IL34630A - Polypeptides and derivatives thereof,their production and pharmaceutical compositions containing them - Google Patents

Description

ems o.' 'san mnpin *"i»w:sm Polypeptides and derivatives thereof, their production and pharmaceutical compositions containing them SANDOZ A.G.

G: -32861 34630 3 1' - 1 - The present invention relatee to polypeptides or 1 polypeptide derivatives of general formula I, S-CHg-CIiX-CO-Ser-Asn-Leu-Ser-Xiir-Cys-Val-Leu- Gly-Lys-Leu-Ser-Gln-Glu-Leu-His-Lys-Leu-Gln- I Thr-Tyr-Pro-Arg-Thr-Asn-Thr-Gly-SerrrGly-Thr-Y wherein X is -II, -NH2, R-C0-NH-, wherein ia alkyl of 1-4 carbon atoms, phenyl or i benzyl, both the latter optionally substituted with · halogen, lower alkoxy or nitro, or R*-0-C0-NH-, wherein R' is not capable of being split off with trifluoroacetic acid, and is alkyl of 1-4 carbon atoms, alkenyl of 2-4 carbon atoms, benzyl, p-nitrobenzyl, p-chlorobenzyl, p-bromobenzyl, p-phenyl-azobenzyl or p-( '-methoxyphenyl)-azobenzyl, Y is L-prolinamide or L-prolin-L-valinamide , the terminal peptide unit 3-0II2-CBX-C0- having the L-fonn when X is -ϋΗ2, R-CO-M-..or R'-O-CO-NH-, and wherein an L-^seryl radical, may be exchanged for an L-threon l or L-alanyl radical, an L-asparaginyl radical for an L-aspartyl, L-glutaminyl or L-glytamyl radical, an L-threonyl radioal for an L-seryl or L-alanyl radical, an L-lysyl radical for, a BOC-lyeyl, L-ornithyl or L-arginyl radical, an L-glutaninyl radical for ah L-glutamyl, L-asparaginyl or L-aspartyl radical, an L-glutamyl radical for an OTB-glutamyl, L-glytaminyl, L-asparaginyl or L-aspartyl radical, an L-leucyl radioal for an L-tyrosyl radical, an L-tyrosyl radical for an L-pheny alanyl radical and an L-arginyl radical for an L-ornithyl or L-lysyl radical, or a pharmaceutically acceptable aoid addition salt or heavy metal complex thereof, and processes fo their production* •34630/2 As already stated, in the peptide sequence of the general formula I single amino acids may be exchanged for other, natural amino acids or their derivatives.

For example, ' j_n positions 2, 5 , 13 and 29 Ser may be replaced by Thr or Ala, in positions 3 and' 26 Asn may be replaced by Asp, Gin or Glu, in. positions 6, 21, 25 , 27 and 31 Thr may be replaced by Ser or Ala, in positions 11 and 18 Lys may be replaced by BOC-Lys, Orn or Arg, in positions 1 and 20 Gin may be replaced by Glu, Asn or Asp, in position 15 Glu may be replaced by OTB-Glu, Gin, Asn or Asp, in position 19 Leu may be replaced by Tyr, in position 22 Tyr may be replaced by Phe, and in position 2K Arg may be replaced by Orn or Lys.

■ A compound of general formula I may be produced by a process comprising Joining two peptide units by an amide linkage, the peptide units being such that when Joined the amino acid sequence of the compound of general formula I is achieved, and when desired, forming a pharmaoeutically acceptable acid addition salt or heavy metal complex of the resulting compound. - 3 - 100-3077 Preferably one of the peptide units incorporates the group of formula II, S-CH2-CHXC0-Ser-Asn-Leu-Ser-Thr-Cys- II wherein X has the above significance, wherein an S-S bridge exists between the terminal amino acid and L-cysteinyl, and wherein an L-seryl radical may be exchanged for an L-threonyl or L-alanyl radical, an L-asparaginyl radical for an L-aspartyl, L-glutaminyl or L-glutamyl radical, an L-leucyl radical for an L-tyrosyl radical, and an L-threonyl radical for an L-seryl or L-alanyl radical, and preferably also, the S-S bridge is formed in the last stage In the production of the peptide unit.

It is to be understood that the expression "peptide unit" includes within its meaning a single amino acid, and a plurality of amino acids joined by amide linkages In the form of a peptide.

The peptide units comprising more than two amino acids may be produced by generally known methods for the synthesis of compounds of this type, it being possible to join together the amino acids in the desired order one at a time or by first forming smaller peptide units and joining these together. The peptide units' may, for example, - 100-3077 be joined by reacting an amino acid or a peptide having a protected a-amino group and an activated terminal carboxyl radical with an amino acid or a peptide having a free a-amino group and a free or protected terminal carboxyl radical, or by reacting an amino acid or a peptide having an activated a-amino group and a protected terminal carboxyl radical with an amino acid or a peptide having a free terminal carboxyl radical and a protected a-amino group.

The carboxyl radical may, for example, be activated by conversion into an acid azide, anhydride, imidazolide, isoxazolide or an activated ester, or by reaction with a carbodiimide or Ν,Ν' -carbonyl-diimidazole . The preferred condensation methods are the carbodiimide method, the azide method, the method of the activated esters, the anhydride method and the Merrifield method.

The introduction of a desired protective radical on the terminal L-hemicystin molecule of the final product may be effected in any desired stage before the last stage.

In the last stage of the condensation it is convenient to employ methods avoiding or minimizing racemization, preferably by using the azide or activated ester method, whereby activation is conveniently effected with N-hydroxysuccinimide. - 5 - 100-3077 The free functional groups which do not participate in the reaction may be protected as follows during the building up of the peptides: With reference to partial sequence B, described hereinafter, a suitable radical for blocking the guanido radical of the arginin radical is the nitro radical, but other suitable protective radicals, such as the tosyl radical, the p_-nitrocarbobenzoxy radical or the 2-(isopropyloxycarbonyl)-3, ,5»6-tetrachlorobenzoyl radical, may likewise be used. It is also possible to use the protective effect of the protonization of the guanido radical during the synthesis.

With reference to partial sequence D, described hereinafter, a suitable radical for blocking the ^-carboxyl radical is the tert.butyloxy radical, but other protective radicals, such as the methoxy, ethoxy, tert.amyloxy, the amide or the benzyloxy radical, may likewise be used.

With reference to partial sequence C, described hereinafter, a suitable radical for blocking the ttT-amino radical of the lysyl radical is a carbo-tert.alkoxy radical, preferably the carbo-tert.butoxy radical.

With reference to partial sequences EP and EFJ, described hereinafter, the benzyl or trityl radical is preferably used as mercapto-protective radical. When benzyl or trityl radicals are used for the protection of SH radicals, it is usual to split them off at the end of the synthesis by treatment with sodium in liquid ammonia. It has now been found that the splitting off of the benzyl or trityl-protective radicals and the formation of the S-S bridge at the stage before the last stage of the build up lead to particularly high yields of final product.

The conversion of a protected mercapto or amino radical into a free radical, as well as the conversion of a functionally modified carboxyl radical into a free carboxyl radical during the course of the process for the production of the polypeptides is effected in conventional manner by treatment with hydrolyzing or reducing agents.

The starting materials for producing the polypeptides or polypeptide derivatives, insofar as they were hitherto unknown, may be obtained by methods for the synthesis of peptides in actual use or described in the literature, whereby the amino acids may be joined one at a time or after the formation of smaller peptide units.

The polypeptides or polypeptide derivatives may likewise be obtained or used in the form of their salts. Examples of salts are those with organic acids, e.g. acetic, lactic, succinic, benzoic, salicylic, methanesulphonic or toluenesulphonic acid, as well as polymeric acids, such as tannic acid or carboxymethyl cellulose, and salts with inorganic acids, such as hydrohalic acids, e.g. hydrochloric acid, or sulphuric acid or phosphoric acid. An example of a heavy metal complex is a complex formed with zinc ®® .

The compounds produced in accordance with the invention represent an important therapeutic principle. They lower the content of calcium in- plasma and produce a positive calcium balance in the bones as antagonists of the parathormone. Biologic testing of the new compounds on rats indicates a value of between 5000 and 5000 MRC units per mg of peptide for the lowering of the calcium content in the blood. - 7 - 100-3077 The dose to be administered depends on the desired effect as well as the mode of application. However, satisfactory results are generally obtained with a single daily dose of 1 to 10 MRC units per kg of animal body weight. For larger mammals the daily dose amounts to about 70 to 700 MRC units, which may be administered in a single dose or in several portions. For intramuscular administration a suitable application form contains about 70 to 700 MRC units of peptide as an active agent, mixed with a liquid. carrier. A suitable indicated unit dosage form comprises between 20 and 700 MRC units of active agent.

The compounds produced in accordance with the invention are therefore indicated for use in the treatment of conditions in which a lowering of the calcium content in plasma or an influence on the bone metabolism is desired, e.g. hypercalcemias produced by endogenic thyrocalcitonin deficiency, caused by dysfunction of the thyroid gland tissue or by hyperfunction of the parathyroid glands. They are furthermore indicated for use in the treatment of bone diseases based on an increased decomposition, or in which a fixation of calcium in the bones is desired, e.g. osteoporosis of various origins (e.g. postclimacteric, posttraumatic, caused by corticosteroid therapy or inactivity), fractures, osteomalacia, rickets,' renally conditioned osteodystrophy, and especially for a combined therapy with calcium or phosphate.

An advantage of the polypeptides or polypeptide derivatives of the general formula I, produced in accordance with the invention, wherein X is > R-CO-NH- (R = alkyl, optionally substituted aralkyl, optionally substituted aryl) or R'-O-CO-NH- (R1 = alkyl, alkenyl, optionally substituted aralkyl, optionally substituted aryl), is that they are not affected by aminopeptidase degradation. _ 8 . 100-5077 The compounds produced in accordance with the invention may be used as medicaments, e.g. in the form of pharmaceutical preparations. These may contain the said compounds in mixture with an organic or inorganic carrier material which is suitable for parenteral administration. They may likewise be administered in the form of a depot preparation. - 9 - 100-3077 The following abbreviations are used: z Carbobenzoxy (Benzyloxycarbonyl ) Bzl Benzyl BOC tert. Butyloxycarbonyl Trt = Trityl = Triphenylme hyl 0TB zz tert.: Butyloxy ONP = £-Nitrophenyl ester OCP = 2,4,5-Trichlorophenoxy OMe = Methoxy OEt = Ethoxy 02 Nitro Ser r= L-Seryl Asn - L-Asparaginyl Leu = L--Leucyl Thr L-Threonyl Val = L-Valyl Tyr L-Tyrosyl Arg — L~Arginyl Gin - ■ L-Glutaminyl Glu - L-Glutamyl His = L-Histidyl Pro L-Prolyl Gly = Glycyl Lys L-Lysyl -10 - 100-5077 L-Cystelnyl Pivaloyl (3-Mercaptopropionyl N-OxySliceinimlde Ethoxycarbonyl Dicyclohexylamine In the following non-limitative Examples all temperatures are indicated in degrees Centigrade. The value c for the optical rotation amounts to 1.

Amino acid analysis in the following Examples showed that the single amino acids occur in the expected proportions. - 11 - 100-3077 ^rMi-f-9H£?≤S_^ : H-Ser-Gly-Thr-Pro-NH£ a) H-Thr-Pro-NH . HC1 13^ g of Z-Thr-NHNH are dissolved at -5° in 2 liters of 1 N hydrochloric acid, and 0.55 liters of IN sodium nitrite are. added. After 5 minutes potassium carbonate is added until the pH of the solution amounts to 9, the resulting azide is extracted with ethyl acetate, and a solution of 80 g of H-Pro-NH^ . hydrochloride in 100 cc of water, 500 cc of dimethyl for amide and 77 cc of triethyl amine is added. The ethyl acetate is evaporated at 20° in a vacuum and the reaction mixture is allowed to stand over night at 25°. The remaining solution is evaporated in a vacuum, the residue is dissolved in ethyl acetate, the solution is washed with water, dilute hydrochloric acid and an aqueous calcium carbonate solution and is dried over sodium sulphate. The reaction mixture is concentrated by evaporation in a vacuum, the residue is dissolved in warm ethyl acetate and is cooled. Z-Thr-Pro-NHg, having a M.P. of ]A8°, [α]β = -72° in 95 % acetic acid, is obtained. 90 g of Z-Thr-Pro-NH, are subsequently dissolved in 2 liters of dioxane and 260 cc of IN hydrochloric acid, and hydrogenation is effected at 20° and normal pressure in the presence of a palladium catalyst. Filtration is effected, the solution is- evaporated in a vacuum and the crystalline residue is washed with ethyl acetate, whereby H-Thr-Pro-NH^ . HC1 , having a M.P. of 216°, [ j^ = -6 ° in 95 ace ic acid, is obtained. 14.5 g of Z-Ser-OH are dissolved in 100 cc of chloroform and 6.1 g of -methyl morpholine are added to the solution; 8.2 g of chloroformic acid isobutyl ester are subsequently added dropwise. - 12 - 100-3077 After 10 minutes a solution of 6.6 g of glycine ethyl ester in 50 cc of chloroform is added, and the mixture is stirred at room temperature for 1 hour. The reaction mixture is washed with dilute ammonia, subsequently with hydrochloric acid solution, is dried over sodium sulphate, and the organic phase is removed by evaporation. The resulting Z-Ser-Gly-OEt is recrystallized from ethyl acetate. M.P. 103° , 20 [ JD = +3° in dimethyl formamide. 19.4 g of Z-Ser-Gly-OEt are dissolved in 270 cc of ethanol, 27 cc of hydrazine hydrate are added and the mixture is allowed to stand at room temperature for 2 days. The crystalline mass is filtered off, washin with methanol and drying are effected. Z-Ser-Gly-NHNH^, having a M.P. of l80° , [a]D = +2° in dimethyl formamide, is obtained. c) H-Ser-Gly-Thr-Pro-NH2 cc of a h- N hydrogen chloride solution in ethyl ether are added to 200 cc of dimethyl formamide, and 11 g of Z-Ser-Gly-NHNH^ are dissolved therein. 5 A cc of tert.butyl nitrite are subsequently added dropwise at -10° , l8 cc of triethyl amine are added to the solution, and filtration is subsequently effected. 12 g of H-Thr-Pro-NH^ · HC1 are simultaneously dissolved in 100 cc of dimethyl formamide, 6.2 cc of triethyl amine are added, the precipitated triethyl amine hydrochloride is filtered off, and the dipeptide azide solution obtained above is added to the filtrate. The reaction mixture is allowed to stand at 0° for 2 hours, is evaporated to dryness, the residue is dissolved in a mixture of 500 cc of methanol and 100 cc of water, and the resulting solution is filtered through a 100 cc Dowex-50 column (H+ form). The solution is concentrated by evaporation and the residue is crystallized from methanol/ethyl acetate.

Z-Ser-Gly-Thr-Pro-NH2, having a M.P. of 152° (decomp.), [a] = -22° in dimethyl formamide, is obtained. 22 g of Z-Ser-Gly-Thr-Pro-NH2 are dissolved in a mixture of 440 cc of methanol and 88 cc of water, and hydrogenation is effected at 20° and normal pressure in the presence of palladium charcoal.

Filtration is subsequently effected, the filtrate is evaporated to dryness, the residue is washed with ether and dried.

H-Ser-Gly-Thr-Pro-NH2, having a M.P. of 95° (decomp.), [aJD = -27° in dimethyl formamide, is obtained. a) Z-Asn-Thr-Gly-OEt g of Z-Thr-Gly-OEt are dissolved in 500 cc of dimethyl formamide, and hydrogenation is effected at normal pressure and room temperature in the presence of palladium charcoal. Filtration is effected, 60 g of Z-Asn-OCP are added, the mixture is allowed to stand at 25° for 16 hours, is concentrated to about 250 cc, ethyl acetate is added, the reaction mixture is washed with dilute hydrochloric acid, dried over sodium sulphate and evaporated to dryness. The residue is washed with ethyl acetate and dried.

Z-Asn-Thr-Gly-OEt, having a M.P. of 225° , [α]β = -4° in dimethyl formamide, is obtained. b) Z-Thr-Asn-Thr-Gly-OEt 29 g of Z-Asn-Thr-Gly-OEt are dissolved in 700 cc of dimethyl formamide, hydrogenation is effected at normal pressure and room temperature in the presence of palladium charcoal, filtration is effected, and Z-Thr-N.,, produced from 50 g of Z-Thr-NHNH2 [see partial sequence A, a)], is added to the filtrate. After allowing to - 1 - 100-5077 stand at 0° for 12 hours the reaction mixture is evaporated to dryness, the residue is washed with water, ether and ethanol.

Z-Thr-Asn-Thr-Gly-OEt, having a M.P. of 230°, [a]D = -8° in dimethyl formamide, is obtained. c) B0C-Arg(N02)-Thr-Asn-Thr-Gly-:NHNH2 22 g of Z-Thr-Asn-Thr-Gly-OEt are dissolved in 700 cc of dimethyl formamide, hydrogenation is effected at normal pressure and room temperature in the presence of palladium charcoal and filtration is effected. 23 g of B0C-Arg(N02)-0H are dissolved in 500 cc of dimethyl acetamide, 12 cc of triethyl amine are added, the solution is cooled to -10°, 9A cc of chloroformic acid isobutyl ester are added, the mixture is allowed to react for 10 minutes, and the tetrapeptide solution obtained above is added. After standing at 250 for 0 minutes 100 cc of water are added, the resulting solution is treated with amberlite-IRA-4l0 (OH form) until a negative chlorine reaction is obtained, filtration and evaporation to dryness are effected. The residue is washed with chloroform, ethyl acetate and ether and is dried. B0C-Arg(N02)-Thr-Asn-Thr-Gly-0Et, having a M.P. of 100°, [α]β = -4° in dimethyl formamide, is obtained. 12 g of B0C-Arg(N0 )-Thr-Asn-Thr-Gly-0Et are dissolved in a mixture of 200 cc of acetic acid and 40 cc of water, hydrogenation is effected in the presence of palladium charcoal at 20° and normal pressure, filtration and evaporation to dryness in a vacuum at 20° are effected. The residue is dissolved in dimethyl formamide and is evaporated. This treatment is repeated until no acetic acid can be detected. The residue is dissolved in 120 cc of dimethyl formamide, 6 cc of hydrazine hydrate are added, the solution is allowed to stand ° - 15 - 100-30.77 methanol/ether (1:1) and then again with ether, and drying is effected.

BOC-Arg-Thr-Asn-Thr-Gly-NHNH^, having a M.P. of 175° (decomp.), [ ]D = - 2° in dimethyl formamide/l N HC1 (1:1), is obtained. are dissolved in 270 cc of dimethyl formamide/water (8:2) , cooling is effected to -15°, 40 cc of a N solution of hydrochloric acid in dioxane and 2.5 cc of tert.butyl nitrite are added. The solution is stirred at -15° for 10 minutes, 28 cc of triethyl amine are added, filtration is effected, 10.5 S of H-Ser-Gly-Thr-Pro-NH_ (partial . sequence A) are added and stirring is effected at 0° for 16 hours. The reaction solution is subsequently evaporated to dryness, the residue is washed with ether and dried. The residue is dissolved in methanol, one tenth by volume of water and a sevenfold quantity of chloroform are added, and the resulting solution is filtered through a column of 00 g of silica gel. After elution with an increasing amount of methanol and concentrating by evaporation, BOC-Arg-Thr-Asn-Thr-Gly-Ser-Gly-Thr-Pro-NH2 is obtained. This is suspended in 200 cc of a N solution of hydrochloric acid in dioxane and the solution is stirred at 25° for 2 hours. The solution is evaporated to dryness, the residue is dissolved in 0.2 N acetic acid, the solution is treated with amberlite-IRA- lO (acetate form), and the aqueous solution is lyophilized. The residue is subsequently washed with ether, ethyl acetate and again with ether and drying is effected over sodium hydroxide shavings. H-Arg-Thr-Asn-Thr-Gly-Ser-Gly-Thr- 20 Pro-NH . 2 CILCOOil , having a M.P. of.1§5° (decomp. ) , [a] = -64° dimethyl formamide/l N HC1 (1:1), is obtained. - 16 - 100-3077 Partial sequence C: Z-Hls-Lys(BOG)-Leu-Gln-Thr-Tyr-Fro-OH a) H-Thr-Tyr-Pro-OH 176 g of Z-Tyr(Z)-Pro-CMe are dissolved in 1000 cc of methanol/1 N HC1, hydrogenation is effected in the presence of palladium charcoal at 20° and 1 atmosphere, and the solution is concentrated by evaporation. The residue is dissolved in 400 cc of dimethyl formamide, 100 cc of triethyl amine are added at 0°, the triethyl amine hydrochloride which crystallizes is filtered off, and Z-Thr-N-. [produced from 87 g of Z-Thr-NHNH^, see partial sequence A, a)], is added to the filtrate. The reaction mixture is subsequently allowed to stand at 0° for l6 hours, is evaporated to dryness, the residue is dissolved in ethyl acetate, the solution is successively washed with dilute hydrochloric acid and dilute ammonia, is dried and concentrated by evaporation. The residue is pulverized in heptane, washing with petroleum ether and drying are effected.

Z-Thr-Tyr-Pro-OMe, having a M.P. of 80° (decomp.), [ ]ρ = -23° in dimethyl formamide, is obtained. 53 S of Z-Thr-Tyr-Pro-OMe are dissolved in 530 cc of methanol, 100 cc of a 2 N caustic soda solution are added, the solution is allowed to stand at 25° for 1 hour, 0 cc of 2 N hydrochloric acid are added, the solution is concentrated to about 200 cc, 100 cc of water are added, the pH value of the solution is adjusted to 10, the aqueous solution is washed twice with ethyl acetate, the pH value of the solution is subsequently adjusted to 1 with N hydrochloric , acid, the precipitated tripeptide Z-Thr-Tyr-Pro-OH is extracted with ethyl acetate, is dried and concentrated by evaporation. M.P. 8o°, 20 [ajp = -23° in dimethyl formamide. The residue is dissolved in a mixture of 500 cc of dioxane and 100 cc of water, and hydrogenation - 17 - 100-3077 is effected at 20° and normal pressure in presence of a palladium catalyst. Filtration is effected, the filtrate is evaporated to dryness, the residue is washed with ether and dried.

H-Thr-Tyr-Pro-OH, having a M.P. of l80° (decomp.), [a]^ = -51° in 95 % acetic acid, is obtained. b) Z-Leu-Gln-0Me 70 g of Z-Gln-OH are dissolved in 1.5 liters of dioxane, and a solution of diazomethane in ether is allowed to flow in until the solution remains yellow coloured. The solution is subsequently concentrated by evaporation and the white residue is treated with ether. Z-Gln-OMe, having a M.P. of 136°, [ ]D = -21° in dimethyl formamide, is obtained. 71 S of the ester are dissolved in 1.7 liters of methanol. 5 S of 10 % palladium on active charcoal are subsequently mixed by stirring with 59 cc of 4 N hydrochloric acid, and this is added to the solution. The entire material is subjected to hydrogenation at room temperature and normal pressure for 3 hours, whereby about 83 of the theoretic amount of hydrogen are used up. The catalyst is filtered off and the filtrate is completely concentrated by evaporation, whereby H-Gln-OMe . HC1 is obtained in the form of a viscous oil. 64 g of Z-Leu-OH are dissolved in 200 cc of acetonitrile, 27.7 g of N-hydroxysuccinimide are added and the solution is cooled to -10° . A solution of 0 g of dicyclohexyl carbodii ide in 100 cc of acetonitrile is added.thereto. After about 30 minutes the dicyclohexyl urea is filtered off. 7.3 S of H-Gln-OMe . HC1 , dissolved in dimethyl formamide, are added to the filtrate while simultaneously adding one equivalent of N-methyl morpholine. The reaction mixture is allowed to stand for 4 hours and is concentrated by evaporation. The - 18 - 100-5077 residue is dissolved in ethyl acetate and the solution is washed with a 5 % sodium bicarbonate solution, 1 N hydrochloric acid and water.

The organic phase is dried over sodium sulphate and concentrated by evaporation in a vacuum. After recrystallization from methanol and ether Z-Leu-Gln-OMe, having a M.P. of 179° , [ ]D = -16° in dimethyl formamide, is obtained. c) Z-Lys(BOC)-Leu-Gln-O e 42 g of Z-Leu-Gln-OMe are dissolved in 1500 cc of methanol. 16 g of 10 palladium on active charcoal are subsequently mixed by stirring with 25.8 cc of N hydrochloric acid, and this is added to the solution. The entire material is subjected to hydrogenation at room temperature and normal pressure for 8 hours, whereby 95 of the theoretic amount of hydrogen are used up. The catalyst is filtered off and the filtrate concentrated by evaporation. H-Leu-Gln-OMe . HC1 is obtained in the form of a foam. H-Leu-Gln-OMe . HC1 is subsequently dissolved in 300 cc of dimethyl formamide, 34 g of Z-Lys(BOC) -CCP and 8.4 cc of triethyl amine are added. After shaking the reaction mixture is allowed to stand for l6 hours. The reaction mixture is subsequently concentrated by evaporation in a vacuum. The residue is treated with water and filtered off from the water. The residue is subsequently dissolved in a small amount of methanol and precipitation is effected with ether. After iltration and drying Z-Lys(BOC)-Leu-Gln-OMe, [a]D - -21° in dimethyl formamide, is obtained. d) Z-His-Lys(BOC)-Leu-Gln-NHNH2 38 g of Z-Lys(BOC)»Leu-Gln-OMe are dissolved in 300 cc of dimethyl formamide, 15 g of 10 palladium charcoal are added, hydrogenation is effected at normal pressure and room temperature, - 19 - 100-3077 and the catalyst is filtered off. Z-His-N^ produced from 5-2 g of Z-His-NH H^, is subsequently added, the reaction mixture is allowed to stand at 20° for 5 hours, is evaporated, the residue is dissolved in ethyl acetate, the solution is washed with water, dried over sodium sulphate, evaporated, and the residue is washed with ether.

Z-His-Lys(BOC)-Leu-Gln-OMe, having a .P. of 120° (decomp.), [a]p = -19° in dimethyl formamide, is obtained. 33 g of Z-His-Lys(B0C)-Leu-Gln-0Me are dissolved in 330 cc of methanol, 7·5 cc of hydrazine hydrate are added, the reaction mixture is allowed to stand at 25° for 2 days and is subsequently evaporated to dryness. The residue is subsequently dissolved in dimethyl formamide, is concentrated by evaporation and this treatment is repeated until no traces of hydrazine can be detected. The residue is washed with ether, water and ether, and drying is effected, whereby Z-His-Lys(B0C)-Leu-Gln-NHNH2, having a M.P. of 199°, [ ]p = -22° in methanol, is obtained. e) Z-His-Lys(BOC)-Leu-Gln-Thr-Tyr-Pro-OH 0 g of Z-His-Lys(B0C)-Leu-Gln-NHNH2 are dissolved in 400 cc of dimethyl formamide, the solution is cooled to -20°, 75 cc of dioxane/2 N HC1 are added and subsequently 6 cc of tert.butyl nitrite are added, the mixture is stirred at -20° for 10 minutes, 30 cc of .triethyl amine and 20 g of H-Thr-Tyr-Pro-0H are added and the mixture is allowed to react at 25° for l6 hours. The reaction mixture is subsequently evaporated to dryness, the residue is washed with ether, dilute acetic acid, ether and subsequently with hot ethyl acetate. Drying is subsequently effected in a high vacuum, whereby Z-His-Lys(B0C)-Leu-Gln-Thr-Tyr-Pro-0H, having a M.P. of 21 ° (decomp. / [a = -23° in dimethyl formamide, is obtained. - 20 - 100-3077 ial_se¾uence_Cl : Z-His-Lys(BOG)-Tyr-Gln-Thr-Tyr-Pro-OH a) H-Thr-Tyr-Pro-OH See partial sequence C, a) . b) Z-Gln-Thr-Tyr-Pro-OH 40.1 g of Z-Gln-ONP and J>8A g of H-Thr-Tyr-Pro-OH are dissolved in 500 cc of dimethyl formamide and 12.2 cc of N-raethyl morpholine are added to the solution. The solution is allowed to stand at room temperature for 16 hours and is subsequently completely concentrated by evaporation in a vacuum. The evaporation residue is taken up in ethyl acetate/n-butanol (10:1) and is washed with 1 N sulphuric acid/30 % sodium chloride solution (1:1) and then with 15 % sodium chloride, solution. After drying over sodium sulphate the organic phase is completely concentrated by evaporation in a vacuum. The residue is dissolved in chloroform/methanol (1:1) and filtration is effected through a silica gel column (20-fold quantity).

Z-Gln-Thr-Tyr-Pro-OH is obtained in the form of a foam. [a]p = -19° in dimethyl formamide. c) z-Ij s(BOC)- yr-0Me 1*5.9 g of Z-Lys(B0C)-0H . DOHA are dissolved in 600 cc of warm dimethyl formamide and after cooling the solution to 35° a solution of 65.I g of H-Tyr-OMe . HC1 in 200 cc of dimethyl formamide is added, whereby DOHA . HC1 crystallizes quantitatively. After filtration 30 g of N-hydroxysuccinimide are added to the filtrate, the mixture is cooled to -15° and a solution of 53.6 g of dicyclohexyl carbodiimide in 200 cc of acetonitrile is added. The mixture is shaken at 0° for 15 hours, the dicyclohexyl urea is filtered off and the filtrate is concentrated by evaporation in a vacuum. The residue - 21 - 100-3077 is dissolved in ethyl acetate, the solution is washed with 5 % sodium bicarbonate solution, water, 1 N sulphuric acid and water, is dried over sodium sulphate and concentrated by evaporation to a foam, whereby Z-Lys(B0C)-Tyr-0 e, [ ]D = -6° in methanol, is obtained. d) Z-His-Lys(B0C)- yr-NHNH2 II3.5 g of Z-Lys(B0C)-Tyr-0Me are dissolved in 2 liters of methanol, a suspension of 10 g of 10 % palladium on active charcoal in 50 cc of 4 N hydrochloric acid is added to the solution, and hydrogenation is effected at room temperature and normal pressure.

After hydrogenating for 3 hours 90 % of the theoretic amount of hydrogen has been used up. The catalyst is filtered off and the filtrate is completely concentrated by evaporation in a vacuum. The foamy H-Lys(B0C)-Tyr-0Me . HC1 is uniform in accordance with thin layer chromatography and is joined with Z-His-N_, as follows: 88.5 g of Z-His-NH H^ are dissolved in l800 cc of dimethyl formamide, the solution is cooled to -20°, 405 cc of a 3·7 N solution of HC1 in ether are added and 36.6 cc of tert.butyl nitrite are added dropwise. After stirring at -20° for 10 minutes 20 cc of triethyl amine and a solution of H-Lys(B0C)-Tyr-0Me . HC1 in 600 cc of dimethyl formamide, already containing 27.3 cc of triethyl amine, are added. After 5 hours the triethyl amine . HCl which crystallizes is filtered off. The filtrate is completely concentrated by evaporation in a high vacuum. The residue is dissolved in ethyl acetate/water (2:1), the organic phase is washed 4 times with a 15 sodium chloride solution, is dried over sodium sulphate and concentrated by evaporation. After dissolving the residue in chloroforra/methanol (98:2) and filtering is obtained over a silica gel column (25-fold quantity) Z-His-Lys(B0C)-Tyr-0Me/ in amor hous form, a = -11° in dimeth l formamide. 90.4 g of - 22 - 100-5077 Z-His-Lys(BOC) -Tyr-OMe are dissolved in 300 cc of methanol and 20 cc of hydrazine hydrate are added. After allowing to stand for 2 days the resulting hydrazide crystallizes. 300 cc of ether are added, filtration with suction, washing with ether and drying in a vacuum are effected. Z-His-Lys(B0C)-Tyr-NH H2, having a M.P. of l87° , [a]p = -24° in dimethyl formamide, is obtained. e) Z-His-Lys(B0C)- yr-Gln-Thr-Tyr-Pro-OH 36.7 g of Z-Gln-Thr-Tyr-Pro-OH are dissolved in 500 cc of dimethyl formamide, 15 g of 10 % palladium on active charcoal are added, hydrogenation is effected at room temperature and normal pressure, and the catalyst is subsequently filtered off. The filtrate contains H-Gln-Thr-Tyr-Pro-OH, which is joined with Z-His-Lys(B0C)-Tyr-N5 as follows: kO. k g of Z-His-Lys(BOC)-Tyr-NHNH^ are dissolved in 400 cc of dimethyl formamide, the solution is cooled to -20° , 88 cc of dioxane/2N HC1 are added and subsequently 7· 3 cc of tert.butyl nitrite are added. After stirring at -20° for ten minutes 37 cc of triethyl amine and the dimethyl formamide solution containing H-Gln-Thr-Tyr-Pro-OH are added. After standing at 20° for hours the reaction mixture is completely concentrated by evaporation in a vacuum, the residue is dissolved in ethyl acetate/n-butanol (5:1)* the solution is washed with dilute acetic acid with the addition of sodium chloride solution, is dried over sodium sulphate and concentrated by evaporation. Ether is added to the oil which has not been entirely concentrated by evaporation, the mixture is mixed well and filtered with suction. After drying in a high vacuum Z-His-Lys(BOC) -Tyr-Gln- 20 Thr-Tyr-Pro-OH, having a M.P. of 193° , [α]β = -33° n dimethyl formamide, is obtained. - 23 - 100-3077 Partial_sequence_ABC : H-His-Lys(BOC) -Leu-Gln-Thr-Tyr-Pro-Arg-Thr- Asn-Thr-Gly-Ser-Gly-Thr-Pro-NH^ ♦ 3 CH^COOH g of heptapeptide (partial sequence C) are dissolved in 100 cc of dimethyl formamide, the solution is concentrated by evaporation, the residue is dissolved in 100 cc of dimethyl formamide, g of N-hydroxysuccinimide are added, the solution is cooled to 0° , 5 g of dicyclohexyl carbodiimide are added, the reaction mixture is allowed to react for 3 hours, the precipitated dicyclohexyl urea is filtered off, the filtrate is concentrated to 3 cc and the resulting heptapeptide oxysuccinimide ester Z-His-Lys(BOC)-Leu-Gln-Thr-Tyr-Pro-OSu is precipitated by the addition of ether. After washing out with ether the residue is dissolved in 100 cc of dimethyl formamide, 10 g of nonapeptide amide (partial sequence AB) are added and the mixture is allowed to react for 16 hours. 500 cc of ethyl acetate are added and filtration is effected. The residue is dissolved in dimethyl formamide, 10 cc of acetic acid are added and precipitation is again effected with ethyl acetate. Filtration, washing with ethyl acetate and ether and drying are effected. Z-His-Lys(BOC)-Leu-Gln- Thr-Tyr-Pro-Arg-Thr-Asn-Thr-Gly-Ser-Gly-Thr-Pro-NH2 . 2 AcOH , [a)D = -19° in dimethyl formamide, is obtained and is directly dissolved in 80 acetic acid. 5 S of palladium charcoal are subsequently added and hydroge ation is effected at normal pressure and room temperature. Filtration is effected, the filtrate is evaporated to dryness at 20° in a high vacuum and the residue is washed with ether. After drying over KOH shavings partial sequence ABC, having a M.P. of 170° (decomp.), [a] = -53° in 1 N acetic acid, is obtained. - 24 - 100-3077 '" : H-Hls-Lys(BOC)-Tyr-Gln-Thr-Tyr-Pro-Arg-Thr-Asn- Thr-Gly-Ser-Gly-Thr-Pro-NH^ . 3 CH^COOH Partial sequence ABC1, having a M.P. of 166° (decornp.), [ jp = -50° in 1 N acetic acid, is obtained by a process analogous to that described for the production of partial sequence ABC, from 10 g of heptapeptide (partial sequence Cl) and 10 g of nonapeptide (partial sequence AB) .

Trt-Gly-Lys(BOC) -Leu-Ser-Gln-Glu(0TB)-Leu-NHNH£ a) Z-Glu(0TB)-Leu-0Me 151 g of Z-Glu(OTB)-0H and 54 g of N-hydroxysuccinimide are dissolved in 700 cc of acetonitrile and the solution is cooled to -20°. 96.5 S of dicyclohexyl carbodiimide, dissolved in 350 cc of acetonitrile, are subsequently added. After allowing the solution to stand for about 3 minutes the resulting dicyclohexyl urea is filtered off. 71.2 g of H-Leu-OMe are added to the filtrate, the mixture is allowed to stand for 4 hours and is concentrated by evaporation in a vacuum. Ethyl acetate/water (2:1) is added to the solid residue. The organic phase is subsequently washed with % sodium bicarbonate solution, water and dilute sulphuric acid (pH 3) > is washed until neutral, dried over sodium sulphate and concentrated by evaporation. The residue is dissolved in chloroform containing 1 % of methanol, and filtration is effected over a silica gel column (10-fold quantity). Z-Glu(OTB)-Leu-0Me, having a M.P. of 31°, [a]p = -12° in dimethyl formamide, is obtained. b) H-Gln-Glu(OTB)-Leu-0Me US g of Z-Glu(0TB)-Leu-0Me are dissolved in 2000 cc of methanol. 15 g of 10 % palladium on active charcoal are subsequently - 25 - 100-3077 mixed by stirring in 63 cc of 4 N hydrochloric acid, and this is added to the solution. The entire material is subjected to hydrogenation at room temperature and normal pressure for 2 hours, whereby about 85 of the theoretic amount of hydrogen are used up.

The catalyst is filtered off and the filtrate is completely concentrated by evaporation, whereby H-Glu(0TB)-Leu-OMe . HC1 is obtained in the form of an amorphous foam. 91. g of Z-Gln-ONP and 93 - 8 g of H-Glu(OTB)-Leu-QMe . HC1 are dissolved in 500 cc of dimethyl formamide, and 51 cc of N-methyl morpholine are added. The solution is allowed to stand at room temperature for 15 hours and is subsequently completely concentrated by evaporation in a vacuum. The resulting residue is mixed >well with water and is then filtered off from the water. Recrystallization from methanol is subsequently effected, whereby Z-Gln-Glu(CTB)-Leu-0Me, (decomp. ), 20 having a M.P. of 210V MD = - 0° in dimethyl formamide, is obtained.

KkA g of Z-Gln-Glu(OTB)-Leu-OMe are dissolved in 1200 cc of methanol/dimethyl formamide (l:l). 15 g of 10 % palladium on active charcoal are subsequently mixed with 0 cc of water, and this is added to the solution. After hydrogenation for about 2 hours an almost quantitative amount of hydrogen is used up. The catalyst is filtered off and the filtrate is concentrated by evaporation in a vacuum. The residue is dissolved in ethyl acetate and washed thrice with a saturated sodium chloride solution. The organic phase is dried over sodium sulphate and the filtrate is concentrated by evaporation to about one third of its original volume. After the addition of petroleum ether the solid tripeptide can be filtered off.

H-Gln-Glu(OTB)-Leu-0Me, having a M.P. of 123° , [a] = -JA° in dimethyl formamide, is obtained. - 26 - 100-5077 c) Z-Lys(B0C)-Leu-Ser-Oe g of H-Leu-Ser-OMe . HC1 and 80 g of Z-Lys(B0C)-0CP are dissolved in 400 cc of dimethyl formamide, 22 cc of triethyl amine are added, the solution is allowed to stand at 25° for 25 hours, the precipitated triethyl amine is filtered off and the filtrate is evaporated to dryness. The residue is dissolved in ethyl acetate, the solution is washed with dilute hydrochloric acid and dilute potassium bicarbonate solution, drying over sodium sulphate, filtration and concentration by evaporation are effected. The residue is re-crystallized from ethyl acetate/ether, whereby Z-Lys(B0C)-Leu-Ser-0Me, having a M.P. of 110°, = -14° in dimethyl formamide, is obtained. d) Trt-Gly-Lys(30C)-Leu-Ser- HNH2 158 g of Trt-Gly-OH are dissolved in 2C00 cc of dichloromethane and cc of triethyl amine, the solution is cooled to -5 50 cc of chloroformic acid ethyl ester are added and the mixture is stirred at -5° for 10 minutes. 500 g of Z-Lys(BOC)-Leu-Ser-0Me are simultaneously dissolved in 5000 cc of dioxane/water (8:2), hydrogenation is effected at room temperature and normal pressure in the presence of palladium charcoal, filtration and evaporation are effected. The residue is dissolved in chloroform and the mixed anhydride obtained above is added to the solution. The reaction solution is allowed to stand at 0° for 1 hour, the chloroform solution is washed with dilute acetic acid and subsequently with dilute potassium bicarbonate solution, is dried and evaporated to dryness. The residue is dissolved in 1250 cc of methanol, 175 cc of hydrazine hydrate are added and the reaction mixture is allowed to stand at 25° for l6 hours. The reaction mixture is concentrated to - 27 - 100-5077 with water and dried over phosphorus pentoxide.

Trt-Gly-Lys(BOC)-Leu-Ser-NHNH^, having a M.P. of 204°, [ ]ρ = -6° in dimethyl formamide, is obtained. e) Trt-Gly-Lys(BOC)-Leu-Ser-Gln-Glu(0TB)-Leu-NHH^ 76 g of Trt-Gly-Lys(B0C)-Leu-Ser-NH H2 are dissolved in 300 cc of dimethyl formamide, the solution is cooled to -20°, 75 cc of dioxane/4 N HC1 are added and subsequently 11.6 cc of tert.butyl nitrite are added, the mixture is stirred at -1 ° for 10 minutes, 70 cc of triethyl amine and 9.5 g of H-Gln-Glu(0TB)-Leu-0Me are added. The reaction mixture is subsequently stirred at 0° for 16 hours.

The precipitated triethyl amine hydrochloride is filtered off, the filtrate is concentrated to about 100 cc and water is added to the residue. The precipitated residue is filtered off, is then washed thoroughly with water and subsequently dried in a high vacuum at 50° . The residue is boiled with ethyl acetate, washed with ether and dried.

The resulting heptapeptide ester Trt-Gly-Lys(BOC)-Leu-Ser-Gln-Glu(0TB)- 20 Leu-OMe, having a M.P. of 234° , [a]^ = -20° in dimethyl formamide, is dissolved in 200 cc of dimethyl formamide, 4 cc of hydrazine are added and the mixture is allowed to stand at 25° for 24 hours. After concentrating to about 100 cc 00 cc of water are added, the precipitated material is filtered off, washing is effected with water until a neutral reaction is obtained and drying is effected over phosphorus pentoxide at 50° . Trt-Gly-Lys(BOC)-Leu-Ser-Gln-Glu( 0TB)-Leu-NHNH , having a M.P. of 265°·, [a] = -l8° in dimethyl formamide/water (8:2), is obtained. - 28 - 100-3077 ί' ΓΪί^--_?-:9^ί5ί1------Α^5?: H-Gly-Lys(B0C)-Leu-Ser-Gln-Glu(0TB)-Leu-His- Lys(BOC)-Leu-Gln-Thr-Tyr-Pro-Arg-Thr-Asn-Thr- 3 g of heptapeptide hydrazlde (partial sequence d) are dissolved in 30 cc of dimethyl formamide, the solution is cooled to -20°, cc of dioxane/4 N HC1 are added and subsequently 0.3 cc of tert.butyl nitrite are added. The reaction mixture is stirred at -20° for 10 minutes, 2 cc of triethyl amine and 3 g of hexadecapeptide amide (partial sequence ABC) are added, the mixture is stirred at 0° for l6 hours and is evaporated to dryness. The residue is washed at 0° with dilute acetic acid and subsequently with v/ater, filtration is effected, the residue is dissolved in methanol/water (9:1)* the solution is filtered over silica gel, the residue is evaporated, washed with ethyl acetate, dissolved in 200 cc of 80 % acetic acid, the solution is allowed to stand at 25° for 4 hours, is evaporated to dryness at 20°, the residue is washed with ether, ethyl acetate and ether, filtration and drying are effected. Partial sequence ABCD, having a .P. of 240° (decomp.), [ ]β = -38° in acetic acid, is obtained.

P^ tial_sequence_Dl : Trt-Gly-Lys(BOC)-Leu-Ser-Gln-Gln-Leu-NHNH a) Z-Gln-Leu-0 e • 40.1 g of Z-Gln-ONP and 19.1 g of H-Leu-OMe . HC1 are dissolved in 300 cc of dimethyl formamide, 3^.1 cc of N-methyl morpholine are added and the solution is allowed to stand at 20° for 15 hours with occasional shaking. The reaction solution is subsequently completely concentrated by evaporation in a high vacuum. The residue is mixed well with water and filtered off. After crystallization from methanol Z-Gln-Leu-OMe, having a M.P. of 168° , [a]D = -11° in dimethyl formamide, is obtained. - 29 - 100-5077 b) H-Gln-Gln-Leu-CMe 27.8 g of Z-Gln-Leu-OMe are dissolved in 1J0 co of glacial acetic acid and 220 cc of 40 % hydrogen bromide in glacial acetic acid are added. The entire material is allowed to stand for 1 hour while shaking occasionally. After concentrating completely by evaporation in a vacuum, the residue is pulverized in anhydrous ether. After filtration the strongly hygroscopic H-Gln-Leu-OMe . HBr is dried in ether-moist condition at room temperature in a drying chamber. 77- 5 S of Z-Gln-ONP and QlA g of H-Gln-Leu-Oi e . HBr are dissolved in 750 cc of dimethyl formamide and 66.5 cc of N-methyl morpholine are added. The reaction mixture is allowed to stand at room temperature for 15 hours and is subsequently completely concentrated by evaporation in a vacuum. The resultin residue is mixed well with water and filtered off from the water. The suction filter residue is treated with acetone and separated from the acetone phase by means of a press. After drying in a vacuum Z-Gln-Gln-Leu-OMe, having a M.P. of 2380 , [ ]D = -16° in dimethyl formamide, is obtained. 54.8 g of Z-Gln-Gln-Leu-OMe are dissolved in 1200 cc of dimethyl formamide. A suspension of 10 g of 10 palladium on active charcoal in 70 cc of water is subsequently added to this solution. After hydrogenating at room temperature and normal pressure for about 1 hour the quantitative amount of hydrogen is used up. The catalyst is filtered off and the filtrate completely concentrated by evaporation in a vacuum. The residue is recrystallized from methanol/ether.

H-Gln-Gln-Leu-OMe, having a M.P. of 151°, = -10° in dimethyl formamide, is obtained. c) Z-Lys(B0C)-Leu-Ser-0Me See partial sequence D, c). - 30 - 100-3077 d) Trt-Gly-Lys(BOC) -Leu-Ser-NHNH^ See partial sequence D, d) . e) Trt-Gly-Lys(BOC)-Leu-Ser-Gln-Gln-Leu-NHNH^ Trt-Gly-Lys(BOC) -Leu-Ser-Gln-Gln-Leu-NHNH^* having a M.P. of 270° , [a]D = -12° in dimethyl formamide/water (8:2), is obtained by a process analogous to that described for partial sequence D, e), whereby H-Gln-Glu(OTB)-Leu-OMe is replaced by the same amount of H~Gln-Gln-Leu-0Me. -.^lii^i^^Sy.:?^,^?1 : H-Gly-Lys(BQC)-Leu-Ser-Gln-Gln-Leu-?Iis- Lys(BOC)-Leu-Gln-Thr-'ryr-Pro-Arg-Thr-Asn-Thr- Gly-Ser-Gly-Thr-Pro-NH^ . 3 AcOH . H„0 Partial sequence ABCD1, having a M.P. of 2^5° (decomp.), [o;Jp = - 0° in acetic acid, is obtained by a process analogous to that described for the production of partial sequence ABCD, whereby partial sequence D is replaced by the same amount of partial sequence Dl.

Partial_sequence_ ABCID : H-Gly-Lys(B0C)-Leu-Ser-Gln-Glu(OTB)-Leu-His- Lys(BOC) -Tyr-Gln-Thr-Tyr-Pro-Arg-Thr-Asn-Thr- Partial sequence ABCID, having a M.P. of 230° (decomp.), [ajp = -40° in acetic acid, is obtained by a process analogous to that described for the production of partial sequence ABCD, whereby partial sequence ABC is replaced by the same amount of partial sequence ABCl. - 51 - 100-3D77 ^Σ^ϊ^ -.3^Ώ^.^1Ώ1 : H-Gly-Lys(BOC)-Leu-Ser-Gln-Gln-Leu-His- Lys(BOC)-Tyr-Gln-Ttir-Tyr-Pro-Arg-Thr-Asn- Thr-Gly-Ser-Gly-Thr-Pro-NH . 3 AcOH . 3 H^Q Partial sequence ABCIDI, having a M.P. of 255° (decomp.), [ajp = -41° in acetic acid, is obtained by a process analogous to that described for the production of partial sequence ABCD, whereby partial sequence ABC is replaced by the same amount of partial sequence ABC1, and partial sequence D is replaced by the same amount of partial sequence Dl.

: H-Thr-Cys(Bzl)-Val-Leu-OH a) H-Cys(Bzl)-Val-Leu-OMe . HBr 21.0 g of Z-Cys(Bzl)-OCP and 12.3 S of H-Val-Leu-OMe . HC1 are dissolved in 120 cc of dimethyl formamide. 5·9 cc of triethyl amine are subsequently added, the reaction mixture is allowed to stand - 32 - 100-3077 at 25° for l6 hours, ethyl acetate is added, washing with dilute hydrochloric acid, drying over sodium sulphate and evaporation to dryness are effected, and the residue is crystallized from ethyl acetate/diethyl ether. Z-Cys(Bzl)-Val-Leu-OMe, having a M.P. of l60°, 20 [ ]^ = -28° in dimethyl formamide, is obtained and is dissolved in 210 cc of a 40 % solution of hydrogen bromide in glacial acetic acid.

The solution is allowed to stand at 25° for 1 hour, is evaporated to dryness and the residue is recrystallized from isopropanol/diethyl ether. H-Cys(Bzl)-Val-Leu-OMe . HBr, having a M.P. of 168°, [a] = +14° in dimethyl formamide, is obtained.

H-Thr-Cys(Bzl)-Val-Leu-OMe . 1.3 HBr are dissolved in 350 cc of dimethyl formamide, the solution is cooled to -20°, 100 cc of a solution of 2 N hydrochloric acid in dioxane are added and subsequently 10 cc of tert.butyl nitrite are added. The reaction mixture is allowed to stand at -20° for 10 minutes, whereupon 5 cc of triethyl amine and 5. g of H-Cys(Bzl)-Val-Leu-0Me . HBr are added and the resultin mixture is shaken at 0° for l6 hours. The reaction mixture is evaporated to dryness, the residue is dissolved in a mixture of ethyl acetate/ water, the organic phase is washed with dilute hydrochloric acid, dried over sodium sulphate, concentrated by evaporation, and the residue is recrystallized from ethyl acetate. Z--Thr-Cys(Bzl)-Val-Leu-OKe, having a M.P. of 208°, [a] = -27° in dimethyl formamide, is. obtained. g of the .tetrapeptide obtained above are dissolved in 200 cc of a mixture of trifluoroacetic acid/ethyl acetate (1:1), a stream of gaseous hydrogen bromide is passed through the solution at 0° for 1 hour, the solution is subsequently concentrated by evaporation and the residue is recrystallized from methanol/diethyl ether. - 33 - 100-3077 H-Thr-Cys(Bzl)-Val-Leu-OMe . 1.3 HBr, having a M.P. of 202°, [a] = -10° in dimethyl formamide, is obtained. c) H-Thr-Cys(Bzl)-Val-Leu-0H 372 g of H-Thr-Cys(Bzl)-Val-Leu-OMe . 1.3 HBr are dis-solved in l800 cc of methanol, 900 cc of a 2 N caustic soda solution are added, the mixture is allowed to stand at 25° for 1 hour, 240 cc of glacial acetic acid are added and the mixture is allov/ed to stand at 0° for 2 hours. The precipitated crystalline mass is filtered off, is washed first with 1 N acetic acid and subsequently with water, and drying is effected at 50° in a high vacuum.

H-Thr-Cys(Bzl)-Val-Leu-0H, having a M.P. of 219°, [ajp = -53° in 1 N ammonia, is obtained. a) H-Asn-Leu-Ser-OMe . HC1 3 g of H-Leu-Ser-OMe . HC1 and 53 g of BOC-Asn-ONP are dissolved in 400 cc of dimethyl formamide, 22 cc of triethyl amine are added, the solution is allowed to stand at 25° for 16 hours, is evaporated to dryness and the residue is recrystallized from methanol.

BOC-Asn-Leu-Ser-OMe, having a M.P. of 190°, [a] = -24° in dimethyl formamide, in obtained and is dissolved in 500 cc of a 4 N solution of hydrochloric acid in methanol. The solution is allowed to stand at ° for 1 hour, is evaporated to dryness, the residue is dissolved in methanol and precipitation is effected with diethyl ether.

H-Asn-Leu-Ser-OMe . HC1, having a M.P. of l80°, [α3 = "23° in dimethyl formamide, is obtained. - 5 - 100-3077 b) H-Ser-Asn-Leu-Ser-OMe . HC1 39.5 g of BOC-Ser-NHNH^ are dissolved in 500 cc of dimethyl formamide, the solution is cooled to -20°, 200 cc of a 2 N solution of hydrochloric acid in dioxane are added and subsequently 20 cc of tert.butyl nitrite are added. After allowing the reaction mixture to stand at -20° for 10 minutes 90 cc of triethyl amine and 38.0 g of H-Asn-Leu-Ser-OMe . HC1 are added, the mixture is stirred at 0° for 16 hours, is evaporated to dryness, and the residue is recrystallized from chloroform/diethyl ether. BOC-Ser-Asn-Leu-Ser-OMe, having a M.P. of 135°, [a]D = -22° in dimethyl formamide, is obtained and is dissolved in 420 cc of a 4 N solution of hydrochloric acid in methanol.

The solution is allied to stand at 25° for 1 hour, is evaporated to dryness, and the residue is recrystallized from methanol/ethyl acetate.

H-Ser-Asn-Leu-Ser-OMe . HC1, having a M.P. of 155° (decomp.), = ~^-5° i-n dimethyl formamide, is obtained. c) BOC-Cys(Bzl)-Ser-Asn-Leu-Ser-NHNH I8.5 g of H-Ser-Asn-Leu-Ser-OMe . HC1 and l8 g of BOC-Cys(Bzl )-0NP are dissolved in 100 cc of dimethyl formamide, 10 cc of water, 3·5 cc of acetic acid and 5·6 cc of triethyl amine are added, the reaction mixture is allowed to stand at 25° for l6 hours, is evaporated to dryness and the residue is recrystallized from methanol. BOC-Cys(Bzl)-Ser-Asn-Leu-Ser-OMe, having a M.P. of 182°, 20 = -17° in dimethyl formamide, is obtained and is dissolved in 200 cc of dimethyl formamide with slight heating. 200 cc of methanol and 20 cc of hydrazine hydrate are added, the mixture is allowed to stand at 50° for l6 hours, precipitation is effected with diethyl ether, the precipitate is washed with diethyl ether/methariol (1:1) and the resulting BOC-Cys(Bzl)-Ser-Asn-Leu-Ser-NHNH , having a M.P. of 224°, - 35 - 100-5077 --^ i--^-.^----^-;-?? : BOC-Cys-Ser-Asn-Leu-Ser-Thr-Cys-Val-Leu-OH 18.4 g of BOC-Cys(Bzl)-Ser-Asn-Leu-Ser-NHNH , (partial sequence F) are dissolved in 150 cc of dimethyl formamide, the solution is cooled to -20°, and 0 cc of a 2 N solution of hydrochloric acid in dioxane and 15 cc of tert.butyl nitrite are added. After allowing the reaction mixture to stand at -20° for 10 minutes, 28 cc of triethyl amine and 16.2 g of H-Thr-Cys(Bzl)-Val-Leu-OH (partial sequence E) are added and the mixture is stirred at 25° for 16 hours. Filtration is subsequently effected, the solution is concentrated by evaporation and the residue washed thoroughly with 1 N acetic acid.

BOC-Cys(Bzl)-Ser-Asn-Leu-Ser-Thr-Cys(Bzl)-Val-Leu-OH having a M.P. of 217°, [a]D = -17° in dimethyl formamide, is obtained.

The resulting product is dissolved in 5000 cc of dried ammonia, sodium metal is added with stirring and while the ammonia boils, until a dark blue colouration is obtained. Ammonium chloride is added for purposes of decolouration. The solution is evaporated to dryness and the residue is washed with 1 N acetic acid and acetone.

After drying BOC-Cys-Ser-Asn-Leu-Ser-Thr-Cys-Val-Leu-OH, having a M.P. of 248° (decomp.), (α]β = - l° in dimethyl formamide/water (3:1), is obtained.

The resulting nonapeptide is dissolved in 000 cc of 0.01 N ammonia, 1 N hydrogen peroxide is added with stirring, until a negative nitroprussiate reaction is obtained, 200 cc of glacial acetic acid are added, filtration and lyophilization are effected.

I 1 BOC-Cys-Ser-Asn-Leu-Ser-Thr-Cys-Val-Leu-OH, .having a M.P. of 238° (decomp.), [a] = -18° in dimethyl formamide/water (3:l)» is obtained. - 36 - 100-3077 ?^ii i_-53H£i£5_-??i: Piv-Cys-Ser-Asn-Leu-Ser-Thr-Cys-Val-Leu-OH BOC-Cys-Ser-Asn-Leu-Ser-Thr-Cys-Val-Leu-OH (partial sequence EF) is dissolved in 300 cc of trifluoroacetic acid, the solution is allowed to stand at 25° for 30 minutes, is evaporated, the residue is washed with ethyl acetate and dissolved in 300 cc of dimethyl formamide. g of pivalic acid jD-nitrophenyl ester, produced as described hereinafter, as well as Ik cc of triethyl amine are added to this solution, the mixture is allowed to stand at 25° for 16 hours, is evaporated, and the residue is washed with chloroform. The residue is dissolved in 300 cc of dimethyl formamide, 50 cc of water and 50 g of Dowex-50 are added, stirring is effected for 15 minutes, filtration is effected, the resin is washed with dimethyl formamide and the filtrate is evaporated to dryness. The residue is washed with chloroform and subsequently with ethyl acetate and drying is effected.

Piv-Cys-Ser-Asn-Leu-Ser-Thr-Cys-Val-Leu-OH, having a M.P. of 243°, [dijp = -17° in dimethyl formamide/water (3 : 1 ) , is obtained.

Pivalic acid p-nitrophenyl ester 95 g of pivalic acid and 131 g of p-nitrophenol are dissolved in 1000 cc of ethyl acetate, 197 g of dicyclohexyl carbodiimide are added, the mixture is stirred at 25° for 2 hours, filtration is effected, the filtrate is concentrated by evaporation and the residue is recrystallized from a mixture of ethyl acetate/petroleum ether.

Pivalic acid p-nitrophenyl ester, having a M.P. of 104° , is obtained. t » ]_-?---i:i---i_--!lu£B5?_l?§: EOC-Cys-Ser-Asn-Leu-Ser-Thr-Cys-Val-Leu-OH Partial sequence EF2 is obtained by a process analogous to that described for the production of partial sequence EF1, except that partial sequence EF and carbonic acid £-nitrophenyl ethyl ester, - 37 - 100-3077 produced as described hereinafter, are used as starting materials.

M.P. 263° , [ct]D = -21° in dimethyl formamide/water ( 3:1).

Carbonic acid p-nitrophenyl ethyl ester 200 g of jD-nitrophenol and 113 cc of pyridine are dissolved in 1200 cc of ethyl acetate, the solution is cooled to 0° , 156 cc of chloroformic acid ethyl ester are added with stirring, the mixture is stirred at 0° for a further 30 minutes, washing with water, drying over a^SO^ and evaporation to dryness are effected. The residue is dissolved in diethyl ether and carbonic acid £-nitrophenyl ethyl ester is crystallized by the addition of petroleum ether.

M.P. 65° .

PsLTtial_sequerice_F3 : Bzl-MCP-Ser-Asn-Leu-Ser-NHNH£ a) H-Asn-Leu-Ser-OMe . HC1 See partial sequence F, a) . b) H-Ser-Asn-Leu-Ser-OMe . HC1 See partial sequence F, b) . c) Bzl-MCP-Ser-Asn-Leu-Ser-NHNH Bzl-MCP-Ser-Asn-Leu-Ser-NH H2, having a M.P. of 251°, [a]D = -16° in dimethyl formamide, is obtained by a process analogous to that described for the production of partial sequence F, c), except that 18.5 g of H-Ser-Asn-Leu-Ser-OMe . HC1 and 13.5 g of Bzl-MCP-CNP are used as starting material. t t MCP-Ser-Asn-Leu-Ser-Thr-Cys-Val-Leu-NH H 16.2 g of Bzl-MCP-Ser-Asn-Leu-Ser-NHNH (partial sequence F3) are dissolved in 150 cc of dimethyl formamide, the solution is cooled to -20°, 40 cc of a 2 N solution of hydrochloric acid in - 38 - ICO- 3077 dioxane and 15 cc of tert.butyl nitrite are added. The solution is allowed to stand at -20° for 10 minutes, whereupon 28 cc of triethyl amine and 24.8 g of H-Thr-Cys(Bzl)-Val-Leu-OMe . 1.3 HBr . [partial sequence E, b)] are added and the mixture is stirred at 25° for l6 hours. Filtration is subsequently effected, the solution is concentrated by evaporation and the residue washed thoroughly with hot methanol. Bzl-MCP-Ser-Asn-Leu-Ser-Thr-Cys(Bzl)-Val-Leu-OMe, having a M.P. of 2W5 , [ ] = -22° in dimethyl formamide, is obtained and is dissolved at 60° in 100 cc of dimethyl formamide. 10 cc of hydrazine hydrate are added and the solution is allowed to stand at 60° for two hours. The resulting nonapeptide hydrazide crystallizes. Washing with diethyl ether, water and methanol and drying over phosphorus pentoxide in a high vacuum are effected. The nonapeptide hydrazide, having a M.P. of 2650 , = "55° in dimethyl formamide/water (3 : 1) * is obtained.

The resulting product is dissolved in 5000 cc of dried ammonia, sodium metal is added with stirring and while the ammonia boils, until a dark blue colouration is obtained. Ammonium chloride is added for purposes of decolouration and subsequently a stream of air is passed through the solution until a negative nitroprussiate reaction is obtained. The solution is evaporated to dryness and the residue is washed with water and acetone. After drying MCP-Ser-Asn-Leu-Ser-Thr-Cys-Val-Leu-NHNH , having a M.P. of 278° , 20 [a]., = -19° in dimethyl formamide/water (3 :1 ) * is obtained. - 39 - 100-3077 ' I 1 EXAMPLE 1; BOC-Cys-Ser-Asn-Leu-Ser-Thr-Cys-Val-Leu-Gly-Lys(BOC)-Leu- Ser-Gln-Glu(OTB)-Leu-His-Lys(BOC)-Leu-Gln-Thr-Tyr-Pro-Arg- Thr-Asn-Thr-Gly-Ser-Gly-Thr-Pro-NH^ . 2 CH,COOH . 5 RLO * 2 ; j 2— 1.0 g of nonapeptide (partial sequence EF) is dissolved in 10 cc of dimethyl formamide, 1.5 g of N-hydroxysuccinimide and 0.52 g of dicyclohexyl carbodiimide are added, the mixture is stirred at 2 ° for 6 hours, filtration is effected and the filtrate is evaporated to dryness. The residue is washed with ethyl acetate and diethyl ether and drying is effected. BOC-Cys-Ser-Asn-Leu-Ser-Thr-Cys-Val-Leu-OSu, having a M.P. of 242°, is obtained and is dissolved in 10 cc of dimethyl formamide. 3.1 g of tricosapeptide diacetate (partial sequence ABCD) and 1.2 g of N-hydroxysuccinimide are added to this solution and the mixture is stirred at 25° for l6 hours. The reaction mixture is evaporated to dryness, the residue is washed with diethyl ether., chloroform and acetone. The crude, protected dotriacontapeptide is obtained and is dissolved in 50 cc of a mixture of chloroform/ methanol/water (70:30:5). The resulting solution is arranged in layers on a silica gel column (5 x 100 cm), which has been balanced with the mixture indicated above. Elution is effected with an increasing concentration of methanol. The combined fractions which contain the pure peptide are evaporated, subsequently washed with diethyl ether and dried over potassium hydroxide in a high vacuum.

The title compound, having a M.P. of 230° (decomp.), [ j^ = -4 ° in 50 % acetic acid, is obtained. - 0 100-3077.

EXAMPLE 2 ; H-Cys-Ser-Asn-Leu-Ser-Thr-Cys-Val-Leu-Gly-Lys-Leu-Ser-Gln- Glu-Leu-His-Lys-Leu-Gln-Thr-Tyr-Pro-Arg-Thr-Asn-Thr-Gly- Ser-Gly-Thr-Pro-NH^, . hexaacetate ♦ decahydrate 3 · 3 S of the protected dotrlacontapeptide . diacetate . trlhydrate (Example 1) are dissolved under an atmosphere of nitrogen in 100 cc of trifluoroacetlc acid, the solution is allowed to stand at 20° for 15 minutes and is evaporated to dryness. The residue is dissolved in 300 cc of 0.2 N acetic acid, the solution is treated with 20 cc of amberlite-IRA-^lO (acetate), lyophilization is effected, the residue is washed with diethyl ether and dried over potassium hydroxide in a high vacuum. The title compound, having a M.P. of 220° (decomp.), [α]β = -46° in 50 % acetic acid, is obtained.

The following compounds are produced by a process analogous to that described in Example 1 or 2 : EXAMPLE 3 : Piv-Cys-Ser-Asn-Leu-Ser-Thr-Cys-Val-Leu-Gly-Lys(BQC)-Leu- Ser-Gln-Glu(OTB)-Leu-His-Lys(B0C)-Leu-Gln-Thr-Tyr.-Pro-Arg~ Thr-Asn-Thr-Gly-Ser-Gly-Thr-Pro-NH . diacetate . trihydrate Starting materials :,' artial sequences EF1 and ABCD.

Process in accordance with Example 1.

M.P. 220° (decomp.), [a]_ = - 5° in 0 % acetic acid.

EXAMPLE : Piv-Cys-Ser-Asn-Leu-Ser-Thr-Cys-Val-Leu-Gly-Lys-Leu-Ser- Gln-Glu-Leu-?Iis-Lys-Leu-Gln-Thr-Tyr-Pro-Arg-Thr-Asn-Thr- Gly-Ser-Gly-Thr-Pro-NH ♦ hexaacetate ♦ octahydrate Starting material: dotriacontapeptide of Example 3 · Process in accordance with Example 2.

M.P. 216° (decomp.), [a] = -49° in 50 % acetic acid. - 41 - 100-3077 EXAMPLE 5 : E0C-Cys-Ser-Asn-Leu-Ser-Thr-Cys-Val-Leu-Gly-Lys(BOC)-Leu- Ser-Gln-Glu(OTB)-Leu-His-Lys(B0C)-Leu-Gln-Thr-Tyr-Pro-Arg- Thr-Asn-Thr-Gly-Ser-Gly-Thr-Pro-NH^ . dlacetate . trihydrate Starting materials: partial sequences EF2 and ABCD.

Process in accordance with Example 1.

P.F. 230° (decomp.), [aJD = -45° in 50 acetic acid.

EXAMPLE 6 : EOC-Cys-Ser-Asn-Leu-Ser-Thr-Cys-Val-Leu-Gly-Lys-Leu-Ser- Gln-Glu-Leu-His-Lys-Leu-Gln-Thr-Tyr-Pro-Arg-Thr-Asn-Thr- Gly-Ser-Gly-Thr-Pro-NH^ . hexaacetate . octahydrate.

Starting material: dotriacontapeptide of Example 5 · Process in accordance with Example 2.

M.P. 240° (decomp.), [a]D = -52° in 50 % acetic acid.

EXAMPLE 7: MCP-Ser-Asn-Leu-Ser-Thr-Cys-Val-Leu-Gly-Lys(BQC)-Leu-Ser- Gln-Glu(OTB)-Leu-His-Lys(BOG)-Leu-Gln-Tlu'-Tyr-Pro-Arg-Thr- Asn-Thr-Gly-Ser-Gly-Thr-Pro-NH^ . diacetate . trihydrate 1.05 E of octapeptide-hydrazide (partial sequence EF3) are dissolved in a mixture of 50 cc of dimethyl formamide and 1.2 cc of dioxane/2N HC1 at -20°. 0.116 cc of tert.butyl nitrite are added, stirring is effected for 10 minutes at -20°, 1.4 cc of triethylamine and 3·1 g of tricosapeptide diacetate (partial sequence ABCD) and cc of water are added and stirring is effected for 16 hours at 0° .

The reaction mixture is evaporated to dryness and the residue is washed with diethyl ether, chloroform and acetone. The resulting crude peptide is dissolved in 100 cc of 0.3 N acetic acid, is treated with 20 cc of amberlite-IRA-410 (acetate) and 0 cc of 0.6 N ammonium hydroxide are - 42 IOO-5 77 added. The pH value is adjusted to 6.5 and the resulting solution is placed in a carboxymethyl cellulose column ( 10 x 100 cm) which has been treated with a 0.15 N ammonium acetate buffer solution. The elution is effected with an increasing concentration and pH gradient (0.15 N to 0.4 N; pH 6.5 to pH 7.0) . The combined fractions which contain the pure peptide are lyophilized thrice, the residue is washed with ethanol and subsequently with diethyl ether and is dried over potassium hydroxide in a high vacuum. The title compound, having a .P. of 230° (decomp.), [a]_. = -45° in 50 % acetic acid, is obtained.

EXAMPLE 8: MCP-Ser-Asn-Leu-Ser-Thr-Cys-Val-Leu-Gly-Lys-Leu-Ser-Gln- Glu-Leu-His-Lys-Leu-Gln-Thr-Tyr-Pro-Arg-Thr-Asn-Thr-Gly- Ser-Gly-Thr-Pro-NH ♦ hexaacetate . octahydrate Starting material: dotriacontapeptide of Example 7· Process in accordance with Example 2.

M.P. 220° (decomp.), [ ]β = -60° in 0 % acetic acid.

EXAMPLE 9 : BQC-Cys-Ser-Asn-Leu-Ser-Thr-Cys-Val-Leu-Gly-Lys(BCC)-Leu- Ser-Gln-Gln-Leu-His-Lys(BOC)-Leu-Gln-Thr-Tyr-Pro-ArK-Thi'- Asn-Thr-Gly-Ser-Gly-Thr-Pro-NH2 . 2 CH^COOH . 3 ^O Starting materials: partial sequences EF and ABCD1.

Process in accordance with Example 1.

M.P. 235° (decomp.), [a] = -46° in 50 % acetic acid.

EXAMPLE 10 : H-Cys-Ser-Asn-Leu-Ser-Thr-Cys-Val-Leu-Gly-Lys-Leu-Ser-Gln- Gln-Leu-His-Lys-Leu-Gln-Thr-Tyr-Pro-Arg-Thr-Asn-Thr-Gly- Starting material: dotriacontapeptide of Example 9· Process in accordance with Example 2. ?0 M.P. 2300 (decomp.), [<*]* = -48° in 0 % acetic acid. 43 - 100-3077 EXAMPLE 11; B0C-Cys-Ser-Asn-Leu-Ser-Tir-Cys-Val-Leu-Gly-Lys(BOC)-Leu- Ser-Gln-Glu(0TB) -Leu-His-Lys(BOC)-Tyr-Gln-Thr-Tyr-Pro-Arg Thr-Asn-Thr-Gly-Ser-Gly-Thr-Pro-NH^ . 2 CK,C00H . 3 H 3 c. — Starting materials: partial sequences EF and ABC1D.

Process in accordance with Example' 1.

M.P. 230° (decomp.), [α] = - 5° in 50 % acetic acid.

EXAMPLE 12: H-Cys-Ser-Asn-Leu-Ser-Thr-Cys-Val-Leu-Gly-Lys-Leu-Ser-Gln- Glu-Leu-His-Lys-Tyr-Gln-Thr-Tyr-Pro-Arg-Thr-Asn-Thr-Gly- Ser-Gly-Thr-Pro-NH^ . hexaacetate . decahydrate Starting material: dotriacontapeptide of Example 11.

Process in accordance with Example 2.

M.P. 216° (decomp.), [a].. = - 3° in 50 % acetic acid.

EXAMPLE 13 : BOC-Cys-Ser-Asn-Leu-Ser-Thr-Cys-Val-Leu-Gly-Lys(BOC) -Leu- Ser-Gln-Gln-Leu-His-Lys(BOC)-Tyr-Gln-Thr-Tyr-Pro-Ars-Thr- Asn-Thr-Gly-Ser-Gly-Thr-Pro- Hg . diacetate . trihydrate Starting materials: partial sequences EF and ABC1D1.

Process in accordance with Example 1.

M.P. 235° (decomp.), [a] = -43° in IN acetic acid/methanol (2 :1 ) .

EXAMPLE 14: H-Cys-Ser-Asn-Leu-Ser-Thr-Cys-Val-Leu-Gly-Lys-Leu-Ser-Gln- Gln-Leu-His-Lys-Tyr-Gln-Thr-Tyr-Pro-Arg-Thr-Asn-Thr-Gly- Ser-Gly-Thr-Pro- H^ . hexaacetate . decahydrate Starting material: dotriacontapeptide of Example 13 · Process in accordance with Example 2.

M.P. 216° (decomp. ) [ ]D = -43° in IN acetic acid.

Claims (3)

1. CLAIMS 1. A process for the production of a compound of general formula I, S-CHg-CHX-CO-Ger-Asn-Leu-Ser-I'hr-C s-Val-Leu- - G-ly-Lys-Leu-Ser-Gln-Olu-Leu-tlis-Lya-Leu-ttln- I Thr-Tyr-Pro-Arg-Tlr-Aan-Thr-Gly-Ser^aiy-Thr-Y wherein X is -H, -B¾2, R-C0-NH-, 1 wherein R is alkyl of 1-4 carbon atoms, phenyl or benzyl, both the latter optionally substituted with halogen, lower alkoxy or nitro, or R'-O-CO-NH-, wherein R* is not capable of being split off with trifluoroacetic acid, and is alkyl of 1-4 carbon atoms, alkenyl of 2-4 carbon atoms, benzyl, p-nitrobenzyl, p-chlorobenzyl , p-bromobenzyl, p-phenyl-azobenzyl or p-(p'-methoxyphenyl)-azobenzyl, Y is L-prolinamide or L-prolin-L-valinamide, the terminal peptide uni S-CHg-CHX-CO- having the L-forra when X is -NH2, R-CO-NH- or R'-O-CO-NH-, and wherein an L-seryl radical may be exchanged for an L-threonyl or L-alanyl radical, an L-asparaginyl radical fo an L-aspartyl, L-glutaminyl or L-glutamyl radical, an L-threonyl radical for an L-seryl or L-alanyl radical, an L-lysyl radical for a BOC-rlyeyl, L-omithyl or L-arginyl radical, an L-glutaminyl radical for an L-glutamyl, L-asparaginyl or L-aspartyl radical, a L-glutamyl radical for an OTB-glutamyl, L-glutaminyl, L-asparaginyl or L-aspartyl radical, an L-leucyl radical for an L-tyrosyl radical, an L-tyrosyl radical for an L-phenylalanyl radical and an L-arginyl radical for an L-omithyl or L-lyeyl radical, or a pharmaceutically acceptable acid addition salt or heavy metal complex thereof, comprising joining two peptide units by an amide linkage, the peptide units being such that when joined; the amino acid sequence of the compound of general formula I is achieved, and when desired, forming a pharmaceuticall acceptable acid addition salt or heavy metal complex of the resulting compound .

2. A process accordin to Claim 1, wherein one of the peptide units incorporates the group of formula II, 8-GH2-GHXC0-Ser-Asn-leu-Ser-ihr-C s- II therein X has the significance stated in Claim 1, wherein an 3-3 bridge exists between the terminal peptide unit and the L-cysteinyl radical, and wherein an L-seryl radical may be exchanged for an L-threonyl or L-alanyl radical, an L-asparaginyl radical for an L-aspartyl, I-glutaminyl or L-glutamyl radical, an L-leucyl radical for a.n L-tyroayl radical, and an L-threonyl radical for an L-seryl or L-alanyl radical. _ 46 - 100-3077 4. A process according to Claim 5, wherein in the production of the peptide unit incorporating the group of formula II, the S-S bridge is formed during the last stage. 5· A process for the production of a compound of general formula I, stated in Claim 1, or a pharmaceutically acceptable acid addition salt or heavy metal complex thereof, substantially as described herein with reference to any one of Examples 1 to l4. 6. A compound of general formula I, stated in Claim 1, or a pharmaceutically acceptable acid addition salt or heavy metal complex thereof, produced by a process according to any one of the preceding Claims. 7. A compound of formula I, stated in Claim 1, or a pharmaceutically acceptable acid addition salt or heavy metal complex thereof. 8. BOC-Cys-Ser-Asn-Leu-Ser-Thr-Cys-Val-Leu-Gly-Lys(BOC)-Leu-Ser-Gln-Glu(OTB)-Leu-His-Lys(BOC)-Leu-Gln-Thr-¾rr-Pro-Arg-Thr-Asn-Thr-Gly-Ser-Gly-Thr-Pro-NH .. t r 9. H-Cys-Ser-Asn-Leu-Ser-Thr-Cys-Val-Leu-Gly-Lys~Leu- Ser-Gln-Glu-Leu-His-Lys-Leu-Gln-Thr-Tyr-Pro-Arg-Thr-Asn-Thr-Gly-Ser-Gly-Thr-Pro-NH2 . . . . 10. Piv-Cys-Ser-Asn-Leu-Ser-Thr-Cys-Val-Leu-Gly-Lys(BOC). Leu-Ser-Gln-Glu(CTB) -Leu-His-Lys (BOC) -Leu-Gln-Thr-Tyr-Pro-Arg-Thr-Asn-Thr-Gly-Ser-Gly-Thr-Pro-NH . - 7 - ΪΟΟ-3077 I t 11. Piv-Cys-Ser-Asn-Leu-Ser-Thr-Cys-Val-Leu-Gly-Lys-Leu Ser-Gln-Glu-Leu-His-Lys-Leu-Gln-Thr-Tyr-Pro-Arg-Thr-Asn-Thr-Gly-Ser-Gly-Thr-Pro-NH . 12. EOC-Cys-Ser-Asn-Leu-Ser-Thr-Cys-Val-Leu-Gly-Lys(BOC)-Leu-Ser-Gln-Glu(OTB)-Leu-His-Lys(BOC)-Leu-Gln-Thr-Tyr-Pro-Arg-Thr-Asn-Thr-Gly-Ser-Gly-Thr-Pro-NH . : 15 j EOC-Cys-Ser-Asn-Leu-Ser-Thr-Cys-Val-Leu-Gly-Lys-Leu-Ser-Gln-Glu-Leu-His-Lys-Leu-Gln-Thr-Tyr-Pro-Arg-Thr-Asn-Thr-Gly-Ser-Gly-Thr-Pro-NH . l . ■ MCP-Ser-Asn-Leu-Ser-Thr-Cys-Val-Leu-Gly-Lys(BOC)-Leu-Ser-Gln-Glu(OTB) -Leu-His-Lys(BOC)-Leu-Gln-Thr-Tyr-Pro-Arg-Thr-Asn-Thr-Gly-Ser-Gly-Thr-Pro-NH . t I 15. MCP-Ser-Asn-Leu-Ser-Thr-Cys-Val-Leu-Gly-Lys-Leu-Ser-Gln-Glu-Leu-His-Lys-Leu-Gln-Tl;ir-Tyr-Pro-Arg-Thr-Asn-Thr-Gly-Ser- Gly-T r-Pro-NH. 16. BOC-Cys-Ser-Asn-Leu-Ser-Thr-Cys-Val-Leu-Gly-Lys(BOC). Leu-Ser-Gln-Gln-Leu-His-Lys(B(X)-Leu-Gln-Thr-Tyr-Pro-Arg-Thr--Asn-Thr-Gly-Ser-Gly-Thr-Pro-NH . . .. ■ 17. H-Cys-Ser-Asn-Leu-Ser-Thr-Cys-Val-Leu-Gly-Lys-Leu-Ser-Gln-Gln-Leu-His-Lys-Leu-Gln-Thr-Tyr-Pro-Arg-Thr-Asn-Thr-Gly- Ser-Gly-Thr-Pro-NH . - - 100-3077 18.' BOC-Cys-Ser-Asn-Leu-Ser-Thr-Cys-Val-Leu-Gly-Lys(BOC)-Leu-Ser-Gln-Glu(CTB)-Leu-His-Lys(BOC)-Tyr-Gln-Thr-Tyr-Pro-Arg-Thr- Asn-Thr-Gly-Ser-Gly-Thr-Pro-NH . 19. H-Cys-Ser-Asn-Leu-Ser-Thr-Cys-Val-Leu-Gly-Lys-Leu Ser-Gln-Glu-Leu-His-Lys-Tyr-Gln-Thr-Tyr-Pro-Arg-Thr-Asn-Thr-Gly-Ser-Gly-Thr-Pro-NH . 20. BOC-Cys-Ser-Asn-Leu-Ser-Thr-Cys-Val-Leu-Gly-Lys(BOC)-Leu-Ser-Gln-Gln-Leu-His-Lys(BCC)-l r-Gln-Thr-Tyr-Pro-Arg-Thr-Asn-Thr-Gly-Ser-Gly-Thr-Pro-NH . t t 21. H-Cys-Ser-Asn-Leu-Ser-Thr-Cys-Val-Leu-Gly-Lys-Leu-Ser- Gln-Gln-Leu-His-Lys-Tyr-Gln-Thr-Tyr-Pro-Arg-Thr-Asn-Thr-Gly-Ser-Gly-Thr-Pro-NH2 . 22. A pharmaceutical composition comprising a' compound of general formula I, stated in Claim 1, or a pharmaceutically acceptable acid addition salt or heavy metal complex thereof as active agent, in association with a pharmaceutically acceptable diluent or carrier. 2

3. A pharmaceutical composition according to Claim 22, in unit dosage form comprising between 20 and 700 MRC units of the active agent. 37OO/RO/ D/HR