Patent Publication Number: US-3880823-A

Title: Process for preparing peptides using N-hydroxy succinimide mono-and dichloroacetate esters

Description:
United States Patent [1 1 Hagitani et al.  
  1*Apr. 29, 1975 [73] Assignee: Toyo Jozo Kabushiki Kaisha,  
 &#39; Ohhito, Shizuoka, Japan The portion of the term of this patent subsequent to Sept. 14, 1987, has been disclairned.  
  22 Filed: Aug. 24, 1970 [211 App]. No.: 66,613  
 Related US. Application Data [63] Continuation-impart of Scr. No. 667.662, Sept. 14,  
 1967, Pat. No. 3,541,084.  
 [ Notice:  
 [30] Foreign Application Priority Data Sept. 16. 1966 Japan 41-60768 [52] US. Cl ..260/112.5; 260/326.14 T;  
 260/326.44; 260/471 A; 260/48] R; 260/482 R; 260/557 R; 260/558 A,260/558 S; 260/559 T, 260/559 A, 260/561 A, 260/561 S, 260/561 B, 260/562 N, 260/562 S, 260/262 K, 260/562 P,  
  260/562 A 51} 1nt..C1..... C07c 103/52; C07g 7/00; C08h H00 [58} Field of Search 260/1 12.5  
 [56] References Cited UNITED STATES PATENTS 3,317,559 Anderson 2610/1125 3.541.084 11/1970 Hagitani et al. 260/l12.5  
 OTHER PUBLICATIONS Sakakibara et al., Bull. Chem. Soc. Japan, 37, 1231 Primary Examiner-- Lewis Gotts Assistant E.raminerRegina1d J. Suyat Attorney, Agent, or FirmSughrue, Rothwell, Mion, Zinn &amp; Macpeak [57] ABSTRACT I. A group of new compounds having a general formula:  
 wherein R and R represent various radicals to be disclosed herein and adapted for use as medicine.  
 2. A process for the manufacture of said new compounds starting from the material v co-on R -co-o-u CO-Cl-l R standing for hydrogen, monochloro loweralkyl or dichloro loweralkyl group, through the successive reaction with the compounds having general formula:  
 2 Claims, No Drawings The present application is a continuation-in-part of i Ser. .-No. 667,662 filed Sept. 14, 1967, now Pat. No.  
 I This invention relates to a process for the manufacture of acid amides. It particularly relates to the process for -the production of the compound having the acid amide linkage; said, compound having the general formula:  
 wherein R stands for a member selected from the group consisting of. alkyl, cycloalkyl, alkenyl, cycloalkenyl, aryl, aralkyl,:-arylalkenyl and heterocyclic carboxyl group; these radicals hereinabove defined are substituted or unsubstituted by at least a member selected from the group consisting of halogen, nitro, nitrose, amino, imino, amidino, hydroxy, cycloalkyl, alkoxy, aryloxy, alkoxycarbonyl, alkylcarbonyl, arylcarbonyl, alkylamide, arylamide, alkylthio, alkenyl, cycloalkenyl, alkenylthio, alkenylcarbonyl, aryl, aralkyl, arylalkenyl andheterocyclic group; and R stands for a member selected from the group consisting of alkyl, cycloalkyl, aryl, aralkyl, amino, alkylamino, cycloalkylamino, arylamino, aralkylamino, alkylamide, cycloalkylamide, alkenylamide, arylamide, aralkylamide, cyclohexylcarboxyamide and cyclohexylamino group; these radicals are substituted or unsubstituted by at least a member selected from the group consisting of halogen, nitro, alkyl, alkoxy, carboxy, alkylcarbonyl, alkylamino, alkylamide, cycloalkyl, alkoxycarbonyl, cycloalkylamino, cycloalkylcarbonyl, cycloalkylamide, alkenyl, cycloalkenyl, alkenylacarbonyl, alkenylamino, alkenylamide, aryl,- aralkyl, arylalkenyl, arylamino, arylamide, arylcarbonyl, aralkylcarbonyl, aralkylamino, aralkylamide and heterocyclic group.  
  It has been proposed previously to produce aforementioned compounds having the general formula 1 especially to produce peptide compounds. As examples of the process for the production of the said compounds, there may be mentioned, for example, a method of using p-nitrophenylester of acyl amino acids [M. Bodanszky, Ann. NY. Acad. Sci., 88, 655 (1960)], N-hydroxyphthalimides method [G.H.L. Nefkens, et al., J. Am. Chem. Soc., 83, 1263 (1961)], and Andersons method [6. W. Anderson, et al.: J. Am. Chem Soc.; 86(9), 1839 (1964)]according to which an amino acid, protected in its amino group, is reacted with N-hydroxy succinimide in the presence of dicyclohexyl carbodimide as a dehydrating agent to a succinimide ester:  
 which was then reacted with amino acid to the peptide compound. Among others is known a method by Sakakibara et al. [Bull. Chem. Soc. Japan, 38, 1979 1965)]. According to this method, trifluoroacetic acid anhydride was reacted with N-hydroxy succinimide to give the ester, i.e. trifluoroacetoxy succinimide, being followed by a synthesis to provide the aforementioned succinimide ester, which was then reacted further with amino acid to the peptide compound.  
  This last mentioned prior process has a number of grave disadvantages, including that the intermediate product or trifluoroacetoxy succinimide is almost impossible to crystallize and further that this compound and the starting material, the latter being highly poisonous trifluoroacetic acid, are substantially difficult to handle. Also these substances are highly expensive.  
  The main object of this invention is to provide a process for the production of the compound having acid amide linkage which is useful for a physiologically active agent such as pharmaceuticals and the like.  
  It is another object of the present invention to provide a process of the above kind, capable of obviating the above mentioned various and grave disadvantages.  
  In order to realize the above objectives, the process according to this invention in its broadest aspect comprises: reacting a compound having a general formula:  
 C0-CH wherein R stands for a member selected from the group consisting of hydrogen, monochloro lower alkyl and dichloro lower alkyl group, with a compound having the following general formula:  
 wherein R has the same meaning as set forth hereinbefore, to provide the succinimide ester compound having the general formula:  
 CO--CH wherein R has the meaning set forth above, and subsequently reacting the said succinimide compound with a compound of the following general formula:  
  R;,NH  
 wherein R has the same meaning as set forth hereinbefore, thereby to prepare the desired productexpressed in the general formula (I).  
  Comparing with these conventional methods, the process according to the present invention provides a number of industrial advantages; i.e., succinimide ester as expressed in the general formula:  
 O-CH  
 is easily synthesized from N-hydroxy succinimide ester of the general formula:  
 CO-CH 1 II) 5 the reaction may be conducted at a roomor reduced temperature, as may be required. Generally, the temco ca o 2 perature may vary between to 20 C. Succimmide wherein R and R have respective same meaning as beester as expressed by general formula (II) resulting fore&#39; I from the reaction between the said compound (III) and The Stamng T Sable m the novel Process the said compound (IV) is separated therefrom and of easy to t nelther hazardous f course can be used in the next following reaction step.  
  There also no 88 for speclfically P F It is not always necessary, however, to isolate the comthe intermediate succinimide ester from reaction mixpound (n) and so aforementioned Compound (I) so the memloned COmPOPQdI R3 NH2 ing the acid amide linkage may be obtained without any can be added dlrecFly for Obta,mmg the sald separating steps and thus by reacting the said reaction final compound havmg the amlde linkage mixture of the said compound (IV) directly with the The compound set forth heremabove, represented by compound expressed generally as R3 NH2. general formula (III) such as formate ester of N- Generally, from 1 to 2 moles of compound (I!) is hydroxy succinimide, monochlorofatty acid ester of N- acted per 1 mole of the compound R3 NH2, at a hydroxy succinimide may be used as starting material. perature of from to room temperature (Len E of these mammals y be prepared by conven&#39; 20C). The reaction may be conducted in the presence tional processes, or alternatively by the novel process of from 1 to 3 moles of a tertiary organic amine, per as dlsclosed herem&#39; mole of the compound R NH For carrying out the invention, the process can be F rth is of cou Se necessar to rotect or block carried into effect, for instance, in such a way that an u 1 r y p the radicals R and R in aforementioned compounds, ether solution of formic, monochlorofatty or dlh] f u man or com late] Saturated if such radlcals should be decomposed or affected by Oro a y is pa y y undesired change. Amino, hydroxy and the like radiwith ketene to prepare mixed anhydrlde of acetic acid I cals may be mentioned in this case.  
 formic acid, -monochlorofatty acid or -dichlorofatty acid, which is then reacted with N-hydroxy succinimide T compound (4) P p y f Senes of the to prepare the starting material. Ordinarily the said N- actlor} p f y be extracted If deslred at Controlled hydroxy succinimide-formate ester, N-hydroxy succini- P Wlth a Sultable Organic as y acetate, mide-monochlorofatty acid ester or N-hydroxy succinimethyl lsobutylketone like be$1de 1t be mide-dichlorofatty acid ester produced by the aforecovered by Vanous CQmemIOnaI Pfocesses, Wllwh are mentioned process may be used even in their crude Commonly employed I the Separation of T&#34;- form non recrystallized pounds, such as drying-up n vacuo, crystallizat on The reaction between the Said N hydmxy succinifrom solvents, chromatographic processing and the like mide ester (Ill) and the said compound: R COOH techmque- (IV) may be conducted, preferably in an inert organic Examples of compounds which correspond to formusolvent, such as tetrahydrofuran, dimethyl formamide 40 las (III) and (IV) and their reaction product succinior the like. Generally, l to 2 moles of compound (Ill) mide ester (II), forming part of the instant invention, are reacted with every mole of compound (IV). It may are enlisted in the following Table 1:  
 Table l P (&#34;5 Compound (IV) C m d (1]) CO-CH 2 CO-CH2 n -CO-0-N I R -O0OH R co-cn 2 2 C0 0 CO-CH N-dichloroacetoxy succinimide acetic acid 8 N-acetoxy succinimide acrylic acid N-acrylyloxy succinimide crotonic acid cyclohcxanccarboxylic acid ,Bcyclohcxylacrylic acid carbobcnzoxyglycinc carbobenzoxyvalinc a-cthyl-phenoxyacetic acid 2-cthoxyl -naphthoic acid thiophcnc-Z-acctic acid a-mcthylphcnoxyacctic acid p chlorophcnylacctic acid p-methylbcnzoic acid p-naphthoic acid B-S-nitro-Z-furfuryl acrylic carbobenzoxy-L-phcnylalanine carbobcnzoxy-D-phcnylglycinc a-methoxy-3, 4-dichlorophcnylacctic acid 2,4-dichlorophcnoxyacctic acid N-crotonyloxy succinimide N-cyclohexanoyloxy succinimide N-Bcyclohcxylacrylyloxy succinimide N-carbobcnzoxyglycyloxy succinimide N-carbobenzoxylvalyloxy succinimide N-carbobenzoxy-L-phcnylalanyloxy succinimide N-carbobenzoxy-D-phenylglycyloxy succinimide N-a-methoxyisobutyloxy succinimide N-phcnylacctoxy succinimide N-capryloxy succinimide N-stcaryloxy succinimide N-cinnamoyloxy succinimide N-isonicotinoyloxy succinimide N-benzoyloxy succinimide N-carbobenzoxy-L-lcucyloxy succinimide N-phenoxyacctoxy succinimide N-a-methoxy-3, 4-dichlorophenylacctoxy succinimide N-a-ethyl-phenoxyacctoxy succinimide N-Z-cthoxyl-naphthoyloxy succinimide N-thiophene-2-acetoxy succinimide N-a-methyl-phcnoxyacctoxy succinimide N-2, 4dichlorophenoxyacetoxy succinimide N-p-chlorophenylacetoxy succinimide N-p-methylbenzoyloxy succinimide N-Z-naphthoyloxy succinimide N-B-S-nitrolfurfuryl acrylyloxy The following preparations illustrate several modes of preparing succinimide ester (11) in the present invention.  
 PREPARATION 1 N-acetoxy succinimide (145.9971. H;4.63%, N;8.96% theoretical as based upon C H NO C;45.8671, H;4.49%, N;8.91%  
 found;  
 PREPARATION 2 N-carbobenzoxyglycyloxy succinimide 2 ml of tetrahydrofuran were added to a mixture of 1.05 g of carbobenzoxyglycine (5 m mole) and 2.26 g of N-dichloroacetoxy succinimide m mole). To this solution 1 ml of triethylamine was added with stirring. The mixture was stirred for a further 1 hour at room temperature. After finishing the reaction, water was added, cooled, then filtered, and solid material recovered was washed with water. This material was dissolved in methylenechloride, and dried with anhydrous sodium sulfate. After removal of the drying agent by filtration, an addition of diethylether resulted in the precipitation of N-carbobenzoxyglycyloxy succinimide as crystal.  
 Yield: 1.10 g(recovery: 73%) m.p.: found; 1 C;  
 reference; 114C Elemental analysis:  
 found;  
 theoretical PREPARATION &#39;3 N-carbobenzoxyglycyloxy succinimide To a solution of 1.05 g of carbobenzoxyglycine (5 m. mole) and 2 g of N-monochloroacetoxy succinimide (ca. 10 m mole) in 1 ml of tetrahydrofuran was added dropwise 1 ml of trimethylamine (7.2 m mole) with stirring. The reaction was allowed to continue for 2 hours. Following the reaction, the precipitated reaction product was filtered after addition of ml of water under cooling, then washed with water. The precipitated material was recrystallized from ethanol-ether mixture, yielding N-carbobenzoxyglycyloxy succinimide in white crystals.  
 Yield: 750 mg (recovery: 49%) m.p.: found; 1l2.5113.5C;  
 reference: 113-114C PREPARATION 4 N-carbobenzoxyglycyloxy succinimide The triethylamine in Preparation 3, was replaced by pyridine to produce N-carbobenzoxyglycyloxy succinimide as white crystals.  
 Yield: 342 mg (recovery: 23%) PREPARATION 5 8.4 ml of triethylamine (60 m mole) were added dropwise to a cooled, stirred solution of 4.08 g of phenylacetic acid (30 m mole) and 10.2 g of N- dichloroacetoxy succinimide (45 m mole) in 6 ml of di&#39; methylformamide. The solution was stirred for about 2 hours and then an excess amount of water was added. The mixture was allowed to stand overnight. After cooling the thus precipitated crystals were separated by filtration, washed with water repeatedly, yielding 5.55 g of crude crystal of N-phenylacetoxy succinimide (recovery: 79%), which was recrystallized as colorless crystals from ethylether.  
 Yield: 4.15 g (recovery: 60%) m.p.: 118119C Elemental analysis:  
 PREPARATION 6 N-capryloxy succinimide 1.05 ml of triethylamine were added dropwise with stirring to a cooled solution of 0.86 g of n-capric acid (5 m mole) and 1.7 g of N-dichloroacetoxy succinimide (7.5 m mole) in 2 ml of dimethylformamide. The mixture was then stirred in an ice bath for about 1 hour, and then 15 ml chilled water was added, the reaction mixture was then cooled and the precipitated crystals were separated. This precipitated material was recrystallized from isoprophylalcholethylacetate mixture.  
 Yield: 500 mg (recovery: 37%) Elemental analysis:  
 - found;  
 PREPARATION 7 N-carbobenzoxy-L-phenylalanyloxy succinimide 1.05 ml of triethylamine were added dropwise with cooling to a mixture of 1.2 g of carbobenzoxy-L- phenylalanine (4 m mole) and 1.36 g of N- diehloroacetoxy succinimide (6 m mole) in 2 ml of dimethylformamide. The mixture was stirred with cooling, followed by addition of 15 ml of water, then the sedimented crystals were separated therefrom. N-car bobenzoxy-L-phenylalanyloxy succinimide was recrystallized from isopropyl alcohol.  
 Yield: 670 mg (recovery: 42%) m.p.: 133C 9 PREPARATION x N-stearyloxy succinimide 1.05 ml of triethylamine were added dropwise to a cooled, stirred mixture of 1.42 g of stearic acid m PREPARATION 10 N-benzoyloxy succinimide 1.05 ml of triethylamine were added dropwise to a 5 solution of 0.61 g of benzoic acid (5 m mole) and 1.7 mole) and 1.7 g of N-dichloroacetoxy succinimide (7.5 g of N-dichloroacetoxy succinimide (7.5 m mole) in 2 m mole) in 6 ml of dimethylformamide. The solution ml of dimethylformamide. The solution was stirred for was stirred for about 1 hour with cooling, and then about 2 hours with cooling, adding ml of water, added with 24 ml of water. The precipitated crystals of standing in an ice bath, and then separating the precipi- N-stearyloxy succinimide were separated, which were 10 tated crystals of N-benzoyloxy succinimide. finally recrystalized from isopropyl alcohol. Yield: 500 mg (recovery: 46%) Yield: 1.16 g (recovery: 61%) m.p.: 133l35C m.p.: 88-90C t Elemental analysis: Elemental analysis:  
  found; C;60.27 H4. 19%, 6579.  
  theoretical as based upon CUHMNO4 found: (269.6271. mosz /l. N;3.94/1 Q6027? HAM? M6397 theoretical as based upon C- ,2H;,,,NO4  
 C;69.257, H&#39;,l0.30 /l. N;3.67&#39;/z 2O PREPARATION 11 N-carbobenzoxy-D-phenylglycyloxy succinimide 1.05 ml of triethylamine were added dropwise with stirring to a cooled solution of 1.43 g of carbobenzoxy- PREPARATION 9 D -phenyIgIyCine (5 m mole) I and 1.7 g of N- dlchloroacetoxy succinimide (7.5 m mole) in 2 ml of N&#39;cmnamoyloxy Succmlmlde dimethylformamide. The solution was then stirred for 1.43 ml of triethylamine were added to a mixture of 2 hours in an ice bath, adding an equal volume of 7 g f i &#39;i id 5 m mole) d 1 7 g f N- chilled water and then precipitated crystals thus sepadichloroacetoxy i i id (75 m l i 2 f rated. The recrystalization was performed from isoprodimethylformamide,stirring for about 1 hour, followed Py alcohol, thus Obtained N&#39;carbobenzoxy&#39;D&#39; by. addition of 10 m1 of water. The solution was neutralphenylglycyloxy succinimide as white needle crystal. ized with 1 N-HCl and cooled, thereby separating the Yieldi 610 g (recovery? sedimented crystals and washed with water. The pre- &#39;P- cipitated material was recrystalized from isopropyl al- Elemental&#39;analysisi coholethyl acetate.  
 held: 450 mg (recovery: 37%) found; C;63.317(.H;4.86/,N:7.45 /z m.p.: 173l 5 theoretical as based upon C H N O Elemental analysis: 062.82%. H;4.75%, N;7.33%  
  Compounds, which are represented by formula (11), f d 067 407 H4 79% N5 47% formula R NH and compound having acid amide theoretical z t h &#39;l linkage as formula (1) are shown in the following Table C;63.6771. H;4.5372, N;5.717z 2,  
  Table 2 i i im ester compound having acid amide linkage compound (11) compound (1) a -co-o-N CH2 R NR (JO-CH2 glycine ethyl ester 8-aminovaleric acid methyl ester carbobenzbxyglycylglycine ethyl ester B-carbobenzoxy aminovaleryl- -amino-valerie acic methyl ester N-carbobenzoxy-L-phenylalanyloxy N-carbobcnzoxy-L-leucyloxy succinimide N-dichloroacetoxy succinimide N-propionyloxy succinimide glycine ethyl ester glycine ethyl ester L-phenylalanyl glycylethyl ester -aminopenicillanie acid o-aminopenicillanic acid o-aminopenicillanic acid 6-aminopenicillanic acid 7-aminocephalosporanic acid oaminopenicillanic acid G-aminopenicillanic acid n-hexylaminc bcnzylamine allylamine Z-aminopyridine 2-amino-5 nitrothiazole phenylhydrazine p-chloroaniline aniline carbobenzoxy-L-phenylalanyl-glycine ethyl ester &#34;carbobenzoxyL-leucylglycine ethyl ester carbobenzoxy-L-leucyl-L-phenylalanylglycine ethyl ester benzyl penicillin phenoxypropyl penicillin 3,4-dichloro-a-methoxybenzyl penicillin Z-ethoxyJ-naphthyl penicillin cephalotin phcnoxyethyl penicillin a-aminobcnzyl penicillin N-n-hexylacetamide N-benzylacetamide N-allylacetamidc N-Z-pyridylacetamidc N-S-nitro-Z-thiuzolyl acetamide N-phcnylaminoucetamide N-p-chlorophenyl dichloroacetamidc N-phcnylpropionamide TABLE 2 Continued succinimtde eater compound (I!) OO-CH a -co-o-u 2 R3 NH:  
 co-cn compound having acid amide linkage compound (1) N-eyclohexanecarbonyloxy succinimidc N-phcnylacetoxy succinimide N-benzoyloxy succinimidc N-phcnylacetoxy succinimide N-2,4-dichlorophcnoxyacetoxy succinimidc cyclohcxylamine bcnzylaminc p-anisidinc cyclohcxylamine p-chloroaniline p-toluidinc n-butylamine N-p-chlorophcnylacctoxy succinimidc N-p-mcthylbcnzoyloxy succinimidc N-2-naphthoyloxy succinimide N-Z-naphthoyloxy succinimide N-phcnoxyacetoxy succinimide N-acrylyloxy succinimide N-isonicotinoyloxy suceinimide NB-S-nitro-Z-furfurylacrylyloxy succinimide Nphcnylacctoxy succinimide N-isonicotinoyloxy succinimide N-p-chlorophcnoxyacctoxy succinimide N-acetoxy succinimide N-hippuropyloxy succinimidc N&#39;B-chloropropionyloxy succinimidc benzylaminc isopropylaminc 2-aminothiazolc benzylamine morpholine phenylhydrazine aniline benzylamine 3,4-dichloroaniline B-phcnylethylamine isopropylhydrazine p-aminobenzoic acid ethyl ester The compounds produced by the present process are useful as therapeutic agents for the treatment of vari-&#39; ous types of bacterial infections. Specifically, the pres ent process is useful for the production of oxytocin, which is known as a stimulant of uterine contraction (See, for example, US. Pat. Nos. 2,938,891 and 3,076,797). The production of oxytocin by the present process is as follows:  
 EXAMPLE OF PREPARATION OF OXYTOClN 25 ml of a 25%-solution of hydrogen bromide in acetic acid was added to 7.0 g of N-carbobenzoxy leucyl glycine ethyl ester (hereinafter abbreviated to Cbz-Leu-GlyOEt) and the mixture was reacted for 30 minutes at room temperature, then 450 ml of dry ethyl ether was added thereto.  
  The precipitated oil was separated by decantation and then dissolved in 40 ml of CHCl with 6 ml of triethylamine (referred to as TEA&#34; hereinafter).  
  To the solution, 7.0 g of N-carbobenzoxyproline N- oxysuccimide ester (to be abbreviated hereinafter to Cbz-Pro-NHS) were added and left overnight for reaction at room temperature. After completion of the reaction, 150 ml of CHCl were further added thereto and the reaction mixture was washed successively with lN-NaHCO H O, lN-HCl and H 0, respectively. After removal of the solvent by drying, the reaction products were recrystallized from ethyl acetate, to obtain carbobenzoxyprolyl leucyl glycine ethyl ester. Yield: 7.0 g; Recovery: 76%; mp. 15 l152C; [04],, 82 8 (C 2.5 EtOH).  
  90 g of N-carbobenzoxyprolyl leucyl glycine ethyl ester (hereinafter referred to briefly as Cbz-Pro-Leu- CilyoEt) were dissolved in 1.5 lit. of methanol and NH gas was introduced into the solution at C for 2 hours. The reaction mixture was left stationary at room temperature for hours, the solvent was distilled off and the reaction products were recrystallized from ethyl acetate to provide carbobenzoxy prolyl leucyl glycinamide (hereinafter referred to briefly as &#34;Cbz- Pro-Leu-Gly-NH Yield: 76.5 g; Recovery: 91.571; m.p. l62-164C.  
  7.0 g Cbz-Pro-Leu-Gly-NH was added to 25 ml of a 25%-solution of HBr in acetic acid and reacted at room temperature for 30 minutes, thereafter 450 ml of dry ethyl ether was added and the resulting oil was removed from the solvent by decantation, CHCl and 5 ml of TEA were added to the separated oil.  
  To the solution thus obtained, 7.5 g of N-carbobenzoxy-S-benzyl cystein N-oxysuccinimide ester (hereinafter referred to as were added and the mixture was left stationary overnight at room temperature for reaction. After the reaction, CHCl was additionally introduced and the mixture was washed successively with lN-NaHCO H O; lN-HCl and H 0. Then, the solvent was distilled off to provide N-carbobcnzoxy-S-benzyl cysteinyl proly leucyl glycinamide (to be referred to hereinafter briefly as BZ &#39;Cbzys-ProLeuGlyNH Yield: 126g; Recovery 12.0 g of were suspended in 25 ml of acetic acid and 50 ml of a 25%-solution of HBr in acetic acid, and then the reaction mixture was left stationary at room temperature for an hour. Then, 400 ml of dry ethyl ether were added. The sedimented products were washed carefully with ethyl ether and dissolved in dimethylformamide (referred to hereinafter as &#34;DMF), TEA was added and the resulting mixture was reacted with 7.7 g ofcarbobenzoxy asparagine N-oxysuccinimide ester (to be called briefly as by agitating at room temperature for 2 days and then adding ethyl acetate. The separated sedimentation products were filtered. The reaction product amounted to 13 g. m.p. 2l32l4C. [a],, 60 (C l, DMF).  
 2.9 g of dissolved in 30 ml of acetic acid was added thereto along with 30 ml of a 25%-sol ution of HBr in acetic acid and reacted at room temperature for an hour, and then 450 ml of dry ethyl ether was added.  
  The separated sedimentation was dissolved in 9 ml of DMF and 2.6 ml of TEA was added and then reacted with 2g of H7 Cbz-Elti-NHS.  
  The mixture was reacted overnight under stirring at room temperature, and then ethyl acetate was added. The thus precipitated product was filtered and washed with ethanol and ethyl acetate, respectively to obtain 3g (recovery; 90%) of the precipitate.  
 7.0g of the thus-obtained EH H s-Bz Cbzlu-Asp-Cys-Pro-Leu- GLl.y--NH- acetate were added to obtain the precipitate. Yield: 8.0 S  
 g (Recovery: 90%).  
 were dissolved in 50 ml of acetic acid. and 50 ml of a 25% solution of HBr in acetic acid was added thereto. The solution was reacted for 2 hours at room temperature. and then dry ethyl ether was added thereto.  
  The thus-obtained precipitate was dissolved in 85 ml of dimethylformamide, and 9 ml of triethylamine were added thereto and reacted with 6.0 g of After the reaction continued for hours at room temperature, 450 ml of ethyl acetate were added. Thus, 1 1 g (Recovery 91%) of precipitate were obtained.  
  The precipitate was suspended in ml of acetic acid, and 60 ml of 25% HBr in acetic acid was added; the reaction was continued for 2 hours at room temperature. After the reaction, dry ethyl ether was added to produce a precipitate.  
  The precipitate was dried a short time over CaCl, and NaOH, and then dissolved in 60 ml of dimethylformamide; 5-6 ml of triethylamine was added and the mix- 0 ture was reacted with The reaction was continued for 3 days with stirring.  
  After completion of the reaction, 200 ml of ethyl acetate were added The produced precipitate was washed with 400 ml of ethanol and ml of ethyl acetate respectively. Yield: 7.3 g (Recovery: 92%). m.p. 23724lC. 100 mg of and 0.1 ml of anisol were dissolved in 2 ml of anhydrous hydrogen fluoride. The solution was held for 1 hour at room temperature, and then hydrogen fluoride was removed in vacuo and dried with methanol. The thus obtained substance was dissolved in ml of H 0, and the pH was adjusted to 6.5 with aqueous ammonia for 2 hours. Next, the pH of the substance was further adjusted to 4 with acetic acid.  
  The solution showed an oxytocin property ranging from 13,000 to 14,000 units, in uterine contraction experiments with mice.  
  When the solution was passed through an ionexchange resin (Dow 50) to freeze-dry the peptidefractions, 20 mg of solid were obtained.  
  The following examples are set forth for purposes of illustration only and are not to be construed as limiting the scope of the invention.  
 EXAMPLE 1 Carbobenzoxyglycylglycine ethyl ester 1 ml of triethylamine (7 m mole) was added dropwise I to a cold solution of 1.05 g of carbobenzoxyglycinc (5 m mole) and 1.55 g of N-dichloroacetoxy succinimide (7 m mole) in 3 ml of dimethylformamide. After 2 hours 0.07 ml of water was added. stirring for several minutes. and then a mixture of 0.7 g og glycine ethyl ester hydrochloride (5 m mole 0.7 ml of triethylamine m mole), 3 ml of dimethylformamide and 0.55 ml of water was added thereto. The reaction was allowed to continue for 1 hour with stirring. After adding 20 ml of chilled water, allowed to stand in an ice bath, the solution was extracted twice with ethyl acetate. This ethyl acetate extract was combined with additional ethyl acetate extract and the combined extracts containing the product were washed with solution of sodium bi carbonate. 1 N-hydrochloric acid and water respectively, and dried over anhydride sodium sulfate. After removal of the drying agent by filtration, the extract was dried up in vacuo. The residue was recrystallized from water containing ethanol to obtain crystals of carbobenzoxyglycine ethyl ester.  
 Yield: 1.08 g (recovery: 73%) m.p.: found; 8l82C; reference; 80-8lC Elemental analysis:  
 found;  
 EXAMPLE 2 The procedure of Example 1 was again followed substantially in every detail except that the dimethylformamide was replaced by tetrahydrofuran to obtain the same product as crystals in Example 1.  
 Yield: 1.05 g (recovery: 71%) EXAMPLE 3 fi-carbobenzoxy aminovaleryl aminovaleric acid methyl ester 1 ml of triethylamine was added dropwise with stirring to a cooled solution of 1.26 g of S-carbobenzoxy aminovaleric acid (5 m mole) and 1.58 g of N- dichloroacetoxy succinimide (7 m mole) in 3 ml of dimethylformamide. The mixture was stirred in an ice bath for about 4 hours, followed by an addition of 0.07 ml of water, and then kept in cooled condition for further 40 minutes. Next, the mixed solution of 0.85 g of 8-aminovaleric acid methylester hydrochloride (ca. 5 m mole), 0.7 ml of triethylamine (5 m mole) and 4 ml of dimethylformamide were added thereto with cooling, while stirring for 2.5 hours. The mixture was allowed to stand overnight, then added with 30 ml of water and extracted with ethyl acetate. The ethyl acetate extract was washed with sodium bicarbonate solution, diluted with hydrochloric acid and water respectively and dried over anhydrous sodium sulfate. After removal of the drying agent by titration, the extract was condensed in vacuo. The sedimented material was separated, washed with ether, dried in desiccator, thus obtaining the product in white crystals.  
 Yield: 1.37 g (recovery: 75%) Elemental analysis:  
 found;  
  EXAMPLE 4 Carbobenzoxy-L-phenylalanylglycine ethyl ester 1.07 ml of triethylamine were added dropwise with 5 cooling to a mixture of 1.5 g of carbobenzoxy-L- phenylalanine (5 m mole) and 1.96 g of N- dichloroacetoxy succinimide (7.5 m mole) in 3 ml of dimethylformamide. After 2 hours, 0.07 ml of water was added with stirring for few minutes, further a found; reference;  
 Elemental analysis:  
  EXAMPLE 5 Carbobenzoxy-L-leucylglycine ethyl ester 2.28 ml of triethylamine were added dropwise with stirring to a cold mixture of 2.65 g of carbobenzoxy-L- leucine (10 m mole) and 3.39 g of N-dichloroacetoxy succinimide (15 m mole) in 3 ml of dimethylformamide. The mixture was stirred for 3 hours. and then added with 0.14 ml of chilled water with continuous cooling and stirring. To this solution, a mixture of 1.39 g of glycine ethyl ester hydrochloride 10 m mole). 14 ml of dimethylformamide and 1.43 ml of triethylamine was added. This reaction mixture was then stirred for further 3 hours. After adding chilled water, the solution was in vacuo. The residue was recrystallized from water containing ethanol to obtain crystals of carbobenzoxyglycine ethyl ester.  
 Yield: 1.08 g (recovery: 73%) m.p.: found; 8182C; reference: 808lC Elemental analysis:  
 found;  
 EXAMPLE 2 The procedure of Example 1 was again followed substantially in every detail except that the dimethylformamide was replaced by tetrahydrofuran to obtain the same product as crystals in Example 1.  
 Yield: 1.05 g (recovery: 7171) mixed solution of glycine ethyl ester hydrochloride (5 l EXAMPLE 3 S-carbobenzoxy aminovaleryl aminovaleric acid methyl ester 1 ml of triethylamine was added dropwise with stirring to a cooled solution of 1.26 g of -carbobenzoxy aminovaleric acid (5 m mole) and 1.58 g of N- dichloroacetoxy succinimide (7 m more) in 3 ml of dimethylformamide. The mixture was stirred in an ice bath for about 4 hours, followed by an addition of 0.07 ml of water, and then kept in cooled condition for further 40 minutes. Next, the mixed solution of 0.85 g of 8-aminovaleric acid methylester hydrochloride (ca. 5 m mole), 0.7 ml of triethylamine (5 m mole) and 4 ml of dimethylformamide were added thereto with cooling, while stirring for 2.5 hours. The mixture was allowed to stand overnight, then added with 30 ml of water and extracted with ethyl acetate. The ethyl acetate extract was washed with sodium bicarbonate solution, diluted with hydrochloric acid and water respectively and dried over anhydrous sodium sulfate. After removal of the drying agent by filtration, the extract was condensed in vacuo. The sedimented material was separated, washed with ether, dried in desiccator, thus obtaining the product in white crystals.  
 Yield: 1.37 g (recovery: 75%) Elemental analysis:  
 found;  
 &#39; EXAMPLE 4 Carbobenzoxy-L-phenylalanylglycine ethyl ester 1.07 ml of triethylamine were added dropwise with cooling to a mixture of 1.5 g of carbobenzoxy-L- phenylalanine (5 -m mole) and 1.96 g of N- dichloroacetoxy succinimide (7.5 m mole) in 3 ml of dimethylformamide. After 2 hours, 0.07 ml of water was added with stirring for few minutes, further a mixed solution of glycine ethyl ester hydrochloride (5 m mole), 7 ml of dimethylformamide and 0.7 ml of triethylamine was added thereto and the mixture was stirred for further 2 hours.  
  After the reaction, 20 ml of water was added and the mixture was extracted with ethyl acetate. The ethyl acetate phase was separated, washed with a %-solution of sodium bicarbonate, and diluted with hydrochloric acid and water, then dried with anhydrous sodium sulfate, followed by condensation in vacuo to obtain the product in white crystals.  
 Yield: 1.55 g (recovery: 81%) m.p.: found; l081 l 1C; reference; ll0l l 1C Optical rotation:  
 found; reference;  
 Elemental analysis:  
 found;  
  EXAMPLE 5 Carbobenzoxy-L-leucylglycine ethyl ester amide. The mixture was stirred for 3 hours, and then i added with 0.14 ml of chilled water with continuous cooling and stirring. To this solution, a mixture of 1.39 g of glycine ethyl ester hydrochloride 10 m mole), 14 ml of dimethylformamide and 1.43 ml of triethylamine was added. This reaction mixture was then stirred for further 3 hours. After adding chilled water, the solution was extracted twice with ethylacetate and the extract containing the product was washed with diluted hydrochloric acid, sodium bicarbonate solution and water successively, and dried over anhydrous sodium sultate. After removal of the drying agent by filtration, the ethyl acetate layer was evaporated at a reduced pressure so as to precipitate the product which was finally recrystallized from ethyl acetate/petroleum ether.  
 Yield: 1.60 g (recovery: 55.1%)  
 Optical rotation: [0:1 25.5(c=2.08, ethanol) Elemental analysis:  
 found:  
 EXAMPLE 6 Carbobenzoxy-L-phenylalanylglycine ethyl ester N-dichloroacetoxy succinimide in Example 4 was replaced by N-monochloacetoxy succinimide to produce crystals of carbobenzoxy-L-phenylalanylglycine ethyl ester.  
 Yield: 34%  
 Optical rotation: [01],, =l6.l (c=l.646, ethanol) EXAMPLE 7 Benzylpe nicillin procaine 0.4 ml of triethylamine (3 m mole) was added to a cooled solution of 0.23 g of N-phenylacetoxy succinimide (l m mole), prepared in accordance with the aforementioned preparation 5, and 0.21 g of 6- aminiopenicillanic acid 1 m mole) in 3.6 ml of dimethylformamide. The solution was stirred for about 3 hours at room temperature, and then poured into ice water, adjusting the pH to 2 by addition of l N- hydrochloric acid. This solution was extracted with methyl isobutyl ketone and the methyl isobutyl ketone layer was separated. This extract was washed with 0.01. N-hydrochloric acid, then extracted twice with 1 ml of 1 mole sodium bicarbonate solution and once with 0.5 ml of the said solution. To the extracts combined together containing the product, benzylpenicillin, there was added 0.27 g of procaine hydrochloride and cooled in an ice water bath, to precipitate benzylpenicillin procaine in white crystals, which were washed then with ethanol and diethyl ether. dried in vacuo, and found to have an activity of 1000 units/mg. Yield: 400 mg (recovery:  
 m.p.: 124125C (dec.)  
 lemental analysis:  
 found; 0.64.2271. H;6.077c, N;9.02%  
 theoretical as based upon (C H N O S) .C zu z C;63.68/z. H;5.80%, N;9.29%  
 found C;59.2271, H;6.92%, N;9.90&#39;/r, S5217:  
 thcorctical as based upon C H ,,N O S.C H N O .H O  
  ,;-2,;5.&#39; c.9174 H6851N95 /S 54/( EXAMPLE N, N-dibenzylethylenediamine phenoxymethyl EXAMPLE 8 penicillin Benzylpemclllm procamc 10 16.8 ml of triethylamine (0.12 mole) were added 0.28 ml of triethylamine was added dropwise with dropwise to an ice cold solution of 9.12 g of phenoxystirring to a cold solution of 0.34 g of N- acetic acid (0.06 mole) and g of N dichloroacetoxy succinimide (1.5 m mole) and 0.136 dichloroacetoxy succinimid mole) ill 50 ml of g of phenylacetic acid (1.0 m mole) 3 ml of dimethyll5 dimethylformamide. The mixture was stirred for 1.5 formamide. The mixture was stirred for about 2 hours hours and then the solution was extracted with 100 ml in an ice water bath and then added with 0.04 ml of wa of ethyl acetate, which is followed by successive washter, followed by continuous agitation. Next, a solution ing ith 1 N-sodium bicarbonate Solution, 1 N- of 0.15 g of 6-aminopenicillanic acid (0.7 m mole) i hydrochloric acid and water. After the ethyl acetate 0.28 ml of triethylamine and 1.5 ml of dimethylform- 20 layer was dried, it was concentrated and e residue amide was added thereto. The mixture was stirred at was recrystallized from P PY alcohol to g g room temperature for 3.5 hours. After adjusting the H of -p y y suecinimide y: to 2.0 and pouring into ice water containing 3 m mole Next, the reaction Was nt nu d und r Cooling for of hydrochloric acid, the solution was extracted with abOut hours with cll&#39;opwlse addltlol&#39;l 0f g of t methyl isobutyl ketone. The methyl isobutyl ketone ethylamine mole) to a Solution of g of N- layer was washed with diluted hydrochloric acid and phenoxyacetoxy succlnlmlde mole) and g of extracted with 1 mole of aqueous sodium bicarbonate 6-amll&#39;lopenlclllflnlc acid mole) in 100 ml of l ti Addi 0,19 g f procaine h d hl id to methylformamide. The solution was adjusted to pH 2.0, this extract, the precipitated product formed by cooling eXtl&#39;acted with 150 ml Of methyl isobutyl ketone, and was filtered, washed with water, ethanol and diethyl followed y exlfactlon with l N-sodlum a nat ether, respectively, dried and thus obtained crystals of Sllltl0ll- The extract was adjusted to P 7 and y p b l i illi procaine, lized. 10 g of N, N&#39;-dibenzylethylenediamine diacetate Recovery; 63% solution were added to 1 1.7 g of this lyophilized powder and filtered and the precipitate was washed with EXAMPLE 9 water, dried in vacuo to give N, N- wcarbobenzoxy aminobenzyl penicillin dibenzylethylenediamine phenoxymethyl penicillin.  
 dibenzylethylenediamine (recovery:  
 m.p.: 9597C 11.2 ml of triethylamine (0.08 mole) were added dropwise to an ice-cold solution of 13.6 g of N- dichloroacetoxy succinimide (0.06 mole) and l 1.4 g of 40 a-carbobenzoxyamino phenylacetic acid (0.04 mole) found;h l b 5.59.0178 l-li i5.9g.SN-.t2i%N in 30 ml of dimeth lformamide. The reaction contincomm use Pmll 2 5 1 ued for about 3.5 h urs at room temperature, then the H5967 Nls&#39;ssqr reaction mixture was extracted with 250 ml of ethyl acetate and washed successively with water, lN-sodium EXAMPLE 11 bicarbonate solution, IN-hydrochloric acid and water.  
  The ethyl acetate layer was dried with anhydrous sodium sulfate. After drying agent was removed by filtration, the ethyl acetate layer was evaporated under reduced pressure. The residue was dissolved in 30 ml of dimethyl formamide, followed by addition of 8.64 g of 6-amino-penicillanic acid (0.04 mole) and further-by dropwise addition of 16.8 ml of triethylamine (0.12 mole) with cooling in ice. The resulted clear solution was allowed to warm up to room temperature and to Elemental analysis:  
 N, N&#39;-dibenzylethylenediamine phenoxyethyl penicillin 1n the process of Example 10, the phenoxyacetic acid was replaced by phenoxypropionic acid to produce N, N&#39;dibenzylethylenediamine phenoxyethyl penicillin.  
 (recovery: 62.5%) m.p.: 93-100C Elemental analysis:  
 continue the reaction for 2.5 hours. then adjusted the theoretical as based upon (C,,H. ,N O,,S) .C.,H N, pH to 2 and extracted with 300 ml of methyl isobutyl C160fi3q-H1-l9%\N=8-67% ketone. The extract was washed with water, followed by ad usting of the pH to 7 by addition of lN-sodium bi- 6O EXAMPLE l2 carbonate solution and of dropwise addition of 7.2 g of dibenzylethylenediamine diacetate in water. After pre- Nl&#39;dlbenzylethylenediamlne phenoxypropyl cipitated the product was recovered by filtration, pemclnm washed with water, dried over phosphorus pentoxide in In an Example 10 the phenoxyacetic acid was re vacuo to give N, N&#39;-dibenzylethylenediamine salt of placed by phenoxybutyric acid to produce N, N&#39;- a-carbobenzoxy aminobenzyl penicillin. dibenzylethylenediamine phenoxypropyl penicillin.  
 m.p.: l27-158C (dec.) (recovery: 50.3%) Elemental analysis: Elemental analysis:  
 Example 13 In the process of Example 10, the dichloroacetoxy succinimide and phenoxyacetic acid were replaced by monochloroacetoxy suceinimide and phenoxybutyric acid respectively to produce the same product as obtained in Example 12.-  
 Example 14 N,N&#39;-dibenzylethylenediamine-3-(o-chlorophenyl)-5- methyl-4-isoxazolyl penicillin 14.0 ml of triethylamine (0.1 mole) were added dropwise under ice cooling to a solution of 11.85 g of 3-(o-chlorophenyl)-5-methyl-4-isoxazol carboxylic acid (0.075 mole) in 30 ml of dimethylformamide. The mixture was reacted with stirring in an ice water bath for 1 hour and then at room temperature for 3 hours. The solution was extracted twice with total 300 ml of ethyl acetate, subsequently washed with water, 1 N- sodium bicarbonate solution, 1 N hydrochloric acid and water, respectively. The washed solution was dried over anhydrous sodium sulfate.  
  After removal of the drying agent by filtration, the ethyl acetate layer was evaporated in vacuo and the residue was dissolved in 25 ml of dimethylformamide, to which is added then 6.5 g of -amino-penicillanic acid (0.03 mole) and 2.45 g of Z-methylimidazole under cooling to react with each other for 3 hours. The reacted solution was allowed to stand overnight at room temperature, adjusted to pH 2.0 in an ice water bath and extracted with 100 ml of diethyl ether. The ether solution was then extracted with 30 ml of IN- sodium bicarbonate solution and 5.4 g of N,N- dibenzylethylenediamine diacetate in water was added thereto. After cooling off the reaction mixture, the filtered precipitate was washed with water and dried to obtain N, N&#39;-dibenzyl-ethylenediamine-3-(o-chlorophenyl)-5-methyl isoxazolyl penicillin.  
 Yield: 7.7 g (recovery: 45%) Elemental analysis:  
 found: (158.25%, H&#39;,5.46&#39;7(, N;9.557(, Cl;6.34%  
 theoretical as based upon (C H N fi )zui zn z EXAMPLE l5 N,N&#39;-dibenzylethylenediamine oz-methoxy-3, 4-dichlorobenzyl penicillin In the process of Example 10, the phenoxyacetic acid was replaced by a-methoxy-3, 4-dieholorophenylaeetic acid to produce N, N&#39;dibenzylethylenediamine a-methoxy-3. 4-dichlorobenzyl penicillin. (recovery: 45%) Elemental analysis:  
 ethoxyl -naphthoie EXAMPLE l6 Example 17 N,N&#39;-dibenzylethylenediamine 6-(carbobenzoxyglycly) amino-penicillanic acid In the process of Example 10, the phenoxyacetic acid was replaced by carbobenzoxyglycine to produce N,N dibenzylethylenediamine-6-( earbobenzoxyglycyl aminopenicillanic acid. (recovery: 70.7%)  
 Elemental analysis:  
 EXAMPLE l8 N,N&#39;-dibenzylethylenediamine 6-(carbobenzoxyphenylalanyl)-aminopenicillanic acid In the process of Example 10, phenoxyacetic acid was replaced by carbobenzoxyphenylalanine to produce N,N-dibenzylethylenediamine-6-(carbobenzoxyphenylalanyl)-aminopenicillanic acid. (recovery: 72.3%)  
 Elemental analysis:  
 EXAMPLE 19 N,N-dibenzyethylenediamine-6-(carbobenzoxyseryl)- aminopenicillanic acid In the process of Example 10, the phenoxyacetic acid was replaced by carbobenzoxyserine to produce N,N- dibenzylethylenediamine-6-( carbobenzoxyseryl aminopenicillanie acid. (recovery: 60.7%)  
 Elemental analysis:  
 EXAMPLE 20 N,N-dibenzylethylenediamine-6-( carbobenzoxyalanyl)-aminopenicillanic acid In the process of Example 10, the phenoxyacetic acid was replaced by carbobenzoxyalamine to produce N,N-  
 &#39;-dibenzylethylenediamine-6-( carbobenzoxyalanyl aminopenicillanic acid. (recovery: 75.8%)  
 Elemental analysis:  
 EXAMPLE 21 NN &#39;-dibenzylethylenediamine-o-myristoylaminopenicillanic acid In the process of Example 10, the phenoxyacetic acid was replaced by myristic acid to produce N,N&#39;- dibenzylethylenediamine-6-myristoylaminopenicillanic acid. (recovery: 55.4%)  
 Elemental analysis:  
 EXAMPLE 22 N,N &#39;-dibenzyethylenediamine-6-lauroylaminopenicillanic acid In the process of Example 10, the phenoxyacetic acid was replaced by lauric acid to produce N,N- dibenzylethylenediamine-6-aminopenicillanic acid. (recovery: 62.0%)  
 Elemental analysis:  
 EXAMPLE 23 NN -dibenzylethylenediamine-b-caproylaminopenicillanic acid In the process of Example 10, the phenoxyacetic acid was replaced by caproic acid to produce N,N&#39;- dibenzylethylenediamine-o-caproly-aminopenicillanic acid. (recovery: 58.0%) Elemental analysis:  
 Various changes and modifications of the invention can be made and, to the extent that such variations incorporate the spirit of this invention, they are intended to be included within the scope of the appended claims.  
  What is claimed is: l. A process for the production of a deptide having the general formula:  
 wherein:  
 R is a member selected from the group consisting of a-lower alkyl-, phenyland benzyla-amino acetic acid residues N-protected with carbo-benzoxy groups; and  
  R is an amino acid residue, said process comprising:  
 reacting in an inert organic solvent at a temperature of from about 5C to room temperature, a first compound having the general formula:  
 /CO-CH2 R1 -co -O-N CO-CHZ wherein:  
 R is a member selected from the group consisting of a monochloromethyl and a dichloromethyl group, with a second compound having the following general formula:  
 wherein:  
 R has the same meaning as hereinbefore described, wherein the molar ratio of said first compound to said second compound is from about 1.0 to about 2 moles of said first compound per 1 mole of said second compound, said reaction being conducted in the presence of from about 1.0 to about 3.0 moles of a tertiary organic amine per 1 mole of said second compound to yield a succinimide ester compound having the general formula:  
 R2 -CO-O-N wherein:  
 R has the meaning hereinbefore described; and subsequently reacting said succinimide compound with an amino compound in a ratio of from 1 to 2 moles of said succinimide compound per 1 mole of said amino compound, said amino compound being represented by the following general formula: