CA2005281A1 - Tnf peptides - Google Patents

Abstract

- 33 - O. Z . 0050/40383 ABSTRACT OF THE DISCLOSURE:

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

Description

~OQS;~l O.Z. 0050/40383 NOVEL TNF PEPTIDES

The present invention relates to novel peptides derived from tumor necrosi~ factor (TNF), the preparation thereof and the use thereof as drugs.

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 necrosi~ in various mouse tumors. This activity was ascribed to tumor necrosis factor. TNF also has a cytostatic or cytotoxic effect on a large number of tran~formed cell lines in vitro, wherea~ normal human and animal cell lines are unaffected (Lymphokine Report~ Vol.

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

It is possible to deduce from this data the following protein structurs for mature human TNFs V IAqO~ f ~ g-~--e~--~ysprovaL~sHisvalvaLuJA~o ValGl--~ u ~valv~rpn~incluGly~o~r~aale~br GlnV~L~ eLysGly~ yCy~o6e~n~f~JV~LL~ulhdOk~SrIle S~gIleAlaVal~5DG~nirLysVa~ b~leLy~Pro Cyd~nxq~uThrknx~uGlyA~uAlaLyu?~nrprpcluproIle~leu GayG~yValPhoG~IK~luLysGlyAypb~euS3LU4~luIleA~YrgPn~sp Tyri~4pPh~ u&~yGlnValTyrPh~yIleIle~k~eu The TNF genes of cattle, rabbits and mice have also been de~cribed (Cold Spring Harbor Symp. Quant. Biol. 51 (1936) 597~.

Besides its cytotoxic properties, TNF i~ one of the main 2(~0~2~1.
- 2 - O.Z. 0050/40383 subctances involved in inflammatory reaction-q (Pharmac.
R~ . 5 (1988) 129). AnLmal models have shown that TNF i8 involved in septic shock (Science 229 (1985) 869) and graft-versus-ho~t disease (J. Exp. Med. 166 (1987) 1280).

we have now found that peptide~ with a con~iderably lower molecular weight have beneficial propertie~.

The present invention relates to peptides of the formula I
X-A-Asn-B-Y I, where A is Asp or Asn, B i~ Gln or Ser, X iY G-NH-CHM-CO-, G-NH-CHM-CO-W-, G-R-NH-CHN-CO- or G-R-NH-CHM-CO-W- and 5 Y i8 -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 i8 OH or NH2 or a carboxyl-protective group or G and Z together are also a covalent bond or -CO-(CH2),-NH-, where a i8 from 1 to 12, R, U, V and W are peptide chains composQd of 1-4 natu-rally occurring ~-amino acids and N and Q aro hydrogens or one of the following groups CH(CH3) 2, -CH~CH3)-C2H5, -C5H~, -CH(OH)-CH3, --CH2~ CH2~ or --ICH2)b--T

(with b being from 1 to 6 and T being hydrogan or OH, CH30, CH3S, (CH3)2CH, C6H5~ p-HO-C~H~, HS, H2N, HO-CO, H2N-CO, H2N-C(=NH)-NH or M and Q together are a -(CH2)c-S-S-(CH2) d- ~ - ( CHz).-CO-NH-(CH2)r~ or -(CHZ).-NH-co-(cH2)s-NH-co-(cH2)r- 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), Z005Z~l - 3 - O.~. 0050/40383 a well as the salt~ thereof with physiologically tolera-ted acids.

The peptides of the formula I are constructed of L-amino acids, but they can contain 1 or 2 D-amino acid~. The side-chain3 of the trifunctional amino acids can carry protective groups or be unprotected.

Particularly preferred physiologically tolerated acids are: hydrochloric acid, citric acid, tartaric acid, lactic acid, phosphoric acid, methanesulfonic acid, acetic acid, formic acid, maleic acid, fumaric acid, malic acid, succin~c 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 paptides can be open-chain (G = H, amino-protective group; Z = OH, NH2, carboxyl-protective group, and Q not connected together) and, in particular, have a disulfide bridge (G = H, amino-protective group;
Z = OK, NH2, carboxyl-protective group; M + Q = -(CH23c~
S-S~(CH2)d-) or a side-chain bridge (G = H, amino-protec-tive group, Z = OH, NH2~ carboxyl-protective group, M + Q
5 - ( CH2 ) ~-NH-CO- (CH2)~- or -(CH2).-NH-CO-(CH2)s-N~-CO-(CH2)~-) or be linked head-to-tail (G + Z = covalent bond or -CO-(CH2),-NH-).

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

Thus, the peptides can be con~tructed sequentially from a~ino acid~ or by linking together suitable smaller peptide fragments. In the ~equential construction, the peptide chain i8 extended stepwise, by one amino acid each tlme, starting at the C terminus. In the ca~e of fragments coupl;ng, it i8 possible to link together 200~81.

- 4 - O.Z. 0050/40383 fragment~ of different lengths, these in turn being obtainable by sequential con~truction from amino acid~ or coupling of other fragments. The cyclic peptides are obtained, after synthesic of the open-chain peptides, by a cyclization reaction carried out in high dilution.

In the case both of sequential construction and of fragment coupling, it i~ necessary for the building blocks to be linked by formation of an amide linkage.
Enzymatic and chemical methods are suitable for this.

Chemical methods for forming amide linkages are dealt with in detail by M~ller, Methoden der Organischen Chemie (~ethods 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, Rlausner, Ondetti, Peptid~ Synthe0is, pp 85-128, John Wiley & Sons, New York, 1976 and other standard works 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), diisopropylcarbodiimide (DIC), l-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ), l-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCI), n-propanephosphonic anhydride (PPA), N,N-bis(2-oxo-3-oxazolidinyl)amidophosphoryl chloride (~OP-Cl), diphenylphosphoryl azide (DPPA), Ca~tro's re~gont (BOP), O-benzotriazolyl-N,N,N~,N'-tetra-methyluronium salts (HBTU), 2,5-diphenyl 2,3-dihydro-3-oxo-4-hydroxythiophene dioxide (Stoglich~s reagent;
HOTDO) and l,l'-carbonyldiimidazole (CDI). The coupling r~agents can be employed alone or in combination with additive~ such a~ N,N~-dimethyl-4-aminopyridine (DMAP), N-hydroxybenzotriazole (HOBt), N-hydroxybenzotriazine (HOOBt), N-hydroxy~uccinimide (HOSu) or 2-hydroxy-'~05'~8~.

- 5 - O.Z. OOSO/40383 pyridine.

Whereas it is normally possible to dispense with protec-tive groups in enzy~atic peptide synthesis, for chemical synthe~is it is necessa~y for there to be reversible protection of the reactive functional groups which are not involved in the formation of the amide linkage on the two reactants. Three conventional protective group techniques are preferred for chemical peptide qynthQses:
the benzyloxycarbonyl (Z), the t-butyloxycarbonyl (~oc) and the 9-fluorenylmethyloxycarbonyl (Fmoc) technique~.
In each case the protective group on the ~-amino group of the chain-extending building block is identif ied. The side-chain protactive groups on the trifunctional amino acids are chosen 8e that they are not necessarLly elimin-ated together with the ~-amino protective group. A
detailed review of amino acid protective groups is given by Muller, Methoden der Organischen Chemie Vol XV/l, pp 20-906, Thiemo Verlag, Stuttgart, 1974.

The building blocks used to construct the peptide chain can be react2d in solution, in ~uspension or by a method similar to that described by Merrifield in J. Amer. Chem.
Soc. 85 (1963) 2149. Particularly preferred methods are tho~e in which peptides are constructed sequentially or by fragment coupling by u3e of ths Z, Boc or Fmoc protec-tive group technique, in which case the reaction takesplace in solution, as well as those in which, similar to the Nerrifield technique, one reactant is bound to an insoluble polymeric support ~also called resin herein-after). Thi~ typically entails the peptide belng con-structed sequentially on the polymeric support, by use ofthe Boc or Fmoc protective group technique, with the growing pQptide chain being covalently bonded at the C terminus to tha insoluble resin particles (cf. Figures 1 and 2). This procedure allows reagents and byproducts to be removed by filtration, and thus recry~tallization ~005~81.

- 6 - O.Z. 0050/40383 of intermediates i-~ superfluous.

The protected amino acids can be bonded to any ~uitable polymers which merely need to be insoluble in the sol-vent~ used and to have a 3table physical form which allows easy filtration. The polymer must contain a functional group to which the first protected amino acid can be firmly linked by a covalent bond. A wide variety of polymers i3 suitable for this purpose, for example cellulose, polyvinyl alcohol, polymethacrylate, sulfon-ated poly~tyrene, chloromethylated copolymer of styreneand divinylbenzene (Merrifield resin), 4-methylbenz-hydrylamine-xesin (MBHA-resin), phenylacetamidomethyl-resin (Pam-resin), p-benzyloxybenzyl alcohol-resin, benzhydrylamine-resin (BHA-resin), 4-hydroxymethyl-benzoyloxymethyl-resin, the re~in u~ed by Breipohl et al.
(Tetrahedron Lett. 28 (1987) 565; from BACHEM), HYCRAM
re~in (from ORPEGEN) or SASRIN resin (from BACHEM).

Solvents suitable for peptide synthesis in solution are all tho~e which are inert under the reaction conditions, in particular water, N,N-dimethylformamide (DMF), dimethyl sulfoxidQ (DNS0), acetonitrile, dichloromethane (DC~), 1,4-dioxane, tetrahydrofuran (lnr), N-methyl-2-pyrrolidone ~N~P) and mixture~ of the said solvents.
Peptide syntheai3 on polymeric supports can be carried out in all inert organic solvent~ which dissclve the amino acid derivatives used; however, solvents which also have resin-swelling properties are preferred, such as DMF, DCN, NNP, acetonitrile and DMS0, as well as mixtures of the~e solvents.

After the peptide has been synthesized it i8 cleaved off the polymeric support. The cleavage conditions for the various types of resins are disclosed in the literature.
The cleavage reactions most commonly use acid and palladium catalysis, in particular cleavagQ in anhydrous 2 0 0 S ~

- 7 - O.Z. 0050/40383 liquid hydrogen fluoride, in anhydrous trifluoromethane-sulfonic acid or in dilute or concentrated trifluoro-acetic acid, or palladium-catalyzed cleavage in THF or THF-DCM mixtures in the presence of a weak base ~uch as morpholine. 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 is to carry out certain derivatization reactions or a cyclization.

Some of the novel peptides have good cytotoxic proper-ties. Some others of the peptide~ have high affinity for the cellular TNF receptor without, however, having cytotoxic activity. They are therefore TNF antagonists.
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 diseases a~
well as for controlling and preventing infections, inflammations and tran~plant re~ection reactions. Simple experLments can be used to elucidate the mode of action of the individual peptides. The cytotoxicity of the peptide i8 determined by incubating a TNF-sensitive cell line in the pre~ence of the peptide. In a second experi~
mental approach, the cell line iB incubated with the relevant peptide in the presence of a lethal amount of TNF. It is possible in this way to detect the TNF-antagoni~tic effect. In addltion, the affinity of the psptide for the cellular TNF receptor is determined in an in vitro binding experiment.

The following te~t ~y~t~ms were used to characterize the agoni~tic and antagonistic effects of the novel peptides:

I. Cytotoxicity test on TNF-~ensitive indicator cells, II. Cytotoxicity antagonism test on TNF-sensitive ~0~;~81.

- 8 - O.Z. 0050/40383 indicator cells, III. Competitive receptor-binding test on indicator cells expressing TNF receptor.

I. Cytotoxicity test The agonistic effect~ of the novel peptide~ are assessed on the basis of their cytotoxic effect on TNF-sQnsitive cell~ (e.g. L929, MCF-7, A204, U937).
The test with L929 and MCF-7 was carried out as follow8 5 1. 100 ~1 of culture medium containing 3 to 5 x 103 freshly tryp~inized, exponentially growing, L929 cell~ (mouse) or MCF-7 cells (human) were pipetted into the wells of a 96-well flat-bottom culture plate. The plate wa~ incubated at 37C
overnight. The air in the incubator was ~aturated with water vapor and contained 5% CO2 by volume.

The L929 culture medium contained 500 ml of lx Earle~s M~M (Boehringer Mannheim), 50 ml of heat-inactivated (56C, 30 min) fetal calf serum (FC5), 50 ml of L-glutamine (200 mM), 5 ml of lOOx non-es~ential amino acids, 3 ml of 1~ HEP~S
buffer pH 7.2,and 50 ml of gentamicin (50 mg/ml).

The NCF-7 culture medium contained 500 ml of lx Dulbecco~s ME~ (Boehringer Mannheim), 100 ml of heat inactivated (56-C, 30 min) FCS, 5 ml of ~-glutamine and 5 ml of lOOx non-essential amino acids ~

2. The next day 100 ~1 of the peptide solution to be te~t~d ware added to the cQll culturQs and sub~ected to serial 2-fold dilution. In addition, qom~ cell controls (i. Q . CQll culture~ not treated with peptide dilution) and some rhu-TNF

~0~
~ 9 - O.Z. 0050/40383 controls (i.e. cell cultures treated with recom-binant human TNF) were also made up. The culture plate waB incubated at 37C in an atmosphera of air saturated with water vapor and containing 5%
C02 by volume for 48 h.

3. The percentage of surviving cells in the cultures treated with peptide dilution was determinad by staining with crystal violet. For this purposQ, the liquid was removed from the wells of the test plate by tapping it. 50 ~1 of crystal violet solution were pipetted into each well.

The composition of the crystal violet ~olution was as followss 3.75 g of cry~tal violet lS 1.75 g of NaC1 161.5 ml of ethanol 43.2 ml of 37% formaldehyde water ad 500 ml The crystal violet solution wa~ left in the wells for 20 min and then likewise removed by tapping.
~he plate~ were then wa~hed 5 times by immersion in water in order to remove dye not bound to the cell~. The dye bound to the cells was e~tracted by adding 100 ~1 of reagent solution (504 eths-nol, 0.1% glacial acetic acid, 49.9~ water) to each well.

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

Z~05~8~
- 10 - O.Z. 0050/40383 5. Subs~quently, by relating to the cell control, the 50% cytotoxicity value was defined, and the reciprocal of the ~ample dilution which resulted in 50~ cytotoxicity was calculated as the cyto-toxic activity of the te~t sample.

II. Cytotoxieity antagoniqm test The antagonistic effect of the peptides was a~se~ed on the basis of their property of antagonizing the cytotoxie effeet of rhu-TNF on TNF-sensitive cells (~.q. L929, MCF-7, A204, U937). Th~ cytotoxicity antagonism teat with L929 and MCF-7 eells was earried out as follow~t 1. 100 ~1 of eulture medium eontaining 3 to 5 x 103 freshly trypsinized, exponentially growing, L929 cells (mou~e) or MCF-7 eell~ (human) were pipatted in~o the wells of a 96-well flat-bottom eulture plate. The plate was ineubated at 37C
overnight. The air in the ineubator was saturated with water vapor ~nd eontained 5% CO2 by volume.

The L929 elllture medium eontained 500 ml of lx Earle's M~M (Boehringer Mannheim), 50 ml of heat-inaetivated (56C, 30 min) PCS, 5 ml of L-gluta-mine (200 m~), 5 ml of lOOx non-es~ential amino aeids, 3 ml of lM HEPES buffer pH 7.2, and 500 ~1 of gentamiein (50 m~/ml).

The NCF-7 eulture medium eontained 500 ml of lx Dulbeeeo~s NEM (Boehringer Mannheim), 100 ml of heat inaetivated (56C, 30 min) FCS, 5 ml of L-glutamine (200 mM) and S ml of lOOx non-essen-tial amino aeids.

2. ~he next day 100 ~1 of the peptide solution to be teEted were added to the cell eulture~ and sub~eet2d to ser~al 2-fold dilution. Then, 100 ~1 ~00~81.

~ O.Z. 0050/40383 of a rhu-TNF 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 cultures. In addition, some cell controls (i.e. cell culture~ not treated with peptide solution or with rhu-TNF solution) and some rhu-TNF controls (= cell cultures treated only with rhu-TNF solution) were also made up. The culture plate was then incubated at 37C in an atmosphere of air saturated with water vapor and containing 5% C02 by volume for 48 h.

3. The percentage of surviving cells in the culture~
treated with sub~tance solution was determined by ~taining with crystal violet. For this purpose, the liquid wa~ removed from the well~ of the test plate by tapping it. 50 ~1 of crystal violet solution were pipetted into each well.

The crystal violet solution had the composition specifi~d in I.3 The crystal violet solution was left in the well~
for 20 min and then likewise removed by tapping.
The plate~ were then washed 5 times by immersion in water in order to remove dye not bound to the cell~. The dye bound to the cells was extracted by adding 100 ~1 of reagsnt 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 301ution of uniform color in each well. The sur~iving cells were determined by measuring the extinction at 540 nm of the colored solution in the individual wells.

zoos~
- 12 - O.Z. 0050/~0383 5. Subsequently, by relating tO the cell control and the rhu-TNF control, the 50~ antagonism value was defined, and the ~ample concentration which regulted in 50~ antagonism of rhu-TNF cytotox-icity at the rhu-TNF concentration used was calculated as antagonistic activity of the sample te~ted.

III. Competitive receptor-binding test Both the agonistic and antagonistic effect~ of peptides are conditional on the iatter binding to the TNF receptor. Thi~ means that peptides with an agonistic or antagonistic effect compete with rhu-TNF for binding to the TNF receptor on TNF-sensitive indicator cells (e.g. U937). The competi-tive receptor-binding te~t was carried out as followss 1. 100 ~1 of medium containing various concentra-tions of the peptide to be te~ted and of rhu-TNF
(= control) were pipatted into the reaction ve~sels. The medium comprised 500 ml of PBS
(Boehringer Mannheim), 10 ml of heat-inactivated (56UC, 30 min) FCS and 100 mg of sodium a2~ide.

2. Subsequently, 100 ~1 of medium containing 1 ng of 1Z5I-labsled rhu-TNF (Bolton lactoperoxida~e method) were placed in the reaction vessels and mixed. The non-specific b~nding (NSB) was deter-mined by mixing in the reaction vessel~ the 125I-laboled rhu-TNF (1 ng of125I-rhu-TNF in 100 ~1 of medium) with a 200-fold excess of unlabaled rhu-TNF ~200 ng of rhu-TNF in 100 ~1 of medium)~

3. Then 100 ~1 of medium contsining 2 x loB U937 cells (human) wsre pipetted into the reaction ve~8el8 and mixed. The reaction ves~al~ (test - 13 - O.Z. 0050/40383 volume 300 ~1) were incubated at 0C for 90 min.
The reaction mixture~ were remixed after 45 min.

4. After the incubation, the cell~ were centrifuged at 1800 rpm and 4C for 5 min, wa~hed 3 times with medium and tran~ferred quantitatively into cou~ting vials, and the cell-bound radioactivity was determined in a Clini gamma counter 1272 (LRB Wallac).

5. After the measurement~ had been corracted for the non-~pecific binding, the 50~ competition value was defined by relating to the overall binding, and the sample concentration which led to 50%
competition ofl25I-rhu-TNF binding at the125I-rhu-TNF concentration used wa~ calculated as the competitive activity of the sample tested.

The Examples which follow ar~ intended to explain the invention in more detail. The proteinogenous amino acids are abbreviated in the Examples u~ing the conventional three-letter code. Other meanings ares Abs = 4-æminobutyric acid, Ac = acetic acid, Ahp - 7-aminoheptanoic acid, Ahx = 6-aminohexanoic acid, Bal = ~-alanine, Hcy = homocysteine, Hly = homolysine, Orn - ornithine.

A. General procedures I. The peptides claimed in claim 1 were synthesized u~ing standard methods of solid-phase peptide synthe~is in a completely automatic model 430A
peptide synthesizer from APP~IED BIOSYSTEMS. The apparatus use~ different ~ynthesis cycles for the Boc and Fmoc protective group techniques.

a) Synthesis cycle for th~ Boc protective group ~0~
- 14 - O.Z. 0050/40383 technique 1. 30% trifluornacetic acid in DCM 1 x 3 min 2. 50% trifluoroacetic acid in DCM1 x 17 min 3. DCM washing 5 x l min 4. 5% dilsoprcpylethylamine in DCM l x 1 min 5. 5% di~3aprcpylethylamine in NMP 1 x 1 min 6. NMæ wzshing 5 x 1 min 7. A~dition of preactivated pGctocted amino acid (activation by 1 equival~L of DCC
and 1 Q~ivalent of HOBt in NMP/DCM);
peptide coupling (lst part) 1 x 30 min 8. A~dition of DMSO to the reacticn nixturs u~rtil it ccntains 20% ~90 }~y vellm~

9. Peptide co~pling (2~ part) 1 x 16 min 10. A~iti~ of 3.8 ffa!liV~ of ~ii~
p~letl~ylan~ to tho ~n nL~

11. P~e ca~pling (3rd part) ~ x 7 min 12. DCM washirbg 3 x 1 min 13. If reactian i~ i~lote, rE~etiticsn of ca~lir~ (retu~n to 5. ) 14. 10% acetic ar~ride, 5% dii~pyl-~ ylamine in DCM 1 x 2 min 15. 10S acetic ar~ydride in DC~ 1 x 4 min 16. DCM wa~hing 4 x 1 min 17 Reburn to 1.

b) SyntheBiR cycle for the Fmoc protective group techndquo 1. ~IP wa~ 1 x 1 min 2. 20~ piperidine im NMP 1 x 4 min 3. 20% piperidine in N~P 1 x 16 n~n 4. N~P w ~ g 5 x 1 m~n 5. A~dition of preactivated proeeltod amino acid (activatian ky 1 eq~ivalent of DOC
and 1 equivalent of HLBt in NMP/DCM);
~ e ooupling 1 x 61 n~n 6. NNP washing 3 x 1 min 7. I~ reacti~n i8 inocmple~e, repekiti~n of Z ~ 0 S~ 8~
- 15 -O.Z. OOS0/4~383 o~ling (return to 5.~
8. 10~ ic ~ride in NMP1 x 8 min 9. NMP washing 3 x 1 min 10. R~l~n to 2.

II. Working up of peptide-resins obtained as in Ia The peptide-resin obtained as in Ia wa~ dried under reduced pressure and transferred into a reaction vessel of a Teflon HF apparatus (from PENIN~ULA).
Addition of a scavenger, preferably anisole (1 ml/g ld of resin), and of a thiol in the case of tryptophan-containing peptides, to remove the indole formyl group, preferably ethanedithiol (O.S ml/g of resin), was followed by condensation in of hydrogen fluoride (10 ml/g of resin) while cooling with liquid N2. The lS mixture wa8 allowed to warm to O-C, and was stirred at this temperature for 45 min. The hydrogen fluor-ide was then ~tripped off under reduced pressure and the residue was wa~hed with ethyl acetate in order to removs remaining scavenger. The peptide was extracted with 30% strength acetic acid and filtered, and the filtrate was freeze-dried.

To prepare peptide hydrazides, the peptide-resin (Pam- or ~errifield resin) was suspended in DME
(15 ml/g of resin), hydrazine hydrate (20 equiva-~5 lents) w~s added, and the mixture was stirred at room temperature for 2 days. To work up, the resin wa~ filtered off and the filtrate was evaporated to dryness. The re~idue was crystallized from DME/Et20 or Me0H/Et20~

III. Working up of the peptide-resins obtained as in Ib The peptide-resin obtained as in Ib was dried under reduced pressure and subsequQntly sub~ectad to one ~005~81.
- 16 - O.Z. 0050/40383 of the following cleavage procedures, depending on the amino acid composition (Wade, Tregear, Howard Florey Fmoc-Workshop Manual, Melbourne 1985).

Peptide containing Cleavage conditionY
__ _ _ Arg(Mtr~ Met Trp TFA Scavenger Reaction tLme . . _ _ no no no 95% 5% H2O 1.5 h yes no no 95% 5% thioani~ole 2 3 h no yes no 95% 5% ethyl methyl 1.5 h sulfide no no yes 95% 5% ethanedithiol/ 1.5 h anisols (1:3) no yes yas 95~ 5~ ethanedithiol/ 1.5 h anisole/ethyl methyl sulfide (1:3sl) yes yes yes 93% 7% ethanedithiol/ 2 3 h anisole/ethyl methyl sulfide (1:3:3) The ~uspension of the peptide-resin in the suitable TFA mixture was stirred at room temperature for the stated time and then the re~in was filtered off and washed with TFA and with DCM. The filtrate and the washings were extensively concentrated, and the peptide was precipitated by addition of diethyl ether. The mixture was cooled in an ice bath, and tha precipitate wa~ filtered off, taken up in 30%
acetic acid and freeze-dried.

IV. Purification and characterization of the peptides Purification was by gel chromatography (SEPHADEX~
G-10, G-15/10% HQAc; SEPHADEX~ LH20/MeOH) and sub-Z~O~X81 - 17 - O.Z. 0050/40383 ~equent medium pressure chromatography (~tationary pha~e: HD-SIL C-18, 20-45 ~, 100 A; mobile phases gradient with A = 0.1% TFA/MeOH, B = 0.1% TFAtH2o).

The purity of the final products was determined by analytical HPLC (~tationary phase: 100 x 2.1 mm YYDAC C-18, 5 ~, 300 A; mobile phase = CH3CN/HzO
gradient buffered with 0.1% TFA, 40C). Charac-terization was by means of amino acid analy~i~ and fast atom bombardment ma~s spectrometry.

B. Specific procedures H-Arg-A8p-A~n-Gln-Leu-NH2 1.1 g of Boc-Leu-p-MBHA-resin (substitution 0.45 mmol/g), corresponding to a batch size of 0.5 mmol, were reacted a~ in AIa with 2 mmol each of Boc-Gln-OH
Boc-A~p(OChx)-OH
Boc-Asn-OH
Boc-Arg(Tos)-OH

After the ~ynthe~is was complete, ths peptide-resin underwent N-terminal deprotection (steps 1-3 as in AIa) and subsequent drying under reduced pre~sure; the yield was 1.45 g.

0.73 g of the resin obtained in this way was sub~ected to HF cleavage a~ in AII. The crude product (125 g) was purified by gel filtration (SEPHADEX G-10) and medium pressure chromatography (cf. ~IV; 10-25 ~ A; 0.25 %
min~1). 89 mg of pure product wsrQ obtained.

~ 5æI~ a 2005~1 Ac~eu~ A8p-~n~ ln~u-0~

3.43 g o~ u-p-alkox~an~yl a~ohol-~e~ln (suhs~i-~u~on ~.5g mms:~l/g~ ~orresponding ~o a batch ~i~e ~f S 0.25 ~nol, wa~ ~eaotsd a~ in ~Ib with 1 mmol each of Fmoc--t ;ln-OK E~oc -Arg ( Mtr ) -O~
Fmoc-A4n-O~ Fmoc-L~u~
Fmoc~ 0t~u ) -0~1 Aft~sr the ~ he~i~ w~ co~ te, the N te~minu~ waQ
ac~tyl~ted ~te~s ~-J, ~d 8-9 as in AIb)~ T~e re~ultir~g ~eptide-re~in wa~ d uz~Ldar ~educed pressure; the yielci wa8 0-6 ~T-~Q c:rude ~ptLdo ~143 ~g) sbtained after ~FA cleavage as in AIII wa~ pu~ ed ~y ~el ~iltrat~on (5~P~E~ 10) and m~diu3n pre~ure ch~omatography (ef. AIV; 15-25 ~ At O.25 96 m~n ~j . 107 mg o~ pure product were obtained~

~h~ ~ollow~n~ e ~rQp~r~ Ln a ~nilar ~a~uter to ~x~nples 1 and 2 *~ GESRI`lTSE I TEN 0132 `~"

Z005~81.

- 19 - O.Z. 0050/40383 3. H-Arg-Asp-Asn-Gln-Leu-OH
4. Ac-Arg-Asp-Asn-Gln-Leu-OH
5. Ac-Arg-Asp-Asn-Gln-Leu-NH2 6. H-Leu-Ar9-Asp-Asn-Gln-Leu-oH
7. ~-Leu-Arg-ASp-ASn-Gln-Leu-NH2 8. Ac-Leu-Arg-Asp-Asn-Gln-Leu-NH2 9. H-Glu-Leu-Arg-Asp-Asn-Gln-Leu-OH
10. Ac-Glu-Leu-Arg-Asp-Asn-G1n-Leu-OH
Il. H-Glu-Leu-Arg-Asp-Asn-Gln-Leu-NH2 12. Ac-Glu-Leu-Arg-Asp-Asn-Gln-Leu-~H2 13. H-Glu-Leu-Arg-Asp-Asn-Gln-Leu-Val-OH
14. Ac-Glu-Leu-Arg-Asp-Asn-Gln-Leu-Val-NH2 15. ~-Val-Glu-~eu-Arg-Asp-Asn-Gln-Leu-Val-OH
16. Ac-Val-Glu-Leu-Arg-Asp-Asn-Gln-Leu-Val-NH2 17. H-Gly-Val-Glu-Leu-lrg-Asp-~Sn-Gln-L~u-Val-Val-OH
18. Ac-Gly-Val-Glu-Leu-Arg-Asp-Asn-Gln-Leu-Val-Val-NH2 19. Ac-Leu-Arg-Asn-Asn-G~n-Leu-NH2 20. Ac-Glu-Leu-Arg-~sp-Asn-Ser-Leu-NH2 21. ~-Val-Glu-L~u-~rg-~sn-Asn-Gln-Lou-Val-OH

22. H-61y-~hr-Glu-Leu-~rg-Asp-Asn-Gln-Leu-Val-Val-OH

EX~PL~ 23 Ac -Cys -Arg-A~p-Asn-Gln-Cys -NH2 1.16 g of Boc-Cys (pMB) -p-MBHA-resin ( substitution 0 . 43 mmol~g), corresponding to a batch size of 0 . 5 mmol were reacted as in AIa with 2 mmol each of Boc-Gln-OH Boc-Arg ( Tos ) -OH
Boc-Asn-OH Boc-Cys ( pMB ) -OH
Boc-Asp ( OChx) -OH

The resulting peptide-resin was dried under reduced pressure; the yield was 1.45 g.

0.73 g of the resin obtained in this way was ~ub~ected to HF cleavaga as in AII. The freeze-dried crude product wa~
talcen up in 2 1 of 0.195 strength acetic acid, and the pH
wa~ then ad~usted to 8 . 4 with aqueou~ ammonia . Under an ZOO~
- 20 - O.Z. 0050/40383 argon atmosphere, 0.01 N K3rFe(CN)6] solution was slowly added dropwise until the yellowish-green color per~i~ted for at lea~t 15 min. The mixture was then stirred for 1 h and then acidified to p~ 4.5 with glacial acetic acid, and 15 ml of an aqueou~ ~u~pen~ion of an anion exchanger (BIORAD~ 3 x 4A, chloride form) were added. After 30 min, the ion exchanger resin was filtered off, and the filt-rate was concentrated to 100 ml in a rotary evaporator and subsequently freeze-dried.

All the solvent~ used had previously been saturated with nitrogan in order to prevent any oxidation of the free cysteine residues.

The crude product was purified by gel chromatography (SEPHADEX G-15) and medium pressure chromatography (cf.
AIV; 10-30 % A; 0.25 ~ min1). 71 mg of pure product were obtained.

The following can be prepared in manner similar to Example 23 (Pam-recin was used to prepare the peptide acid~)s 24. H-Cys-AFg-Asp-Asn-GIn-Cjs-OH
25. Ac-Cys-Arg-Asp-Asn-Gln-Cys-OH
26. AC-CyS-Arg-O-Asp-Asn-Gln-Cjs-NH2 27. H-cys-Ar9-Asp-Asn-Gln-cys-NH2 28. Ac-CjS-Glù-ASp-ASn-Gln-CyS-NH2 29. H-Cys-Glu-Asp-Asn-Gln-Cys-OH
30. H-Cys-Thr-Asp-Asn-Gln-Cys-OH
31. Ac-Cys-Thr-Asp-Asn-Gln-Cys-NH2 ;~OSZ~l.
- 21 - O.Z. 005~/40383 32. H-Cys-Ly5-A5p-Asn-Gln-Cj5-OH
r 33. Ac-Cys-Lys-Asp-Asn-Gln-Cy5-NH2 34. H-Cys-Ser-Asp-Asn-Gln-Cys-OH
35. Ac-Cys-Ser-Asp-Asn-Gln-Cys-NH2 36. Ac-Hcy-Arg-Asp-Asn-Gln-Cys-NH2 37. Ac-Cys-Arg-Asp-Asn-Gln-Hcy-NH2 38. Ac-Hcy-Arg-Asp-Asn-Gln-Hcy-NH2 39. Ac-Cys-Leu-Arg-~sp-Asn-Gln-Cys-NH2 40. Ac-Hcy-Leu-Arg-Asp-Asn-Gln-Cys-NH2 41. Ac-Cy5-Arg-A5p-Asn-Gln-Leu-Cjs-NH2 42. Ac-Hcy-Arg-Asp-Asn-Gln-Leu-Cjs-NH2 43. H-Cys-Leu-Arg-Asp-Asn-Gln-Leu-Cys-OH
44. Ac-Cys-Leu-Arg-Asp-Asn-Gln-Leu-Cys-OH
45. H-Cy5-Leu-Arg-Asp-~sn-Gln-Leu-Cys-NH2 46. Ac-Cys-Leu-Arg-Asp-Asn-Gln-Leu-Cys-NH2 47. Ac-Hcy-Leu-~rg-Asp-~sn-Gln-Leu-Cys-NH2 48. Ac-CyS-Leu-Arg-Asp-Asn-GI n-Leu-Hcy-NH2 49. Ac-Hcy -Leu-Arg-Asp-ASn-Gln-Leu-Hcy-NH2 50. H-Cys-Arg-Asp-Asn-Gln-Leu-Cys-NHz 51. Ac-Cy5-Arg-Asn-Asn-Gln-Cys--~H2 52. Ac-Cys-Arg-D-Asp-Asn-Gln-CyS-NH2 53. Ac-Val-Glu-Cjs-Arg-Asp-Asn-Gln-Cjs-Val-Val-Pro-NH2 54. Ac-Cys-Lys-Asp-Asn-Ser-Cys-NH2 55. Ac-Asn-Gly-Yal-Cys-Leu-Arg-Asp-Asn-Gln-Leu-CjS-Val-Pro-NH2 OS~81 - 22 - O.Z. 0050/40383 Ac-Glu-Arg-Asn-Asn-Gln-Ly~-NH2 1.16 g of Boc-Lys(Cl-Z)-MBHA-resin (substitution 0.43 mmol/g), corre~ponding to a batch size of 0.5 mmol, were reacted as in AIa with 2 mmol each of Boc-Gln-OH Boc-Arg( TOB ) -OH
Boc-Asn-OH Boc-Glu(OBzl)-OH
Boc-Asn-OH

After the synthesi~ was complete, the N terminus was acetylated (steps 1-6 and 14-16 as in AIa). The resulting peptide-re~in was dried under reduced pressure; the yield was 1.6 g.

~ he crudo product (317 mg) obtained after HF cleavage a~
in AII was dissolved in 500 ml of degassed DNF, and 0.2 ml of triethylamine and (at -25C) 0.20 ml of di-phenylphosphoryl azide were added. The mixture was stirred at -25C for 2 h, ~tored at -20C for 2 ~ays~ at 4C for 2 days and at room tamperature for 2 day~ and ~ubsequently evaporated to drynes~. The crude peptide was purified by gel chromatography (SEPHADEX LH 20) and medium pre~ure chromatography (cf. AIV; 10-25% A;
0.25% min~l). 101 mg of pure product were obtained.

Examplo 57 Ac-Orn-Arg-Asp-Asn-Gln-ABp-NH2 2.5 g of reQin de~cribed by Breipohl et al. (from BACHEM), corresponding to a bstch size of 1.25 mmol, was reacted a~ in AIb with 5 mmol each of 200$Z~

- 23 - O.Z. 0050/40383 Fmoc-A8p(Ot~u)-OH Fmoc-Asp(OChx)-OH
Fmoc-Gln-OH Fmoc-Arg(~os)-OH
Fmoc-Asn-OH Fmoc-Orn(Boc)-OH

After the synthesis was complete, the N terminus was acetylated (step~ 2-4 and 8-9 as in AIb). The peptide-resin was dried under reduced pressure; yield 1.4 g.

The crude product (1.22 g) obtained after TFA cleavage as in AIII was purified as in AIV (414 mg). 200 mg were dissolved in 250 ml of degassed DMF. 0.24 ml of NEt3 and, at -25C, 0.24 ml of diphenylphosphoryl azide were added and then the mixture was stirred at -25C for 2 h. It was subsequently stored at -20C for 2 days, at 4C for 2 days and at room temperature for 2 day~. It was then evaporated to dryness, and ths crude peptide was purified by gel chromatography (SEPHADEX LH 20). The i~olated monomer (143 mg) was deprotected with HF as in AII and purified by medium pressure chromatography (cf. AIV;
5-25~ A; 0.25% min1). 89 mg of pure prod~ct werQ
obtained.

~xample 58 Ac-Glu-Leu-Arg-Asp-A~n-Gln-Leu-Lys-OH

3.2 g of Fmoc-Lys(Boc)-Merrifield resin (substitution 0.38 mmol/g), corresponding to a batch si~Q of 1.0 mmol, were reacted as in AIb with 4 mmol each of Fmoc-Leu-OH Fmoc-Arg(To~)-OH
Fmoc-Gln-OH Fmoc-Leu-OH
Fmoc-Asn-OH Fmoc-Glu(OtBu)-OH
Fmoc-Asp(OChx)-OH

Subsequently N-terminal deprotection and acetylation (step~ 2-4 and 8-9 as in AIb) were carried out, and the X~OS~81.

- 24 - O.Z. 0050/40383 t-butyl and Boc protective groups were cleaved off (~teps 1-6 as in AIa). The cycliz~tion on thr re~in took place in NMP with addition of 1.77 g of BOP and 1.74 ml of diisopropylethylamin~ (40 h). The peptide-resin was dried under reduced pressure. Thr yield was 3.g5 g. The crude product obtained after HF cleavage a~ in AII was purified by gel filtration (Sephadex G-15) and medium pre 3ure chromatography twice (cf. AIV; 5-25% A; 0.25% min~').
17 mg of pure product were obtained.

The following can be prepared in a sLmilar manner to Examples 56, 57 and 58:
59. Ac-GIu-Asp-Asn-GIn-LyS-NH2 60. bc-GIu-Arg-Asp-Asn-GIn-Lys-NH2 61. Ac-Asp-Arg-Asp-Asn-GIn-Lys-NH2 62. H-ASp-Arg-Asp-Asn-GIn-Ljs-NH2 63. Ac-Asp-Arg-Asp-ASn-Gln-Orn-NH2 64 . AC -Lys-Arg-Asp-Asn-Gln-G~u-OH
65. H-Lys-Arg-Asp-Asn-Gln-Glu-OH
66. Ac-G~u-Arg-Asp-Asn-Gln-Hty-NH2 67. H-0rn-Arg-A5p-ASn-GlU-NH2 68. Ac-Giu-Asp-Asn-Ser-Lyjs-NH2 69. Ac-Lys-Arg-Asp-Asn-Gln-Asp-NH2 70. Ac-Lys-Arg-Asp-Asn-Gtn-~sp-OH
71. Ac-Glu-Arg-Asp-Asn-Gln-LjS-OH
72. H-G~u-~rg-Asp-Asn-Gln-Ljs-OH
73. Ac Lys-Arg-A5p-~sn-Gln-Glu-N~2 Z0052~

- 25 - O.Z. 0050/40383 74. Ac-Glu-Leu-Arg-Asp-Asn-Gln-Leu-Lys-OH
75. Ac-Glu-Arg-Asp-Asn-Gln-Leu-Lys-NH2 ~6. Ac-Glu-Arg-Asp-Asn-Gln-Leu-Orn-OH
1-- ~
77. Ac-Orn-Arg-Asp-Asn-Gln-Leu-Asp-NH2 78. Ac-Glu-Leu-Arg-Asp-Asn-Gln-Leu-Lys-NH2 79. Ac-G~u-Leu-Arg-Asp-A5n-Gln-Leu-Hiy-NH2 80. Ac-Asp-Leu-Arg-Asp-Asn-Gln-Leu-Lys-NH2 81. Ac-Asp-Leu-Arg-Asp-Asn-Gln-Leu-Orn-NH2 82 . AC -Orn-Lcu-Arg-Asp-Asn-61n-~eu-Asp-NH2 83. Ac-Lys-L~u-Arg-Asp-Asn-Gln-Leu-Asp-NH2 84. Ac-Lys-Leu-Arg-Asp-Asn-Gln-Leu-Asp-OH
85. Ac-Lys-Leu-Arg-Asp-Asn-Gln-Leu-Glu-NH2 86. Ac-Glu-Asp-Asn-61n-Leu-Ljs-NH2 87. ~-G1u-Asp-~sn-Gln-Leu-Lys-OH

88. Ac-Lys-Asp-Asn-Gln-Leu-G u-NHz r 89. Ac-Gly-Val-61u-Orn-~rg-Asp-~sn-Gln-~sp-Val-Val-NH2 90. ~c-Asp-~5-Asp-~n-Gln-L~s-NH2 1. ~c-Gtu-Leu-~rg-D-~sp-~sn-Gln-L~u-Lys-Nt~2 2. ~c-~n-~ly-Val-Orn-Leu-~rg-~sp-~sn-Gln-Leu-~sp-Val-Pro-NH

~0~

- 26 - O.~. 0050/40383 Exampl~ 93 IBal-Arg-Asn-Asn-Gln-Leul 0.56 ~ of Boc-Leu-Merrifield re~in (Yubstitution 0.9 mmol/g), corre~ponding to a batch size of 0.5 mmol, wa~ reacted as in AIa with 2 mmol each of Boc-Gln-OH Boc-Arg(To~)-OH
Boc-Asn-OH Boc-Bal-OH
Boc-Asn-OH

After the synthesis was complete, the peptide-resin underwent N-terminal deprotection (steps 1-3 as in AIa) and subsequent dryinq under reduced pressure. The yield was 0.85 g.

The crude product (267 mg) obtained after HP cleavage a~
in AII was di3solved in 500 ml of degassed DNF. 210 mg of NaHCO3 and, at -25C, 0.20 ml of diphenylphosphoryl azide were added and then the mixture wa~ ~tirred at -25C for 2 h and at room temperature for 2 days. It was then evaporated to dryness, and the crude peptide was purified by gel chromatography (SEPHADEX0 LH 20) and medium pres-sure chromatography (cf. AIV; 20-50~ A; 0.25% min~l).
75 mg of pure product were obtained.

Example 94 ~Glu-Leu-Arg-Asp-Asn-Gln-Leu-Val 0.88 g of Fmoc-Glu(OtBu)-p-alkoxybenzyl alcohol-re~in (substitution 0.57 mmol/g), corre~ponding to a batch sizQ
of 0.5 iol, wa3 reacted as in AIb with 2 mmol ~ach of ;~005;~

- 27 - O.Z. 0050~403~3 Fmoc-Val-OH Fmoc-Asp(OBzl)-OH
Fmoc-Leu-OH Fmoc-~rg(Tos)-OH
Fmoc-Gln-OH Fmoc-Leu-OH
Fmoc-As~-OH

After the ~ynthesi~ was complete, the peptide-re-~in underwent N-terminal deprotection (steps 2-4 a~ in AIb) and subsequent drying under r~duced pressure. The yi~ld was 1.2 g.

The crude peptide obtained after TFA cleavag~ a~ in AIII
was diRsolved in 500 ml of degassed DMF 210 mg of NaHCO3 and, at -25C, 0.24 ml of diphenylphosphoryl azide were added, and the mixture was stirred at -25C for 2 hours and at room temperature for 2 days. It was then evapora-ted to dryness, and the crude peptide was purified by gel chromatography (SEPHADEX LH 20). The isolated monomer (87 mg) was deprotected with HF as in AII and purified by medium pre~sure chromatography (cf. AIV; 25-45% A; O.25%
min~~. 49 mg of pure product were obtained.

The following can be prepared in a similar m~nner to Examples 93 and 94s -95. rArg-Asp-Asn-Gln-Ahx 96. Asp-Asn-Gln-Leu-~h~
r _ l 5~. rArg-asp-Asn-Gln-~hp '?3. rArg-Asp-~sn-GIn-Leu-3al 99. ~y-~rg-~sp-Asn-Gln-Leu lOO.rLeu-Arg-ASp-Asn-Gln-LCU-Ah 101.rGlu-L~u-Arg-Asp-~sn-Gln-Leu-Bal 102. rLeu-Arg-D-Asp-Asn-Gln-L
103. r~eu-Lys-Asp-~sn-Gln-Leu-~h 104 . r~bs-~rg-~p-~ n-61n-Leul ;~oos~

- 28 - O.Z. ~050/40383 lQ5.rLeu-~rg-~sp-Asn-GIn-D-Leu 106. rLeu-Arg-~sp-Asn-GIn-0-Pro 107. rLeu-Arg-Asp-Asn-GIn-GIy 108. rLeu-Arg-Asp-Asn-Gln-D-Ala

Claims (8)

1. A peptide of the formula I

X-A-Asn-B-Y I, where A is Asp or Asn, B is Gln or Ser, X is G-NH-CHM-CO-, G-NH-CHM-CO-W-, G-R-NH-CHM-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, U, V and W are peptide chains composed of 1-4 natu-rally occurring .alpha.-amino acids and M and Q are hydrogens or one of the following groups -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, 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)?-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 tolera-ted acids.

2. 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 are not - 30 - O.Z. 0050/40383 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 qroup, 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 -(CH2)?-NH-CO-(CH2)f- or -(CH2)?-NH-CO-(CH2)g-NH-CO-(CH2)f.

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

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

7. The use of a peptide 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.