BE675694A - - Google Patents

Description

  

   <Desc / Clms Page number 1>
 



  New peptides having the effect of ACTH and process for their preparation
The present invention relates to peptides in which the arginyl residues at position 17 and 18 of peptides having the effect of ACTH, for example peptides with the sequence of the first 19 to 28 amino acids of the ACTH molecule , are replaced by remainders. ornithine, as well as the acid addition salts, the derivative and the complexed, in particular the sine complexes, of the indicated peptides, and to a process for their preparation.



   To the peptides indicated having the effect of ACTH, also belong those in which the different amino acids of the natural sequence, in particular those of positions 1 to 9, counting from the amino end, are exchanged for other acids. natural amines, for example the first amino acid, namely serine, against glycine, or the second amino acid, to

 <Desc / Clms Page number 2>

 
 EMI2.1
 know the 8ine. against phenylalanine, or the fourth amino acid, namely methionine, against norvaline, leucine or a-amin0but7riq.ua acid, or the fifth amino acid, namely glutamic acid, against gluta- aine, as well as the sequences in which the first
 EMI2.2
 amino acid, namely serine, is lacking.



   It was found that the indicated peptides, in which the two arginina residues in position 17 and 18 are replaced by ornithine residues, also have a very strong adrenocorticotropic effect and in addition a strong lipolytic effect, but have the advantage,
 EMI2.3
 vla-A # vis peptides containing arginine in position 17, 18, to be. of easier synthesis and
 EMI2.4
 âtra obtained with a better yield.

   from this point of view, 'lys instead of highlighting particularly the tetrecosapeptide which is L-a6r, rl-L-tyroayl-L-a6rylL-8éth1onTl-L-glutamyl-L-histidyl-L-phenylalanyl-t-arg1nylL -tryptophyl-glycyl-L-lyayl-L-prolyl-L-valyl-glycyl-t-1Y8ylsyl-L-ornith11orn1th11-L-prolyl-t-valyl-t-1YBylL-val71-L-tyrosyl-L-proline, as well that the corresponding compound which, instead of the glutamyl residue, presents the remainder of the glutamine, as well as the acid addition salts, derivatives and complexes of these tetracosapeptides.



   As acid addition salts, mention should be made in particular of the salts of acids which can be used therapeutically, such as hydrochloric acid, acetic acid, as well as hardly soluble salts such as sulphates, phosphates or sulphonates.



   By derivatives, it is meant above all esters, for example lower alkyl esters

 <Desc / Clms Page number 3>

 such as methyl, ethyl, propyl, tert-butyl esters, or benzyl esters which are unsubstituted or substituted, for example, by the NO 2 group, by halogen atoms, by lower alkyl groups or by lower alkoxy groups, as well as amides and hydrazides.



   By complexes, it is necessary to understand the compounds of complex type whose structure is not yet clarified, which form when certain inorganic or organic substances are added to peptides with adrenocorticotropic offet, and especially those giving them a prolonged effect. Such inorganic surfaces are compounds derived from metals such as calcium, magnesium, aluminum, oobalt and in particular zinc, above all hardly soluble salts such as phosphates and pyrophosphates, as well as the hydroxides of these metals.



  Organic substances causing prolongation of the effect are, for example, non-antigenic gelatins, for example hydroxypolygelatin, polyvinyl-pyrrolidone and oarboxymethyl-cellulose, as well as sulfonic or phosphoric esters of alginic acid, dextran, polyphenols and polyalcohols, especially polyphloretin phosphate and phytinic acid, as well as polymerization products and copolymerization products of amino acids which have a predominant fraction of α-amino acids like glutamic acid or aspartic acid.



   The new compounds have an adreno effect. corticotropic and lipolytic high and can correspondingly be used in human and veterinary medicine, for example instead of the natural hormone.

 <Desc / Clms Page number 4>

 



   The new peptides are prepared according to the methods known for the preparation of peptides having a long chain length. In this case, the amino acids are linked in the order indicated, individually or after prior formation of rather small peptide units. In particular, the so-called solid support substance synthesis will also be emphasized (cf. Merrifield, "J. Am. Chem. Soc.", 85, 2.149 [1963]), during which the peptide is built up from the carboxylated end. ester-linked to a polymer, the amino acids being in order reported pure condensation.



   The condensation is carried out either using a condensing agent, or by activating the [alpha] -amino-group or the terminal carboxyl group. As the condensation methods, there may be mentioned, for example, the carbodiimide method, the azide method, the activated ester method and the anhydride method.

   For example, one of the amino acid or peptide molecules in the form of an ester can be linked with another amino acid or peptide molecule containing a protected aminogenic group, in the presence of a condensing agent such as phosphorous acid carbodiimide or halide, or it is possible to react with the amino acid or with the peptide ester having a free amino group an amino acid or a peptide having an activated carboxyl group (and a group protected aminogen), eg, acid halide, azide, anhydride, imidazolide, isoxazolide (eg from N-ethyl-5-phenyl-isoxazolium 3'-sulfonate, see Woodward et al., " J. Am. Chem.



    Soc. "89, 1.001 [1961]), or an activated ester such as the ester

 <Desc / Clms Page number 5>

   oyanomethyl, carboxymethyl thiol ester or nitrophenyl ester. Conversely, it is also possible to react an amino acid or a peptide comprising a free carboxyl group (and a protected amino group) with an amino acid or with a peptide comprising an activated amino group (and a protected oarboxyl group), for example on a. phosphitamide.



   The free functional groups not participating in the reaction are advantageously protected, in particular by residues which can be easily split by hydrolysis or by reduction, the carboxyl group preferably by esterification, for example with methanol,
 EMI5.1
 tertio-bx.anol, benzyl alcohol, p-nitrobenzyl alcohol, the aminogenic group, for example by introducing tosyl, trityl, formyl, trifluroacetyl, phtalyl or carbobenzoxy groups, or colored protective groups such as p-phenylazo-benzyloxy- group
 EMI5.2
 carboayl or the p- (p¯methoxyphenylezo) -benzyloxycarbonyl group, or in particular the tert-butyloxycarbonyl residue.

   To protect the aminogenic group in the guanidino group of arginine, the NO2 group is suitable; However, the indicated aminogenic group of arginine does not have to be protected during the reaction. The lminogenic group of histidine can also be protected by the benzyl residue or the trityl residue,
The transformation of a protected aminogenic or iminogenic group into a free group, as well as the transformation during the process of a functionally modified carboxyl group into a free carboxyl group, takes place according to methods known per se, by treatment with of

 <Desc / Clms Page number 6>

 
 EMI6.1
 hydrolysis agents and reducing agOntOs.



  In order to prepare the tetracosapaptide mentioned above, it is possible, for example, to oondenaer the decapeptide of formula L-seryl-L-tyrosyl-L-s'ryl-'th1oU11wL-glt & m11.



  (or Lglutaminyl) h.-histidyl..L.phenylalanyl-h..arginyl- L-tryptophyl-glycine, or an activated ester, for example the p-nitrophenyl ester of this deaapeptide, in which the a-aminogenic group of the seryl residue and optionally the -carboxyl group of the glutamyl residue are protected, with
 EMI6.2
 an ester of the tetradecapeptide of the formula L-lysyl-L-prolyl L-valyl-glycyl-L-lysyl-L-lyayl-L-ornithyl-L-ornithylL prolyl-L-valyl-L-lysyl-L-valyl-L- tyrosyl-L-proline, in which the amino groups of the side chain of the lysine residues and of the ornithine residues are protected, then the protecting groups of the tetracosapeptide derivative are removed.

   As protecting groups for the α-aminogenic group of serine and for the aminogenic groups of the side chains of the residues of lysine and ornithine, the tert-butyloxycarbonyl group, the -carboxyl group of glutamic acid and the terminal carboxyl group of proline are advantageously protected by the tert-butyl ester group. All of these protecting groups can be cleaved simultaneously by acid hydrolysis, for example with trifluoroacetic acid. The indicated decapeptide can, for example, be prepared according to the methods described in French patent N 1.310.513 of May 31, 1961, in the first certificate of addition N 85.657 of February 20, 1964 and in the second certificate of addition N 85.759 of February 20, 1964.

   Tetradecapeptide is

 <Desc / Clms Page number 7>

 advantageously prepared according to scheme N 1 illustrated below in which:
 EMI7.1
 30C denotes the tert-butyloxy-carbonyl residue Z "" "carbobanzoxy
 EMI7.2
 tohu "" "tert-butyl NP" "" nitrophenyl Tos "" "tosyl.

 <Desc / Clms Page number 8>

 
 EMI8.1
 



  Null Scheme
 EMI8.2
 BOC BOC BOC BOC BOC BOC 7 .... Lys Pro Val Gly Lys Lys-f-NH-NH3 Zj-Ornj-QNP Z- [àON2 L ra ONP H7EOtBu BOC Z o Val Lys Val Tyr Prc) 4-OtBu BOC H Pro Val Ls Val Tyr Pro OtBu BOC BOC Z-On Pro Val Lys Val Tyr Pro -} - OtBu c BOC BOC H - (- 0n Pro Val Lys Val Tyr Fro4-OtBu BOC BOC BOC Z - {- 0n or'n Pro Val Lys Val Tyr Pro OtBu BOC BOC BOC H - Orn Orn pro Val Lys Val Tyr Fro OtBu BOC BOC BOC BOC BOC Z4 Lys Pro Val Gly Lys Lys Om pin Pro V.al 1 L s Val Tyr Py OtBu BOC BOC BOC BOC BOC BOC H Lys Pro Val Gly Js Lys Orn on Pro Val L Val 2'yr Pro OtBu

 <Desc / Clms Page number 9>

 
Instead of the tert-butyl ester of proline, it is also possible in the above process to use the amide of proline.

   If another lower alkyl ester or a benzyl ester is used instead of the tert-butyl ester of proline, one obtains
 EMI9.1
 then the lower alkyl esters or the corresponding benzyl esters of the tetracosapeptide. The methyl ester can in a manner known per se be converted into the hydrazide with the aid of hydrazine hydrate.



   For example, the hydrazide of the tetracosapeptide can also be obtained by condensing the indicated decapeptide, in which the α-aminogenic group of serine
 EMI9.2
 is protected by the tert-butyloxycarbonyl group and dan [which optionally the -carboxyl group of glutamic acid is protected by the tert-butyl ester group:

  .
 EMI9.3
 with 11 - p.phenyl3zobenzyl aster of the indicated tetradecapeptide in which the amino groups of the side chains of the lysine residues and the ornithine residues are protected by the phtalyl residue, by cleaving the protective amino groups and the tert-butyl ester group with an acid , for example with trifluoroacetic acid, then by converting the ester p-nitrophenylazobenzy-
 EMI9.4
 lique of tetracosapeptide, to hydrazide using hydrazine monc-acetate. The mentioned tetradecapeptide derivative is obtained, for example,

   by condensing the ester
 EMI9.5
 p-phenylazobenzyl Ne-phthalyl-L-lysyl-Ne-phthalylL-lysyl-Nb-phthalyl-L-ornithyl-Nb-phthalyl-L-crnithylL-prolyl-L-valyl-N e -phthalyl-L-lysyl- L-valyl-L-tyrosylL-proline ± -Prepared in a manner analogous to the decapeptide derivative indicated in French patent N * 1.343.58?

 <Desc / Clms Page number 10>

 
 EMI10.1
 of September 11, 1962, by replacing BOO-Are (N02) -QS by 30C-Orn (ht) -OH with BOCLvalyl-glyoiaa in the presence of iacbutyl chlorocarbonate, by splitting the BCG group with trifluoroacetic acid , condensing the dodecapeptide ester with BOC - phthalyl-L-11811L-proline in the presence of doisebutyl chlorocarbonate and cleaving the BOC group of the tetradecapeptide with trifluoroacetic acid.



   According to another method for the preparation of the tetracosapeptide, the tetrapeptide can be condensed.
 EMI10.2
 what is L-3eryI-L-tyrosyl-L¯seryl-l-raethioninef in which the α-aminogen group is protected, for example by the tertio-but110xyc.rbonyl group, with the dicosapoptide that e3t la -glutamy1 - (or glutaatnyl) - L hiatidyl L-r'ény.alanyl-L-arginyl-7r-typtoph-, J; -glycyl-L-lysyl ..



  L-prolyl-L-va: ylglycyl-L-lysyl - lY8yl-L-ornithyl- 1-ornithyl-L-polyl-L-valyl-L-1Y9y: -L-T81yl-L-tyrosyl- L-proline, in which the aminogenic groups of the side chains of the restea lysine and of the ornithine zest are protected, for example by the tert-butyloxycarbo- group
 EMI10.3
 nyl, and in which the 1-cbonyl group of glutamic acid is optionally esterified, for example with the tartio-butyl ester group, preferably according to the azide method.



   The invention also relates to the embodiments of the process in which one starts from a compound obtained as an intermediate product at any stage of the process and carries out the phases still missing from said process, or interrupts the latter at any one of its stages, as well as the intermediate products that are then obtained.

 <Desc / Clms Page number 11>

 



   Depending on the procedure chosen, the new compounds are obtained in the form of bases or of their salts. Bases can be obtained from the salts in a manner known per se. From the latter, it is possible, by reaction with acids suitable for the formation of
 EMI11.1
 therapeutically usable salts, obtaining salts such as, for example, those with inorganic acids such as halogenated hydraciies, for example hydrochloric acid or hydrobromic acid, perchloric acid, nitric acid with thiocyanic acid, sulfuric acids
 EMI11.2
 or phosphoric, or organic acids such as formic, Í.J '; ic, prop1oniq, \. o glycolic, lactic, pyruvic, oxalic, malonic, succinic, maleic, fumaric, q: ic, tartaric, citric, ascorbic, hydroxy - :; que, dihydroxy-ma:

  iqueJ benzoic, phenylacetic, #amino-benzoque, 4-hydroxy-benzoic, anthranilic, 1.: '- "'.: lami.que, mandelic, 3alicylic, 4-aminosalicylic, 2-phenoxy.benzorque 2-acetoxy-benzolque, methane-auifonic, ethanesulfonic, droxyethane aulfoniqua, benzene-sulfonic, p-toluenesulfonic, naphthalenesulfonic or sulfanilic.



   The peptides obtained according to the process can be used in the form of pharmaceutical preparations. These contain the peptides in admixture with a pharmaceutical, organic or inorganic carrier material suitable for enteral or parenteral application. For the formation of this carrier material, substances which are not reactive with polypeptides, such as, for example, elatin, lactose, glucose, sodium chloride, starch, magnesium stearate, talc, oils, are contemplated. vegetable, alcohols

 <Desc / Clms Page number 12>

 
 EMI12.1
 benzyls, gums, polyalkylene glycol, petrolatum, cholesterin or other known excipients.



  The pharmaceutical preparations can be presented, for example, in the form of lyophilizate, or in liquid form in the form of solutions, suspensions or emulsions. Where appropriate, are they sterilized and / or contain auxiliary substances such as preservatives, stabilizers, contaminants? or emulsifiers. They may also contain still other therapeutically valuable substances.
 EMI12.2
 



  This as well as a cheerful one brings them to combine with the adjuvants, uauals in the therapy with ACTH, to prolong the effect obtained, for example with hydroxypolygelatine, phosphate of polyphlo-
 EMI12.3
 retina, c8rboxymethyl-celJloae, polyglutamic acid or the sparingly soluble metal compounds which have been mentioned above, in particular the phosphates, pyrophosphates or hydroxides of zinc.



   The invention also relates, as new industrial products, to the compounds obtained by carrying out the process defined above.



   The invention is described in more detail in the non-limiting examples which follow, in which the temperatures are indicated in degrees centigrade.
 EMI12.4
 



  Thin layer chromatographic systems are designated as follows;

 <Desc / Clms Page number 13>

 System 43A: mixture (100; 40: 10) of part amyl alcohol, iaopropanol and water.



  System 430: mixture (100: 40: 55) of cer- tiary amyl alcohol, isopropanol and water.



  System 52: mixture (100: 10:30) of normal butanol, glacial acetic acid and water.



  System 1C0: mixture (62: 21s6: 11) of ethyl acetate, pyridina, glacial acetic acid and water.



  System 101; mixture (30: 20: 6: 24) of normal butanol, pyridine, glacial acetic acid and water.

 <Desc / Clms Page number 14>

 
 EMI14.1
 



  BXBCL3 1. Z-prQ-Val-Ly8 (BOC) -Val-Tyr-? Ro-OtBu a) Dana 10 cm3 of di # athylforttaalde, 5.14 g (6.75 m.molea) of the ester are dissolved pentapeptide H-ValLy3 (aOC) ¯Val-Tyr-Pro-OtBu, prepared for example according to the process described in the first certificate of addition
 EMI14.2
 French? 84. 45 of July 30, 1963, 3.01 9 (8.15 m.moles) of p-nitrophenyl ester of bensylox7carbonyl-L-proline is then added. After having allowed to stand for 20 hours at room temperature, the mixture is then added. treat the reaction mixture by diluting it with 100 car of ethyl acetate, then washing the solution once with 50 cm3 of water,
 EMI14.3
 four times with 25 car-me molar solution of potassium carbonate and twice with 25 cm3
 EMI14.4
 of 6 ..

   The ethyl acetate layer is then cooled to 0 "and washed twice with 25 cm3 each time of a 1% aqueous solution of citric acid, then with water until a reaction has taken place. neutral, and dry with sodium sulfate.
 EMI14.5
 



  The protected hexapeptide ester is obtained in powder form by diluting the concentrated solution with dry ethyl ether; the yield is 97%. The product is unitary by thin layer chromatography on silica gel in the following solvent systems; Methanol: Rf = 0.97 Mixture of chloroform and methanol (95: 5): Rf = 0.26 Mixture of benzene and acetone (1: 1): Rf = 0.47.

 <Desc / Clms Page number 15>

 



   The product is not crystalline; it melts at 156-180 and has a specific rotatory power:
21
 EMI15.1
 , Ccg-.7D -5 (c 1.09 in methanol); ('at73 -46.4 (c 1.01 in dimethylformamide),) In 15 cm' of dimetay: tormamida, 7t6l s (10 a. moles) of free pentapeptide ester H = Jal- :: m are dissolved , 30C) -Jaβ-Tfr-'ro-vtu 3t adds 1.4 cc of tri4t: ln8. The -salange 1 -10 is cooled and 4.73 S,. - n.mo'¯as) ester :: td ¯ o: cy-phtg? isl3i: ue of carbc't '.' n V / loxy-L "proline.! # reaction mixture 3rd color -0n in dark red.> st processed when tdut is <1:;." 3 ':, ::: 3. The ester dissolution and copulation last only 15 minutes.

   The treatment takes place by diluting with ethyl acetate, extracting with 60 cm of water to remove the dimethylformamide, extracting four times with 25 cm3 each time of a 5% bicarbonate solution, extracting with taken up with a 1% citric acid solution and washed until neutral. The solution was then dried with sodium sulfate, filtered and, while stirring vigorously, poured into 500 cm3 of petroleum ether. The suspension obtained is further diluted with half a liter of petroleum ethereal ot filter. After drying, 9.83 g (99%) of a product are obtained which turns out to be pure on chromatography and which is identical to the product obtained in Example 1.
 EMI15.2
 



  2. H-P-1al-tys (B9C) -Val-Tr-Pro-OtBu. a) Free hexapeptide ester
In 150 cm3 of methanol, 6.5 g (6.55 m.moles) of protected hexapeptide ester Z-Pro-Val- are dissolved.
 EMI15.3
 I <ya (30C'-Val-Tyr-ro-Ot3u and hydrogen for 2 hours

 <Desc / Clms Page number 16>

 and a half in the presence of 1.3 g of a carbon containing 10% palladium.



   The methanolic solution is then filtered and the filtrate is concentrated in vacuo.



   A sample is left in the open air and at room temperature, which causes oriatals to form. The latter are used to initiate the crystallization of the crude product. After 20 hours in a cooler, the aigu-Iles (1.41 3) are separated by filtration, washed with a mixture of methanol and ether and finally with ether solicitation. From the mother liquor, a further 3.5 grams of the peptide ester are obtained with the ether. In total, 4.66 g (83%) of a product melting at 174-176 are thus isolated. and exhibiting a specific rotational power [[alpha]] D20 - 91 + 0.5 (c. 1.87 in methanol).



   The peptide ester is soluble in methanol, acetone, dimethylformamide, sparingly soluble in ethyl acetate, ether and petroleum ether. b) Mono-acetate:
In a mixture (4: 1) of glacial acetic acid and water, in the presence of 1.3 g of a palladium carbon, 2.5 g (2.52 m.moles) of the hexapeptide ester protected above. The absorption of hydrogen * is completed after 2 and a half hours. The catalyst is filtered off, the filtrate is concentrated in vacuo at a bath temperature of 40 and the evaporation residue is dried under high vacuum over sodium hydroxide.



  2.34 g (100%) of a colorless reain are obtained which is found to be unitary in a thin layer chromatogram. ;

 <Desc / Clms Page number 17>

 
 EMI17.1
 The compound shows the values of Hf su1va.3 (development with Reindel-Hoppe reagent) on silica gel plates:
 EMI17.2
 
<tb>
<tb> <SEP> system made up of <SEP> by <SEP> a <SEP> mixture
<tb> 8: 2) <SEP> of <SEP> chloroform <SEP> and <SEP> of <SEP> methanol <SEP>; <SEP> Rf <SEP> = <SEP> 0.61
<tb> System <SEP> 43A <SEP> Rf <SEP> = <SEP> 0.50.
<tb>
 
 EMI17.3
 3. N-benzvloxycarbonyl-N -tertio-butyloxycarbonyl-L-orni''hne.



  26 (112 m.moles) of N 6 -tertio-butyloxycarbonyl-L-ornithi (prepared according to the process described in French patent No. 3. ^. Als No. 1,297,720 of August 22, 1960) are carbobenzoxylated in a usual manner. To acidify the alkaline reaction 80lu., Extracted with ether, a pH of 3 is adjusted with citric acid to 0, extracted to no.
 EMI17.4
 calf:. '3t: r, then washed with water and dried, After --r 6limia the 3clvant by distillation, we obtain 3? e8 g (92%) of a colorless uil,
One gram of this oil is dissolved in 3 on
 EMI17.5
 ether 3 is criafcalliated with 600 mg of dioyclohexy. amine.

   The ether obtained is washed; the yield .-. of 1.28 3 (35%) with a melting point of 13'j and a specific optical rotation al7 D3 ... j 2 (c. 2.4 i in absolute methanol).



   Most of the product crystallizes afterwards. dissolving in 100 µl of hot di-isopropyl ether and cooling, white crystals are obtained which melt. After recrystallization from 100 cm3 of di-iso- ether
 EMI17.6
 hot prapylic acid with the addition of 20 cm3 of e! ::,; - '¯e acetate, the melting point is 101. The special rotary power
21 cific is [[alpha]] D = -3.2 (c -2.45 in absolute methanol).

 <Desc / Clms Page number 18>

 
 EMI18.1
 



  4. Ester p xxlt; rophenyliqua of -ben.loX7car¯o -tert1o-butYloxYcarbon, yl-t-crnith1.



  14r6 g (40 .ol.8) of -Z-N6¯BOC- Orn-OH and 6.72 g (48 m.moles) of p-nitrophenol are dissolved in 100 cm3 of ethyl acetate, cooled to 5 and add to
 EMI18.2
 mixture 8.24 g (40 m.molea) of dicjoloB8X11-carbodi1m14. in 50 cm3 of ethyl acetate. After standing for 2 hours at 0 and for 20 hours at room temperature, filtered, the filtrate evaporated to seo, the residue dissolved in 75 because of ethanol and cooled to 0. When crystallization is complete, filtered and washing with di-iaopro- ether
 EMI18.3
 pylic. 1314 g of pure analytical water are obtained, which melts 104 and exhibits specific optical rotation Z-ct-727 * - -26t? <1 (c. 5.6 iana methanol).



  From the mother liquor, we still obtain, after concentration. Another 2.5 grams (19%) of this ester.
 EMI18.4
 



  Aster is soluble in dimethylformamide, acetone, methylene chloride, ether, ethyl acetate, ethanol and acetonisriia, almost insoluble in di-leopropyl ether and di-leopropyl ether. oil.



  5. AYdrox-phthalimidic ester of ', 4 -benzYlocarbon: l N -tertio-butloxYcarbOnyl-t-ornlth1De.



  In 5 cm of dry pyridine, 3.66 g (10 m.moles) of Nn-Z-Nb-BOC-L-Orn-OH are dissolved, cooled to -10 and a solution of 2.06 g (10 m .moles) of dicyclohexyl-carbodiimide in 5 cm3 of pyridine. At the end of
 EMI18.5
 5 minutes, 1.96 g of N-hydroxy-phtaliside are added and the mixture is left to stand for 16 hours at 0 °. Then filtered and then washed with pyridine. The filtrate is freed from pyridine at 40 * under vacuum * added

 <Desc / Clms Page number 19>

 secondary butanol and evaporates again to dryness, then takes up again in a little ether. After 48 hours at -8, the crystals obtained are isolated (4.5 g, ie 88%); they melt at 125-127.



   The crude crystallizate is dissolved by heating in acetonitrile, freed by filtration of the dicyclohexylurea which remains and of the hydroxy-phthalimide, then the filtrate is brought to crystallize by addition of diisopropyl ether. 3.12 g (51%) of pure product are then obtained. After recrystallization from iaopropyl alcohol, the * sien point is 129 * and the specific optical rotation is 72 ,? -25.7 (c = 1 in methanol).



  6. Z-Orn (BOC) -Pro-Val-Lys (BOC) -Val-Tyr-Pro-OtBu. a) In 5 cm3 of dimethylformamide, 2.3 g (2.5 m.moles) of H-Pro-Val-Lys (BOC) -ValTyr-Pro-Otbu acetate are dissolved and 1.71 g (3 , 5 m.moles) of Z-Om (BOC) - ONP. After 24 hours, while stirring vigorously, the reaction mixture is diluted with 200 cm3 of ether.



  During 15 minutes, a white precipitate forms. It is filtered, washed with ether and dried. According to thin layer chromatography, the heptapeptide derivative obtained is unitary in the following solvent systems
 EMI19.1
 
<tb>
<tb> Mixture <SEP> (1: <SEP> 1) <SEP> of <SEP> benzene
<tb> ot <SEP> of acetone <SEP>: <SEP> Rf <SEP> = <SEP> 0.37
<tb> Mixture <SEP> (9: 1) <SEP> of <SEP> chloroform <SEP>: <SEP> Rf <SEP> = <SEP> 0.47
<tb> and <SEP> of <SEP> methanol
<tb> System <SEP> 43A <SEP>: <SEP> Rf <SEP> - <SEP> 0.77
<tb> System <SEP> 52 <SEP>: <SEP> Rf <SEP> = <SEP> 0.92.
<tb>
 



   After drying under vacuum for 48 hours, 2.73 g (90%) of a white powder are obtained which melts at 142-145 and exhibits specific optical rotation.

 <Desc / Clms Page number 20>

 
 EMI20.1
 7J5 - 0 -38 0.5 (c. 2.061 in dimethyltormamide); [[alpha]] D20 = -90 + 1 (c. 0.970 in methanol).



   The product is soluble in dimethylformamide, acetic acid, alcohols and acetone. It dissolves
 EMI20.2
 more difficult in acetonitrile and ast insoluble in ethyl acetate, ether, benzene and petroleum ether.
 EMI20.3
 b) To 429 mg (0.5 m.mola) of H., P: O-V33-Ly9C30C) Val-Tr-Pro-OtBu in solution in 2 cl of di.nthyltormaide, 305 mg (0.60 a.mola) of -benzyloxycarbonyl-N .tertio-butyloX1carbo hydroxy-phttlimidique asser. nyl-L-omithine. A total of 0.14 cm 3 of triethylamine is added, which causes the reaction mixture to turn red, and then treat after a quarter of an hour. While stirring constantly, about 50 cm3 of ether is added to the solution and filtered after half an hour.

   We
 EMI20.4
 obtains in this oas 536 mg (89%) of the protected hoptapeptide ester which turns out to be pure on analysis and is identical to the product obtained under a).
 EMI20.5
 7. H-Orn14Cβ-Pro-Val-eBOC-Val-Tyr-Pro-OtBu. a) Acetate:
In 25 cm3 of 80% acetic acid, one gram (0.83 m.mole) of the protected heptapeptide ester which is described under b) is dissolved, then hydrogen as indicated under 2. When the hydrogenation is complete, the catalyst is separated by filtration, the extract is well extracted by washing and the filtrate is evaporated to dryness.

   There remains 956 g (100%) of a pure material which, according to a thin layer chromatogram, is indistinguishable from the free base (see below) and has specific optical rotation:

 <Desc / Clms Page number 21>

 
 EMI21.1
 a / p0 -874 Z 1 (c. 1.028 in methanol) #. -27 + I (e 1.095 in acetic acid at 809i. B) Free heptapeptide ester:
In methanol, one gram (0.83 m.mole) of the protected heptapeptide ester and hydrogen is dissolved as described under 2.

   The product is not obtained in the crystalline state, may3 no longer contain any starting material and is unitary in a thin layer chromatogram (on silica gel plates); it presents the following values of Rf:
 EMI21.2
 
<tb>
<tb> System <SEP> 52 <SEP>; <SEP> Rf <SEP>. <SEP> = <SEP> 0.68
<tb> System <SEP> 43A <SEP>: <SEP> Rf <SEP> = <SEP> 0.22
<tb> System <SEP> made up <SEP> by <SEP> a
<tb> mixture <SEP> (90: <SEP> 10) <SEP> of <SEP> chloroform
<tb> and <SEP> da <SEP> methanol <SEP>: <SEP> Rf <SEP> = <SEP> 0.13
<tb> System <SEP> made up <SEP> by <SEP> a
<tb> mix <SEP> (1:

   <SEP> 1) <SEP> of <SEP> benzene <SEP> and
<tb> acetone <SEP> Rf <SEP> = <SEP> 0.00.
<tb>
 
 EMI21.3
 From total apragil hydrolysis and from the analysis of amino acids according to Stein and Moore, we find a proportion between amino acids:
 EMI21.4
 Pro: Val: Tyr: Orn: Lys. 204: 2OOt098t1.06t108.



  8. Z-Crn (BOC) -Orn (30C) -Pro-Val-Lys (BOC) -Val-Tyr-Pro-OtBu. In 5 cm3 of dimethylformamide, dissolve
 EMI21.5
 2.69 g (2.38 m.moles) of the acetate of the he ptapeptidic ester obtained according to 7. a) and add 1.4 g of Z-Orn (BOC) -
 EMI21.6
 WE P. The mixture is allowed to stand overnight at room temperature and then concentrated under high vacuum. A brown resin remains which, on addition of 100 cm3 of hot acetonitrile, gives a white precipitate. After 3 hours at 0, this precipitate is separated by fil-
 EMI21.7
 tration, then wash the powder obtained with acetonitr11

 <Desc / Clms Page number 22>

 
 EMI22.1
 and with ether. After s 4 hostage, the octapeptid ester obtained weighs 3.15 g (93%); it melts at 152-154 and pceseate a specific optical rotation 'Ct¯7' -8.3 ± Oo5 (c - 1.046 in methanol).

   The product is unitary in a chromator9.mme in a thin layer (3ur of silica gel) and has the following R ... values. System consisting of a mixture (95: 5) of chloroform and methanol: Rf. 0.54 System consisting of a mixture (1: 1) of benzene and acetone: Rf = 0.56.



   The octapeptide ester dissolves in the
 EMI22.2
 acetic acid, methanol and ethanol dimethylformami3at dissolves less well in dGtonitri18 and is insoluble in ethyl acetate on the other.



  9.i-Orn (BOC) -Or-.i (130C) -Pro-Val-Lys (BOC '-' 7al-Tyr-Pro-OtBue a) Acetate:
The protected octapeptide ester according to 7. is dissolved in 80% acetic acid and hydrogenated as indicated under 2. After having separated the catalyst by filtration, it is evaporated at a bath temperature of at most 30. The product obtained is unitary in a thin-layer chromatogram and has the following Rf values: System consisting of a mixture (95: 5) of chloroform and methanol; Rf = 0.06 System consisting of a mixture (9: 1) of chloroform and methanol: Rf = 0.22 System consisting of a mixture (85:15) of chloroform and methanol: Rf = 0.26 System consisting of a mix
 EMI22.3
 (1: 1) benzene and acetone; rif 0.0.03.



  -5. -49.4 1 (c - 0.942 in dimethylfonaamid) j --76. -85o6o 1 (c - 1.266 in methanol). ---

 <Desc / Clms Page number 23>

 
 EMI23.1
 10. -Lys- (BCC) -Pro-Val-Gly-I.ys (3CC) -Lya (BOC) -Orn (BOC) Orn HOC -ro-Val-Ls (90C -Va '-T; rr o- OtBu.



  In 9 cm of pure dimethylformamide, 900 mg (0,: C Tt <-aole) of heptapeptide hydrazida Z-Lya- (HOC) -? R. = .'-'... ly..Ly9 ( BOC) -Lys (30C) -V'd-Ni2 prepared, for example, according to the process described in the 1st certificate of addition: -runçal3 NI 84.745 of 3 May 1962, then cooled the solution -10. After addition of 1 # 6 cm3 of aida ahlorhydriq ': binormâl and 0.13 cm * of a solution of pfdnanOr!: A' ,: -: li trita of sodium, the mixture is stirred for 10 ainuted 3 -.¯ .. t then adds 1. '. 4. :: out 900 mg (0.67 mol) x: :: "4 ':. B1; ate ie l oc .¯ -.ep ;, ide which is described under 9. 1 :. solution in 1-wu cm, of dilnethyformamide.



  We add i-c-.i1; at all 0.53 cn3 (. ', 0 z.mole3) of triethylamine a: iā: .- # - # stand for 18 hours in a cooler.



  The solution is then evaporated to dryness under a -3cuva4- vacuum: the residue is axed with acetonitrile.



  The acetonitrile insoluble -, 1--1 - ecapeptidi-derivative is filtered 3 times washed with 25 cm 3 of acetonitrile.



  Cr, obtains 1.554 g (98%) of a product of slightly yellowish tint which, in an a layer chromatogram, shows only a trace of an impurity.



  You can neither of the two starting materials.



  The i3 3f values (on silica gel) are as follows i

 <Desc / Clms Page number 24>

 in a system consisting of a mixture (95: 5) of chloroform and methanol: Rf = 0.31 in a system consisting of a mixture
 EMI24.1
 (95: 5) of dioxane and water: R .. 0.76.



   The product is amorphous, but exhibits a distinct melting point at 192-194 and specific optical rotation.
 EMI24.2
 a 75 e -37.8 l (c. 1050 in dimethylformamid. As elemental analysis shows, the peptide derivative is in the hydrate state.
 EMI24.3
 11. Toaylate da H-Lys (30C) -Pro-la.-Gl, y-Lys (3GC; -Lws ", 90C Orn3CC) -OrnHOC Pru-Va 1 - :. st C; -7 3 ¯-: r - '^ oJ you.



  The protected tetradtcapeptide ester (one gram, 0.42 mmol) is suspended in 150 µl of methanol and shaken at 40 until everything has gone into solution. 400 mg of a carbon containing 10% palladium and 0.4 cm3 of glacial acetic acid are then added, then the solution is shaken for 8 hours at 40 in a hydrogen atmosphere and filtered. After evaporation of the solvent, 793 mg (84%) of the ester are obtained.



   To transform it into the tosylate, the product is dissolved in 5 cm3 of pyridine, a solution of 76 mg of p-toluene-sulfonic acid-monohydrate in 2 cm3 of pyridine is added, evaporated to dryness under vacuum and the residue washed several times. taken up with ether, mixing and decanting. After drying, 729 mg of tetradecapeptide ester tosylate are obtained in the form of an amorphous powder which melts at 179-181. It is unitary in a thin layer chromatogram (on gel
 EMI24.4
 silica) and pre 'jnta the following values of al. t System 43C. Rf = 0.49 System 52: Rf = 0.61.

 <Desc / Clms Page number 25>

 
 EMI25.1
 ac> D2. -654 (c 1.980 in pyridine).



  12. BOC8érTyrSér-ltét-Glu (OtBu) -Hïs-Phé-Ar-Try-Gly-'Ly3 (BOG.) - Pro-Val-Gly-Lys (BOC) -Lya (BOe) -Lya (BC ') Tosylate Orn (BOC) -Orn (BOC) -Pro-Val-s (BOC2-Val-Tyr-Prc-OtBu. In 6 cm3 of absolute pyridine, the mixture is stirred.
 EMI25.2
 one hour 720 mg (0.3 a.mole) of decapeptide head ester tosylate with 480 mg (0.33 m.mole) of BCG-Ser-TyrSer-Met-Glu (OtBu) -His-Phe-Arg -Try-Gly-OH, then add 220 ag (1.07 m.mole) of dicyclohoxyl-carbodiimide. The mixture is stirred for 3 days at room temperature under a nitrogen atmosphere. During this time, the solution becomes very viscous. For filtration, another 10 cubic centimeters of pyridine is added.

   In this oaa, 65 mg of dicyclohexylurea are separated. The filtered solution is concentrated under vacuum and the residue is kneaded with about 100 cm 3 of ethyl acetate, the suspension is kept in an icebox overnight and filtered. 1.180 mg of a crude product of yellowish tint is obtained.



   To purify it, 1.060 g of this crude product are placed in 3 distributing elements of a Craig apparatus with each time 10 cm3 of upper phase and 10 cm3 of lower phase, then subjected to a multiplicative distribution in 300 stages in a system. solvent
 EMI25.3
 consisting of a mixture (10: 3; 6t4) of methanol, a buffer solution, chloroform and carbon tetrachloride (the buffer solution consists of a mixture of 28.5 cm3 of glacial acetic acid and 19.25 g of ammonium acetate, made up to one liter with water).

   The
 EMI25.4
 position of the maximum distribution is finally determined; L, with samples of the lower phase, using

 <Desc / Clms Page number 26>

 
 EMI26.1
 Pauly's reaction (#a: t # * 75 K - m Dt3), The protected tetracosapeptide, chromatographically on, is obtained by evaporating to dryness the overall content of the distribution elements 63 to 89 and removing the acetate from ammonium by sublimation at 45 under high vacuum, in the form of a powder
 EMI26.2
 white which melts at 195-200 * when decanting. It shows the following Rf values in a thin layer chromatogram
 EMI26.3
 System + 3c; sillce gel: ir. 0, ô4 System 52; silica gel: 3 0.54 System 100; o- of aluminum; Rf 4 ?.



  13, H-6er-Tyr-SérM hexa-acetto: Jl-H1s-é r.



  Try-Gly Ly3 Pro-Val-Gl7-L78 "; 0rn O> .- n-Pro-yaj," LyaVal-Tyr-? Ro-OH.



  In 10 cm of tri.t'l! .4 -: '. ¯'l' ..! "; Here at 90 96 we dissolve 420 mg of the tetra = sapept4 - c protected and let the clear colorless solution stand p <3 sawing 30 minutes at 0, Finally we concentrate in: -r¯ro a Turkish bath temperature of 40 up to about 2 cm3, add 10 cm3 of water, concentrate again to 5 cm3 and freeze-dry
 EMI26.4
 so. 510 mg of trifluorsc-'tate are obtained which, in order to be converted into acetate, is dissolved in 5 am3 of water and filtered through a column of a weakly basic ion-changer (Merck II) in the form acetate.

   By concentrating the eluate to a volume of about 5 cm3, and then lyophilizing, one obtains, in the form of a colorless powder easily soluble in water, 390 mg of hexa-acetate of the tetracosapeptide, which s 'turns out to be pure
 EMI26.5
 chromatography and electrophoresis-2. In thin layer chromatography on

 <Desc / Clms Page number 27>

 of the aluminum oxide in system 101, it has an Rf value of 0.43. During electrophoresis on
 EMI27.1
 paper, 2CO voila-, pH 5.2, 2 hours), it migrates from 11.5 µm 'r>:. ::' s the cathode (development with ninhydri na v. 1.? reactive the? auly) .



  In the following test Saffran and Schally, and in vivo (;> ## subcutaneous uLl ^ the rat), 13 product shows * 7ort corticourope effect.



     EXAMPLE 2 Two bulks of solution containing
 EMI27.2
 the following c \ ;.) '; 3ats:.) 2 cm3 ampoule ie solution:
 EMI27.3
 Hexa-.ii: - du - :; ét :: '' lCù3.: 1pe :: ': .i., Ie
 EMI27.4
 
<tb>
<tb> (Exempt * <SEP> 1, <SEP> 13) <SEP> 0.25 <SEP> mg
<tb>
 
 EMI27.5
 ZnSC - H2O 9.58 mg Manr-xV ;! 33.0 mg "T, .wf2en -0" 2.0 mg Aged water, Up to 0.5 cm3 2) Bulb of one cubic centimeter of solution!
 EMI27.6
 Na3? Oa r 12H2O 9.5mg
 EMI27.7
 
<tb>
<tb> Distilled <SEP> water, <SEP> up to <SEP> 0.5 <SEP> cm3
<tb>
 
Before use, mix the contents of the two ampoulos. The suspension obtained has a pH value of approximately 7.5.



   EXAMPLE
A dry ampoule is prepared containing the following components i

 <Desc / Clms Page number 28>

 
 EMI28.1
 
<tb>
<tb> Tetracosapeptide <SEP> Hexa-acetate <SEP>
<tb> (Example <SEP> 1, <SEP> 13) <SEP> 0.5 <SEP> mg
<tb> ZnSO <SEP> + <SEP> 7 <SEP> H20 <SEP> 1.23 <SEP> mg
<tb> Na3P04 <SEP> + <SEP> 12 <SEP> H2O <SEP> 1.48 <SEP> mg
<tb> Mannitol <SEP> 25.0 <SEP> mg
<tb> <SEP> sodium chloride <SEP> 3.8 <SEP> mg
<tb>
 
Before use, the contents of the dry bulb are suspended in a cubic centimeter of water. The suspension is isotonic and has a pH value of 7.6.



   EXAMPLE 4
To 1.0 cm3 of an aqueous solution containing 0.001 g of tetracosapeptide hexa-acetate (example 1, 13), 0.02 g of zinc acetate [Zn (CH3COO) 2, 2 H20¯7, 0, 0015 g of sodium chloride, 0.02 g of benzyl alcohol and 0.005 cm3 of normal hydrochloric acid are added at room temperature, with stirring, 1.0 cm3 of a 2.16% aqueous solution of sodium pyrophosphate [Na4P2O7, 10 H2O].



   The suspension of zinc pyrophosphate which arises during this reaction is isotonic and has a pH value of 6.0. The sterilized suspension also has a pH value of 6.0. The suspension contains per cubic centimeter 0.5 mg (50 international units) of tetracosapeptide, 6.94 mg of zinc pyrophosphate, 7.47 mg of sodium acetate, 0.75 mg of sodium chloride and 10.0 mg benzyl alcohol.

 <Desc / Clms Page number 29>

 



   EXAMPLE 5
An ampoule of solution is prepared from the following components;
 EMI29.1
 
<tb>
<tb> Tetracosapeptide <SEP> Hexa-acetate <SEP>
<tb> (Example <SEP> 1, <SEP> 13) <SEP> 0.5 <SEP> mg
<tb> Gelatin <SEP> 280.0 <SEP> mtPhenol <SEP> 5.0 <SEP> mg
<tb> Distilled <SEP> water, <SEP> up to <SEP> 1.0 <SEP> cm3
<tb>
 
EXAMPLE 6
A sterile, filtered aqueous solution of tetracosapetin hexa-acetate (Example 1, 13) is mixed under aseptic conditions with 3-polyphloretin sodium phosphate sodium chloride, then placed in the vials and lyophilized, so that obtains a via, dry containing the following components:

   
 EMI29.2
 
<tb>
<tb> Tetracosapeptide <SEP> hexa-acetate <SEP> <SEP> 0.5 <SEP> mg
<tb> Sodium <SEP> <SEP> phosphate
<tb> polyphlorethylene <SEP> (at <SEP> 86.5 <SEP>%) <SEP> 23.20 <SEP> mg
<tb> <SEP> sodium <SEP> chloride <SEP> 12.28 <SEP> mg
<tb>
 
Before use, mix the contents of the via! dry with 2 cm3 of distilled water enclosed in an ampoule of solution.



   EXAMPLE 7
It is prepared in a suspension from the following components:

 <Desc / Clms Page number 30>

 
 EMI30.1
 Tetracosapeptidd hexa-acetate
 EMI30.2
 
<tb>
<tb> (Example <SEP> 1, <SEP> 131 <SEP> 1.0 <SEP> mg
<tb>
 
 EMI30.3
 zncl2 1085 mg
 EMI30.4
 
<tb>
<tb> Na2HP04 <SEP> 1.7 <SEP> mg
<tb> Benzyl alcohol <SEP> <SEP> 17.0 <SEP> mg
<tb> NaCl <SEP> 2.5 <SEP> mg
<tb> NaOH, <SEP> up to <SEP> a <SEP> pH <SEP> of <SEP> 8.0
<tb> Distilled <SEP> water, <SEP> up to <SEP> 2 <SEP> cm3
<tb>
 
 EMI30.5
 EXAMPLE r.



   In about 5.7 cm 3 of a 10% sodium hydroxide solution, the acid is dissolved.
 EMI30.6
 poly-L-glutawi with an average a # -: ilar weight of about 11,000, so that the: ':: 3 the solution is 7.4. In this solution, oc 9: .3scut 5.0 mg of hexa-aceta, - of tetracosapeptide (example 1, 13) and 0.2 mg of merthiolate, then made up to 10 cm3 with distilled water. The solution is filtered to sterility.

   It contains per cubic centimeter:
 EMI30.7
 Hexacetate iu te4racosapeptil .--, 0.5 mg
 EMI30.8
 
<tb>
<tb> <SEP> poly-L-glutamic acid <SEP> 200.0 <SEP> mg
<tb> Sodium <SEP> <SEP> hydroxide <SEP> solution,
<tb> until <SEP> obtaining <SEP> of a <SEP> pH <SEP> of a
<tb> <SEP> value of <SEP> 7.4
<tb> Merthiolate <SEP> 0.02 <SEP> mg
<tb> Distilled <SEP> water, <SEP> up to <SEP> 1.0 <SEP> cm3
<tb>


 

Claims (1)

Claims. 1, A process for the preparation of novel peptida3 having the effect of ACTH, characterized by the fact EMI31.1 that -> r-? pir <3 of peptides having the effect of ACTH, in lasqu! i 1 j arginine remains in position 17 and 18 are filled because ornithine remains, as well as their aela EMI31.2 of add¯: -1 acids, derivatives and complexes, by binding the amino di: ¯ :: - .cidea or partial peptide sequences', Edified from said acids, according to the metbo .. ¯èuee for the 3ynt3e of the peptides , protected; ¯ - ,,,,,!, Evenly the 3rou; 3 aminogens and the groups ci ''. #>: F: es not participating in condensation reactions ..,; ¯, .t using condensing agents or acar3 .; the α-aminogen group or the carboxyl group terminated 2., The present process can be further characterized by the following points: 1) The various amino acids or partial peptide sequences constructed from said icides are linked, according to the carbodiimide method, the mixed anhydrides method, the activated esters method or the azide method. 2) The α-aminogen group and the 3-aminogen groups are protected by the tert-butyloxycarbonyl group, and the terminal carboxyl group and the groups EMI31.3 carboxyls.3 possibly present in the side chains, by the tert-butyl ester group. 3) Peptides are prepared containing the first 19 to 28 amino acids of the ACTH molecule and in which the arginine residues at position 17 and 18 are <Desc / Clms Page number 32> replaced by ornithine remains, 4) Peptides are prepared in which the fifth amino acid is exchanged for glutamic acid. 5) Tetracosapeptidea are prepared. 6) The tetracoaapeptide of formula is prepared EMI32.1 L-aeryl-L-tyroayl-L-aeryl-L-methionyl-L-glutamyl-L-hiatidylL-phenylananyl-L-arginyl-L-tryptophyl-glycyl-L-lysylLprolylIvalyl-glycyl-L-lys3.-L.lysy : -L-ornithyl L-ornithyl-L-prolyl-Lvnlyl-L-lysyl-L-valyl-L-tyrosyl-L-proline or the corresponding compound which, in place of the glutamyl residue, presents the rest of the glutamine. 7) Complexes of the peptides are prepared with zinc or sparingly soluble salts or hydroxides of zinc. of a 8) In the presence / carbodiimide, the decapeptide of formula L-seryl-L-tyrosyl-L-seryl- L-methionyl-L-glutamyl- (or L-glutaminyl) -L-histidyl- is condensed. EMI32.2 L-phenylalanyl-L-arginyl-L-tryptophyl-glycine, in which the α-aminogen group of the seryl residue and the -oarboxyl group of the glutamyl residue are protected, with an ester of the tetradecapeptide of the formula L-lysyl-L -prolyl-L-valyl-glycylL-lysyl-L-lysyl-L-ornithyl-L-ornithyl-L-prolyl-L-valylL-lysyl-L-valyl-L-tyrosyl-L-proline, in which the amino groups side chains of the lysine residues and of the ornithine residues are protected, then the protecting groups of the tetracosapeptide derivative obtained are eliminated. 9) Protect the α-aminogenic group and EMI32.3 γ-amiaogenic groups by the tert-butyloxycarbonyl group, the terminal carboxyl group and optionally the -carboxyl group of glutamic acid by the ester group <Desc / Clms Page number 33> terio-butyl, 10) The tetrapeptide of formula is condensed EMI33.1 L-6eryl-tyrosyl-L-seryl-L-methione, in which the 2-aminogen group is protected, with the ercosapeptide of the formula L-glutamyl- (or glutaminyl) -L-hi8tidyl-L-phenylany 1-L- arginyl-L-tryptophyl-glycyl-L-lysyl -L-prolyl-L-valy Lglycyl. L-1Y9yI-L-lY8yl-L-ornithyl-L-ornithyl-Lprolyl- ¯valyl-L-lysyl-L-valyl-L-tyrosyl-L-proline, in which the amino groups of the side chains of lysine residues and ornithine residues are protected and in which optionally the -carboxyl group of glutamic acid is esterified, then optionally eliminates the protecting group of the tetracosapeptide obtained, 11) The α-aminogenic group of the tetrapeptide and the α-aminogenic groups of the eicosapeptide are protected by the tert-butyloxycarbonyl group and, if necessary, the -carboxyl group of glutamic acid by the tert-butyl ester group. 12) Condensation is carried out according to the azide method. 13) Complexes of the peptides are prepared with gelatin, polyphloretin phosphate or polyglutamic acid. <Desc / Clms Page number 34> It * - As a new industrial product - peptides in which the arginine residues, located at position 17 and 13, of peptides having the effect of ACTH are replaced by ornithine residues, their acid addition salts, derivatives and complex.