DE2416048A1 - NEW PEPTIDES WITH BIOLOGICAL EFFECT - Google Patents

Description

New peptides with biological effects

The structure of the hypothalamic peptides TRH and LH-RH are distinguished by a high constitutional specificity ,

Surprisingly, it has now been found that this for another hypothalamic factor that has only recently been discovered is the somatostatin (GIF, SRIF) of the formula I.

H-Ala-Gly-Cys-Lys-Asn-Phe-Phe-Trp-Lys-Thr-Phe-Thr-Ser-Cys-OII (l) 1 2 3 4 5 6 7 8 9 10111213H

does not apply in this hatred, so can the amino acids in this peptide 1-25 4, 5 »9 and 14 can be changed without affecting the biological Effect of pre-binding decreases. By suitable substitution be, new peptides with even stronger and most importantly get prolonged effect.

5 098 /. W 11 0.5

The invention thus relates to peptides in general Formula II

X-CH-CO-W-V-Phe-Phe-Trp-tt-Thr-Phe-Thr-Ser-NH-CH-Y

CH-S S- CH ₂ ( ^Ττ

in which X stands for H, ß-Ala-NH, (<£ -Methyl) -AIa-NH, D-AIa-NH, D-AIa-GIy-NH ₅ ß-Ala-Gly-NH, Ala-D-Ala -NH, where 6L -amino groups present can carry an alkanoyl or alkyloxycarbonyl radical with 1-6 carbon atoms in the alkyl moiety, V can be asparagine or alanine, ¥ lysine or ornithine and Y is H, COOH, CONH_, CONHR or CONR where R is represents an alkyl radical with 1-6 carbon atoms or a cycloalkyl radical with 5 or 6 carbon atoms and both substituents R can also jointly represent tetramethylene or pentamethylene.

The invention also relates to preparations for parenteral use or intranasal administration of the peptides of the general formula II.

The invention also relates to a method for production these peptides, which is characterized in that one protected peptide of formula III

. .- X-CH-CO-W- V- Phe-Phe-OH .... ·

CH ₂ -SB

with a -protected peptide of the formula IV ■ ::. H-Trp-W-Thr-Phe-Thr-Ser-NH-CH-Y

-CH-Y k-S-B

where in formula III and IV B stands for benzyl or 4-methoxybenzyl, ^ hydroxyl groups are completely or partially protected by benzyl radicals and amino groups by optionally chlorine-substituted ben?; yloxycarbonyliteste are condensed, the protective groups by treatment with hydrogen fluoride or with sodium in liquid v ^ -a ggi '. existing carboxyl group by benzyl

509844/1105 ' _{/ Ί}

Eliminates ammonia and the resulting bis-raercapto compound the cyclic disulfide of the general formula II by Q..W Oxidation or hydrogenation with air at alkaline pH or with Produces potassium ferricyanide.

Bicyclohexylcarbodiimide is advantageously used as the condensing agent (DCCl) in the presence of an N-hydroxy compound. as such are e.g. N-hydroxysuccinimide (HONSu), 1-hydroxybenzotx-iazole (HOBt) or 3-hydroxy ~ 4-oxo-3, 4-dihydro-1, 2, 3-benzotriazine (HOÖBt) in question, but also other N-hydroxy compounds described in the literature on peptide syntheses as additives to DCCI are suitable. The solvents used are e.g. dimethylformamide, Dimethylacetamide, N-methylpyrrolidone, phosphoric acid, tris-dimethylamide or dimethyl sulfoxide.

One goes ζ, ιΒ. so that you can put the two protected peptides in one of the solvents mentioned dissolves, about 1 to 2 minor, of the N-hydroxy compound and 1.0 to about 1.5 minor. DCCI adds. The reaction temperature is not critical, but it is advantageous to use DCCI in a known manner at a low temperature admit to maintain this temperature for some time and the reaction at room temperature or slightly elevated temperature break up.

In the production of the peptides according to the invention, of course other fragments can also be produced and condensed with one another in a manner known per se. The inventive Process is only one particularly advantageous and economical variant. The known repetitive procedures can also be used be applied, the C-terminal amino acid cysteine or possibly serine is bound to a crosslinked polystyrene carrier like benzyl ester or with the hydroxyl groups of a polyethylene or polypropylene glycol is esterified. After the synthesis has ended, the cleavage from the carrier takes place either together with the cleavage the other protective groups «or by hydrolysis, aminolysis or optionally animonolysis. As removable N-protective groups in the case the cleavage with hydrogen fluoride come tert-alkyloxycarbonyl, especially tert-butyloxycarbonyl protective groups.

5098AA / 110B _{/ h}

The bis-mercapto compounds are oxidized to the cyclic disulfides of formula II in a known manner in the most dilute solution possible, either by air, which is passed through during 2- component doughs with an alkaline reaction, preferably about pH 7-5-9, or at about neutral Reaction by potassium ferricyanide,

Both methods are known from the synthesis of oxytocin. In the present case, too, in addition to the monomers, dimers and polymers are formed, which in a known manner by chromatographic methods, such as those in Biochem, Biophys. Res.Commun.j ^ i (i973) $ page 2jk are to be removed.

The peptides according to the invention inhibit the release of growth hormone from the pituitary gland. To a lesser extent will too the release of other hormones, e.g. glucagon, is inhibited. The peptides according to the invention have approximately the same spectrum of activity as the natural somatostatin.

Since growth hormone and also glucagon are antagonists of insulin, the peptides of general formula II are particularly suitable for the treatment of diabetes in cases in which the blood level of growth hormone is increased. Compared to the natural somatostatin, the compounds have the advantage of a stronger effect and, above all, the longer duration of action. This is reflected, for example, that of the inventive compounds 20 - 6o ° / o of the weight of the natural peptide needed who ^ to obtain the about the same biological effect.

The preparations contain, depending on the type of application about 0.1 to 20 mg of one of the substances according to the invention per dose Links.

Preparations for parenteral administration also contain a buffer, e.g. an ion phosphate buffer, which adjusts the pH between should hold about 3'j 5 and 7, further sodium chloride, mannitol cd-? r Sorbitol to adjust the isotonicity. The preparations can in gofriorget rookne tor or dissolved form vorlifi ^ on. solutions

5098 ^ / 1105 BATH ORIGINAL

contain an antibacterial additive, e.g. 0.2 0.3 ° h ^ -hydroxybenzoic acid ester,

If the already prolonged effect of the compounds according to the invention is to be increased still further, a depot additive can be used. Suitable additives are, for example, up to about 5 percent by weight of polyphloretin phosphate, prepared according to German patent specification 929 66k, Examples 1, 4, and 5> calculated on the final volume of the solution. If necessary, a. Gelatin derivative mixed with hexamethylene diisocyanate, prepared according to DBP 1,118,792 or 1,155,134, Example 1, can also be added up to about 5% by weight, furthermore 0.1-1% zinc in combination with 0.5 are suitable - 5 ° fi protamine (eg as sulfate), the latter preparation as a solution from pH 3 »5 to about 6.5 or as a suspension from about pH 7» 5 to 8.0.

An intranasally acting preparation is obtained, for example, by mixing peptides of the formula II or their salts with physiologically harmless Dissolves acids in an aqueous buffer solution of pH 4 to 7.2 and adds an isotonicity-producing substance. Of the' A polymeric adhesive and / or a preservative is expediently added to the aqueous solution.

Examples of suitable physiologically compatible salts are Acetate, citrate, halides, phosphate or sulfate.

Phosphate, acetate or citrate buffers, for example, can be used as buffers. Pointed adhesives are polyvinylpyrrolidone, polyvinyl alcohol or Ce.ll.uloseath.er. It depends on the dosage of these adhesive substances whether the preparation is obtained as an aqueous solution or as a jelly. The viscosity of the nasal drops should be between 30 and 300 cP. This is achieved, for example, by adding 5 to 15 ° / ° polyvinylpyrrolidone with a K-value of approx. 85 to ^^ or 0.5 to 1.5 "Ό methyl cellulose with an average degree of substitution of approx. 1.5 and a viscosity from 2700 to 4000 cP. If the aqueous preparation of peptides of formula II or their salts are to be used as jelly,

098A4 / 1105/6

for example, the content of the above-mentioned methylcellulose is 1.5 to 3 #.

Quaternary ammonium compounds, trichloroisobutanol, benzyl alcohol or p-hydroxybenzoic acid esters, in each case alone or combined with one another, can be used as preservatives. The dosage for the preservatives for benzyl alcohol is 1 to 2? Ö, p-hydroxybenzoic acid ester, for example. 0.2 to 0.3 ° and chlorobutanol about 0.4 ^c / o.

In addition to the buffers, substances that generate isotonicity are also suitable for example mannitol, sorbitol and sodium chloride.

0.1 mg to 20 mg of peptide, preferably 0.5 to 5 mg, are used as a single dose. This single dose is about 0.05 ml for aqueous solutions and about 0.05 S for jellies.

The aqueous solutions are filled into glass or plastic bottles with a pipette or dropper insert, and also into spray bottles, so-called plastic squeeze bottles. The preferred form of administration is a pressurized gas pack. A small pressure vessel made of glass or metal is 10 to 80 ° o of its volume with; filled with the preparation according to the invention, closed with a suitable valve in a manner known per se and overlaid with the inert gas such as nitrogen or argon up to a pressure of about 8 kg / cm.

If a metering valve is used as the extraction valve, a specific, constant Amount to be withdrawn. The valve can be used for better air handling with a nose applicator or a "mechanical break up" nozzle will provide.

Ge.1 nos verrlen ::. B. in plastic tubes or in protective lacquer-r-ri Tubes to the Re.i rial iimini um or Reinzimi filled. Sensible are there also small plastic containers?,. B. made of polyethylene for taking single doses of the solutions or dos jellies.

5098 ^ / 1105

/ 7 '

- γ -

Intranasally acting preparations can also contain the peptides according to the invention in the form of an oily suspension or a fatty ointment.
Such preparations are obtained by

a) a peptide of the formula II or its salts in an oil, optionally with the addition of surface-active substances with an HLB value (HLB = hydrphilic-lipophilic-balance) suspended under 10 and possibly with the addition of aluminum stearate as a swelling agent or

b) peptides of the formula II or their salts in a soft, Suspended spreadable fat base and, if necessary, adding a surface-active substance with an HLB value below 10 or

c) from a soft, spreadable fat base, a surface-active one Substance with an HLB value below 10 and the solution of peptides of the formula II or one of their salts in Water to make an emulsion ointment.

The preparations can also contain a preservative.

Acetate, halides, citrate, phosphate and sulfate, for example, are suitable as physiologically tolerable salts.

As a vehicle for nasal drops containing oily peptides of the formula II pharmacologically harmless, non-aqueous suspending agents are used for the nasal mucosa, Esters of saturated or unsaturated fatty acids with 8 to 18 carbon atoms with monohydric and polyhydric alcohols are suitable, liquid fatty alcohols and esters of fatty alcohols with fatty acids, as well as, natural vegetable oils. Liquid paraffins can also be used.

As a borderline active? Substances with an IILB-Vert below 10 come into consideration, for example, glycerol, sorbitan or pentaerythritol fatty acid esters. The Ilaf i ffibig-kcit of the oily, peptides of the formula II

5 0 9 B LUI 1 1 0 5, <

BATH ORIGINAL

containing nasal drops on the Fasenschleimliaut can be improved by swelling agents such as aluminum stearate. The viscosity of these nasal drops containing thickened peptides of the formula II should be between 50 and 300 cP. This is achieved, for example, by adding 2 to 5% of a mixture of aluminum di- VLTide monostearates (eg Alugel kh M, Bärlocher GmbH, Munich).

Suitable bases for nasal ointments containing peptides of the formula II are phai-macologically harmless, soft, easily spreadable anhydrous and possibly mixtures of solid and liquid paraffins, esters of saturated and unsaturated fatty acids with polyhydric alcohols, fatty alcohols, esters of fatty alcohols, which are compatible with the nasal mucosa Physiological acids such as citric acid and axes with one another vLxid with one another. Hydrogenated and acetylated fats in the broadest sense can also be used. Here, too, the addition of surface-active substances with an HLB value below 10, such as, for example, sorbitan fatty acid esters or lanolin and its derivatives, has proven itself.

Emulsion nasal ointments containing suitable peptides of the formula II are produced from the same raw materials and emulsifiers as the anhydrous fatty ointments. But they contain up to 70 / o, preferably up to ^ O ⁰ Jo water.

Quaternary ammonium compounds, benzyl alcohol or p-hydroxybenzoic acid esters, in each case alone or combined with one another, can be used as preservatives. The dosage for the preservatives is 1 to 2 ° p for benzene alcohol and about 0.1 to 0.3% for p-hydroxybenzoic acid ester .

The single dose of peptides of the formula II is between 0.1 mg and 20 mg, preferably between 0.5 and 5 wt. This single dose is contained in 0.05 to 0.1 g of the preparation according to the invention.

5 0 9 8 LUI 1 1 0 5. _{/ c)}

The oily suspensions can be filled into glass or plastic bottles with a pipette or dropper insert. Sensible are also small plastic pots or gelatin capsules for taking a single dose. Preferred form of administration for an oily nasal preparation containing peptides of formula II is a pressurized gas pack. In a 12 ml pressure vessel between 0.5 and 5 »0 g of an oily peptide of the formula II containing suspension filled and either in the cold filling process before closing the container with a suitable valve or in the pressure filling process according to the Close the container with a suitable valve with 5.0. up to 9.5 g of a liquefied propellant mixture which is miscible with the oily carrier is added. Preferred propellants are trichlorofluoromethane (Frigen ^ 11), dichlorofluoromethane (Frigen ^ '12), 1,1,2,2-tetrafluorodichioethane

11M and 1,2,2-trifluorotrichloroethane (Frigen (r) 113). FrigenV 11 and Frigen ^ ' 1i4 cannot be used alone. Frigen 12 must be added to the pressure increase.

(r)

Chlorodifluoromethane (Frigen 22) is also suitable for this. A priority as used blowing agent mixture consists of hO to 60 parts by weight of Freon ^ * '12 and 60 to ho parts by weight of Freon 114. If, as Entnahmeventd1 a metering valve, so the valve may be a certain, constant amount to be removed per Betätigting respectively. To make it easier to use, the valve is fitted with a suitable nasal applicator.

The ointments are made in pure tubes with a protective coating on the inside. aluminum, pure tin or a suitable plastic.

The following examples illustrate the preparation of those of the invention Peptides and their preparations for parenteral or intornasal administration.

5098AA / 1105

Examples

For amino acids and protective groups are in the peptide chemistry common abbreviations are used. Further abbreviations:

DCCI Dic3> -clohexy3 carbodiimide

HOBt T-Hydroxybeuzotriazole

HOOBt 3-Hydroxy-4-oxo-3,4-dihydro-1,2,3-benzotriazine

Ibu £ j £ -aminoisobutyric acid (j £ -methyl alanine)

OPcp pentachlorophenyl ester

OTcp trichlorophem ^r lester

5098U / 1 105

example 1

ß-Ala-Gly-Cys-Lys-Asn-Phe-Phe-Trp-Lys-Thr-Phe-Thr-Ser-Cys-NHg

a) Z-Cys (BzI) -OITb

203 g (0.6 mol) of Z-Cys (BzI) -OH, 194 g (0.9 mol) of 4-nitrobenzyl bromide and 126 ml (0.9 mol) of triethylamine are refluxed in ethyl acetate with stirring for 14 h and filtered while hot. The Piltrat is successively with water, 1N HCl 2N KHCO ^. and water washed, dried over NapSOh and in vacuo. constricted. 200 ml of ligroin are added, the precipitate is filtered off and recrystallized from methanol. After drying over P ₀ O ^ yield. 164 g (57 #). 98-99 °.

b) H-Cys (BzI) -ONb * HBr

48.15 g (0.1 mol) of Z-Cys (BzI) -ONb are dissolved in 350 ml of HBr / glacial acetic acid. After about 25 minutes, the product partially precipitates. After a further 30 minutes, it is precipitated with 1 liter of ether, the crystals are filtered off, washed with ether and dried over PpOj-. Yield 37.3 g (88 fo). M.p. I56 ⁰ .

° = -24.9 ^C (c = 1, methanol)

c) Boc-Ser (BzI) -Cys (BzI) -ONb

23.6 g (80 mmol) Boc-Ser (BzI) -OH, 36.2 g (80 mmol) H-Cys (BzI) -ONb «HBr, 10.8 g (80 mmol) HOBt and 9.85 ml (80 mmol) N-Ethylmorpholine are dissolved in 350 ml of dimethylformamide. Man add at 0 ° I6.6 g (80 mmol) of DCCI, allow it to come to room temperature and, after 15 h, filter off the precipitated urea. Then you narrow in the vac. to dryness, takes the residue in

5 098ΛΑ / 1Τ05

24Ί60Λ8

500 ml of ethyl acetate and washes one after the other with ice-cold 5 percent. KHSO ^ solution, 2N KHCO-, and water, dried over ITap, evaporated and the residue recrystallized from methanol. Yield 40.5 g (78 fo)

ξ ° = -25.5 ° (c = 1, dioxane)

d) H-Ser (BzI) -Cys (BzI) -ONb'HCl

7.0 g of the Boc compound are dissolved in 50 ml of J5N HCl in dioxane. After 1 h, the reaction product is precipitated with 500 ml of ether and filtered off. After washing with ether and drying in Vac. Yield 5-2 g. The compound is almost chromatographically pure.

e) Boc-ThI (BzI) -Ser (BzI) -Cys (BzI) -ONb

5.1 g (9.1 mmol) H-Ser (BzI) -Cys (BzI) -ONb-HCl, 5.9 g (10 mmol) Boc-Thr (BzI) -OH, 1.55 g (10 mmol) HOBt and I.17 ml ( 9.1 mmol) N-ethylmorpholine v / earth in 40 mlT at 0 ° with 2.2 g (10.6 mmol) DCCI. The mixture is stirred for 6 hours at room temperature and the urea is filtered off. brings i.Vak. to dryness, the residue is taken up in ethyl acetate and the solution is washed successively with 5 percent strength. KHSOj, (ice cold), 1N NaHCC ₂ and V / water, dries over NapSOh, brings to dryness and recrystallizes the residue from isopropanol. Yield 6.4 g (87 ^), Sehmp. 106 °, / ⁰ ^ p

= 4.8 ° (c = 1, chloroform)

f) H-Thr (BzI) -Ser (BzI) -Cys (BzI) -ONb-HCl

3.0 g of the Boc compound are in J> 0 ml ^ N. HCl dissolved in dioxane. After 1 h it is precipitated with ether, the precipitate is filtered off, washed with ether and dried in vacuo. via PpOp-. Yield 2.7 g m.p. 179-180 °. Chromatographically pure.

* Dimethylformamide

5 0984WM05

g) Z-Thr-Phe-OMe

25.3 S (0.1 mol) Z-Thr-OH, 21.6 g (OI mol) H-Fhe-OMe'HCl and 15.5 g HOBt are dissolved in 500 ml DMP. 12.8 ml (0.1 mol) of N-ethylmorpholine and 22 g (I.06 mol) of DCCI are added, the mixture is stirred for 5 h, the urea is filtered off and the filtrate is concentrated in vacuo. to dry one. The residue is taken up in ethyl acetate, washed as above, dried over sodium sulfate and the ethyl acetate is distilled off. The residue is triturated with ether. Yield 39.5 g (83 fo) m.p. 104-106 °.

h) H-Thr-Phe-OMe tosylate

35 g of the Z compound are dissolved in 350 ml of methanol with the addition of 2N methanol. Toluenesulfonic acid at constant pH (4.5) catalytically hydrogenated on palladium. When the hydrogenation has ended, the catalyst is filtered off and the methanol is distilled off in VaIc. away. Resinous residue.

i) Boc-Lys (Z) -Thr-Phe-OMe

36 g (80 mmol) of H-Thr-Phe-OMe tosylate, 30.5 g (80 mmol) of Boc-Lys (Z) ~ 0H and 10.8 g (80 mmol) of HOBt are dissolved in 350 ml of DMF. 11 ml (86 mmol) of N-sthylmorpholine and 17.6 g (86 mmol) of DCCI are added and the mixture is stirred overnight at room temperature. Then .vom urea is sucked off and the solvent in the; Vac. distilled off. The further work-up follows example 1e). Recrystallization from 120 ml of methanol + 100 ml of water. Yield 36. ^ g (71 %), m.p. 86-. 87 ⁰ . '_:

k) H-Lys (Z) -Thr-Phe -OMe · HCl · '. : ■.:.

The ^{BOc-Gr 1} UPPe is split off from 34.6 g of the Z compound analogously to Example 1d. Yield 29.4 g. Almost uniform in terms of chromatography; in addition to a little urea, there is 1-2 % of a compound with

5Q98AW1105

lower RF-Fourth, possibly also the Z group was split off.

1) Boc-Trp-Lys (Z) -Thr-Phe-OKe

17.3 g (57 mmol) Boc-Trp-OH, 29.4 g (51 mmol) H-Lys (Z) -Thr-Phe-OMe, HCl and 7.7 g (57 mmol) of HOBt are dissolved in 280 ml of DMF. 7.25 ml (56 mmol) of n-ethylmorpholiri and 12.3 g (60 mmol) are added DCCI and works as described under Example 1i). Yield after recrystallization from ethanol / water 37.5 (89 f &). M.p. 105-115 °.

m) Boc-Trp-Lys (Z) -Thr-Phe-OH

16.6 g (20 mmol) of the methyl ester are in a mixture of 80 ml of dioxane. Dissolved 50 ml of methanol and 20 ml of water. Then, at constant _{temp F} (12.5), a total of 21.9 nil IN NaOH is added dropwise, after the saponification is complete _{, the solvent is first brought to p TT} 7.4 with 1N HCl, filtered and the solvent is distilled

Xl

in the vac. The residue is taken up in water. It is acidified with ice-cold 1N HCl to p, _T 2.2, and the precipitate is filtered off

χι - - ■ ■

and washes it with water. Yield after drying in the yak. via PpOi-I ² I-.6 g (90 ^), chromatograph, almost uniform, only 1-2 % unsaponified ester can still be recognized.

n) Boc-Trp-Lys (Z) -Thr -Phe-Thr (BzI) -Ser (BzI ) -Cys (BzI) -ONb

8.3 g. (10 mrtol), Boc-Trp-Lys (Z) -Thr-Phe-OH, 7.5 S (10 mmol) H- ■

Thr (BzI) -Ser- (Bzl) -Cys (BzI) -Nb-HCl and 2.7 S (20 mmol) HOBt are dissolved in 50 ml DMF. One gives 1.5 ml (10 mKol) N-iithylmorpholin and J>. 1 g (15 mmol) of DCCI are added and the mixture is stirred overnight at room temperature. The urea which has precipitated is then suctioned off and the peptide is precipitated from the filtrate with 500 ml of ether. It is boiled with a little ethanol, filtered off after cooling, with

5098Λ4 / 1105

Washed a little cold ethanol and dried in vacuo, over Pp ° c. Yield 12 g (79 } o). Chromatographically uniform.

o) Boc-Trp-Lys (Z) -Thr-Phe ~ Thr (Bzl) -Ser (Bzl) -Cys (BzI) -NHp

12 g of the finely ground nitrobenzyl ester are poured into 200 ml flow. Dissolved ammonia and stored in a pressure vessel overnight. The ammonia is allowed to evaporate and the residue is triturated with Ether. The conversion into the amide can be recognized by the decrease in polar extinction at 275 nm to the value of tryptophan.

p) H / Trp-Lys (Z) ~ Thr-Phe-Thr (BzI) ~ Ser (BzI) -CyS- (BzI) -NH ₂ * HCl

10 g of the Boc compound + 3 g of thiophenol are dissolved in 80 ml of 6N HCl / dioxane and the solvent is distilled after standing for 15 minutes at room temperature in vac. almost completely off. Isopropyl ether is added to the residue, suction filtered and washed with ether. • Education 8.7 g

q) Z-Phe-Phe-OB ^

79.2 g (0.2 mol) of Z-Phe-ONSu are added to 48.6 g (0.22 mol) of H-Phe-OBu * in 300 ml of DMP, while cooling to 0 °. After 3 hours of standing at room temperature, the solvent is distilled in vacuo. from, dissolves the residue in ethyl acetate, washes one after the other with ice-cold 5 percent. KHS0 | ,, 1N NaIiCO-, and water, dries over Na _o S0 |, and distills off the ethyl acetate. The residue is over P ₂ O ₅ in a vacuum. dried. Yield 84.5 g (84 %) / pCj ^ = -17.5 ° (c = 1, methanol)

r) H-Phe-Phe-OBi ^ · HCl

49 g of Z-Phe-Phe-OBu are dissolved in 300 ml of methanol in the presence of palladium black at p, _T 5 with the dropwise addition of methanol. HCl

5098A kl 1105

catalytically hydrogenated. The reaction has ended after 2 hours. The catalyst is filtered off, evaporated to dryness and the residue is triturated with ether.
Yield 35.8 g m.p. 152 - 154 ° (decomp.)

■ s) Z-Asn-Phe-Phe-OB ^

40.5 g (0.1 mol) of H-Phe-Phe-OBu ^ HCl, 38.7 g (0.1 mol) of Z-Asn-ONp and 1.35 S (10 mmol) of HOBt are stirred in 250 ml of DMF for 5 hours. You narrow in the vac. to dryness, rub the residue with ice-cold 5 percent. KHSO ^, 1N NaHCO-, and water, boils with Diiscpropyläther and recrystallizes from methanol / water. Yield 48 g (78 %). M.p. 184 °

= -6.6 ° (c = 1.90% acetic acid).

t) H-Asn-Phe-Phe-OBu ^ HCl

50 g of the Z compound are hydrogenated analogously to 1r) and obtained after working up 39 x 8 g of the chromatographically pure compound.

u) "Boc-Lys (z) -Asn-Phe-Phe-OBu

20.3 g (55 mmol) Boc-Lys (Z) -OH, 25.9 S ■ (50 mmol) H-Asn-Phe-Phe-OBu ¹ ^ -HCl and 8.4 g (55 mmol) HOBt are in 250 ml DMF with 6.4 ml (50 mmol) of N-ethylmorpholine and 11.3 g of DCCI (50 mmol) were added. After stirring overnight, the urea is filtered off and the filtrate in vacuo. concentrated to dryness. The residue is taken up in methylene chloride and washed successively with ice-cold 5 percent strength. KHSO ^, 1N NaHCO, and water, dried over Na ₂ SO ^ and the methylene chloride is distilled off. The solid residue is recrystallized from methanol / water and over P-Otim Vak. dried. Yield 32.1 g (76 #). 147-149 °. f = - *. 9 ° (O-1, CHCl)

50.9844 _/ 1 _105/17

ν) H-Lys (Z) -Asn-Phe-Phe-OH

25% of the compound obtained according to 1u) is dissolved in 250 ml of 6N HCl / dioxane and precipitated after 1 h with ether. The dried precipitate is triturated with 1H NaHCO, and water and made from 80 percent. Recrystallized ethanol.
Yield 17 g (83 fo)
Shrap. 218 -220 ° (dec.)

15 S (21.7 mmol) H-Lys (Z) -Asn-Phe-Phe-OH, 12.8 g (24.6 mmol). Boc-Cys (BzIOMe) -OTcp (m.p .: 124-125 °, βij _D = -41.9 (c = 1, DMF)), 3.12 g (24.6 mmol) of HOBt and 2.78 ml (21.7 mmol) of N-ethylmorpholine are in 200 ml of dimethylformamide were stirred at room temperature for 2 h. Then with ice cooling with 5 percent. KHSOn solution acidified and diluted with water. The precipitate is filtered off, washed with water and in vacuo. is dried over P ₂ ^{0 C.} The dried product is boiled with ethyl acetate and reprecipitated from methanol / water. Yield 18.9 g (85.7 fo), m.p. 232 - 2 ^ 5 ° (decomp.) Ä7 _D = -16.9 ° (c = 1, DMF)

x) H-Cys (BzIOMe) -Lys (Z) -Asn-Phe-Phe-OH

17 g of the Boc compound are dissolved in 25 ml of trifluoroacetic acid containing 1 % mercaptoethanol, while cooling, and the mixture is stirred for 25 minutes at room temperature. Then, while cooling with ether, a precipitate is precipitated out, which is filtered off and washed with ether. It is then dissolved in a little dimethylformamide. It is made neutral with 1N NaHCO 3 and mixed with water. After drying, the precipitate is boiled with ethyl acetate. Yield I3.6 g (88.7 fo).

509844/1105 _{/ ia}

- 18 Y) Z-β-Ala-Gly-OPcp

2.8 g of Z-ß-Ala-Gly-OH (1 cM) are dissolved in 50 ml of tetrahydrofuran solved. 2.9 g (1.1 cMol) of pentachlorophenol and 2.2 g of DCCI are added in the cold, and the mixture is stirred for 1 h at 0 ° and 4 h at room temperature. The urea is filtered off, brought in vacuo. to dryness and boil the residue with isopropanol. Yield after the residue has dried, 4.3 g * m.p. 179 - 180 °,

2) Z-3-Ala-Gly-Cys (BzIOMe) -Lys (Z) -Asn-Pfte-Phe-OH

5.0 g (5.5 mmol) H-Cys (BzIOMe) -Lys (Z) -Asn-Phe-Phe-OH, "3.17 g (6 mmol) Z-ß-Ala-Gly-OPep, 0.74 g HOBt and Ο.71 ml N-Ethylmorpholine is stirred in 40 ml of dimethylformamide for 2 hours. It is acidified with 5% aqueous KHSOh solution, precipitated with water, the precipitate is filtered off and washed with water. The dried product is washed with methanol / ethanol ( 1: 1) boiled and reprecipitated from dimethylformamide / ether, washed with ether.
Yield 5.6 g (86.7 ? O), m.p. 208-210 ° (decomp.)

aa) Z-i3-Ala-Gly-Cys (BzIOMe) -Lys (Z) -Asn-Phe-Phe-Trp-Lys (Z) -Thr- Phe-Thr (BzI) -Ser (BzI) -Cys (BzI ) -

5.2 g (4.5 mmol) Z-ß-Ala-Gly-Cys (BzIOMe) -Lys (Z) -Asn-Phe-Phe-OHi 6.0 g (4.55 mmol) H-Trp-Lys (Z) -Thr-Phe- Thr (BzI) -Ser (BzI) -Cys. (BzI) -NHp.HCl and 1.25 S (9.1 mmol) HOBt are dissolved in 150 ml DMF. 1.28 ml (10 mmol) of N-fithylmorpholine and 1.0 g (4.8 mmol) of DCCI are added and the mixture is stirred over power at room temperature. Then, the precipitated urea is filtered off and the piltrate in vac. brought to dryness. The residue is with ether, 5 percent. KHSOh, 1N NaHCO ^ and water triturated and boiled with 50 ml of methanol. Yield after drying 8.25 g (74 %), the verb, is chromatographically uniform.

509844/1105

bb) ß-Ala-Gly-Cys ^ Lys-Asn-Phe-Phe-Trp-Lys-Thr-Phe-Thr-Ser-Cys- NHp-triacetate

Dissolve 2.0 g of the protected compound in 100 ml of flow. Ammonia and, at the boiling point of ammonia, adds enough sodium while stirring vigorously that "the blue color takes 20-30 seconds. remains. Then it is decolorized by adding ammonium chloride and the ammonia is evaporated off. The dehydration and the Purification takes place in a known manner analogous to Biochem. Biophys. Res. Commun. _54 (1973) * page 234. After the desalination on Sephadex G-15 will ion exchange on Amberlite IR-45 B turned on in the acetate form.

Yield after freeze-drying: 0.24 g as triacetate Amino acid analysis: Asp 1.08, Ser O.83 ^x ), Thr I.58 ^x \ GIy 1.00, Cys I.57 ^x \ Phe 2.93, ß-Ala 0.96, Lys 2.03, Trp (spectrophotometric): 0.94

' not corrected

Example 2

ß-Ala-Gly-Cys-Lys-Asn-Phe-Phe-Trp-Lys-Thr-Phe-Thr-Ser-Cys-n butylamide

a) Boc-Trp-Lys (Z) -ThrrPhe-Thr (BzI) -Ser (Bzl) -Cys (BzI) -ΝΗ-

3 g (2 mmol) of the Boc-heptapeptide nitrobenzyl ester prepared according to Example 1n) are boiled in 50 ml of methanol with 0.6 ml of n-butylamine for 2 h. The methanol is distilled off and the triturated Thoroughly remove residue with A'ther and boil it out with ethanol. Yield 2.4 g. Analogously to example 1o) the conversion is by decrease the absorbance is controlled.

509844/1105 / ₂₀

b ) H-Trp-Lys (Z) -Thr-Fhe-Thr (BzI) -Ser (BzI) -Cys (BzI) -NHC ^ H ₀ - HCl

The Boc group is cleaved analogously to Example 1p) from 2 g of the Boc Compound and receives 1.67 g of heptapeptide hydrochloride.

c) H-ß-Ala-Gly-Cys-Lys-Asn-Phe-Phe-Trp-Lys-Thr-Phe-Thr-

-Ser-Cys-HH-C ^ Hq

5.2 g Z-heptapeptide, prepared according to Example 1z), v / ground analogously to Example 1aa) with 6.25 g of H-heptapeptide hydrochloride, prepared according to Example 2b), 1.23 g of HOBt and 1.0 g of DCCI in 150 ml of dimethylformamide in the presence of 1.28 ml of N-ethylmorpholine implemented and worked up analogously to Example 1aa). Yield 8.38 g.

The protective groups are split off in a known manner for 60 minutes, storage in flow. HF / anise oil, dehydration through air and purification through gel filtration on Sephadex G and partition chromatography on Sphadex G 25 in the n-butanol-acetic acid-water (4: 1: 5) system according to Biochem. Biophys. Res. Commun. jj4 (1973), page 234. I.19 g of a chromatographically pure compound after freeze-drying. Amino acid analysis: Asp I.07, Ser 0.79 > Thr 1.66 ^x ', ■ GIy 1.00, Cys 1.68 ³ ^, Phe 3.01, β-Ala 0.98, Lys I.99 Trp. (Spectrophotometric): O.96

¹ not corrected

example

Boc-Cys-Lys-Asn-Phe-Phe-Trp-Lys-Thr-Phe-Thr-Ser-Cys-lIHp

a) BoC-CyS (BzIOHe) -LyS (Z) -ASn-PhC-PhB-TrP-LyS (Z) -Thr- Phe-Thr (BzI) -Ser (BzI) -Cys (BzI) -HH ₂

5098AA / _1105/21

1.0 g (ImMoI) Boe pentapeptide, prepared according to Example 1w), 1.51 g (1 mmol) H-heptapeptide hydrochloride, prepared according to Example 1p), 270 mg (2 mmol) HOBt and 0.13 ml (1 mmol) II-ethylmorpholine v; ground in 20 ml of dimethylformamide with the addition of 250 mg of DCCI, stirred overnight. It is precipitated with ether and the precipitate is digested with hot methanol. Yield 1.59s ($ 71) · Th ^e connection is chromatographically uniform and free of starting materials.

b) Boc-Cys-Lys-Asn-Phe-Phe-Trp-Lys-Thr-Phe-Thr-Ser-Cys-lIHg

1 g of the protected dodecapeptide is freed from the protective groups analogously to Example 1 bb), dehydrated to the disulfide and "purified. Yield 124 mg as diacetate. Amino acid ^{analysis: Asp 1.05, Ser 0.81 x} ', Thr I.70 ^x ', Cys 1.72 ^x \ Phe 3.00, Lys 2.02
Trp (spectrophotometric): 0.95

' not corrected

BeisDiel 4

H-D-Ala-Gly-Cys-Lys-Asn-Phe-Phe-Trp-Lys-Thr-Phe-Thr-

-Ser-Cys-NHp

a) Z-D-Ala-Gly-OTcp

28.Ο g Z-D-Ala-Gly-OH, prepared according to J. Biol. Chem. J [09 (1935), Page 325, and 19.7 g of 2.4.5-trichlorophenol are ± 1,300 ml 20.6 g of DCCI are added to dimethylformamide at 0 °. The mixture is stirred for 1 h at 0 ° and 18 h at room temperature, filtered and the distilled Solvent in vac. Crystallization from isopropanol / i'ithanol (1: 1), yield. 37.7 g (82 #), m.p. 160-162 °

509844/1105

2 ^ 16048

b) "Z-D-Ala-Gly-CysCBzlOI-? e) -Lys (Z)" Asn-Phe-Phe-OH

5.Og pentapeptide, prepared according to Example 1x), 2.Sg ZD-Ala-Gly-OTcp., 0.7 ² I-g HOBt and 0.71 ml II-ethylnorpholine are stirred in ^ O ml dimethylformamide for 2 hours. Work-up and isolation as in Example 1z). Yield 5.6 (87 p), m.p. 203-210 °.

c) ZD-Ala-Gly-Cys (BzIOMe) -Lys (Z) -As.n-Phe-Phe-Trp-Lys (Z) - Thr-Phe-Thr (BzI) -Ser (BzI) -Cys (BzI ) -i: H ₂

5 × 2 g of the Z-heptapeptide prepared according to b) are added to 6.0 g of H-heptapeptide, prepared according to Example 1p) with the addition of I.23 g of HOBt, 1.28 ml of N-ethylmorpholine and 1.0 g of DCCI in 150 ml of diethylforman ; id * analogous to example laa) with each other and works on as described. Yield 8.5 g (76 p). The compound is chromatographically uniform and free from starting materials.

d) H-D-Ala-Gly-Cys-Lys-Asn-Phe-Fhe-Trp-Lys-xhr-Phe-Thr-

-Ser-Cys-NHp

1 g of the protected peptide prepared according to c) are freed from the protective groups analogously to Example 1bb), for cyclic disulfide dehydrated and purified. Yield 1j52 mg as triacetate.

Amino acid analysis: Asp 1.02, Ser Ο.79 ', Thr 1.66' GIy 1.00,

AIa 1.02 Cys 1.82 ^X ^, Phe 2.99, Lys 2.02 Trp (spectrophotometric.) 0.9β

^ not corrected

50984A / 1105/23

Example 5

S-CH ₂ -CH ₂ -C0-Lys-Asn-Phe ~ Phe-Trp-Lys-Thr-Phe-Thr-Ser-Cys-iIH ₂

a) (S-BzlOMe) -ß-mercaptopropionic acid 2.4.3-trichlorophenyl ester

4.52 g (20 mmol) (S-BzlOMe) -ß-mercaptopropionic acid and 3-94 g (20 mmol) 2.4.5-trichlorophenol v / earth in 15 ml methylene chloride reacted with 4.12 g (20 mmol) DCCI. After 4 h, the urea is filtered off and the filtrate is evaporated. The residue crystallizes after standing for a short time. Yield 5 x 7 S m.p. 44-46 °

b) (BzIOMe) -S-CH ₀ -CH ^ -CO-LyS (Z) -Asn-Phe-Phe-OH

2.6 g (6.4 mmol) of the trichlorophenyl ester obtained according to a) is put in 10 ml of dimethylformamide with 5.2 g (4.5 mmol) H-Lys (z) -Asn-Phe-Phe-OH, obtained according to Example 1v), in the presence of O.85 g (6.4 mmol) (4l-Ethylmorphclin um. After 5 min. The reaction time is the reaction products by adding 5 percent. KHSOii solution and water precipitated from dimethylformamide / water reprecipitated and after drying boiled with methanol / ether. Yield 3.4 g, m.p. 213-218 ° (decomp.)

c) (BzIOMe) ~ S-CH ₂ -CH _r -C0-Lys (Z) -Asn-Phe-Phe-Trp-Lys (Z) -Thr-Phe-Thr (Bzl) -Ser (Bzl) -

0.9 g (ImMoI) tetrapeptides produced according to b), 1.J1 g (1 mmol) H-heptapeptide hydrochloride, prepared according to Example 1p), 270 mg (2 mmol) of HOBt and 0.13 ml (1 mmol) of N-ethylmorpholine become stirred in 20 ml of dimethylformamide with the addition of 250 mg of DCCI overnight. You fall with ether and digest with hot Methanol. Training in 3.g

The compound is chromatographically uniform and free from starting materials.

* HOBT and 0.81 ml {6.U mmol)

5098 ^ 4/1105

/ 2k

d) S- (CHpJo-Cö-Lys-Asn-Phe-Phe-Trp-Lys-Thr-Phe-Thr-Ser-Cys-l · ^

1 g of the protected undecapeptide are analogous to Example 1bb) freed from the protective groups, dehydrated to disulfide and purified. Yield 119 mg as diacetate.

Amino acid analysis: Asp 0.99, Ser 0.79 * K Thr 1.77 * K Cys O.87 Phe 3-CO, Lys. 1.97

Trp (spectrophotometric) 0.96

¹ not corrected
Example 6 D-Ala-Gly-Cys-Orn-Asn-Phe-Phe-Trp-Lys-Thr-Phe-Thr-Ser-Cys-OH

a) Z-0rn (Tos) -Asn-Phe-Phe-0Bu

3.0 g (5.8 mmol) H-Asn-Phe-Phe-OBu ^ HCl, 5.0 g (8.3 mmol Z-Orn (Tos) -OTcp, Ο.78 g (5.8 mmol) HOBt and 0.75 ml (5.8 mmol) N- Ethylmorpholine is stirred in 10 ml of dimethylformamide for one hour. The reaction product is precipitated with ether and reprecipitated from dimethylformamide / 1% acetic acid. From 3.4 g (67 %), melting point 169-170 °

b) Z-Orn (Tos) -Asn-Phe-Phe-OH

3.0 g of the tert-butyl ester prepared according to a) are dissolved in 15 ml of trifluoroacetic acid for 1 h at room temp. touched. It is precipitated with ether, the precipitate is washed with ether and precipitated from dimethylformamide / ether and dimethylformamide / water. Yield 2.6 g, m.p. 173-178 ⁰ (decomp.)

c) H-Orn (Tos) -Asn-Phe-Phe ~ O H

2.6 g of Z compound are dissolved in dimethylformamide / methanol (1: 1)

509844/1105

catalytically hydrogenated on palladium / barium sulfate. The catalyst is filtered off and the filtrate is concentrated in a vacuum. to dry one and rub the residue with ethyl acetate. Yield 1.1 g Sehmp. 205 ° (decomp.)

d) Boc-Cys (BzIOMe) -Orn (Tos) -Asn-Phe-Phe-OH

4.0 g (5.8 mmol) H-Om (ToS) -Asn-Phe-Phe-OH, 3.5 g (6.7 mmol) BOC-CyS (BzIOMe) -OTCp, 0.8 g (5.9 mmol) HOBt and 0.8 ml (6.3 mmol) of N-ethylmorpholine in 20 ml of dimethylformamide for 2 h touched. It is precipitated at 0 ° with KHSO ^ solution and water and washed the precipitate with water and after drying it boils with ethyl acetate and ethyl acetate / ethanol. Yield 5.8 g, m.p. 195 ° (decomp.)

e) H-Cys (BzIOMe) -Orn (Tos) -Asn-Phe-Phe-OH

5.8 g of the Boc compound are in the presence of 1 ml of mercaptoethanol Stirred in 25 ml of trifluoroacetic acid for 30 min. Man falls with ether, dissolves the residue in a little dimethylformamide / water, makes it neutral with NaHCO ^ solution and falls with water. The precipitate is boiled with ethanol. Yield 2.4 g Sehmp. 223 - 224 ° (dec.)

f) Z-D-Ala-Gly-Cys (BzI0Me) -Orn (Tos) -Asn-Phe-Phe-OH

2.0 g (2.2 mmol) of pentapeptide, prepared according to e), 1.12 g (2.4 mmol) of ZD ~ Ala-Gly-OTcp, 0.29 ml (2.3 mmol) of N-ethylmorpholine and 0.32 g (2.4 mmol) of HOBt are in 10 ml Dimethylformamide 3 h at room temp. touched. It is precipitated with KHSO ₂ solution and water, the precipitate is washed with water and digested with methanol. Yield 2.02 g, m.p. 211-213 °

509844/1105

- 26 - (ζ) H-Cys (BzIOMe) -OBzI. H Cl

19-2 g (0.08 mol) of H-Cys (BzIOMe) -OH are dissolved in 16O ml of 0.5 N methanol. KOH dissolved. Freshly distilled acetoacetic ester is added to 11.1 ml, the mixture is refluxed for 10 minutes and the solvent is distilled in vacuo. away. The dried, solid residue is dissolved in dimethylformamide. 9-2 ml (0.08 mol) of benzyl chloride are added and the mixture is left to stand at room temperature for 48 hours. Then stirred into a mixture of 4θΟ ml N NaHCO and 4θΟ ml ethyl acetate ¹ , · the ethyl acetate extracted twice with hydrogen carbonate solution, washed with water, dried over Na SO. and the solvent is distilled off. The oily residue is dissolved in ΐ6θ ml N methanol. HCl dissolved, the solvent in vac. distilled off and the residue digested with ether.
Yield 17 g (58? Έ), m.p. * 125 °. / <* l7 _d = -27.8 ° (c = l, methanol.

h) Boc-Ser (Bzl) -C.ys (BzIOMe) -OBzI

2.9 S (44 mmol) of Boc-Ser (BzI) -OH as dicyclohexylamine salt are dissolved in 100 ml of ethyl acetate with 5 percent strength. watery KHSO. Solution converted into the free acid. The ethyl acetate phase is dried and the solvent is distilled in vacuo. away. The residue is dissolved in 50 ml of dimethylformamide. 5-4 g (4θ mmol) of the compound obtained according to g) are added, 1 equivalent of N-ethylmorpholine is added and 8.25 S DCCI in a little dimethylformamide is added. After 16 hours, the procedure is analogous to Example 1c) and 21.2 g of melting point 03-94 ° / ° ζ ~ / _Ό = -3 ^ -6 ° (c = 1, methanol.

i) H-Ser- (Bzl) -Cys (BzIOMe) -0Bzl ° HCl

20 g of the Boc dipeptide obtained after h) are stored in 4N for 20 min HCl stored in dioxane at room temperature. The solvent is distilled in vacuo. gently and digests the residue with ether / petroleum ether. Yield I6.8 g (93? Ö)> M.p. 14θ °

/ 27

509844/1105

-27- 2416Q48

k) Boc-Thr (BzI) -Ser (Bzl) -Cys (SzlOMe) -OBzI

16.3 S (30 mmol) of the compound obtained according to i), 8 g (30 mmol) of Boc-Thr (BzI) -OH and 4.05 g (30 mmol) of HOBt are dissolved in 100 ml of dimethylformamide. 1 equivalent of N-ethylmorpholine and 1.1 equivalent of DCCI are added and, after 15 h, work up analogously to Example 1c) and 22.6 g ($ 95) of compound with melting point 97-98 ° - / ^ J _D = -22.8 ° (c = 1, methanol)

l) H-Thr (Bzl) -Ser (Bzl) -Cys (BzlOMe) -OBzl 'HCl

22 g of the compound obtained according to k) are treated analogously to i). Yield 18 g (89 ° / o), mp. I65 ^0th / ÖC_7 _D -3 ^ .3 ° (c = 1, methanol)

) Boc-Trp-Lys (Z) -Thr-Phe-Thr (BzI) -Ser (BzI) -Cys (BzIOMe) -OBzI

8.3 S (10 mmol) of the compound obtained according to Example 1m) is reacted with 7-35 S (10 mmol) of the tripeptide obtained according to 1) analogously to Example 1n) and worked up as described there. Yield 11. "45 g, m.p. 190-192 °, /" C_7 π ° = -13 * 9> ( ^c = 1) 95 percent acetic acid).

n) H-Trp-Lys (z) -Thr-Phe-Thr (Bzl) -Ser (BzI) -Cys (BzIOMe) -OBzl »HCl

The Boc protective group is cleaved from 10 g of the Boc compound obtained according to m) analogously to Example 1p) and worked up as described there. Yield 9 · 1 p

509844/1105

ο.) ZD-Ala-Gly-Cys (BzIOMe) -Orn (Tos) -Asn-Phe-Phe-Trp-Lys (Z) -Thr-Phe-Thr (BzI) -Ser (BzI) -Cys (BzI) - OBz I

1.17 g (ImMoI) Z-heptapeptide, produced according to f) and 1.41 g (1 mmol) H-heptapeptide benzyl ester, produced according to n), are reacted and worked up analogously to Example Jaa). Yield 1.5 g (60 %). Free of starting products for chromatographic testing.

ρ) D-Ala-Gly-Cys-Orn-Asn-Phe-Phe-Trp-Lys-Thr-Phe-Thr-Ser-Cys-OH

1 g of the protected tetradecapeptide is freed from the protective groups analogously to Example 1bb), dehydrated to the disulfide and cleaned. Yield 144 mg as triacetate.

Amino acid analysis: Asp 1.01, Ser Ο.83 ^x \ Thr I.78 ^x \ GIy 1.00,

AIa 1.02, Cys I.72 ^X \ Phe 2.96, Lys 2.04 Trp (spectrophotometric) 0.95

' not corrected

Example 7
Ibu-Gly-Cys-Lys-Asn-Phe-Phe-Trp-Lys-Thr-Phe-Thr-Ser-NH-CH,

-NS,

/ 29 509844/1105

a) "BoG-Gly-Cys (BzIOMe) -Lys (Z) -Asn-Phe-Pne ~ OH

9-1.5 g (10 mmol) of pentapeptide, prepared according to Example 1x) are dissolved in 80 ml of dimethylformamide and mixed with 3.9 g (11 mmol) of Boc-Gly-OTcp in the presence of 1.35 g (10 mmol) of HOBt and 1.28 ml for 2 hours (10 mmol) N-fithylmorpholine stirred. Work-up analogous to Example 1z). 8.2 ¹ I- g (77 %), chromatographically uniform, melting point 205 - 210 ° (decomp.)

b) H-Gly-Cys (BzIOMe) -Lys (Z) -Asn-Phe-Phe-OH-HCl

8.0 g of the compound prepared according to a) are 45 min. In 3> N HCl, stored in dioxane. You fall with ether and wash the precipitation with ether. Yield almost quantitative. Lt. Chromatogram free from parent compound. -

c) Z-Ibu-Gly-Cys (BzIOMe) -Lys (Z) -Asn-Phe-Phe-OH

7. 05 g (70 mmol) of the hexapeptide prepared according to b) are in 50 ml of dimethylformamide with Z-Ibu-OTcp, prepared from 2.0 g of Z-Ibu-OH analogously to Example 4a), in the presence of 945 mg (7 mmol) of HOBt and 1.28 ml (10 mmol) of N-ethylmorpholine were stirred for 4 h. Work-up analogous to Example 1z). Yield 6.47 g (80 %) m.p. 209-215 ° (decomp.), Uniform by chromatography.

d) H ₂ N-CH ₂ -CH ₂ -S-BzIOMe

11.5 g (0.1 mol) of cysteamine HCl are dissolved in 10 ml of water. 5.6 g (O.I mol) of KOH are added to 100 ml of methanol and 11.5 ml Acetoacetic ester and refluxed for 10 min. After filtration, the filtrate is in vacuo. brought to dryness, the residue in 100 ml of dimethylformamide with 15.6 g (0.1 mol) of 4-methoxybenzyl chloride stirred overnight. The solvent is distilled off and the residue is partitioned between sodium hydrogen carbonate and ethyl acetate. The ethyl acetate phase is separated off, the

509844/1105 / ₃₀

2416G48

Ethyl acetate in vac. distilled off. The residue is left with Stand 1 ^ 0 ml of 1N HCl in methanol for 10 minutes, the distilled Methanol in vac. and distribute the residue between soda solution and ethyl acetate.

The ethyl acetate phase is again separated off and the ethyl acetate is distilled off. 15% oil is obtained.

e) Boc-Ser (BzI) -NH-CH ₂ -CH ₂ -S-BzIOMe

12 g (60 mmol) of the amine prepared according to d) are reacted in J> 0 ml of dimethylformamide with 18 g (60 mmol) of Boc-Ser (BzI) -OH and the equivalent amount of DCCI and HOBt. After standing overnight, the procedure is analogous to Example 1c) and 23.1 g of oil is obtained, which chromatographically represents an almost uniform substance.

f) H-Ser (BzI) -NH "CH ₂ -CH ₂ -S-Bzl0Me * HCl

The compound obtained according to e) is stored for 20 minutes in 4N HCl in dioxane, then the solvent is distilled off in vacuo. and rub the residue with ether. Yield 17 7 Ss m.p. 91 °, / ° £ 7p ° : + 9.6 ° (c = 1, methanol)

g) Boc-Thr (BzI) -Ser (BzI) -NH-CHg-CH ₂ -S-BzIOMe

8.7 g (28 mmol) of Boc-Thr (BzI) -OH and 11.6 g (28 mmol) of the after f) the compound produced are in the presence of equivalent amounts of N-ethylmorpholine, PIOBt and DCCI in 80 ml of dimethylformamide Stirred for 4 h. The procedure is analogous to Example 1c) and 17.5% of an oily compound is obtained which, by chromatography is almost uniform.

h) H-Thr (BzI) -Ser (BzI) -NH-CH ₂ CH ₂ -S-BzIOxMe.HCl

509844/1105 / ₃₁

The substance obtained according to g) is kept for 20 minutes in 100 ml of 4N HCl in dioxane, and the solvent is distilled in vacuo. and rub the residue with ether. After reprecipitation from isopropanol / ether 13 g of melting point I33-135 ° (decomp.) °. _ ₂₉ . ₇ ° ( _{c =} -j ^ methanol)

i) Boc-Trp-Lys (Z) -Thr-Phe-Thr (BzI) -Ser (BzI) -NH-CH ₂ -CH ₂ -S-BzIOMe

8.3 g (10 mmol) of Boc-Trp-Lys (Z) -Thr-Phe-OH and 5.95 g (10 mmol) of the compound prepared according to h) are dissolved in 80 ml of dimethylformamide in the presence of 2.7 g of HOBt and 1.28 ml of N-ethylmorpholine implemented with 2.2 g DCCI. It is worked up as described under Example 1n) and 9 × 8 g (71 %) of the protected hexapeptide are obtained. The compound is chromatographically uniform.

k) H-Trp-Lys (Z) -Thr-Phe-Thr (BzI) -Ser (BzI) -MI-THCl ^! ——

The compound obtained according to i) is kept for 20 min. In 80 ml of 4N HCl in dioxane which contains 4 g of thiophenol, and is distilled the solvent in vac. and rub the residue with ether * yield. almost quantitative.

l) H-Ibu-Gly-Cys-Lys-Asn-Phe-Phe-Trp-Lys-Thr-Phe-Thr-Ser-NH

I. Q_r> u _m

, -CH ₂

5.75 g (5 mmol) of Z-heptapeptide, prepared according to c), and 7.05 g (5 mmol) of H-hexapeptide hydrochloride, prepared according to k), in the presence of 1.35 g (10 mmol) of HOBfc and 0.64 ml (5 mmol) N-ethylmorpholine in 100 ml of dimethylformamide with 1.55 g (7.5 mmol) of DCCI reacted overnight. Working up according to Example 1aa) gives 8.05 g (63 p) of protected tridecapeptide. The cleavage groups, oxidation and purification are carried out according to Example 1bb).

5 09844/1105

Yield 980 mg of the pure cyelosic disulfide. Amino acid analysis: Asp 1.04, Ser O.85 ^x \ Thr I.69 ^X ', GIy 1-00 Cys 0.86 ^x ), Ibu 1.02, Phe 3.01, Lys 2.02 Trp (spectrophotometric) 0.95

'not corrected

Example 8

D-Ala-Gly-Cys-Lys-Ala-Phe-Phe-Trp-Lys-Thr-Phe-Thr-Ser-Cys-liFip

a) Z-AIa-Phe-Phe-OMe

17.85 g (^ 9 mmol) of H-Phe-Phe-OMe «HCl, 12.2 g (55 mmol) of Z-AIa and 12.2 g (59 mmol) of DCCI are dissolved in 150 ml of dimethylformamide in the presence of 8.0 g (59 mmol) of HOBt and 6.6 ml (5I mmol) of N-ethylmorpholine reacted with one another. After stirring overnight, the urea is filtered off, the filtrate is concentrated to a small volume, precipitated with water, the precipitate is reprecipitated from ethyl acetate / petroleum ether and crystallized from ethanol. Yield 25.9 &, m.p. 195-196 °

b) H-Ala-Phe-Phe-OMe'HCl

The compound obtained according to a) is catalytically analogous to Example 1r) hydrogenated. Yield almost quantitative. M.p. 212-215 °.

c) Boc-Lys (Z) -Ala-Phe-Phe-OMe

11.0 g (29 mmol) of Boc-Lys (Z) -OH and 11. 3 g (26 mmol) H-Phe-Phe-0ME "HCl are dissolved in 100 ml of dimethylformamide in the presence of 5.35 ml (26 mmol) of N-ethylmorpholine and 3.85 g (29 mmol) of HOBt with 5.95 g (29 mmol) of DCCI stirred overnight. One works

509844/1105

analogous to Example 1c) and isolated the initially oily residue by precipitation from methanol / ether. Yield 14.6 g, m.p.

d) Boc-Lys (Z) -Ala ~ Phe-Phe-OH

11.7 S of the methyl ester are dissolved in 150 ml of methanol and saponified by adding 18.5 ml of 1N NaOH. The methanol is then distilled in vacuo. off, the aqueous residue is added with conc. Citric acid solution at 0 °, filter off the crystalline precipitate and wash it with water. Yield 10.8 g, Very np. 14O - 144 °.

e) H-Lys (Z) -Ala-Phe-Phe-OH

10.86 g of the Boc compound are stirred in 50 ml of trifluoroacetic acid at room temperature for 20 minutes. After cooling, it is precipitated with ether, the precipitate is taken up in methanol and precipitated with -0.5 N NaHCO ₃ .

Wash with water and dry. Yield 8.59 S * m.p. 200 202 ° (Decomp.)

f) Boc-Cys (BzIOMe) -Lys (Z) -Ala-Phe-Phe-OH

7.O g (IO.9 mmol) H-Lys (Z) -Ala-Phe-Phe-OH and 6.3 g (12 mmol) Boc-Cys (BzIOMe) -0Tcp are in 40 ml of dimethylformamide in the presence of 1.4 ml (10.9 mmol) of N-ethylmorpholine and 1.47 g (10.9 mmol) HOBt stirred for 3 h. You acidify with 5 percent. KHSO ^ at and falls with water. The dried precipitate is boiled with ethyl acetate and triturated with ether. Yield 7 18 g, M.p. 213-215 °.

g) H-Cys (BzIOMe) -Lys (Z) -Ala-Phe-Phe-OH, CR ₅ COOH

6.7 g of the Boc compound prepared according to f) are 45 min.

509844/1105

2A16048

In 40 ml of trifluoroacetic acid, which contains 1 ml of mereaptoethanol, stirred at room temperature. While cooling with ice, it is precipitated with ether, suctioned off and washed with ether. Yield 6.8 g, m.p. 200-210 ° (decomp.)

h) Z-D-Ala-Gly-Cys (BzIOMe) -Lys (Z) -Ala-Phe-Phe-OH

6.8 g (6.9 mmol) of the pentapeptide prepared according to g) will be in 50 ml of dimethylformamide for 2 h with 5.55 g (7.7 mmol) of Z-D-AIa-Gly-OTcp, prepared according to Example 4a), in the presence of 1.98 ml (15.5 mmol) of N-ethylmorpholine and 0.94 g (6.9 mmol) of HOBt implemented. A crude product is precipitated with ether, which is taken up in dimethylformamide and treated with 5 percent. KHSOu / water like will. The precipitate is washed with water and dried. Yield 6.5 g, m.p.> 213> (Decomp.)

i) H-D-Ala-Gly-Cys-Lys-Ala-Phe-jErp-Lys-Thr-Phe-Thr-Ser-Cys-iTHp

(Phe \

'2 # 5 S (2 mmol) of the heptapeptide prepared according to h) are analogously to Example 1aa) with 2.64 g of heptapeptide, prepared according to Example 1p), in the presence of 2 equivalents each of N-ethylmorpholine, HOBt and O.618 g (3 mmol ) DCCI reacted in 45 ml of dimethylformamide for 18 h at room temperature. Work-up analogous to Example 1aa). Yield 3.8 g (78 %). Almost uniformly chromatographically.

The compound is freed from the protective groups, dehydrated and purified analogously to Example 1bb). Yield 4.6 g as triacetate.

Amino acid analysis / Ser O.87 ^X % Thr 1.77 ^X ', GIy 1.00, Cys 1.63 AIa 2.01, Phe 2.98, Lys 2.02
Trp (spectrophotometric) 0.95

'not corrected

B 0 9 8 4 U 1 1 1 0 5/35

Preparations to be administered parenterally: Example 9

50 mg of a peptide prepared according to the examples are dissolved 1 to 8 ^ in 50 ml redist. ¥ water and mixed with 10 ml of 1N Phosphate buffer pH 4.5j then give the calculated amount NaCl to isotonic and make up to 100 ml with water. After sterile filtration, ampoules of 1 or 2 ml are filled and lyophilized.

Example 10

A solution is prepared according to Example 9, but before filling up with a barrel of 0.25 S ^ -hydroxybenzoic acid methyl ester is added. After sterile filtration, ampoules of 1 or 2 ml are filled.

Example 11

Dissolve 10 g of zinc sulfate in 100 ml of 1 percent. Acetic acid, which is adjusted to pH 5 with sodium hydroxide solution. Separately, 50 g of protamine sulfate and 2 g of methyl hydroxybenzoate are dissolved in 700 ml of water, the pH also being set to 5. The two solutions are combined, the solution of 1 g of a peptide is added; prepared according to Examples 1 to 8 _; in 100 ml of water and make up to 1.0 l with water. After sterile filtration, ampoules of 1 or 2 ml are filled. .

Example 12

A solution is prepared according to Example 11, but the ^ -hydroxybenzoic acid ester is replaced by 15 g of benzyl alcohol and is obtained making up a suspension with water after sterile filtration

5 0 9 8 A 4/1 1 0 5/36

by adding sodium hydroxide solution to pH 7 · 8 with stirring. then Make up to 1 «ο 1 with water and distribute the suspension on ampoules of 1 or 2 ml.

Example 13

3 g of polyphloretin phosphate are dissolved in 50 ml of water while adjusting the pH to 6.8 using 2N NaOH. At the same time, 100 mg of a peptide, prepared according to Examples 1 to 8, 200 mg NaCl and 3 S gelatin derivative, prepared according to German Patent 1,118,792, Example 1 and then lyophilized, are dissolved in 40 ml of water, the solutions are combined and made up to 100 with water ml on. After sterile filtration, ampoules of 1 or 2 ml are filled and, if necessary, lyophilized.

example lh

It is analogous to Example 13, a solution is prepared which g in 100 ml 5.0 Polphloretinphosphat, 5 · 0 g gelatin derivative and 500 ^m S of a peptide prepared according to Examples 1 to 8, contains.

Preparations to be administered intranasally: Example 15

In 8 l of distilled water, without heating, R1.2 g of NaH ₂ PO ^. 2 H ₂ O, 66.29 g Na ₂ HPO ^, 25 g sodium chloride and 100 g benzyl alcohol dissolved Then 500 S polyvinylpyrrolidone with a k value of 85 to 59 (eg Kollldon ^ ^R * 90 from BASF) is added and stirring solved. Finally, 200 g of a peptide, prepared according to Examples 1 to 8, are added and dissolved cold. It is made up to 10 l with distilled water. The solution is filtered.

4-4 / 11 OS

Example i6

The procedure is as in Example 15, but instead of polyvinylpyrrolidone, polyvinyl alcohol with a K value of about 90 (for example Mowiol ^ ^R 'N 90-98 from Hoechst) is used.

Example 17

9 1 distilled lasser "are heated to the boil and in it 20g

Dissolved p-hydroxybenzoic acid methyl ester. The mixture is cooled to about 30 ° C., 89.58 g of Na ₂ HPO ^, 35.1 g of citric acid, 10 g of sodium chloride and 250 g of mannitol are added and 100 g of a peptide prepared according to Examples 1 to 8 are then dissolved therein , on. Make up to 10 l with distilled water and filter the solution.

Example 18

50 S methyl cellulose with an average degree of substitution of approx. 1.5 and a viscosity of 2700 to 4000 cP (Tylose '' MH 4000 ρ der Farbwerke Hoechst Ag). The suspension is allowed to cool to room temperature. This causes d: b methyl cellulose to swell. Then a mixture of 400 ml of η-acetic acid and 282 ml of η-sodium hydroxide solution is added, then 350 g of mannitol, 500 mg of benzalkonium chloride and 200 g of a peptide, prepared according to Examples 1 to 8, are dissolved and then made up to 10 with distilled water. The solution is filtered.

Example 19

The procedure of Example 18 is followed, but instead of methyl cellulose, hydroxyethyl cellulose is used.

50984 ^ / 1105/38

Example 20

The procedure of Example 18 is repeated, but using hydroxypropylmethyl cellulose instead of methyl cellulose.

Example 21

In 9 liters of distilled water, 35 * 44 S citric acid, 32.76 g Na HPO ^, 400 g sorbitol and 40 g chlorobutanol are dissolved without heating. Then 400 g of a peptide prepared according to Examples 1 to 8 / are dissolved. The mixture is made up to 10 l with distilled water and the solution is filtered.

Example 22

In 6 l of distilled water from cd. At 50 ° C., 200 g of methyl cellulose with an average degree of substitution of approx. 1.5 and a viscosity of 2700 to 4000 cP (eg Tylose ' ^R ') are suspended. The suspension is allowed to cool to room temperature with stirring. The methylcellulose swells in the process. 400 ml of η-acetic acid, 282 ml of n-sodium hydroxide solution, 350 S Manit, 500 mg of benzalkonium chloride and finally 100 g of a peptide / prepared according to Examples 1 to 8 are dissolved in 3 l of distilled water. The solution is filtered and combined with the methylcellulose solution. Distilled water is used to make up to 10 liters.

Example 23

400 ml of η-acetic acid, 282 ml n sodium hydroxide solution, 50 g of sodium chloride and 100 g of benzyl alcohol are added. 100 g of a peptide ^ are produced in this mixture after, the Examples 1 to 8 ^ solved. It is distilled with Water filled to 10 barrel, the solution is filtered.

509844/1105 / 39 '

~ 39 -

Example 2k

10Og microfine peptide produced according to Examples 1 to 8, Λΐerden made up to 1 liter with liquid paraffin and homogenized.

Example 25

10g of benzyl alcohol are mixed with 2-octyldodecanol (e.g. eutanol ^ 'G

Henkel u.Cie, Düsseldorf) made up to 0.990 1. To do this there one 10g microfine peptide, prepared according to the examples 1 to 8 / and homogenized.

Example- 26

k0 g microfine peptide, prepared according to Examples 1 to 8 _y and 10 g benzyl alcohol are made up to 1 l with a triglyceride mixture of saturated vegetable fatty acids of medium chain length (e.g. Miglyol '', Dynamit Nobel, Cologne) and homogenized.

Example 27

20 g of microfine peptide, prepared according to Examples 1 to 8,

10 g of benzyl alcohol and 40 g of sorbitan sesquioleate (e.g. Arlacel ^ '83, Atlas Chemie GmbH, Essen) are made up to 1 liter with an oleic acid oleyl ester (e.g. Cetiol'', Henkel and Cie, Düsseldorf) and homogenized.

Bei p iel 28

10 g microfine peptide, produced according to Examples 1 to 8 / and 3 0 g of benzyl alcohol are made up to 1 l with isopropyl myristate and homogenized.

50984A / 1105

Example 29

20 g of a mixture of aluminum di- and monostearates (eg Alugel ^ '44 Μ) are suspended in 0.6 liters of a triglyceride mixture of saturated vegetable fatty acids of medium chain length (eg Miglyol * ¹ ' 812). The suspension is heated to 150 ° C. and kept at this temperature for 15 minutes. Then allowed to cool to room temperature. In a mixture of 0.3 l of the above triglyceride mixture and 10 g of benzyl alcohol, 60 g of microfine peptide / prepared according to Examples 1 to 8 ^ are suspended. Both partial quantities are combined and the final volume is brought to 1 ί with a triglyceride mixture of saturated vegetable fatty acids of medium chain length.

Example 30

.and

100 g of anhydrous lanoliny, 440 g of petroleum jelly are melted together. A suspension of 100 g of microfine peptide prepared according to Examples 1 to 8 in 35Ο g of liquid paraffin is added to the cooled melt. Finally, 10 g of benzyl alcohol are added and the ointment is homogenized.

Example 31

A suspension of 50 g of a peptide ^ prepared according to the Examples 1 to 8 / in 350 g of liquid paraffin is 6Ό0 g Vaseline combined and the ointment homogenized.

Example 32

30 g of wool wax alcohol, 1 60 g of liquid paraffin, 40 g of solid paraffin and 370 g of petroleum jelly are melted together at 75 ° C. until they become clear. 300 g of distilled water are heated to the boil and therein 0.72 g of methyl p-hydroxybenzoate and 0.08 g

5098 44/1105 / 41

Propyl p-hydroxybenzoate dissolved. After the Solution at 80 C it is stirred into the 75 C hot fat melt. The resulting emulsion is stirred while cold. In 97 »2 g of distilled 20 g of a peptide prepared according to Examples 1 to 8 ^ are dissolved in water. The solution becomes the emulsion given and the emulsion homogenized.

Example 33

60 g of sorbitan sesquioleate (e.g. Arlacel ^ '83), 130 g of liquid paraffin, 5OO g of Vaseline and 10 g of cetyl alcohol are melted together at 75 ° C until clear. 200 g of distilled ¥ ater are heated to boiling and is 0, ^ kg methyl p-hydroxybenzoate and dissolved 0.06 g p-Hydroxybenzoesöurepropylester. After the solution has cooled to 80 C, it is stirred into the 75 C hot molten fat. The resulting emulsion is stirred cold. 10 g of a peptide, prepared according to Examples 1 to 8, are dissolved in 98.4 g of distilled water. The solution is added to the emulsion and the emulsion is homogenized.

example

To a suspension of 10 mg of a peptide ^ prepared according to Examples 1 to 8; and 10 mg sorbitan oleate are placed in a pressure vessel either in the cold filling process before closing the container or in the pressure filling process after closing the container, a mixture of 60 mg 1,2,2-trifluorotrichloroethane (e.g. Frigen ^ ' 113), 120 mg trichloromethane (e.g. Frigen ' ^R ' 11) and 800 mg Di chi or di fluorine than / 1, 2, 2-tetrafluoro-dichloroethane (e.g. Frigen ' ^R ' 12/114) such as 60: 4θ.

509844/1105

Example 35

100 g of anhydrous lanolin and 500 g of petroleum jelly are melted together. A suspension of 20 g of microfine peptide prepared according to Examples 1 to 8 is stirred into the cooled melt _; in 380 g of liquid paraffin. The ointment is homogenized.

According to the examples 9-35 corresponding preparations can be prepared for all falling under the general formula II peptides.

50984A / 11G5

Claims (18)

Claims 1.)) Peptides of the general formula II X-CH-CO- ¥ -V-Phe-Phe-Trp- ¥ -Thr-Phe-Thr-Ser-NH-CH-Y in which X stands for H, ß-Ala-NH, (<-Methyl) -AIa-NH, D-AIa-NH, D-Ala-Gly-NH. ß-Ala-Gly-NH, Ala-D-Ala-NH, where existing OL -amino groups can carry an alkanoyl or alkyloxycarbon radical with 1-6 carbon atoms in the alkyl part, V can be asparagine or alanine, ¥ lysine or ornithine and Y is H, COOH, CONH ₂ , CONHR or CONR ₂ , where R is an alkyl radical with 1-6 carbon atoms or a cycloalkyl radical with 5 or 6 carbon atoms and both substituents R can also jointly represent tetramethylene or pentamethylene. 2.) Process for the preparation of the peptides of the general formula II X-CH- ¥ -V-Phe-Phe-Trp- ¥ -Thr-Phe-Thr-Ser-NH-CH-Y OH ₂ -S -: ■ -S-OH ₂ (ii) characterized in that one of the peptides of the general formula V X-CH-CO- ¥ -V-Phe-Phe-Trp- ¥ -Thr-Phe-Thr-Ser-NH-CH-Y CH ₂ -SB CH ₂ -SB (V) 509844/1105 l ^hh in which V, W, X, Y and B are those mentioned in claims 1 and 2 Have meaning, amino groups by removable N-protective groups are closed, OH groups can be etherified with benzyl radicals and any existing ones Carboxyl group is present as a benzyl ester, the protective groups by treating with sodium in flux. Ammonia or hydrogen fluoride splits off and the intramolecular disulfide bridge with oxygen in the case of an alkaline reaction or by Potassium ferricyanide produces in the usual way. 3.) Process for the preparation of peptides of the general formula II X-CH-CO-W-V-Phe-Phe-Trp-W-Thr-Phe-Thr-Ser-NH-CH-Y CH ₂ -S _: S-CH ₂ (II) characterized in that there is an N-terminal amino acid or an N-terminal peptide which corresponds to the amino acid sequence of the peptides of the general formula V X-CH-CO-W-V-Phe-Phe-Trp-W-Thr-Phe-Thr-Ser-NH-CH-Y CH ₂ -SB CH ₂ -SB (V) correspond, but are shortened by one or more amino acid building blocks, with that of the formula V, complementary C-terminal peptide or the C-terminal Amino acid condensed according to the known methods of peptide chemistry, where V, W, X, Y and B are those in claim 1 and Have the meaning mentioned, OH groups can be etherified with benzyl radicals, amino groups can be split off N-protective groups are closed and a possibly 4/1105 / 45 existing carboxyl group is present as a benzyl ester, then from the protected peptide of the general formula V, the protective groups by treatment with sodium in liquid ammonia or hydrogen fluoride is split off and the intramolecular disulfide bridge is split off by oxidation with oxygen with an alkaline reaction or with potassium ferricyanide produces in a manner known per se. H. ) Process for the preparation of the peptides according to Claim 1, characterized in that a protected peptide of the formula III _t X-CH-CO-WV-Phe-Phe-OH CH ₂ -SB (III) with a protected peptide of formula IV H-Trp-W-Thr-Phe-Thr-Ser-NH-CH-Y OH ₂ -SB (IV) ■ where in formula III and IV V, W, X, Y, those mentioned in claim I. Have meaning and B for benzyl or 4-methoxybenzyl is any carboxyl group present through benzyl, hydroxyl groups wholly or partially through benzyl radicals and amino groups are protected by optionally chlorine-substituted benzyl oxycarbonyl radicals, condensed, the protective groups are split off and removed by treatment with hydrogen fluoride or with sodium in liquid ammonia the resulting bis-mercapto compound is the cyclic disulfide of the general formula II by oxidation or Hydrogenation with air at alkaline pH or with potassium ferricyanide. 50 9 844/1105 5.) Ordering medicaments from or containing a peptide according to claim 1. 6.) Process for the production of medicaments consisting of or containing a peptide according to claim 1, characterized that a peptide according to claim 1, optionally together with conventional pharmaceutical excipients, buffer solutions and / or antibacterial additives, in a form suitable for therapeutic purposes brings. 7.) Pharmaceutical preparation for nasal application, marked by a content of a) a peptide according to claim 1 or its physiologically acceptable salt b) an aqueous buffer solution of pH 4 to -7j2 c) Isotonic addition d) if necessary a preservative ^e ) g & f · a polymeric adhesive substance. 8.) Process for the manufacture of a pharmaceutical preparation for nasal application, characterized in that an isotonicity-producing additive, optionally a preservative and / or optionally egg ne polymeric adhesive substance and a peptide in it according to claim 1 or its physiologically acceptable salt dissolves. 9.) Pharmaceutical preparation for nasal application, marked by a content of a peptide according to claim 1 or its physiologically acceptable salt as an active ingredient in the form of an oily suspension or fatty ointment. / 47 509844/1105 - kl - 10.) Preparation according to claim 9, characterized in that the active ingredient is in an oily suspension, which also contains a preservative and / or a surface-active agent Substance with an HLB ¥ ert below 10 and / or aluminum stearate (s) as a swelling agent and / or a propellant the group of halogenated hydrocarbons. 11.) Preparation according to claims 9 and 10, characterized in that that mixtures of fluorinated chlorinated hydrocarbons are used as propellants. 12.) Preparation according to claim 9> characterized in that the active ingredient is present in a fatty ointment that is next to a soft, spreadable, anhydrous fat mixture, possibly preservatives and / or surface-active substances with an HLB value of less than 10. 13.) Preparation according to claim 9> characterized in that the active ingredient is dissolved or suspended in a soft, spreadable water-based emulsion ointment, which possibly contains a preservative and / or surface-active substances with an HLB value below 10. 14.) Process for the production of a pharmaceutical preparation for nasal application, characterized in that one a) an oil, possibly a preservative, a surfactant with an HLB value below 10 and / or contains aluminum stearates as swelling agents, microfine peptide according to claim 1 its physiologically compatible salt adds and optionally with a Filling halogenated hydrocarbons into a pressurized gas pack, or 50984A / 110 b) a soft, spreadable fatty base ointment, which may contain a preservative and / or a surface Substance with an HLB value below 10 contains microfine peptide according to claim 1 or its physiologically tolerable one Salt added and homogenized or c) a soft, spreadable emulsion ointment, which may be contains a preservative and / or a surfactant with an HLB value below 10, a aqueous solution of a peptide according to claim 1 or that of its physiologically acceptable salt added as active ingredient and homogenized. 15.) Pharmaceutical preparation with protracted effect, marked by a content of a) a peptide according to claim 1 b) an aqueous solution of a gelatin derivative crosslinked with diisocyanate, prepared according to the German Patents 1,118,792 or 1,155,13 and c) polyphlorine phosphate 16.) A process for the production of a pharmaceutical preparation with a protracted effect, characterized in that that he a) an aqueous solution of a peptide according to claim b) an aqueous solution of a diisocyanate crosslinked gelatin derivative, prepared according to German patents 1118792 or 1 155 13 ¹ * - and c) an aqueous solution of polyphlorine phosphate combined with one another and the resulting solution lyophilized. 09844/1105 17 ·) Pharmaceutical preparations with a protracted effect, marked by a content of a) a peptide according to claim 1 b) an aqueous solution of water-soluble zinc salt c) an aqueous protamine sulfate solution. 18.) A process for the production of a pharmaceutical preparation with a protracted effect, characterized in that that he a) an aqueous solution of a peptide according to claim 1 b) an aqueous solution of water-soluble zinc salt and c) an aqueous protamine sulfate solution combined with one another and the resulting solution is adjusted to a pH of 3> 5 to> is 6.5. 19 ·) The method according to claim 18, characterized in that according to claim 18 from the three components a) to c) the solution initially obtained is converted into a suspension by adjusting the pH to 7> 5 to 8. 509844/1105