CA2005052A1 - Tnf peptides - Google Patents

Abstract

ABSTRACT OF THE DISCLOSURE:

Peptides of the formula X-A-B-E-Leu-Y, where A, B, E, X and Y are defined in the description, and the preparation thereof are described. The novel peptides are suitable for controlling diseases.

Description

Z0~105~2 O. Z . 0050/40381 NOVEL TNF PEPTIDES

The pre~ent invention relate~ to novel ~eptides derived from tumor necro~is factor (TNF), the preparation thereof and the use thereof as dru~s.

Carswell et al. (Proc. Natl. Acad. Sci. USA 72 (1975) 3666) reported that the serum of endotoxin-treated animals which had previously been infected with the Calmette-Guerin strain of Mycobacteria (BCG) brought about hemorrhagic necro~i~ in variou~ mou~e tumors. This activity was a~cribed to tumor necroYis factor. TNF al~o has a cytDstatic or cytotoxic effect on a large number of tranaformed cell lines in vitro, whereas normal human and animal cell line~ are unaffected (Lymphokine Report~ Vol.

2, pp 235-275, Academic Press, New York, 1981). Recently, the biochemical characterization and tha gene for human TNF have been deqcribed (Nature 312 (1984~ 724, J. Biol.
Chem. 260 tl985) 2345, Nucl. Acid~ Res. 13 (1985) 6361).

It i~ po~ible to deduce from this data the following protein structure for mature human TNF:

V ~ y~nVal~ sValV~U~aAs~o GLnA~u~_~

GlnVa~LæheLysGay~L~yCy~Pn~beqIo~Val~Le~:~h~ le S~gIleAlaVal~noyrG~ f n~ LysV~lA~edle~nt1aIleLyg~Pro Cy~G~yGlulhrP~ yA~ualaLy~el~prpCIuPr~Ile~Ieu GlyGayV~L~YGln~Ju~y~GayA pAn~euS~LU~fluIleA~rgPI~o TylJ~4FPhe~ u~ayGanV~lTyrPb~yIleIle~eu The TNF gene~ of cattle, rabbits and mice have also been de~cribed (Cold Spring Xar~or Symp. Quant . Biol . 51 (1986) 597).

Z01~505Z
- 2 - O.Z. 0050/403~1 Besides it~ cytotoxic properties, TNF is one of the main substances involved in inflammatory reactions (Pharmac.
Res. 5 (1988) 129). AnLmal models have shown that TNF is involved in septic shock (Science 229 (1985) 869) and graft-versus-host disease (3. Exp. Med. 166 (1987) 1280).

We have now found that peptide~ with a considerably lower molecular weight have beneficial propertie~.

The present invention relates to peptides of the formula I
X-A-B-E-Leu-Y I, where A is Glu, Pro or Gln, B is Gly, Glu, A~n or Asp, E is Gln or Ser, 5 X i~ G-NH-CHM-CO-, G-NH-CHM-CO-W-, G-R-NH-CHM-CO- or G-R-NH-CHM-CO-W- and Y i9 -Z, -NH-CHQ-CO-Z, -V-NH-CHQ-CO-Z, -NH-CHQ-CO-U-Z
or -V-NH-CHQ-CO-U-Z, where, in X and Y, G i~ hydrogen or an amino-protective group, Z i~ OH or NH2 or a carboxyl-protective group, or G and Z together are also a covalent bond or -CO-~CH2),-NH- where a i~ from 1 to 12, R and U are peptide chain~ composed of 1 5 naturally occurring ~-amino acidq, W i8 one of the peptide chain~
-~ys-Pro-Val-Ala-His-Val-Val-Ala-Asn-Pro-Gln-Ala-, -Lys-Pro-Val-Ala-Hi~-Val-Val-Ala-Asp-Ile-A~n-Ser-, -Ly~-Pro-Leu-Ala-Hi~-Val-Val-Ala-A~n-Pro-Gln-Val-, -Lys-Pro-Val-Ala-Hi~-Val-Val-Ala-Asn-His-Gln-Val-, -Lys-Pro-Al~-Ala-Hi8-Leu-Ile-Gly-Asp-Pro-Ser-Lys-, -Lyg-Pro-Ala-Ala-His-Leu-Val-Gly-~ p-ProSer-Thr-, -Lys-Pro-Ala-Ala-His-Leu-Val-Gly-Tyr-Pro-Ser-~ys-, a section compo~ed of 5 to 11 2mino acids of one of the abovementioned peptide chain~, or a peptide ,~o~iS()5Z

- 3 - O.z. 0050/40381 chain with 1-4 naturally occurring ~-amino acid~, is one of the peptide chains -Gln-Trp-Leu-Asn-Arg-Arg-Ala-Asn-Ala-Leu-Leu-Ala-, -Arg-Trp-Trp-A~p-Ser-Tyr-Ala-Asn-Ala-Leu-Met Ala-, S -Gln-Trp-Leu-Ser-Gln-Arg-Ala-Asn-Ala-Leu-Leu-Ala-, -Glu-Trp-Leu-Ser-Gln-Arg-Ala-A~n-Ala-Leu-~eu-Ala-, -Leu-Trp-Arg-Ala-Asn-Thr-Asp-Arg-Ala-Phe-Leu-Gln-, -Arg-Trp-Arg-Ala-Asn-Thr-Asp-Arg-Ala-Phe-Leu-Arg-, -Leu-Trp-Arg-Ala-Ser-Thr-Asp-Arg-Ala-Phe-Leu-Arg-, a section composed of 5 to 11 amino acids of one of the abovementioned peptide chain~, or a peptide chain with 1-4 naturally occurring ~-amino acid~, and M and Q are hydrogens or one of the following -CH(C~3)2, -C~(CH3)-C2H5, -CbH5, -CH(OH)-CH3, or (with b being from 1 to 6 and T being hydrogen or OH, CH~O, CH3S, ~CH3)~CH, C6Hs, p-HO-C6H4, HS, H2N, HO-CO, H2N-CO or H2N-C(=NH)-NH) or M and Q together are a -(CH2)c~s-s-(cH2) d- ~ - ( CH2 ) ~-CO-NH-(CH2)~- or -(CH2).-NH-CO-(CH2)~-NH-CO-(CH2)~- bridge (with c and d being from 1 to 4, e and f being from 1 to 6 and g being from 1 to 12), as w~ll a~ the salts thereof with physiologically toler-ated acid~.

2S The peptide~ of the formula I are con~tructed of L-amino acids, but they can contain 1 or 2 D-amino acids. The side-chains of the trifunctional amino acid~ can carry protective groups or be unpro~ected.

Particularly pr~ferred physiologically tolerated acids ares hydrochloric a~id, citric acid, tartaric acid, lactic acid, phosæhoric acid, m~thanesulfonic acid, acetic ac$d, formic acid, maleic acid, fumaric acid, ,~o~3505Z

- 4 - O.Z. 0050/40381 malic acid, succinic acid, malonic acid, sulfuric acid, L-glutamic acid, L-aspartic acid, pyruvic acid, mucic acid, benzoic acid, glucuronic acid, oxalic acid, ascor-bic acid and acetylglycine.

The novel peptides can be open-chain (G = H, amino-protective group; Z = OH, NH2, carboxyl-protective group M and Q not connected together) and, in particular, have a disulfide bridge (G = H, amino-protective group;
Z = OH, NH2, carboxyl-protective group; M + Q = -tCH~)C-S-S-(CH2)d ) or a side chain bridge (G = H, amino-protec-tive group, Z = O~, NH2, carboxyl-protective group, M + Q
= - ( CH2 ) .-NH-CO- ~ CH2 ) r~ or -(CH2)~-NH-CO-(CH2) 3-NH-CO_ (CH2)~-) or be linked head-to-tail (G + Z = covalent bond or -CO-(CH2)~-NH-).

Tha novel compounds can be prepared by conventional methods of peptide chemistry.

Thu~, the peptides can be conatructed sequentially from amino acid~ or by linking together suitable smaller peptide fra$ments. In the sequential con~truction, the peptide chain is extended ~tspwise, by one amino acid each time, 3tarting at tha C terminus. In the ca~e of coupling of fra~ments it is possible to link together fragment~ of different lengths, the~e in turn beinq obtainable by ~equential construction from amino acid~ or coupling of other fragment~. The cyclic peptide~ are obtained, after synthe~i~ of the open-chain peptides, by a cyclization reaction carried out in high dilution.

In the case both of sequential con~truction and of fragment coupling it is necessary for the building block~
to be linked by formation of an amide linkage.
Enzymatic and chemical method~ are 3uitable for this.

Chemical methods for forminq amide linkagas are dealt - 5 - O.Z. 0050/40381 wLth in detail by Muller, Methoden der Organischen Chemie (Methods of Organic Chemistry) Vol. XV/2, pp 1-364, Thieme Verlag, Stuttgart, 1974; Stewart, Young, Solid Phase Peptide Synthesis, pp 31-34, 71-82, Pierce Chemical Company, Rockford, 1984; Bodanszky, Klausner, Ondetti, Peptide Synthesis, pp 85-128, John Wiley & Sons, New Yor~, 1976 and other ~tandard work~ of peptide chemi~try.
Particularly preferred are the azide method, the symmetr-ical and mixed anhydride method, active esters generated in situ or preformed and the formation of amide linkages using coupling reagents (activators), in particular dicyclohexylcarbodiimide (DCC), diisopropylcar~odiimid~
(DIC), 1-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ), l-ethyl-3-(3-dLmethylaminopropyl)carbodiimide hydrochloride (EDCI), n-propanepho~phonic anhydride (PPA), N,N-bi~(2-oxo-3-oxazolidinyl)amidophosphoryl chloride (BOP-Cl), diphenylpho~phoryl azide (DPPA~, Castro'~ reagent (BOP), O-benzotriazolyl-N,N,N',N~-tetra-methyluronium salts (HBTU), 2,5-diphenyl-2,3-dihydro-3-oxo-4-hydroxythiophene dioxide (Steglich' 8 reagent;
HOTDO) and l,l'~carbonyldiimidazole (CDI). The coupling reagents can be employed alone or in combination with additive~ ~uch as N,N'-dimethyl-4-zminopyridine (DMAP), N-hydroxybenzotriazole (HOBt), N-hydroxybenzotriazine (HOOBt), N-hydroxy~uccinimide (HOSu) or 2- ffl droxy pyridine.

Wher~aa it i8 nor~ally possible to diqpen~e with protec-tive group~ in enzymatic peptide synthesi~, for chEmical synthe~i~ it i~ nece~ary for there to be rever~ible protection of the reactive functional group~ which are not involved in the formation of the amide linkaqe on the two reactants. Three conventional protective group techniqu2s are preferred for chemical peptide ~ynthe~es:
the benzyloxycarbonyl (Z), the t-butyloxycarbonyl (Boc) and the 9-~luorenylmethyloxycarbonyl (Fm~c) techniques.
In each cA~e the protective group on the -amino group of 20~505Z

- 6 - O.Z. OOSO/40381 the chain-extending building block i~ identified. The ~ide-chain protective groups on the trifunctional amino acids are choqen so that they are not necessarily elimin-ated together with the ~-amino protective group. A
detailed review of amino acid protective groups i~ given by MUller, Methoden der Organi~chen Chemie Vol XV/1, pp 20-906, Thieme Verlag, Stuttgart, 1974.

The building block~ used to construct the peptide chain can be reacted in solution, in suspension or by a method similax to that described by Merrifield in 3.~mer. Chem.
Soc. 85 (1963) 2149. Particularly preferred method~ are those in which peptide~ are constructed ~equantially or by fragment coupling by use of the Z, Boc or Fmoc protec-tive group technique, in which ca~e the reaction take~
lS place in solution, as well a~ those in which, similar to the Merrifield technique, one reactant i8 bound to an insoluble polymeric support (also called re~in herein-after). Thi~ typically entail~ the peptide beinq con-structed sequentially on the polymeric ~upport, by use of the Boc or Fmoc protective group technique, with the growing peptide chain being covalently bonded at the C terminu~ to the inqoluble re~in particles (cf. Figures 1 and 2). Thi~ procedure allow~ reagents and byproduct~
to be removed by filtration, and thus recry~tallization of intermediate~ i8 superfluous.

The protected amino acid~ can be bonded to any suitable poly~er~ which merely need to be insoluble in th~ ~ol-vents used and to ha~e a stable physical form which allows easy filtration. ~ha polymer must contain a functional group to which the first protected zmino acid can be firmly linked by a covalent bond. A wide variety of polymers i~ ~uitable for this purpose, for example cellulo~e, polyvinyl alcohol, polymethacrylate, sulfon-ated poly~tyrene, chloromethylated copolymer of ~tyrene and divinylbenzene (~errifield resin), 4-methylbenz-20~`~505.'Z

- 7 - O.Z. 0050/40381 hydrylamine-re~in ~MBHA-regin), phenylacetamidomethyl-resin (Pam-resin), p-benzyloxybenzyl alcohol-re~in, benzhydrylamine-re~in (BHA-resin), 4-hydroxymethyl-benzoyloxymethyl-resin, the resin used by Breipohl et al.
(Tetrahedron Lett. 28 (1987) 565; from BACHEM), HYCRAM
resin (from ORPEGEN) or SASRIN resin (from BA~HEM).

Solvents suitable for peptide synthesis in solution are all those which are inert under the reaction condition~, in particular water, N,N-dimethylformamide (DMF), dimethyl sulfoxide (DMS0~, acetonitrile, dichloromethane (DCM), 1,4-dioxane, tetrahydrofuran (THF), N-methyl-2-pyrrolidone (N~P) and mixtures of the said solvents.
Peptide synthesi~ on polymeric supports can be carried out in all inert or~anic solvents which dissolvs the amino acid derivatives used; however, solvents which have re~in-swelling properties are preferred, such a DMF, DCM, NMP, acetonitrile and DMS0, a~ well as mixtures of these 301vent~.

After the peptide has been synthe~ized it i8 cleaved off the polymeric support. The cleavage conditions for the variou~ types of resins are di~closad in the literature.
The cleavnge reaction~ mo~t commonly u~e acid and palladium cataly~i~, in particular cleava~e in anhydrous liquid hydrogen fluoride, in anhydrou~ trifluoromethane-sulfonic acid, Ln dilute or concentrated trifluoroacetic acid or palladium catalyzed cleavage in THF or r~r-DCM
mixtures in the presence of a weak base such aa morpho-line. The protective groups may, depending on the choice thereof, be retained or likewi~e cleaved off under the cleavage conditions. Partial deprotection of the peptide may also be worthwhile if the intention iR to carry out cextain derivatization reaction~ or a cyclization.

Some of the novel peptides have good cytotoxic proper-tis~. Some other8 of the peptides have high af~inity for ~0~50~i~

- 8 - O.Z. 0050~40381 ths cellular TNF receptor without, however, having cytotoxic acti~ity. They are therefore TNF antagonist~.
They compete with natural TNF for binding to the cellular TNF receptor and thus suppress the TNF effect. The novel peptides are valuable drugs which can be employed for treating neoplastic diseases and autoimmune diReases as well a~ for controlling and preventing infectiQns, inflammations and transplant rejection reactions. SLmple experiments can be used to elucidate the mode of action of the individual peptide~. The cytotoxicity of the peptide is determined by incubating a TNF-~en~itive cell line in the presence of the peptide. In a second experi-mental approach, the cell line i~ incubated with the relevant peptide in the presence of a lethal amount of TNF. It is pos~ible in this way to detect the TNF-antagonistic effect. In addition, the affinity of the peptide for the cellular T~F receptor i~ determined in an in vitro binding experiment.

The following te~t system~ were used to characterize the agonistic and antagonistic effects of the novel peptides:

I. Cytotoxicity test on TNF-sensitlve indicator cells t II. Cytotoxicity ~ntagonism te~t on TNF-sensitive indicator cells, III. Competitive receptor-binding test on indicator cells expre~ing TNF receptor.

I. Cytotoxicity test The ~gonistic effects of the novel peptide~ are a3~e~ed on tho ba~i~ of th~ir cytotoxic effect on TNF-sensitive cell~ (e.g. L929, MCF-7, A204, U937).
The test with L929 and ~CF-7 wa~ carried out as follow~s 1. 100 ~1 of culture medium containing 3 to 5 x 103 freshly tryp~inized, exponentially growing, L929 zo~o~z - 9 - 0 . ~ . 0050~40381 cell~ (mou~e) or MCF-7 cell~ (human) were pipetted into the well~ of a 96-well flat-bottom culture plate. The plate wa~ incubated at 37 C
overnight. The air in the incubator was saturated S with water vapor and contained 5~ CO2 by volume.

The L929 culture medium contained 500 ml of lx Earle's MEM (Boehringer Mannheim), 50 ml of heat-inactivated tS6C, 30 min~ fetal calf ~erum (FCS), 50 ml of L-glutamine (200 mM), S ml of lOOx non-e~ ential amino acids, 3 ml of lM HEPES
buf~er pH 7.2,a~d 50 ml of gentamicin (5~ mg/ml).

Th NCF-7 culture medium contained 500 ml of lx Dulbecco's MEM ( Boehringer Mannheim), 100 ml of heat inactivated ~S6C, 30 min) FCS, 5 ml of L-glutamine and 5 ml of lOOx nonessential amino acids.

2. The next day 100 ~1 of the pep~ide solution to bs tested were added to the cell cultures and sub~ected to serial 2-fold dilution. In addition, aome cell controls (i.e. cell culture~ not treated with peptide dilution) and ~ome rhu-TNF
control3 (i.e. cell cultures treated with recom-binant human TNF) were also made up. The culture plate was incubated at 37C in an atmo~phere of air satura~ed with water vapor and containing ~%
CO2 by Yolume for 48 h.

3. The p~rcentage of ~urviving cells in the cultures treated with peptide dilution was detarmined by ~taining with crystal violet. For this purpose, th~ liquid wa~ removed from the well~ of the test plate by tapping it. 50 ~1 of crystal violet solution wers pipetted into each we~l.

'~O~D5052 - 10 - O.Z. noso/403sl The composition of the crystal violet solution was a~ follows:

3.7~ g of cry~tal violet 1.75 g of NaCl 161.5 ml of ethanol 43.2 ml of 37$ formaldehyde water ad 500 ml The crystal violet solution was left in the wells for 20 min and then likewi~e removed by tapping.
The plates were then wa~hed 5 times by Lmmer~ion in wat~r in order to remove dye not bound to the cells. The dy~ bound to the cell~ wa~ extracted by adding 100 ~1 of reagent Yolution (50% etha-nol, 0.1% glacial acetic acid, 49.9% water) to each well.

4. The plate~ were shaken for 5 min o obtain a solution of uniform color in each well. The surviving cell3 were determin0d by mea~uring the extinction at 540 nm of the colored solution in the individual wells.

5. Subsequently, by relating to the cell control, the 50~ cytotoxicity value was defined, and the reciprocal of the 3ampl~ dilution which re~ulted in 50% cytotoxicity wa~ calculated a~ the cyto-toxic activity of the test sample.

II. Cytotoxicity antagoni~m test The sntagonistic effect of the peptid~ was asse~ed on the b sis of their property of antagonizing the cytotoxic effect of rhu-TNF on TNF-~ensitive cells (e.g. L929, MCF-7, A204, U937). The cytotoxicity antagonism test with L929 and MCF-7 cells wa~
carr~d out a~ follow~s Z0~5~35~

- 11 - O.Z. 0050/40381 1. 100 ~1 of culture medium containing 3 to S x 103 freshly trypsinized, exponentially growing, L929 cells (mouse) or MCF-7 cells (human) were pipetted into the wells of a 96-well flat-bottom culture plate. The plate was incubated at 37C
overnight. The air in the incubator was saturated with water vapor and contained 5% CO2 by volume.

The L929 culture medium contained 500 ml of lx Earle's MEM (Boehringer Mannheim), 50 ml ef heat-inactivated (56C, 30 min) FCS, 5 ml of L-gluta-mina (200 mM), 5 ml of lOOx non-essential amino acids, 3 ml of lM ~EPES buffer pH 7.2, and 500 ~1 of gentamicin (50 mg/ml).

The MCF-7 culture medium contained 500 ml of lx Dulbecco's MEM (Boehringer Mannheim), 100 ml of heat inactivated (56C, 30 min) FCS, 5 ml of L-glutamine (200 mN) and 5 ml of 100~ nonessen-tial amino acid~.

2. ~he next day 100 yl of the peptide solution to be tested were added to the cell cultures and sub~ected to serial 2-fold dilution. Then, 100 ~1 of a rhu-~NF dilution in culture medium, which dilution had an 80-100% cytotoxic effect in the final concentration in the cell culture, were added to these cell culture~. In addition, 30me c~ll control~ ~i.e. cell culture~ not treated with peptide ~olution or with rhu-~NF solution) and some rhu-TNF controls (= cell cultures treated only with rhu-TNP qolution) were also made up. The culture plate wa~ then ~ncubated at 37C in an atmo~phere of air ~aturated with water vapor and containing 5% CO2 by volume for 48 h.

3. Tha percentage of surviving cell~ in the culturQs Z0~35()52 - 12 - O.Z. 0050/40381 treated with su~stance dilution was determined by staining with crystal violet. For this p~rpo~e, the liquid wa~ r~mo~ed from the well~ of the test plata by tapping it. 50 ~1 of cry~tal violet solution were pipetted into each well.

The crystal violet solution had the composition specified in I.3 The crystal violet solution wa~ left in the well~
for 20 min and then likewi~e removed by tapping.
The plate~ were then washed 5 times by immer~ion in water in order to remo~e dye not bound to the cell~. The dye bound to the cells was extracted by adding 100 ~1 of reagent solution (50% etha-nol, 0.1% glacial acetic acid, 49.9% water) to each well.

4. The plates were shaken for 5 min to obtain a ~olution of uniform color in each well. The surviving cells were determined by measuring the extinction at 540 nm of the colored Qolution in the individual well~.

5. Subsequently, by relating to the cell control and the rhu-TNF control, the 50% antagonism value was defined, and tha sample concentration which re~ulted in 50% anta~onism of rhu-TNF cytotox-icity at the rhu-TNF concentration u3ed was caleulated a~ antagoniqtic activity of the sample te~ted.

III. Competitive receptor-binding test Both the asonistic and antagoni~tic effect~ of peptide~ are conditional on the latter binding to the TNF receptor. This means that peptide~ with an agon~ stic or antagonistic effect compete with ~0~ 5'~

- 13 - O.Z. 0050/40381 rhu-TNF for binding to the TNF receptor on TNF-sensitive indicator cells (e.g. U937). The competi-tive receptor-binding test was carried out a~
follow~:

S 1. 100 ~1 of medium containing various concentra-tions of the peptide to be te~ted and of rhu-TNF
(= control) were pipetted into the reaction vessels. Ths medium compri~ed 500 ml of PBS
(Boehringer Mannheim~ containing 10 ml of heat-inàctivated (56C, 30 min) FCS and 100 mg of sodium azide.

2. Subsequently, 100 ~1 of medium containing 1 ng of l25I_labeled rhu-TNF (Bolton lactoperoxidase method) were placed in the reaction ve~sel~ and mixed. The non-~pecific binding (NSB) wa3 deter-mined by mixing in the reaction ves~el the l25I-labeled rhu-TNF (1 ng of l25I-rhu-TNF in 100 ~1 of medium) with a 200-fold excess of unlabeled rhu-TNF (200 ng of rhu-TNF in 100 ~1 of medium).

3. TheD 100 ~1 of medium containing 2 x 106 U937 cell~ (human) were pipetted into the reaction vessel~ and mixed. The reaction vessels (te~t volu~e 300 ~1) were incubated at 0C for 90 min.
The reaction mixtures were remixed after 45 min.

4. After the incubation the cell~ were centrifuged at 1800 rpM and 4C for 5 min, wa~hed 3 time~
with medium and transferred quantitatively into counting vial~, and the cell-bound radioactivity wa~ determined in a Clini gamma counter 1272 (L~B Wallac).

5. After the measurements had been corrected for the non-specific binding, the 50~ competition value 5 O 5'~

- 14 - O.Z. 0050/40381 was defined by relation to the overall binding, and the sample concentration which led to 50%
competition of '~5I-rhu-TNF binding at the 125I-rhu-TNF concentration used wa~ calculated aR the competitive activity of the sample tested.

The Examples which follow are intended to explain the invention in more detail. The proteinogenous amino acids are abbreviated in the Example~ u~ing the conventi~nal three-letter code. Other meanings are:
Ac = acetic acid, Ahp = 7-2minoheptanoic acid, Ahx = 6-aminohexanoic acid, Aoc = 8-aminooctanoic acid, Ape - 5-aminopentanoic acid, Hcy = homocy~t~ine, Orn = ornithine, Dap = 2, 3-diaminopropionic acid.

A. General procedure 5 I. The peptides claimed in claim 1 were synthesized u~ing standard methods of ~olid-pha~e peptide synthe~is in a completely automatic model 430A
peptide synthesizer from APPLIED ~IQSYSTEMS. The apparatus uses different synthe~i~ cycle~ for the Boc and Fmoc protective group techniques.

a) Synthesis cycle for the Boc protective group technique 1. 30% trifluor~Y~dic ac~d in DC~ 1 x 3 min 2. 50% triflu~rxYI~ic acid in DCM 1 x 17 min 3. Do~wa~ng S x 1 min 4. 5% diL~qpoQy~ m;nP in ~CM 1 x 1 min 5. 5% diLK~QylekhylAm;nP in NKe 1 x 1 min 6. NMP wad~n4 5 x 1 min 7 . A~ditiQn of pn~ivated pno~x~ed ~D
acid (activa~nn ky l eqyivslent of DOC
and 1 equivalenS of HDBt in NMP/DCM);
p~ e coupling (lst part) 1 x 30 min 5()5Z

- 15 - O.Z. 0050/40381 8. Addition of nMso to the reaction mixture unti1 it contains 20% nMso by vQlume 9. Peptide coupling (2nd part) 1 x 16 min 10. A~dition of 3.8 eqlivalents of diiso-pr~pylethylamine to the reaction mixture 11. Peptide coupling (3rd part) 1 x 7 min 12. DCM wQshing 3 x 1 min 13. if reaction is incomplete, repetition of coupling (neturn to 5.) 14. 10% acetic anhydride, 5% dii~oFropyl-ethylamine in DCM 1 x 2 min 15. 10% acetic anhydride in DCM 1 x 4 min 16. DCM wash ~ 4 x 1 mun 17 Return to 1.

b) Syn~hesi~ cy~le for the E~LC p ~ ~e grnup techniq~e 1. NMP w2shing 1 x 1 min 2. 20% p;peridine in NMP 1 x 4 min 3. 20% pip~ridine in NMP 1 x 16 min 4. NMP woshing 5 x 1 min 5. Addition of ~ iYated prohecbel amino acid (activatiDn ~y 1 equivalent of DoC
anl 1 egyivzlent of HoEt in NMP/DCM);
pepkida coupling 1 x 61 min 6. NMP washing 3 x 1 min 7. if rex~cn i~ i~xDç~ebQ, n~ition ~f ooy~ng (r~lrn to 5.) 8. 10% ~o*~ a~dride in NMP 1 x 8 min 9. NMP wQ~hing 3 x 1 min 10. ~l~n to 2.

30 II. Working up of peptide-resin~ obtained a~ in Ia Th~ peptide-resin obtained as in Ia was dried under reduced pre~sure and tran~ferred into a reaction ve~sl of a Teflon HF apparatu~ tfrom PENINSULA).
~ddition of a ~caveng3r, preferably ani~ole ~1 ml/g of re~in), and of a thiol, in the case of tryptophan-20~3~i05Z
- 16 - O.Z. 0050/40381 containing peptides, to remove the indole formyl group, preferably ethanedithiol (0.5 ml/g of re~in), wa~ followed by condensation in of hydrogen fluoride (10 ml/g of resin) while cooling with liquid N2. The mixture wa~ allowed to warm to 0C, and was Ytirred at this temperature for 45 min. The hydrogen fluor-ide was then stripped off under reduced pre~sure and the re~idue was washed with ethyl acetate in order to remove remaining sca~enger. The peptide wa.~
extracted with 30~ strength acetic acid and filtered, and the filtrate was freeze-dried.

To prepare peptide ~ydrazides, the peptide-resin Pam- or Merrifield re~in) wa3 3uspended in D~F
(15 ml/g of resin), hydrazine hydrate (20 equiva-lents) wa~ added, and the mixture was stirred at room temperature for 2 days. To work up, the resin was filtered off and the filtrate wa~ evaporated to dryneas. The residue was crystallized from DMP~Et2O
or NeOH/Et2O-III. Working up of the peptide-resins obtained a~ in Ib The peptide-resin obtained as in Ib was dried under reduced pre~ure and subsequently sub~ected to one of the following cleavage procedureQ, depending on the amino acid composition (Wade, ~regear, Howard 2S Florey Fmoc-Workshop Manual, Melbourne 1985).

2(~ orj~:
- 17 - O.Z. OOSO/40381 Peptide containing Cleavage condition~

Arg(Mtr) Met Trp ~FA Scavenger Reaction TLme S _ no no no 95~ 5% H20 1.5 h yes no no 95~ 5% thioani~ole 2 3 h no yes no 95~ 5% ethyl methyl 1.5 h ~ulfide no no ye~ 95% 5% ethanedithiol/1.5 h anisole (12 3 ) no yes yes 95% 5~ ethanedithiol/l.S h anisole/ethyl methyl ~ulfide (1:3:1) lS yes ye~ ye~ 93~ 7~ ethanedithiol/~ 3 h anisolQ/ethyl methyl _ sulfide (1:3:3) The su~pension of the peptide-resin in the suitable TFA mixture wa~ stirred at room temperature for the ~tated time and then the resin wa~ filtered off and washed with TFA and with DCM. The filtrate and the washings were extensively concentrated, and the peptid~ waa pr~cipitated by addition of diethyl ether. ~he mixture was cooled in an ice ba~h, and the precipitate wa~ filtersd off, taken up in 30%
acetic acid and freeze-dried.

IV. Purific~tion and characterization of the peptides Purification was by gel chroma~ography (SEPHAD~X~
G-10, G-15/10% HQAc; SEPHADEX0 LH20/MeOH) and 8ub-~equent medium pre~ure chromatography ~tationary pha~es HD-SIL C-18, 20-45 ~, loO-A phases gradisnt w~th A - O.lS TFA/~eOH, B - 0.1~ TFA/H20~.

~20~5~f~

- 18 - O.Z. 0050/40381 The purity of the final products was determined by analytical HPLC (stationary phase: 100 x 2.1 mm VYDAC C-18, 5 ~, 300 A; mobile phase = CH3CN~H2O
gradient buffered with 0.1% TFA, 40C~. Charac-terization wa3 by means of amino acid analy~is and fast atom bombardment mass spectrometry, B. Specific procedures EXA~PLE 1 H-Pro-Gln-Ala-Glu-(~ly-Gln-Leu-NH2 1.2 g of Boc-Leu-MB~A-resin (~ub~itution 0.42 mmol/g), corresponding to a batch ~ize of 0.5 mmol, were reacted as in Ala with 2 mmol each of Boc-Gln-OH Boc-Ala-OH
Boc-Gly-OH Boc-Gln-OH
Boc-Glu(OBzl)-OH Boc-Pro-OH.

After the synthesis wa~ complete, the peptide-resin underwent N-terminal deprotection (Ateps 1-3 a~ in AIa) and subsequent drying under reduced pre~ure; the yield wa~ 1.49 g.

0.75 g of the resin obtained in thi~ way wa~ sub~ected to HP cleavage a3 in AII. The crude product ~141 mg) wa~
purified by g~l filtration (S~PHADEX~ G-10) and medium pre~ure chromatography (cf. AIV; 20-40% A; 0.25% minl).
97 mg of pure product were obtained.

EXAMP~E 2 Ac-Ala-Glu-Gly-Gln-Leu-Gln-OH

O.47 g of Fmoc-Gln-p-alkoxybenzyl alcohol-resin ~sub~ti-tution 0.S3 mmol/g), corresponding to a batch ~ize of 0.25 mmol, was reacted a~ in AIb with 1 mmol each of Z0~3~05Z

- 19 - O.Z. 0050/40381 Fmoc-Leu-OH Fmoc-Glu(OtBu)-OH
Fmoc-Gln-OH Fmoc-Ala-OH
Fmoc-Gly-OH

After the synthesis was complete, the N terminus was acetylated (steps 2-4 and 8-9 a~ in AIb) . The re~ulting peptide-resin was dried under reduced pressure; the yield was 0.57 g.

The crude peptide (137 mg) obtained after TFA cleavage as in A$II was purified by gel filtration (SEPHADEX~ G-10) and medium pre~sure chromatography (cf. AIY; 20-40~ A;
0.25~ min~'). 89 mg of pure product were obtained.

The following can be prepared in a similar manner to Examples 1 and 2:
3. H-Ala-Asn-Pro-Gln-Ala-Glu-Gly-Gln-Leu-Gln-Trp-OH
4. Ac-Ala-Asn-Pro-Gln-Ala-Glu-Gly-Gln-Leu-Gln-Trp-OH
5. H-Ala-Asn-Pro-Gln-Ala-Glu-Gly-Gln-Leu-Gln-Trp-NH2 6. Ac-Ala-Asn-Pro-Gln-Ala-Glu-Gly-Gln-Leu-Gln-Trp-NH2 7. H-Pro-Gln-Ala-Glu-Gly-Gln-Leu-OH
8. Ac-Pro-Gln-Ala-Glu-Gly-Gln-Leu-OH
9. Ac-Pro-Gln-Ala-Glu-Gly-Gln-Leu-NH2 10. H-Ala-Glu-Gly-Gln-Leu-Gln-OH
11. H-Ala-Glu-Gly-Gln-Leu-Gln-NH2 12. Ac-Ala-Glu-Gly-Gln-Leu-Gln-NH2 13. H-Val-Val-Ala-A~n-Pro-Gln-Ala-Glu-Gly-Gln-Leu-Gln-Trp-~eu-Asn-Arg-Arg-Ala-A~n-Ala-OH
14. Ac-Val-Val-Ala-A~n-Pro-Gln-Al~-Glu-Gly-Gln-Leu-Gln-Trp-Leu-Asn-Arg-Arg-Ala-A~n-Ala-OH
15. H-Val-Val-Ala-A~n-Pro-Gln-Ala-Glu-Gly-Gln-Leu-Gln-Trp-Leu-A~n-Al g-Arg-Ala-A~ a-NH2 16. Ac-Val-Val-Ala-A3n-Pro-Gln-Ala-Glu-Gly-Gln-Leu-Gln-Trp-Leu-Asn-Arg-Arg-Ala-A~3n-Ala-NH2 ~0~50~

- 20 - O.Z. 0050/40381 Ac~ al-Ala-Asn-Pn~an-Ala-Gau-Gly~n-Leu~n-~Leu~-N~
0.52 g of Boc-Cys(pMB)-MB~A-re~in (~ub~titution 0.97 S mmol/g), corre~ponding to a batch ~ize of 0.5 mmol, were reacted as in AIa with 2 mmol each of Boc-L~u-OH Boc-Ala-OH
Boc-~rp(CHO)-OH Boc-Gln-O~
Boc-Gln OH Boc-Pro-OH
~oc-Leu-OH Boc-Asn-OH
Boc-Gln-OH Boc-Ala-OH
Boc-Gly-OH Boc-Yal-OH
Boc-Glu(OCh~)-O~ Boc-Cy~(pMB)-OH.

After the ~ynthe~i~ wa~ complete, the N terminu~ wa~
acetylated (3tep~ 1-6 and 14-16 as in AIa?.

The reRulting peptide-resin wa~ dried under reduced preseure; the yield wa~ 1.3 g.

0.65 g of the resin obtained in thi~ way wa~ ~ub~ected to HF cleavage a~ in AII. The freezQ-dried crude product wa~
taken up in 2 1 of 0.1% strength acetic acid, and th~ pH
was then ad~usted to 8.4 with aqueou~ ammonia. Under an argon atmo0phere, 0.01 N R3~Fe(CN)5] solution wa~ ~lowly added dropwiee until the yellowish-green color persi~ted for at lea~t lS min. ~he mixture wa~ then ~tirred for 1 h and then acidified to pH 4.5 with glacial acetic acid, and 15 ~1 of an aqueou~ ~u~pen~ion of an anion exchanger (~IORAD 3 x 4A, chlorlde for~) were added. After 30 ~in, the ion exchanger resin waa filtered off, and the filt-rate was concentrated to 100 ~1 in a rotary ev~porator and ~ubsequently freeze-dried.

20~50~

- 21 - O.Z. 0050/40381 All th~ ~olvents used had previously been saturated with nitrogen in order to prevent any oxidation of the free cysteine residues.

The crude product was purified by gel chromatography (SEPHADEX~ LH-20). 17 mg of pure product were obtained.

The following can be prepared in a similar manner to Example 17 ~Pam-resin was used to prepare the peptide acid~):

-18.H-Cys-Val-Val-Ala-A~n-Pro-Gln-Ala-Glu-Gly-Gln Leu-Gln-Trp-Leu-Asn-Arg-Arg-Ala-A~n-Ala-Cys-OH

l9.Ac-Cys-Val-Val-Ala-Asn-Pro-Gln-Ala-Glu-Gly-Gln Leu-Gln-Trp-Leu-A8n-Arg-Arg-Ala-A~n-Ala-Cy8-N~2 -20.H-Cys-Asn-Pro-Gln-Ala-Glu-Gly-Gln-Leu-Gln-Cy~-OH

21.Ac-Cy~-A~n-Pro-Gln-Ala-Glu-Gly-Gln-Leu-Gln-Cys-OH

22.H-Cys-Val-Ala-Asn-Pro-Gln-Al~-Glu-Gly-Gln-Leu-Gln Trp-Leu-Cys-OH
r 23.Ac-Cys-Pro-Gln-Ala-Glu-Gly-Gln-Leu-Cys-NH2 r - i 24.H-Hcy-Asn-His-Gln-Val-Glu-Glu-Gln-Leu-Glu-Hcy-OH

25.Ac-Hcy-Asn-His-Gln-Val-Glu-Glu-Gln-Leu-Glu-Hcy-NH2 -26. H-Cys-Ala-A~n-Pn~ln-Ala-Glu-Gly-Gln-Leu~k~k~p~Y-oH

27. Ac-Cys-Ala-Asn-Pn~ln-Ala-Glu-Gly-Gan-leu~k~Tqp{~-NH2 ~ - I

28.~-Cy~-Pro-Gln-Ala-Glu-Gly-Gln-Leu-Cy~-OH

~5 05 ~
- 22 - O.Z. 0050/40381 29. H-Ala-His-Val-Val~-Asn-P~}~an-Ala-Gly-Gay~n-Leu~n~--Leu-Asn{~I

30. Ac-Ala-His-~al-Val~ sn-Pn~{an-Ala-Gly ~ y~n-Leu~n~
-~eu-Asn-NH2 31. H-Cys-Asn-Pro-Ala-Gln-Asn-Gln-Leu-Gln-Cys-OH

32.Ac-Cys-Asn-Pro-Ala-Gln-Asn-Gln-Leu-Gln-Cys-NH2 33. H-Cys-Val-Ala-Asn-Pro-Gln-Ala-Glu-Gly-Gln-Leu-Gln-Trp-Leu-A~n-Arg-Ala-Asn-Cys-OH

34. Ac~ A~n-Po~n-Ala-Glu-Gly-Gan-Leu-Gan-T~p~-NH2 -35. Ac-Cys-Val-Ala-Asn-Pro-Gln-Ala-Glu-Gly-Gln-Leu-Gln-Trp-heu-A~n-Arg-Ala-Asn-Cy~-NH2 :

36.H-Cys-His-Val-Val-Ala-A~n-Pro-Gln-Ala-Glu-Gly-Gln-i Leu-Gln-Trp-Leu-Asn-Arg-Ala-A~n-Ala-Leu-CyE-OH

37. Ac-Cys-His-Val-Val-Ala-Asn-Pro-Gln-Ala-Glu-Gly-Gln-Leu-Gln-Trp-Leu-Asn-Arg-Ala-A~n-Ala-Leu-Cy~-NH2 38.H-Cys-L~u-Ile-Gly-Asp-Pro-Ser-Ly~-Gln-Asn-Ser-Leu Leu-Trp-Arg-Ala-Asn-Thr-Asp-Arg-Ala-Cy~-OH

3 9 . Ac -Cy~ -Leu- I le-Gly-A~p-Pro-Ssr-Lys-Gln-Affn-Ser-Leu -- --I
Lau-Trp-Arg-Ala-Asn-'rhr-Asp-Arg-Ala-Cys -NH2 40. H~ x~V l-Ala-Thr-Val-Val-Ala-A~n-PQ~n-Ala-Gau-Gly-~ln-Leu-Gln-$~Leu-Asn-Arg-A~g-ALa-Asn~Ala~-Leu-Ala-Asn-Gay-CH

Z0~5~52 - 23 - O.Z. ~050/40381 _ 41. Ac~ ~Val-Ala-Thr-Val-Val-Ala-A~n-Pn~n-Ala-Glu ~ y~n-i T~ ~ n-T ~ Ieu-Asn-Arg-Arg-Ala-Asn-Ala{~ys-Ieu-Ala-A~n~ -NH2 EX~qE 42 j Ac-Lys-Pro-Gln-Ala-Glu-Gly-Gln-Leu-Asp-NH2 1 g of resin described by Breipohl et al. (from BACHEM), corresponding to a batch ~ize of 0.5 mmol, wa~ reacted a~ in AIb with 2 mmol each of Fmoc-A3p(OtBu)-O~ Fmoc-Ala-OH
Fmoc-Leu-OH Fmoc-Gln-OH
Fmoc-Gln-OH Fmoc-Pro-OH
Fmoc-Gly-OH Fmoc-Lys(Boc)-OH
Fmoc-GlutOBzl)-OH

After the synthesis was complete, the N terminu~ was acetylated (steps 2-4 and 8-9 as in AIb). The peptide-re~in was dried under reduced preq~ure; yield 1.62 g.

The crud~ product (425 mg) obtained after TFA cleavage a~ in AIII was purified a~ in AIV and dis~olved in 500 ml of degassed DNF. 0.24 ml of N~t3 wa~ added and, at -25C, 0.24 ml of diphenylphosphoryl azide, and the mixture wa~
stirred at -25C for 2 h. It was then left to stand at -20~C for 2 day~, at 4C for 2 days and at room tempera-ture for 2 day~. It was then evaporated to drynes~ andthe crude pept$de was pur$fied by gel chromatography (SEPHADEX~ L~ 20). The i~olatsd product (175 mg) was deprotected with HF as in AII and purifi~d by medium pressure chromatography (cf. AIV, 20-35% A, 0.25% min~1).
58 mg of pure product were obtained.

zo~ri(~sz - 24 - O.Z. 0050/40381 _ H-Lys-Asn-Pro-Gln-Ala-Glu-Gly-G1n-Leu-Gln-Asp-OH
2.44 g of Fmoc-Asp(OtBu)-Merrifield re~in (substitution 0.41 mmol/g), corresponding to a batch size of 1 mmol, were reacted as in AIb with 4 mmol each of Fmoc-Gln-OH Fmoc-Glu(OBzl)-OH Fmoc-Asn-OH
Fmoc-Leu-OH Fmoc-Ala-OH Fmoc-Lys(Boc)-OH
Fmoc-Gln-OH Fmoc-Gln-OH
Fmoc-Gly-OH Fmoc-Pro-OH

The t-butyl and Boc protective groups were then cleaved off (step~ 1-6 a~ in AIa). ~he cyclization on the resin took place in NMP with the addition of 1.77 g of BOP and 1.74 ml of diisopropylethylamine (48 h). The peptide-re3in underwent N-terminal deprotection (step~ 2-4 as in AIb) and drying under reduced pressure. ~he yield wa~
3.05 g.

~he crude product obtained after HF cleavage as in AII
wa~ purified by gel filtration ~SEPHADEX~ G-25) and mediulm pressure chromatography (cf. AIV: 20-40% A;
0.25% min~1). 32 mg of pure product were obtained.

The following can be prepared in a ~imilar manner to Exampl~s 42 and 43:

1. . , 44.Ac-Asp-A~n-Pro-Gln-Ala-Glu-Gly-Gln-Leu-Gln-Dap-NH2 45.H-Asp-Aan-Pro-Gln-Ala-Glu-Gly-Gln-Leu-Gln-Dap-HO

46.Ac-Orn-Asn-Pro-Gln-Ala-Glu-Gly-Gln-Leu-Gln-Asp-NH2 47.Ac-Ly~-Asn-Pro-Gln-Ala-Glu-Gly-Gln-Leu-Gln-A~p-OH

~:0~35iO ~
- 25 - O.Z. 0050/40381 48.Ac-Dap-His-Gln-Val-Glu-Glu-Gln-Leu-Glu-A~p-NH~

49. H-Ly~-Hi~-Gln-Val-Glu-Glu-Gln-Leu-Glu-Asp-OH

50. Ac-Asp-Val-Ala-Asn-Pn~4~n-Ala-Glu-Gly-Gln-Leu-Gln~n-NH2 S1. H-Asp-V~l-Ala-Asn-P~ln-Ala-Glu-Gly-Gln-Leu~n~orn-CH

52. Ac-His-Val-Val-AsF-Asn-Pn~n-Ala-Glu-Gay~n-Leu~n~
Ieu-Asn-NH2 -53.Ac-Asp-Val-Val-Ala-Asn-Pro-Gln-Ala-Glu-Gly-Gln-Leu-Gln-Trp-Leu-Asn-Arg-Arg-Ala-Asn-Ala-Lys-NH2 54.Ac-Lys-Pro-Gln-Ala-Glu-Gly-Gln-Leu-A~p-NH2 rPro-Gln-Ala-Glu-Gly-Gln-Leu-Aoc~

1.22 g of Fmoc-Leu-p-alkoxybenzyl alcohol-re~in (substi-tution 0.41 mmol/g), correspondin~ to a bat~h ~ize of 0.5 mmol, were reacted as in AIb with 2 mmol each of Fmoc-Gln-OH Fmoc-Gln-OH
2S Fmoc-Gly-OH Fmoc-Pro-OH
Fmoc-Glu(OBzl)-OH Fmoc-Aoc-OH
Fmoc-Ala-OH Fmoc-Leu-OH

After the synthesis was complete, the peptida-re~in underwent N-terminal deprotection ~teps 2-4 a~ in AIb) and subsequent drying under reduced pre~surQ. The yisld was 1.53 g.

The crude peptide obtained after TFA cleavage a~ in AIII
was di~olved in 505 ml of dega~sed DMF. 210 mg of NaHCO3 21~052 - 26 - O.Z. 0050/40381 and (at -25C) 0.24 ml of diphenylphosphoryl azide were added, and the mixture was then stirred at -25C for 2 h and at room temperature for 2 days. It was then evapor-ated to dryness, and the crude peptide was purified by gel chromatography (SEPHADEX~ LH20). The isolated monomer (138 mg) was deprotected with HF as in AII and purified by medium pressure chromatography (cf. AIV; 25-45% A;
0.25% min~1). 84 mg of pure product were obtained.

The following can be prepared in a manner sLmilar to Example 55:
56.rAsn-Pro-Gln-Ala-Glu-Gly-Gln-Leu-Gln-Ahp 57.rAsn-Pro-Gln-Ala-Glu-Gly-Gln-Leu-Gln-Ahx 58.rAsn-Pro-Gln-Ala-Glu-Gly-Gln-Leu-Gln-Ape 1 59.rVal-Val-Ala-Asn-Pro-Gln-Ala-Glu-Gly-Gln-Leu-Gln-Trp-.
Leu-Asn-Arg-Arg-Ala-Asn-Ala-Ah 60.rVal-Ala-Asn-Pro-Gln-Ala-Glu-Gly-Gln-Leu-Gln-Trp-Leu-Asn-Asn-Arg-Arg-Ala-Asn-Ahxl

Claims (8)

1. A peptide of the formula I, X-A-B-E-Leu-Y I, where A is Glu, Pro or Gln, B is Gly, Glu, Asn or Asp, E is Gln or Ser, X is G-NH-CHM-CO-, G-NH-CHM-CO-W-, G-R-NH-CHN-CO- or G-R-NH-CHM-CO-W- and Y is -Z, -NH-CHQ-CO-Z, -V-NH-CHQ-CO-Z, -NH-CHQ-CO-U-Z
or -V-NH-CHQ-CO-U-Z, where, in X and Y, G is hydrogen or an amino-protective group, Z is OH or NH2 or a carboxyl-protective group, or G and Z together are also a covalent bond or -CO-(CH2).-NH- where a is from 1 to 12, R and U are peptide chains composed of 1-5 naturally occurring .alpha.-amino acids, W is one of the peptide chains , a section composed of 5 to 11 amino acids of one of the abovementioned peptide chains, or a peptide chain with 1-4 naturally occurring .alpha.-amino acids, V is one of the peptide chains , , a section composed of 5 to 11 amino acids of one of the abovementioned peptide chains, or a peptide chain with 1-4 naturally occurring .alpha.-amino acids, and M and Q are hydrogens or one of the following -CH(CH3)2, -CH(CH3)-C2H5, -C6H5, -CH(OH)-CH3, , or -(CH2)b-T

(with b being from 1 to 6 and T being hydrogen or OH, CH3O, CH3S, (CH3)2CH, C6H5, p-HO-C6H4, HS, H2N, HO-CO, H2N-CO or H2N-C(=NH)-NH) or M and Q together are a -(CH2)c-S-S-(CH2)d-, -(CH2)e-CO-NH-(CH2)f- or -(CH2)e-NH-CO-(CH2)g-NH-CO-(CH2)f- bridge (with c and d being from 1 to 4, e and f being from 1 to 6 and g being from 1 to 12), as well as the salts thereof with physiologically toler-ated acids.

2. A peptide as claimed in claim 1, where G ix hydrogen or an amino-protective group and Z is hydroxyl or amino or a carboxyl-protective group, and M and Q are not connected together.

3. A peptide as claimed in claim 1, where G is hydrogen or an amino-protective group and Z is hydroxyl or amino or a carboxyl-protective group, and M and Q together are a -(CH2)C-S-S-(CH2)d- bridge.

4. A peptide as claimed in claim 1, where G is hydrogen or an amino-protective group and Z is hydroxyl or amino or a carboxyl-protective group, and M and Q together are -(CN2)e-NH-CO-(CH2)f- or -(CN2)e-NN-CO-(CN2)g-NH-CO-(CH2)f.

5. A peptide as claimed in claim 1, where G + Z together are a covalent bond or -CO-(CH2)a-NH- .

6. A peptide as claimed in claim 1 to 5 for use for controlling diseases.

7. The use of the peptides as claimed in claims 1 to 5 for controlling neoplastic diseases and autoimmune diseases as well as for controlling and preventing infections, inflammations and transplant rejection reactions.

8. A process for the preparation of a peptide as claimed in claims 1 to 5, which comprises preparation thereof using conventional methods of peptide chemistry.