The process according to claim 1, wherein the reaction medium comprises an organic solvent, preferably selected from the group consisting of alkanols, alkanoic acids, dimethyl sulfoxide, dimethyl formamide, dioxane, tetrahydrofurane, dimethoxy ethane, glycerol, ethylene glycol and polyethylene glycols.

Inventors: BREDDAM KLAUS, BUCHARDT OLE, HENRIKSEN DENNIS, BREDDAM, KLAUS, BUCHARDT, OLE, HENRIKSEN, DENNIS
Applicants: BREDDAM KLAUS, BUCHARDT OLE, HENRIKSEN DENNIS, BREDDAM, KLAUS, BUCHARDT, OLE, HENRIKSEN, DENNIS
Application year: 1991
Main classification: C07K1
IPC codes: C07K1, C07K7, C12N9, C12P21