CA2005057A1 - Tnf peptides - Google Patents

Abstract

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

Description

'~0050~7J
0. Z . 0050/403139 NOVEL TNF PEPTIDES

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

Carswell et al. (Proc. Natl. Acad. Sci. USA 72 (1975) 3666) reported that the serum of endotoxin-treated animal~ which had previou~ly been infected with the Calmette-Guerin strain of Mycobacteria (BCG) brought about hemorrhagic necrosis in various mouse tumor~. This activity was ascribed to tumor necro~is f~ctor. ~NF also has a cytostatic or cytotoxic effect on a large number of transformed cell lines in vitro, whereas normal human and animal cell lines are unaffected (Lymphokine Reports Vol.

2, pp 235-275, Academic Press, New York, 1981). Recently, the biochemical characterization and the g~ne for human TNF have been described (Nature 312 (1984) 724, J. Biol.
Chem. 260 (1985) 2345, Nucl. Acids Re~. I3 (1985) 6361).

It is possible to deduce from this data the following protein structure for mature human TNF:

VaL~x ~ 5e~:~te~pro;~lypLysprovsL~ 3valvaLukA~r~
GlnAlæJu~_~
ValGau~ ~c~ valvarpndb~GluGayL~yr~b~ er ~V lT~P}~Ly~Gly~lyCysR~V~l~am~Ile S~d~gIleAlaV~lionyrG~fSrLysVar~e~ o~leLyES~rPro Cy~nAn~uThrPn~uGlyA~uAlaLysPnf~pCycluProIl~u ~ GlyV~lPh~l ~ ~ GlyA~pr~a ~ uIl ~ a ~
T ~ ~ Ga ~ ~ Valq ~ ?heGayIleIl ~

The TNF gene~ of cattle, rabbits and mice have al80 been de3cribed (Cold Spring Harbor Symp. Quant. Biol. 5 (1986) 597).

Z O O ~ Q ~
- 2 - ~.Z. 0050~403~9 Be~ides its cytotoxic propertie~, TNF is one of the main substances involved in inflammatory reaction~ (Pharmac.
Res. 5 (1988) 129). Animal models have shown that TNF is involved in septic shock (Science 229 (1985) 869) and graft-versus-host disease (J. Exp. Med. 166 (1987) 1280).

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

The present i~vention relate~ to peptides of the formula I
X-A-Y
where A is -Arg-Pro-Asp-Tyr-, -Leu-Pro-Asp-Tyr-, -Gln-Pro-Glu-Tyr-, -Leu-Pro-Lys-Tyr-, -Gly-Ile-Pro-His- or -Gly-Ile-Ser-His-, 15 X is G-, G-NH-CHM-CO-, G-NH-CHM-CO-W-, G-R-NH-CXN-CO-or G-R-NH-CH~-CO-W- and Y is -Z, -NH-CHQ-CO-Z, -V-NH-CHQ-CO-Z, -Nff-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 is OH or NH2 or a carboxyl-protective group or G and Z together are al~o a covalent bond or -CO-(CH2),-NH-, where a i8 from 1 to 12, R, U and W are peptide chain~ compo~ed of 1-4 naturally occurring ~-amino acid~, V i8 a p~ptide chain compo3ed of 1-10 naturally occurring ~-amino acids and M and Q are hydrogen~ or ons of the follo~ing -CH(CH3) 2 ~ -CN(CH3)-C2H~, -CoH~, -CH~OH) -C~3 ~

-ÇH2 ~ -CH2 ~ or -(CH2)b-T
(with b being from 1 to 6 And T beLnq hydrogen or OH, CH30, CH3S, (CH3)2CH, C6H5, p-HO-C6H4, HS, H2N, HO-CO, HzN-CO, HzN-C(-NH)-NH) or M and Q together are a -(cHz)c-s-s-~cHz) d- ~ - ( CH2 ) ~-CO-~H-~0~057 - 3 - o.æ. 0050/40389 ~CH2)~- or -(CH2)e-NH-CO-(CHz)~-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), as well a~ the salt~ thereof with phy~iologically tolera-S ted ac ids .

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

Particularly preferred physiologically tolerated acids are: hydrochloric acid, citric acid, tartaric acid, lactic acid, phosphoric acid, me~hanesulfonic acid, acetic acid, formic acid, maleic acid, fumaric acid, malic acid, ~uccinic acid, malonic acid, sulfuric acid, L-glutamic acid, L-aspartic acid, pyruvic acid, mucic acid, benzoic acid, glucuronic acid, oxalic acid, a cor-bic acid and ace~ylglycine.

The novel peptides can be opan-chain (G 5 H, amino-protective group; Z = OH, N~2~ carboxyl-protectiYe group, M and Q not connected together) and, in particular, have a disulfide bridge (G - H, amino-protective group;
8 5 OH, NX2, carboxyl-protect$ve group; N + Q = -(CH2)C-S-S-(CH2)t-) or a side chair. bridge (G = ~, amino-protective group, Z = OH, NH2, carboxyl-protective group, ~ + Q - -(CH2).-NH-CO-(CH2)~- or -(cN2).-NH-co-(cHz)~-NH-co-(CH~ or be linked head-to-tail (G + Z s covalent bond or -CO-(CHz),-NH-).

The novel compounds can be prepared ~y conventional m~thod~ of paptide chemistry.

Thu~, the peptides can be constructed sequentially from amino acids or by linking together suitable smaller peptide fragments. In the sequential construction, the _ 4 _ o.Z. 0050/403~9 peptide chain is extended stepwise, by one amino acid each time, starting at the C terminu~. In the ca~e of coupling of fragments it is possible to link together fragments of different length~, these in turn being obtainable by sequential construction from amino acid~ or coupling of other fragments. The cyclic peptides are obtained, after synthe~is of the open-chain peptide~, by a cyclization reaction carried out in high dilution.

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

Chemical method~ for forming amide linkages are dealt with in detail by M~ller, Methoden der Organi~chen Chemie (Methods of Organic Chemi~try) Vol. XV/2, pp 1-364, Thieme Verlag, Stuttgart, 1974; Stewart, Young, Solid Phase Peptide Synthe~is, pp 31-34, 71-82, Pierce Chemical Company, Rockford, 1984; Bodanszky, Klau~ner, Ondetti, Peptide Synthe~is, pp 35-128, John Wiley & Sons, New York, 1976 and other ~tandard work~ of peptide chemistry.
Particularly preferred are the azide method~ the symmetr-ical and mixed anhydride method, active e~ters generated in ~itu or preformed and the formation of amide linXages using coupling reagents (activators), in particular dicyclohexylcarbodiimide (DCC), diisopropylcarbodiimide (DIC), 1-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ), l-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCI), n-propanephosphonic anhydride (PPA), N,~-bis(2-oxo-3-oxazolidinyl)amidopho~phoryl chloride (BOP-Cl), diphenylpho~phoryl azide (DPPA), Castro's rea~ent (BOP), O-benzotriazolyl-N,N,N',N'-tetra-methyluronium salt~ tHBTU), 2,5-diphenyl-2,3-dihydro-3-oxo-4-hydroxythiophene dioxide (Steglich's reagent;
HOTDO) and 1,1'-carbonyldiimidazole (CDI). The coupling reagents can be employed alone or in combination with 2C~ i05~7 - 5 - O.Z. 0050/40389 additives such as N,N~-dimethyl-4-amin~pyridine (DMAP), N-hydroxybenzotriazole tHOBt), N-hydroxybenzotriazine (HOOBt), N-hydroxysuccinimide (HOSu) or 2-hydroxy-pyridine.

Whereas it is normally possible to dispense with protec-tive group~ in enzymatic peptide ~ynthesis, for chemical synthesis it is necessary 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 technique~ are preferred for chemical peptide syntheses:
the benzyloxycarbonyl (Z), the t-butyloxycarbonyl (Boc) and the 9-fluorenylmethyloxycarbonyl (Fmoc) techniques.
In each ca~e the protective group on the ~-amino group of the chain-extending building block i8 identified. The side-chain protective groups on the trifunctional amino acids are chosen so that they are not necessarily elimin-ated together with the ~-amino protective group. A
detailed review of amino acid protective group~ is given by M~ller, Methoden der Organi~chen Chemie Vol XV/l, pp 20-906, Thieme Verlag, Stuttgart, 1974.

The building block~ used to construct the pept~d~ chain can be reacted in solution, in ~u~pension or by a method similar to that described by Merrifield in J.Amer. Chem.
Soc. 85 (1963) 2149. Particularly preferred methods are those in which peptides are con~tructed sequentially or by fragment coupling by u~e of the Z, Boc or Fmoc protec-tive group technique, in which case the reaction take place in solution, as well as those in which, similar to the Merrifield technique, one reactant i5 bound to an insoluble polymeric support (also called resin herein-after). Th~s typically entail3 the peptide baing con-~tructed ~equentially on the polymeric qupport, by use of the Boc or Fmoc protective group t~chnique, with the growing peptide chain be~ng covalently bonde~ at the Z ~ ~ ~ O 5~7 - 6 - O.Z. 0050/40389 C terminus to the insoluble resin particles (cf. Figures 1 and 2). This procedure allows reagents and byproduct~
to be removed by filtration, and thuc recrystallization of intermediates is superfluous.

The protected amino acids can be bonded to any suitable polymers which merely need to be insolu~le in the 801-vents used and to have a stable 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 polymer~ i~ suitable for thi~ purpose, for example cellulose, polyvinyl alcohol, polymethacrylate, ~ulfon-ated poly~tyrene, chloromethylated copolymer of ~tyrene and divinylbenzene (Merrifield re~in), 4-methylbenz-hydrylamine-re~in ~MBHA-resin), phenylacetamidonethyl-resin (Pam-re~in), p-benzyloxybenzyl alcohol-resin, benzhydrylamine-re~in (~HA-resin), 4-hydroxymethyl-benzoyloxymethyl-resin, the resin used by Breipohl et al.
(Tetrahedron Lett. 28 (1987) 565; from BACH~M), HYCRAM
resin (from ORPEGEN) or SASRIN resin (from ~ACHEM).

Solvents suitable for peptide synthesis in solution are all those which are inert under the reaction condition~, in particular water, N,N-dimethylformamide (D~F), dimethyl sulfoxide (DNSO), acetonitrile, dichloromethane (DCM), 1,4-dioxane, tetrahydrofuran (THF), N-methyl-2-pyrrolidone (NMP) and mixture~ of the said ~olvents.
Peptide ~ynthe~is on polymeric supports can be carried out in all inert organic eolvents which dissolve the amino acid derivatives used; however, ~olvent~ which have resin-swelling properties are preferr~d, ~uch a3 DMF, DCM, NMP, acetonitrile and DMSO, as well as mixtures of the~e solvents.

After the peptide has been synthe~ized it i8 cleaved off th~ polymeric support. The cleavage conditions for the '~QO~Q5~
- 7 - 0.2. 0~50/40389 various types of resins are di~closed in the literature.
The cleavage reactions most commonly use acid and palladium catalysis, in particular cleavage in anhydrou~
liquid hydrogen fluoride, in anhydrous trifluoromethane-sulfonic acid, in dilute or concentrated trifluoroaceticacid or palladium catalyzed cleavage in THF or THF-DCM
mixtures in the pre~ence of a weak base ~uch a~ morpho-line. The protective groups may, depending on the choice thereof, be retained or likewise cleaved off under the cleavage conditions. Partial deprotection of the peptide may also be worthwhile if the intention i~ to carry out certain derivatization reactions or a cyclization.

Some of the novel peptides have good cytotoxic proper~
tie~. Soms other~ of the peptides have high affinity for the cellular TNF receptor without, however, having cytotoxic activity. They are therefore TNF antagonist~.
They compete with natural TNF for binding to the cellular TNF receptor and thus suppre~ the TNF effect. The novel peptides are valuable drugs which can be employed for treating neoplastic disea~e~ and autoimmune disease~ as well a~ for controlling and preventing infQctions, inflammations and tran~pl~nt re~ection reactions. Simple axperiments can be used to elucidate the mode of action of the individual peptides. The cytotoxicity of the peptide i9 determined by incuba~ing a TNF-~ensitive cell line in the presence of the peptide. In a second experi-mental approach, the c911 line i~ incubated with the rele~ant peptide in tha pre~ence of a lethal amount of TNF. It i~ poa~ible in thi~ way to dstect the TNF-antagoni~tic effect. In addition, the affinity of the peptide for the cellular TNF receptor is determined in an in vitro binding experiment.

The following ta~t ~y~tems were u~ed to characterize the agoni~tic and antagoni~tic effects of the novel peptides:

~,~ O ~ O 5 ~

- 8 - O.Z. 0050/40389 I. Cytotoxicity test on TNF-~en~itive indicator cells, I~. Cytotoxicity antagonism test on TNF-sensitive indicator cells, III. Competitive receptor-binding te~t on indicator cell~
expressing TNF receptor.

I. Cytotoxicity test The agonistic effect~ of the novel peptides are a~sessed on the basis of their cytotoxic effect on TNF-sen3itive cells ~e.g. L929, MCF-7, A204, U937).
The te~t with L929 and MCF-7 was carried out as follows:

1. 100 ~1 of culture medium containiny 3 to 5 x 103 freshly trypsinized, exponentially growinq, L929 cell~ (mouse) or MCF-7 cell~ (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 wa~ saturated with water vapor and contained 5~ COz by volume.

. The L929 culture medium contained 500 ml of lx Earle'3 ME~ tBoehringer Mannheim), 50 ml of heat-inactivated (56C, 30 min) fetal calf serum (FCS), 50 ml of L-glutamine (200 mM), 5 ml of lOOx non-e~sential amino acids, 3 ml of lM HEPES
buffer pH 7.2,and 50 ml of gentamicin (50 mg/ml).

The ~CF-7 culture medium contained 500 ml of lx Dulbecco~s M$M (Boehringer Mannheim), 100 ml of heat-inactivated (55~C, 30 min) FCS, 5 ml of L-glutamine and S ml of lOOx non-essential amino acid~.

2. The next day 100 ~l of the peptide ~olution to be tested were added to the cell culture~ and ~ub~scted to serial 2-fold dilution. }n addition, ~005057 - 9 - o.Z. 0050/40389 some cell contrel~ (i.e. cell culture~ not treated with peptide dilution) and some rhu-TNF
controls (i.e. cell cultures treated with recom-binant human TNF) were also made up. The culture plate was incubated at 37C in an atmosphere of air saturated with water vapor and containing 5%
CO2 ~y volume for 48 h.

3. The percentage of ~urviving cell~ in the cultureY
treated with peptide dilution wa~ determined by staining with cry~tal violet. For this purpose, the liquid wa~ removed from the well~ of the test plate by tapping it. 50 ~1 of crystal violet 801ution were pipetted into each well.

The composition of the crystal violet ~olution wa~ as follows:

3.75 g of crystal violet 1.75 g of NaCl 161.5 ml of ethanol 43.2 ml of 37% formaldehyde water ad 500 ml The cry~tal violet ~olution wa3 left in the wells for 20 min and then likewise removed by tappin~.
The plates were then washed 5 time~ by immer~ion in water ~n ord~r to remove dye not bound to the cells. The dye bound to the cell~ 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. rhe surviving cell~ were determined by measuring the extinction at 540 nm of the colored solution in ., .

05~

- 10 - O.Z. 0050/40389 the individual wells.

5. Subsequently, by relating to the cell contsol, the 50% cytotoxicity value was defined, and the reciprocal of the sample dilution which re~ul~ed in 50~ cytotoxicity was calculated a~ the cyto-toxic activity of the te~t sample.

II. Cytotoxicity antagoni~m test - The antagonistic effect of the peptides was assessed on the basi~ of their property of antagonizing the cytotoxic effect of rhu-TNF on TNF-sen~itive cell~
(e.g. L929, MCF-7, A204, U937). The cytotoxi~ity antagonism te~t with ~929 and MCF-7 cell~ wa~
carried out as follows:
1. 100 ~1 of culture medium containing 3 to 5 x 103 lS fre~hly ~ryp~inized, exponentially growing, L929 cell~ (mouse) or MC~-7 cell3 (human) were pipetted into the wells of a 96-well flat-bottom culture plate. The plate was incu~ated at 37C
overnight. The air in the incubator wa saturated with water vapor and contained 5% CO2 by volume.

The L929 culture medium contained 500 ml of lx Earle'~ MEM (Boehringer Mannheim), 50 ml of heat-in~ctivated (56~C, 30 min) FCS, 5 ml of L-gluta-mine (200 mM), 5 ml of lOOx non-es3ential amino acids, 3 ml of 1~ HEPES buffer pH 7.2, and 500 ~1 of gent~micin (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, S ml of L-glutamine (200 m~) and 5 ml of lOOx non-e3sential amino acid~.

2. Th~ next day 100 ~1 of the peptide solu~ion to ~e 0 5~ 5~

~ O.Z. 005~40389 tested were added to the cell cultures and sub~ected to serial 2-fold dilu~ion. Then, 100 ~1 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 culture~. In addition, some c~ll control3 (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 sulture plate was then incubated at 37C in an atmosphere of air aturated with water vapor and containing 5% CO2 by volume for 48 h.

3. The percentage of ~urviving cell~ in the cultures treated with substance solution wa~ determined by staining with crystal violet. For thi~ purpo~e, the liquid wa3 removed from the well~ of the test plate 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 Th~ crystal violet ~olution wa~ left in the wells for 20 min and then likewi~e removed by tapping.
The plates were then washed 5 times by immer~ion in water in order to remo~e dye not ~ound to the cells. The dye bound to the cell~ waB extracted by adding 100 ~1 of rsagQnt solution (50% etha-nol, 0.1% glacial acetic acid, 49.9% water) to each w~ll.

4. The plates were shaken for S min to obtain a ~olution of uniform color in each well. The surviving cell~ were determined by mea~uring the extinction at 540 nm of the colored solution in )5057 - 12 - O.Z. 0050/40389 the individual well~.

5. Subsequently, by relating to the cell control and the rhu-TNF control, the 50~ antagonism value wa~
defined, and the sample concentration which resulted in 50~ antagonism of rhu-TNF cytotox-icity at the rhu-TNF concentration used wa~
calculated a~ antagonistic actiYity of the sample tested.

III. Competitive receptor-binding test Both the agonistic and antagonistic effects ~f peptide~ are conditional on the latter binding to the TNF receptor. Thi~ mean~ that peptide~ with an agonistic or antagonistic effect compete with rhu-TNF for binding to the TNF receptor on TNF-~enaiti~e indicator cell~ (e.g. U937). Th~ competi-tive receptor-binding te~t was carried out as follows:

1. 100 ~1 of medium contain$ng variou~ concentra-tion~ of the peptide to be te~ted and of rhu-TNF
(= control) were pipetted into the reaction vessel~. The medium compri~d 500 ml of PBS
(Boehringer Mannheim), 10 ml of heat-inactivated (56C, 30 min) FCS and 100 mg of sodium azide.

2. Subsequently, 100 ~1 of medium containing 1 ng of ~I-labeled rhu-TNF (Bolton lactop~ro~ida~e method) were placed in the reàction ve~els and mix~d. The non-specific binding (~SB) was deter-mined by mixing in the reaction ve3~el the l2~I-labeled rhu-~NF (1 ng of~25I-rhu-TNF in 100 ~1 of medium) with a 200-fold excess of unlabeled rhu-TNF (200 ng of r~u-TNF in 100 ~1 of medium).

3. Then 100 ~1 of medium containing 2 x 10~ U937 3~ 5 ~S ~

- 13 - O.Z. ~050/40389 cells (human) were pipetted into the reaction vessels and mixed. The reaction vessels ttest ~olume 300 ~l) were incubated at 0C for 90 min.
The reaction mixtures were remixed after 45 min.

4. After the incubation the cells were centrifuged at 1800 rpm and 4C for 5 min, wached 3 times with medium and transferred quantitatively into counting vials, and the cell-bound radioactivity wa3 determined in a Clini gamma countsr 1272 (L~B Wallac).

5. After the mea~urements had been corrected for the non-specific binding, the 50% competition value was defined by relation to the overall binding, and the s~mple concentration which led to 50%
competition of125I-rhu-TNF binding at the 125I-rhu-TNP concentration used was calculated as the competitive acti~ity of the sample tested.

The Examples which follow are intended to explain the invention in more detail. The proteinogenous amino acids are abbrevia~ed in the Examples using the conventional three-letter code. Other meanings ares Aad = ~-amino-adipic acid, Ac = acetic acid, Ade = 10-a~inodecanoic acid, Ahp - 7-aminoheptanoic acid, Ahx = 6 aminohexanoic acid, Ano ~ 9-aminononanoic acid, Aoc 3 8-aminooctanoic acid, Bal = ~-alanine, Hcy = h~mocysteine, Xly 5 homoly-sin~, Orn = ornithine, Dap = 2,3-diaminopropionic acid.

A. Gsneral procedure~

I. The peptidQs claimed in claim 1 were synthesized using standard methods of ~olid-phase peptide synthesis in a completely automatic model 430A
peptide synthesizer from APPLI~D BIOSYST~S. ~he apparatu~ u~e~ different synthe~i~ cycle~ for the 200505~7 - 14 - 0.2. 0050/40389 ~oc and Fmoc protective group techniques.

a) Synthesis cycle for the Boc protective group technique 1. 30% trifluoroacetic acid in DCM 1 x 3 min 2. 50% trifluon~etic acid Ln DCM 1 x 17 mun 3. DCM washing S x 1 min 4. 5% diix~y~Eylethylanine in DC~ 1 x 1 min 5. 5% diiu~nQylethyla~e in NMP 1 x 1 min ~. N~ wa~hing S x 1 min 7. ~dition of pn~tivated prnl~1ad a~
acid (activation by 1 ff~valent of DOC
and 1 ~ alent of HOBt in NWP/DCM);
~ide coupling (Lst p~rt~ 1 x 30 min 8. Addition of DMSO bo the n~ion mi~h~e until it co~u~ 20% nMs~ by volume 9. ~ide coupli~g (2nd part) 1 x 16 min 10.A~diti~n of 3.8 ~ VQ~ of dikK~
~ylethyl~ to the r#x~lcn mi~l~2 ~ lAP coupli~g (3Dd part) 1 x 7 min 12.DCMwashing 3 x 1 min 13.1f ~ n is ~Yx~plete, rq~ition of ocupli~g (r~h~n bD 5.) 14. 10% ~o~i~ a~ride, 5% dL~nQyl-et~yl~s in DCM 1 x 2 min lS.10% ~o~ dride in DCM 1 x 4 min 16.DCM wa~n~ 4 x l min 17 ~l~n to 1.

b) Syn~is cycle for the Fmoc pn~tive çx~p te~qpe 1. N~æ ~g 1 x 1 m~n 2. 20% piper~e in NMP 1 x 4 min 3. 20~ p~idine in ~MP 1 x 16 min 4. N~æ wa~hing 5 x 1 min 5. Additlon of pn#Y~iva~ed pD~YYeed acid (~lvation ~y 1 ~;valent of D4C

~ 05057 - 15 - O.Z. ~050/40389 and 1 ~valent of HOEt Ln NMP/DCM) ~ide co~pling 1 x 61 min 6. NMP wzshing 3 x 1 mln 7. If n~tion i ~xnplete, nq~ition of S couplLng (return to 5.) 8. 10% acetîc anhydride in NMP 1 x 8 mun 9. NMP w3shing 3 x 1 min 10. ~l~n t~ 2.

II. Working up of peptide-re~ins obtained as in Ia The peptide-resin obtained a~ in la was dried under reduced pressure and transferred into a reaction vessel of a Teflon HF apparatus (from PENINSULA).
Addition of a scavenger, preferably anisole (1 ml/g of resin), and of a thiol, in the case of tryptophan-containing peptide~, to remove the indole formyl group, preferably et~anedithiol ~0.5 ml/g of re~in)~
was 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 stirred at this temperature for 45 min. The hydrogen fluor-ide was then stripped off under reduced pres~ure and the r~sidue wa~ wa~hed with ethyl acetate in order to remove remaining ~cavenger. The peptide was extracted with 30% strength acetic acid and filter~d, and the filtrate was freeze-dried.

To prepare peptidQ hydrazides, the peptide-re~in (Pam- or Nerrifield resin) wa~ ~uspended in DMF
(15 ml/g of resin~, hydrazine hydrate (20 equiva lents) was added, and th~ mixture was stirred at room temperature for 2 day~. To work up, the re~in was filtered off and the filtrats wa~ evaporated to dryness. The re~idue was crystallized from DMY/Bt2O
or MeOH/Et2O.

III. Wor~ing up of the peptide-resins obt~ined a~ in Ib ~c~oso~

- 16 - O.Z. od50~40389 The peptide-resin obtained as in Ib was dried under reduced pressure and subsequently subjected to one of the following cleavage procedures, depending on the amino acid composition (Wade, Tregear, Noward Florey Fmoc-Workshop Manual, Melbourne 1985).

~oalsos7 - 17 - O.Z. 0050/40389 Peptide containing Cleavage conditions Arg(Mtr) Met Trp TFA Sca~enger Reaction TLme no no no 95~ 5% H2O 1.5 h yes no no 95% 5% thioanisole > 3 h no yes no 95% 5% eth~l methyl 1.5 h ~ulfide no no yes 95% 5~ ethanedithiol~1.5 h anisole (1:3) no yes yes 95% 5% ethanedithiol/1.5 h anisole/ethyl methyl sulfide (1:3:1) yes ye~ yes 93% 7% ethanedithiol/~ 3 h anisole/ethyl methyl sulfide (1:3:3) The ~uspension of the peptide-resin in the ~uitable TFA mixture wa~ ~tirred at room te~perature for the ~tated time and then the resin wa3 filtered off and washed with TFA and with DCN. The filtrate and the wa~hing~ were extensively concentrated, and the peptide was precipitated by addition of diethyl ether. The mixture wa~ cooled in an ice bath, and the precipitate wa~ filtered off, ta~en up in 30 acetic acid and freeze-dried.

IV. Purification and characterization of the peptides Purific~tion was by gel chromatography (SEPHADEX~
3 0 G- 1 0, G- 15 /1 0 9~ HOAC; SEPEIADE:X LH2 0 /MeOH ) and ~equent medium pre~ure chromatography (stationary phase: HD-SIL C-18, 20-45 ~, 100 A; mobile pha~e:
gradient with A = 0.1% TFA/MeOH, B z 0.1~ ~FA/H20).

The purity of the final product~ wa determined by - 18 - O.Z. 0050/40389 analytical HPLC (stationary phase: 100 x 2.1 mm VYDAC C-18, 5 ~, 300 ~; mobile phase = CH3CN/H~O
gradient buffered with 0.1~ TFA, 40C). Charac-terization was by means of amino acid analysi~ and fast atom bombardment mass spectrometry.

B. Specific procedures H-Ile-Asn-Arg-Pro-Asp-Tyr-Leu-Asp-NH2 0.98 g of Boc-Asp(OChx)-MBHA-resin (substitution 0.51 mmol/q), corre~ponding to a batch ~ize of 0.S mmol, was reacted as in AIa with 2 mmol each of Boc-Leu-OH Boc-Arg(Tos)-OH
Boc-Tyr(Br-Z)-OH Boc-Asn-OH
Boc-Asp~OChx)-OH Boc-Ile-OH
Boc-Pro-OH

After the synthe3is was complete, the peptide-re~in underwent N-terminal deprotection (steps 1-3 a~ in AIa) and sub~equent drying under reduced pra~ure; the yield was 1.6 g.

0.8 g of the resin obtained i~ thi~ way wa~ sub~ected to a HF cleavage as in AII. The crude product (19~ mg~ wa~
purified by gel filtration (SEPHADEX G-10) and medium pressure chro~atography (cf. AIV; 15 30~ A; 0.25% min~l).
87 mg of pure product were obtained.

Ac-Ser-Ala-Glu-Ile-Asn-Leu-Pro-Ly~-Tyr-Leu-A~p-Ph~-Ala-Glu-OH

~!GtO5o~j7 - 19 - O.Z. 0050/40389 0.46 g of Fmoc-Glu(otBu)-p-alkoxybenzyl alcohol-resin (substitution 0.55 mmol/g), corresponding to a batch size of 0.25 mmol, was reacted as in AIb with 1 mmol each of Fmoc-Ala-OH Fmoc-Lys(Boc)-OH Fmoc-Ile-OH
Fmoc-Phe-OH Fmoc-Pro-OH Fmoc-Glu(OtBu)-OH
Fmoc-Asp(OtBut-OH Fmoc-Leu-OH Fmoc-Ala-OH
Fmoc-Leu-OH Fmoc-Asn-OH Fmoc-SerttBu)-OH
Fmoc-Tyr(tBu)-OH

After the synthesis was complete, the N terminus was acetylated (steps 2-4 and 8-9 as in AIb). The re~ulting peptide-xesin was dried under reduced pre~sure; the yield wa~ 0.7 g.

The crude peptide (268 mg) obtained after TFA cleavage a~
in AIII was purified by gel filtration (SEPHADEX~ G-10) and medium pre~ure chromatoqraphy ~cf. AIV; 40-60% A;
0.25~ min~1). 147 mg of pure produ~t were obtained.

The following can be prepared in a sLmilar manner to Example~ 1 and 2:

3. H-Ala-Glu-Ile-ASn-~rg-Pro-ASp-Tyr-~u-Asp-Ph~-Ala-Gtu-Ser-Gly-Gln-Val-Tyr-OH
4. Ac-Ala-Glu-ll~-Asn-Arg-Pro-Asp-Tyr-Leu-Asp-Phe-Ala-Glu-Ser-Gly-Gln-Val-Tyr-NH2 S. H-~la-~la-Ala-Ata-~la-Ala-Ala-~rg-Pro-~sp-Tyr-Leu-Asp-Phe-Ala-Glu-Ser-Gly-Gln-Val-OH
6. Ac-~la-~la-Ala-Al~-~la-Ala-~1a-Arg-Pro-~sp-Tyr-Lou-Asp-Pha-Ala-Glu-Ser-Gly-Gln-Val -NH2 7. H-ll~-~sn-Arg-Pro-~sp-Tyr-L~u-~sp-OH
8. Ac-ll~-Asn-~rg-Pro-Asp-Tyr-Lou-~sp-OH
9. Ac-Ile-~sn-~rg-Pro-~sp-Tyr-Lou-~sp-NH2 10. H-Ser-~la-61~ sn-L~u--Pro-Lys-~yr-Leu-~sp-Ph~-~la-Glu-OH
Il. H-S-r-~la-Glu-ll--~sn-Leu-Pro-Lys-Tyr-Leu-~sp-Ph~-~la-Glu-NH2 12. ~c-Se~-~la-61u-Ilo-Asn-Leu-Pro-Lys-Tyr-Leu-Asp-P~e-~la-Glu-NH2 13. ~ a-6tu-ll--Asn-~g-pro-~sp-Ty~r-L~u-~sp-p~ ld-Glu-s~r-Gl~-Gln Val-Tgr-OH

~O~S05'7 - 20 - O.Z. 0050/40389 14. Ac-Ala-Glu-~le-Asn-Arg-Pro-Asp-Tyr-Leu-Asp-Phe-~ld-Glu-Ser-Gly-~ln-Val-Tyr-NH2 15. H-Ala-Ala-Ala-Ala-Ala-Ala-Ala-Arg-Pro-Asp-Tyr-Leu-Asp-Phe-Ala-Glu-Ser-GI~-Gln-Val-OH
16. Ac-Ala-~la-Ala-~la-~la-Ala-Ala-Arg-Pro-Asp-Tyr-Leu-Asp-Ph~-Ala-Glu-Ser-Gly-Gln-Val-NH2 17. H-His-rhr-Asp-Gly-lls-pro-His-Leu-val-Leu-s~r-oH
18. Ac-His-Thr-Asp-Gty-ll~-Pro-~is-Leu-Val-L~u-Ser-NH2 Ac-Hcy-Asn-Arg-Pro-Asp-Tyr-Hcy-NH2 O.40 g of Boc-Hcy(pMB)-MBHA-resin (sub~titution 1.24 mmol/g), corre~ponding to a batch size of 0.5 mmol, was reacted a~ in AIa with 2 mmol each of Boc-Tyr(Br-Z)-OH Boc-Arg(Tos)-OH
Boc-Asp(OChx)-OH aoc-Asn-OH
Boc-Pro-OH Boc-Hcy(pMB)-OH.

After the synthe~is wa~ comple~e, the N terminu~ was acetylated (steps 1-6 and 14-16 as in AIa).

The re~ulting peptide-re~in wa~ dried under reduced pre~sure; the yield was 1.1 g.

The resin obtained in this way was ~ub~ected to HF
cleavage as in AII. The freeze-dried crude product was taken up in 1 1 of a 1 molar aqueouQ ~olution of urea~
and the pH was ad~u~ted to 8.4. After the mixture had been stirred at room temperature for 4 days it wa3 acidified to pH 4.5 with glacial acetic acid, and the peptide was adsorbed onto RP Cl8 material and subsequently eluted with methanol, and the eluate wa~ concentrated in a rotary evaporator and then freeze-dried.

All the solvent~ had previously been ~aturated with nitrogen in order to prevent any oxidation of the free cysteine residues.

~3050~7 - 21 - O.Z. 0050~40389 The crude product was rela~ively heavily contaminated and wa~ purified by medium pr~ssure chromatography (cf. AIV;
30-60% A; 0.25% min1). 61 mg of pure product were obtained.

S The following can be prepared in a sLmilar manner to Example 19 (Pam-resin wa~ used to prepare the peptide acids):

20. H-Cys-lle-Asn-Arg-Pro-Asp-Tyr-Cys-OH

r 21. Ac-Cys-lle-Asn-Arg-Pro-Asp-Tyr-Cys-NH2 _ . I
22 . H-Hcy-l I e-Asn-Arg-Pro-Asp-Tyr-Hcy-OH
.
23. Ac-Hcy-lle-Asn-Arg-Pro-Asp-Tyr-Hsy-NH2 24. H-Hcy-Asn-Arg-Pro-Asp-Tyr-Leu-Hcy-OH

25. Ac-Hcy-Asn-Arg-Pro-Asp-Tyr-Leu-Hcy-NH2 26. H-~cy-Ile-Asn-Arg-Pro-AspTyr-Leu-Hcy-OH

27. Ac-Hcy-lle-Asn-Arg-Pro-Asp-Tyr-Leu-Hcy-HH2 I
28. H-Hcy-Thr-Asp-Gly-lle-Pro-His-Leu-Hcy-OH
r ~ 7 29. Ac-Hcy-Thr-Asp-Gly-lle-Pro-His-L~u-Hcy-~H2 30. H-Ala-Glu-Hcy-Asn-~rg-Pro-Asp-Tyr Hcy-Asp-Ph~-Ala-OH
31. Ac-~la-Glu-Hcy-~sn-Arq-Pro-Asp-Tyr-Hc~-~s~-Pha-~la-NH2 32. H-Cgs-IlQ-~sn-~rg-Pro-~sp-Tyr-Lou-~sp-Ph--~la-Glu-Ser-Gty-Gln-Val-Cys-OH
_ 33. Ac-Cys-lle-Asn-~rg-Pro-Asp-Tyr-Leu-~sp-Ph~ a-Glu-S-r-Gly-Gln-Vat-Cys-NH2 34. H-Hcy-~al-~sn-L~u-Pro-~ys-Tyr-Leu-~sp-Pho-~la-Glu-S~r-Gly-61n-~al-Hcy-OH
_ I
35. Ac-Hcy~ Asn-La~-Pro-Lys-Tyr-L~u-~sp-P~ la-61u-S~r-Gly-Sln-Val-Hcy-NH2 .
36. H-Hc~-lt~-Asn-~rg-Pro-~sp-Tyr-Lou-Asp-Ph~-Ala-Glu-Ssr-Gly-Hcy-OH

37. ~c-Hey-ll--~sn-~rg-Pro-Asp-Tyr-Leu-isp-Ph~-~la-61u-Scr-GIg-Hcy-NH2 5 O ~ 7 - 22 - O.Z. 0050/40389 _ 38. H-Cys-Ala-Glu-llo-Asn-Arg-Pro-Asp-Tyr-Leu-Asp-Phe-Ala-Glu-Ser-Gly-Gln-Val-Tyr-P~e-Cys-OH
39. Ac-Cys-A1a-Glu-lle-Asn-Arg-Pro-Asp-Tyr-Leu-Asp-Phe-Ala-Glu-Ser-Gly-Gln-Val-Tyr-Phe-cys-NH2 -40. H-Asp-Arg-Leu-Scr-Cys-Glu-Ile-Asn-Arg-Pro-Asp-Tyr-leu-Asp-Phe-Ala-Glu-Ser-Gly-Gln-~al-Tyr-Cys-Gly-II~ -O~

41. Ac-Asp-Arg-Leu-Ser-Cys-Glu-Ile-Asn-Arg-Pro-Asp-Tyr-Leu-Asp-Phe-Ald-Glu-Ser-Gly-Gln-Val-Tyr-Cys-Gly-Ile-lle-NH2 42. H-Hcy-Asn-Arg-Pro-Asp-Tyr-Hcy-OH
43. Ac-Cys-Asn-Arg-Pro-Asp-Tyr-Cys-NH2 44. Ac-Hcy-Asn-Arg-Pro-Asp-Tyr-CyS-NH2 45. Ac-Cys-Asn-Arg-Pro-Asp-Tyr-Hcy-NH2 46. H-Cys-~lu-Ile-Asn-Arg-Pro-Asp-Tyr-Leu-Asp-Phe-Ala-~lu-Ser-Gly-Gln-Val-Tyr-Cys-OH
47. Ac-Cys-Glu-Ile-Asn-Arg-Pro-Asp-tyr-Leu-Asp-Pho-Ala-Glu-Ser-Gly-Gln-Val-Tyr-cys-NH2 48. Ac-Cys-Ala-Glu-S-r-Gly-Gln-Cys-N~2 49. H-Cys-Ala-Glu-S~r-Gly-61n-Cys-NH2 EXAMPLE SO
Ac-A~p-Asn-Arg-Pro-Asp-Tyr-Orn-NH2 1 g of re~in described by Breipohl et al. (from BACHEM), corresponding to a batch ~ize of 0.5 mmol, was reacted a~ in AIb with 2 mmol each of F~oc-Orn~Boc)-OH Fmoc-Arg(Mtr)-OH

C)5057 - 23 - O. Z . 00S0/40389 E moc - ryr ( tBu ) -Oll E moc -Asn-OH
Fmoc -Asp ( OBz 1 ) ~OH Fmoc -Asp ( Ot3u ) -OH
Fmoc-Pro-OH

After the synthesis was complete, the N terminus was acetylated (steps 2-4 and 8-9 a~ in Alb). The peptide-resin was dried under reduced pres~ure; yield 1.5 g.

The crude product (321 mg) obtained after TFA cleavage as in AIII was dissolved in 500 ml of degassed DMF.
0.2 ml of triethylamine and then, at -20C, 0.2 ml of diphenylphosphoryl azide were added and the mixture was stirred at -25 DC for 2 h. It wa~ ~ubsequently stored at -20C for 2 days, at 4DC for 2 day~ and at room tempera-ture for 2 days. It was then evaporated to dryness, and the crude peptide was purified by gel chromatography ~SEPHADEX~ LH 20). The isolated monomer (128 mg) wa~
deprotected with HF as in AII and purified by medium pressure chromatography (cf. AIV, 25-45~ A, 0.25~ minl).
77 mg of pure product were obtained.

EXAMPL~ 51 Ac-Ser-Ala-Glu-Asp-Asn-Arg-Pro-Asp-Tyr-Orn-Asp-Phe-Ala-OH

2.22 g of ~oc-Ala-Merrifield re~in (substitut~on about 0.27 mmol/g), corresponding to a batch s~ze of 0.6 mmol, w~re reacted as in AIa and AIb with 2.4 mmol each of Boc-Phe-OH Fmoc-Asp(OCHx)-OH Fmoc-A~p(OtBu)-OH
Boc-Asp(OCHx)-OH Fmoc-Pro-OH Fmoc-Glu(OBzl)-OH
Fmoc-Orn(BocJ-OH Fmoc-Arg(Tos)-OH Fmoc-Ala-OH
Fmoc-Tyr(tBu)-OH Fmoc-Asn-OH Fmoc-Ser(tBu)-OH.

The N terminus was acetylated (step~ 2 to 4 and 8 to 9 a~
in AIb).

~05057 - 24 - O.Z. 0050/40389 Su~sequently the t-butyl and Boc protective group~ were cleaved off (step~ 1 to 6 a~ in AIa). The cyclization on the re-~in took place in NMP with the addition of 1.06 g of BOP and 1.05 ml of diisopropylethylamine t24 h). The yield was 2.97 g, The crude product obtained after HF
cleavage as in ~II was purified by gel filtration (SephadexD G-25) and medium pressure chromatography (cf.
AIY 40 to 60~ A; 0.25~ minl). 12 mg of pure product were obtained.

The following can be prepared in a ~imilar manner to Examples 50 and 51:

52. Ac-Glu-Asn-Arg-Pro-Asp-Tyr-Lys-NH2 53. H-Glu-Asn-Arg-Pro-Asp-Tyr-;js-OH
54. Ac-Orn-~sp-Gly-lto-Pro-His-Giu-NH2 55. Ac -Orn-~sp-Gly-ll~-Pro-His-Glu-OH

56. Ac-~p-~sn-~rg-Pro-~sp-Tyr-Leu-Oap-NH2 57. Ac-Aad-Arg-Pro-Asp-Tyr-Hiy-NH2 58. Ac-Ser-~la-GIu-ASp-~sn-~rg-Pro-~sp-Tyr-Orn-~sp-Phc-~la-NH2 59. Ac-~sp-~sn-~rg-O-Pro-~sp-Tyr-Leu-~ap-HH2 :
60. Ac-Asp-61u-11~-Asn-Arg-Pro-Asp-Tyr-L~u-Asp-Pho-Ala-Glu-55r-Gly-Gln-vdl-Tyr-Dap-NH2 61. ~c-Orn-Glu-lto-Asn-~rg-Pro-~sp-Tyr-Leu-Asp-Ph--~la-GIu-5-r-GIy-GIn-~al-T~r-Asp-NH2 62. Ac-Orn-~ sn-Arg-Pro-Asp-Tyr-Lsu-~sp-Pha-~la-~lu-Scr-GIy-~ln-~al-Asp-N~2 3C)5057 - 25 - O.Z. ~050/40389 63. Ac-Asp-AId-GIu-lle-Asn-Arg-Pro-Asp-Tyr-Leu-Asp-Phe-AIa-GIu-Ser-GIy-GIn-i Vdl -Tyr-Phe-Orn-NH2 64. Ac-Arg-Leu-Ser-Asp-Glu-lle-Asn-Arg-Pro-Asp-Tyr-Leu-Asp-P~e-Ala-Glu-Ser-, Gly-Gln-Val-Tyr-Oap-Gly-lle-Ile-NH2 EXANP~E 65 Ile-Asn-Arg-Pro-Asp-D-Tyr-Ahx 1.22 g of Fmoc-Asp(OBzl)-p-alkoxybenzyl alcohol-resin (substitution 0.41 mmol/g), corre~ponding to a batch size of 0.5 mmol, were reacted as in AIb with 2 mmol each of Fmoc-Pro-OH Fmoc-Ile-OH
~mog-Arg(Mtr)-OH Fmoc-Ahx-OH
Fmoc-Asn OH Fmoc-D-Tyr(tBu)-OH.

After the synthesis was complete, the peptide-resin underwent N-terminal deprotection (steps 2-4 as in AIb) and subsequent drying under reduced pras~ure. The yield was 1.5 g.

The crude peptide obtained after TF~ cleavage as in AIII
wa~ dis~olved in 500 ml of degassed DMF. 210 mg of NaHCO3 and, a~ -25-C, 0.2 ml of diphenylphosphoryl azide wer6 added, and the mixture was ~tirred at -25C for 2 h and at room temperature for 2 days. It was then evaporated to dryne~s, and the crude peptide wa~ purified by g~l chromatography (SEP~ADEX' LH 20). The isolated monomer (127 mg) wa~ deprotected with HF a~ in AII and purified by medium pressure chromatography (cf. AIV; 55-75~ A
0.25~ min~1~. 78 mg of pure product were obtained.

The following ^an be prepared in a similar manner to ~IU~50S~

- 26 - O.Z. 005~/40389 Example 65:

66. rAsn-~rg-Pro-Asp-Tyr-Ano 67. rAsn-Arg-Pro-Asp-ryr-Ade 68. rlle-Asn-~rg-Pro-Asp-Tyr-Ahx 69. rAsp-Gty-ll~-Pro-His-70. r~sn-L~u-Pro-Lys-Tyr-Leu-~sp-eal 71 r" ~-Asn-Arg-Pro-Asp-Tyr-Leu-Asp-Phe-Ala-Glu-Ser-Gly-Gln-Val-Tyr-Ahpl~

Claims (8)

1. A peptide of the formula I

X-A-Y I

where A is -Arg-Pro-Asp-Tyr-, -Leu-Pro-Asp-Tyr-, -Gln-Pro-Glu-Tyr-, -Leu-Pro-Lys-Tyr-, -Gly-Ile-Pro-His- or -Gly-Ile-Ser-His-, X is G-, G-NH-CHM-CO-, G-NH-CHN-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)a-NH-, where a is from 1 to 12, R, U and W are peptide chains composed of 1-4 naturally occurring .alpha.-amino acids, V is a peptide chain composed of 1-10 naturally occurring .alpha.-amino acids and N 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, H2N-C(=NH)-NH) or M and Q together are a -(CH2)o-S-S-(CH2)d-, -(CH2)e-, CO-NH-(CH2)f- or -(CH2)e-NH CO-(CH2)g-NHCO(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 tolerated 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 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 -(CH2)e-NH-CO-(CH2)f- or -(CH2)e-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)a-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 prepara-tion thereof using conventional methods of peptide chemistry.