PT100427A - (CCK) analogs containing alpha-substituted aspartame aminosaccharides and pharmaceutical compositions containing the same - Google Patents

Description

4

Description of the WARIOIR-LAMBERT COM-PANY, North American, industrial and commercial patent, established at 201 Tabor Road, Morris Plains, Rew Jersey 07950, United States of America, (inventors: David Christopher Horwell and Reginald Stewart Richardson, residing in the United Kingdom, to " CHEMICAL PROPERTIES " " CHEMISTRY AMENDMENTS "

description

Background of the invention

Agents acting at the receptors of cholecystokinin (CCK) may eventually induce satiety (Schick, Yaksh and Go, Regulatory Peptides 14: 277-291 (1986).

GSP

CCK peptides are widely distributed throughout the various organs of the body including the gastrointestinal tracts, endothermic glands, and nerves of the peripheral and central nervous systems. A number of biologically active forms have already been identified including a hormone with 33 amino acids and various fragments of the

(for example, the CCK26-33 octapeptide and the CCK30-33 tetrapeptide). (G.J. Dookray, Br. Med. Bull., 38 (H2.3): 253-258, 1982).

The various CCK peptides are believed to be involved in the control of smooth muscle contractility, secretion of the extructural and endro-urine glands, transmission of sensory nerves and numerous brain functions. Administration of native peptides leads to gallbladder contraction, amylase secretion, central neuron excitation, inhibition of the urge to eat, anti-convulsive actions and other effects on behavior. (" Cliolecystokinin: Isolation, Struture and Eunotions ", GBJ Glasse, Ed .; Raven Press, New York, 1980, pp. 169-221; J. Ξ Morley, Life Sciences 27: 355-368, 1980; " Cholecysto -kinin in the lervous System, " J. de Belleroche and GJ Dookray, Ed., Ellis Horwood, Chichester, England, 1984, pp. 110-127). The existence of high concentrations of CCK peptides in many areas of the brain also means the existence of fundamental brain functions performed by these peptides (G.J. Dookray, Br. Med. Bull., 38 (12): 253-258, 1982). The most abundant form of cerebral CCKs detected is CCK26-33 although there are also small amounts of CCK30-33 (Rehfeld and Gottterman, J. Neuroohem, 32: 1339-1341 (1979)). Exactly the function of CCK in the central nervous system is known but it is known to be associated with control of the will to eat (Della-Eera and Baile, Science 206: 471-473 (1979)).

Presently available appetite suppressants act peripherally by increasing energy consumption (such as thyroxine) or otherwise acting (such as biguanides), or they still act on appetite or satiety. - 2 -

It has recently been shown that patients with bulimia have lower levels of CCK in their plasmas than Hew England Journal of Medicine 319, 683 (1988)). An additional function of peripheral COKs is to regulate insulin release. COKs have been shown to increase insulin levels when administered to mammals J. Clin. Endoorinol. Metab, 65: 395 (1987)).

Recent studies claim that C-terminal fragments of CCKs function as CCK receptor antagonists (Jansen, et al, Bioohem, Biophys.

Acta 757: 250 (1983); Spanarkel J, Biol Chem, 258: 6746 (1983)) Japanese Patent Application P 70/10506 to Miyao et al. Discloses a tetrapeptide derivative of the carboxy terminal sequence of gastrin (1-Trp-1-Lys -L-Asp-L-Phe-HH 4) which has anti-gastrin activity.

Centrally acting appetite suppressing drugs potentiate the central cetelo-lamin pathways and tend to be stimulants (e.g., amphetamines) or influence serotonergic precursors (for example, fenfluramines). Other forms of drug therapy include volumizing agents which act by filling the stomach thereby inducing a " feeling " of satiation. WO 90/06937 discloses tetrapeptide derivatives as CCK agonists. Such compounds are for the treatment of gastrointestinal disorders, central nervous system disorders, insulin-associated disorders, treatment of pain or serve as appetite regulators.

SUMMARY OF THE INVENTION The present invention relates to novel compounds which are CCK ligands and are mono-, di-, tri-,

compound according to general formula 1 in combination with a pharmaceutically acceptable carrier in unit dosage forms for appetite suppression. The present invention also describes a method for suppressing appetite in mammals, which comprises administering to the mammals in need of such treatment an effective amount of composition described above to suppress appetite. The present invention also describes methods for the treatment of gastrointestinal disorders, 4

central nervous system (CNS) disorders through the use of CNS suppressants which may exhibit antipsychotic, neuroleptic / anxiolytic and anti-convulsive effects. The present invention also describes methods for reducing the secretion of gastric acids and for the treatment of gastrointestinal ulcers. The present invention further describes a blocking reaction caused by the cessation of use of a drug or alcohol and also relates to the potentiation of the effects of morphine and other epoxides used in the treatment of pain.

On the other hand the present invention further provides processes for the preparation of the compounds of formula I.

Finally, the present invention provides novel intermediates useful for the preparation of the compounds of formula I and also provides processes for the preparation of such intermediates,

Figure 1 shows that compound 2 is a partial agonist. Figure 2 demonstrates that compound 4 is an agonist and that compound 5 is a weak agonist. Figure 3 demonstrates that compound 6 is an agonist.

Figures 1 to 4 characterize CCK8s as the reference standard for agonists. 5

Detailed Description The following table provides a dictionary of terms used in the specification of the present invention. TABLE I Short description GLY Glycine p-ALA f-Alanine GABA 4-Aminobutanoic acid DAVA f-aminovaleric acid MET L-Methionine TYR L-tyrosine ASP L-aspartic acid PEH L-phenylalanine TRP L-tryptophan BOC 'Uercbutoxycarbonyl 1-ADOC 1-Adamantiloxycarbonyl 2-ADOC 2-Adamantiloxycarbonyl Z Benzyloxycarbonyl FMOC 9-Fluorenylmethoxycarbonyl Aib α-Aminoisobutyric acid PFP Pen et al.

The compounds of the present invention are analogs of CCK 30-33 and CCK 26-33 and contain at least one residue of an amino acid which is α, α-disubstituted.

Such compounds differ from the genetically encoded natural peptides present in mammals in that the

α-substituents can not all be simultaneously the hydrogen atom.

The compounds of the present invention are represented by the general formula

or are a corresponding pharmaceutically acceptable salt in which R1 is hydrogen, or a BOC # 1-Adoc, 2-Adoc, Z, FMOC, R12 R13 R14 Si (CH3) 2 CO group in each of the symbols R 2, R 3 and R 4 independently represent a lower alkyl group having 1 to 3 carbon atoms and m is an integer from 0 to 3, a group R 0 - or R 4 - a straight or branched chain alkyl group having 1 to 6 carbon atoms or a 1- or 2-adamantyl group; A is a bond or a Gly, a-ALA, GABA, DAVA, Aib, NH in which the symbol p re-

< U is an integer from 1 to 6 NHCO,

Met-Gly,

Tyr-Met-Gly,

Asp-Tyr (OSO 3 H) /

Asp-Tyr-Met-Gly,

Asp-Tyr (SO3 H) -Met-Gly; R 2, R 2, R 3 and R 4 are each independently hydrogen, with the proviso that not all are simultaneously hydrogen, lower alkyl, -CH 2 C H 2 O, -CH -CH 2 CH 2 -CH 2 -CH 2 -CH 2 -CH 2 -CH 2 -CH 2 OR 2 -CH 2 OR 2 -CH 2 OR 2 -CH 2 OR 2 -CH 2 OR 2 -CH 2 OR 2, - (CH 2) g CC> 2 R, - (CH 2) n (R 2) represents an integer from 0 to 3, R 2 is hydrogen or is lower alkyl, Ar is unsubstituted or substituted unsubstituted or substituted heteroatomic or hydrostatic mono- or polycyclic radical; R 2 represents a 2- or 3-indolyl, 1- or 2-naphthyl, 2- or 3-benzothienyl, 2- or 3-benzofuranyl (2- or 3-indazolyl or phenyl group, any of which groups may have between 0 and 3 substituents independently selected from hydrogen, fluorine, chlorine, bromine, iodine or between methyl, methoxy, trifluoromethyl, nitro, hydroxy, NH 2 and OCP 3; (CH2) s (CH) CH wherein R3 is an integer from 1 to 6 and s is an integer from 0 to 6, n is an integer between 1 and 3

R1 is -OH, -COOH, tetrazole, triasol, 16-COOR, 16-CCNR1, R11 represents a lower alkyl group and the symbols R10 and R11 are as defined above for R;

in which R 17 * is a group of the formula or represents a group of the formula CH 2 R 3 or represents the hydrogen atom, wherein each one of R 1 and R 2 is independently hydrogen or is lower alkyl, alkoxycarbonyl, carboxyalkyl or R 10 and R 11 together with the nitrogen atom to which they are attached form a ring having from 3 to 10 carbon atoms; 10 atoms having that ring atoms selected from carbon, nitrogen, oxygen and sulfur; and R4 has the meanings defined for or represents a group R18 and R1 or CH2CO2H or CH3 CONR4 R11 or CONR3 R11; R 2 and R 3, R 4 and R 5, R 6 and R 7, R 8 and R 9 or R 9 and R 9 and R 10 represent radicals which together may form a ring having from 3 and 8 atoms being those atoms selected from carbon, nitrogen, oxygen and sulfur. - 9 -

Preferred compounds of the present invention are compounds of the general formula I wherein R 1 is Fmoc, Z, Boc or Me 2 Si (CH 2) 2 COO; A represents a bond or represents a group Gly, p-ALA, Aifo, Asp-Tyr-Met-Gly, Asp-Tyr (OSO3H) -Met-Gly? 2 4 6 8

Each of R 1, R 2, R 3 and R 4 is independently an alkyl group having 1 to 3 carbon atoms, a carboxy alkyl group having 1 or 2 carbon atoms or a CH 2 OH group; R 2 is 3-indazolyl, 3-benzothienyl, 2-benzofuranyl, 3-benzofuranyl or 2-bromo-3-benzofuranyl; R is an isobutyl, sec-butyl-ethyl, n-propyl or i-propyl group; R 2 is CONH 2, CONMe 2 or OH; 9 does the symbol R have meanings between the 3 that have been defined for the symbol R?

Each of R 10 and R 11 independently represents an alkyl group having 1 to 3 carbon atoms or together R 1 and R 2 form a ring having from 4 to 8 carbon atoms including the nitrogen atom to which they are attached.

The most preferred compounds of the present invention are the compounds of formula I

in which R 2 represents the hydrogen atom or a BOC group; A represents a bond or a β-grupo group;

O A (s Q

When one of R1, R2, R3 and R5 is independently OH or a group O2, R 2 represents a 3-Sadolyl group. 1-naphthyl or 2-naphthyl; R 2 represents an n-butyl, n-pentyl, (SCH) or (CH 2) 2 SCH 2 OH 3 group; R 2 represents an OH, tetrazole, triazole or OOH group; R 2 represents a phenyl, 1-naphthyl or 2-naphthyl group; R 2 and R 3 independently represent the hydrogen atom which represents the CH 2 group or together the hydrogen atom representing the OH group or together R 2 and R 3 form a 5- or 6-membered ring including the nitrogen atom to which they are attached.

The preferred preferred compounds of the present invention are selected from the group consisting of:

consisting of the following compounds: Γ-methyl-DL-tryptophyl-L-methionyl-L-aspartyl-L-phenylalaninamide, Î ± -methyl-D-tryptophyl-L-methionyl-L- aspartyl-L-phenylalaninamide, Methyl [1-methyl-L-tryptophyl-L-methionyl-L-Î ± -aspartyl-L-phenylalaninamide, Methyl [1- (1,1-dimethylethoxy) L-methylethyl-L-aspartyl-L-phenylalaninamide, N-dimethyl-ethoxy] -didyl] -7-d-methyl-DL-tryptophyl-L L-tryptophyl-2-methyl-DL-methionyl-L-aspartyl-L-phenyl-alaninamide, 1 - [(1,1-dimethyl) -ethoxy) -carbonyl] -d-methyl-D-tryptophyl-L-methionyl-L-β-aspartyl-L-phenylalaninamide,

Glycyl-Î ± -methyl-D-tryptophyl-L-methionyl-L- Î ± -aspartyl-L-phenylalaninamide, G-lycylmethyl-DL-tryptophyl-L-methionyl-L-aspartyl- -phenylalaninamide,

Glycyl-Î ± -methyl-L-tryptophyl-L-methionyl-L-Î ±) -aspartyl-L-phenylalaninamide,

Trp-Met-Asp-DL-MePLe-IHo · CP4 COgH, and

Trp-Met-DL-MeAsp-Phe-IHg. Cl,

The D and L configurations of the compounds of formula I are possible at the centers of chirality - 12

θ are considered within the scope of the present invention.

Preferred compounds are those in which the residues of the acid-amines have the β-

The compounds of the present invention may have multiple centers of chirality depending on their structure. There may be centers of asymmetry at carbon atoms which bear substituents represented by R1 in general formula I and in the substituents R2, R1, R2, R3, R4, R8, R8 and R4 in general formula I. In particular, the compounds of the present invention may exist as diastereomers, mixtures of diastereomers or as single optical enantiomers or their possible mixtures. The present invention encompasses all such possible forms of the compounds. Mixtures of diastereomers are typically obtained as a result of the reactions hereinafter described in more detail. The individual diastereomers may be separated from the diastereomeric mixtures by conventional techniques such as column chromatography or successive and repeated recrystallizations.

The compounds of the present invention encompass the solvates and hydrates and also the pharmaceutically acceptable salts of Formula I. The term " lower alkyl " means straight or branched chain alkyl groups having from 1 to 6 carbon atoms unless otherwise specified, the term " alloxycarbonyl " means a group having from 1 to 4 carbon atoms.

Preferred pharmaceutically acceptable salts include benzathine, ooro-prooaine, choline, diethanolamine, ethylene diamine, meglumine, procaine. - 13 -

aluminum, calcium, lithium, magnesium, potassium, sodium, zinc, diethylamine and tromethane.

Especially preferred pharmaceutically acceptable salts are the N-methyl-glucamine and sodium salts derived from acids and the salts of H01, sulfates and trifluoroacetates in the case of the bases.

The individual enantiomers may be separated by conventional methods well known in the art such as the method of conversion to an optically active compound salt, followed by chromatographic or recrystallization separation and subsequent conversion to the non-salt form. The preferred stereochemistry of the compounds of the present invention is that which exhibits the compound of example 2.

The compounds of the present invention may be formed by coupling individual substituted acidamines by methods well known in the art. (For example, the standard synthetic methods described in the textbook H0? He Pepdides, Analysis, Synthesis, Biology, " by Gross and Meinhofer, Academic Press, Hew York.) The starting materials derived from < amino substituted amines are generally known or, if not, can be synthesized and, if desired, resolved by methods known to those skilled in the art. (Synthesis of the racemic form of DL-methyl tryptophan methyl ester - see BraKa, M. 1., et al., J. Heterocyclic Chem., 17: 829 (1980)). The present invention also encompasses novel intermediates which are useful for the preparation of the final products. These intermediates are the α-

the solid phase resin compound: by way of example, it relates to the structure Pmoc-MePhe-Resin.

Rprom-MePhe-RHg, ÂμMoc-Asp (OBu) -MePhe-IH2,

Pmoc-Met-Asp (OBu " * 3) -MePhe-IH2,

Ptno c-Trp-Met-Asp (OBu1 ") -MePhe-M2,

Asp (OPα * 3) -MePhe-M »»

Asp-MePlie-Mg, P-c-Met-Asp-MePlie-JHg,

Met-Asp (OBu " * 3) -MePhe-HH2,

Met-Asp-MePhe-IHg,

Trp-Met-Asp (OBu * 3) -MePhe-lHg, Pmoo-Trp-Met-Asp-MePi-lHg, Pma c-MeAsp (OBu * 3) -Phe-M2,

Pmoc-MeAsp-Phe-IHg,

Pmoo-Met-MeAsp (OBu " * 3) -Plie-BHg,

Pmoc-Prp-Met-MeAsp (OBu " * 3) -Phe-EHg

MeAsp (OBu " * 3) -Phe-Mg,

MeAsp-Phe-EEE2 >

Met-MeAsp (OBu " * 3) -Phe-RHg "

Met-MeAsp-Phe-IBâ,,,

Pmoo-Met-MeAsp-Phe-IHg,

Pmoc-Trp-Met-MeAsp-Phe-RHg,

MeMet-Asp (OBu " * 3) -Plie-MIg,

Pmoc-MeMet-Asp (OBu " * 3) -Phe-HSg > Pmoc-MeMet-Asp-Plie-IHg MeMet-Asp-Phe-Mg, Pmoc-Trp-MeMet-Asp (OBu "

Fmoc-Trp-MeMet-Asp-Plie-Mk ,,

Fmoc-MeTrp-Met-Asp (OBu3) -Phe-HH2,

Fmoc-MeTrp-Met-Asp-Phe-IHg, Fmoc-Gly-Trp-Met-Asp-MePhe-Mg, Fmoc-Gly-Trp-Met-Asp (OFu MePhe-HE, >

Gly-Ile-Met-Asp (OBu15) MePhe-HH2, Fmoo-Gly-Trp-Met-MeAsp-Plie-E4> Gly-Trp-Met-MeAsp (OBu) -Phe-BHg

Gly-Trp-Met-MeAsp (OFu) -Phe-NHg, Fmoc-Gly-Trp-MeMet-Asp-Phe-IEL), Fmoc-Gly-1p-MeMet-Asp (OBu) Gly-Trp-MeMet-Asp (OBu) -Phe-IH2, Fmoo-Gly-MeIrp-MetAAsp-Plie-HG, Fmoc-Gly-MeSp-Met-Asp (OBu) Me-Erp-Met-Asp (OFu " 5) -Phe-IH2,

Fmoc-MeMet-OH,

Fmoc-MeMet-OPEP,

MeAsp (OBu3) OH,

Ftaoo-MeAsp (OBu3) OH and Fmoc-MeAsp (OBu3) OPEP.

For the preparation of pharmaceutical compositions from the compounds of the present invention the usable pharmaceutically acceptable inert carriers may be in the solid state or may be in the liquid state. Solid state preparations include powders, tablets, dispersible granules, capsules, pills and suppositories.

A solid carrier may be one or more substances which also contain

the function of diluents, flavoring agents, solubilizers, lubricants, suspending agents, binders or tablet disintegrating agents; this solid carrier may also be a material for encapsulating. In the case of powders the carrier is a finely divided solid product which is mixed with the finely divided active component. In the case of tablets the active component is mixed with a carrier having the necessary binding properties in suitable proportions and then compacted into the desired size.

For the preparation of suppositories a low melting point wax such as a mixture of fatty acid glycerides and cocoa butter is used which is first melted and then the active ingredient is dispersed in the melt. shaking. This melted homogeneous butter is then poured into molds of suitable dimensions and allowed to cool and solidify.

The tablet powders preferably contain about 5 to 70% active ingredient. Suitable carriers are magnesium carbonate, magnesium stearate, talc, lactose, sugar, pectin, textrin, starch, gum tragacanth, methyl cellulose, carboxymethylcellulose sodium, a low melting wax, cocoa butter and the like. The term " preparation ", when used, is intended to mean the formulation of the active component with the encapsulating material as carrier providing a capsule in which the active component (in the presence or absence of other vehicles) is surrounded by a carrier with the which is thus associated. In the same way, the pills are also included. - 17 -

Tablets, powders, pills and capsules may be used as solid dosage forms suitable for oral administration.

Liquid preparations comprise the solutions, suspensions and emulsions. As examples of liquid preparations suitable for parenteral administration, the solutions may be referred to in sterile water or the solutions in a mixture of water and propylene glycol. Liquid preparations may also be formulated in aqueous solutions containing polyethylene glycol.

Aqueous solutions for oral administration may be prepared by dissolving the active component in suitable odorants, flavoring agents, stabilizing agents and dispersing agents as desired. Aqueous solutions for oral use may be prepared by dispersing the finely divided active component together with the viscous material such as natural synthetic gums, resins, methylcellulose, sodium carboxymethylcellulose and other known suspending agents in the specialty of pharmaceutical formulation.

Preferably the pharmaceutical preparation is in the form of a unit dose. Accordingly, the preparation is divided into unit doses containing suitable amounts of the active component. The unit dosage form may be a packaged preparation, the packet containing such package containing discrete quantities of the preparation, for example tablets, capsules and powders packed in vials or bottles. The unit dosage form may also be the capsule itself, the pill itself or the tablet itself, or may be a suitable number of any such packaged forms. . The biological activity of the compounds of the present invention was evaluated using a test of 18

screening which enabled a rapid and accurate measurement of the binding of the test compound to the known CCK receptor sites. It has been shown that there are specific CCK receptors in the central nervous system. (See Hays et al., Leuropeptides 1: 53-62 (1980); and Satuer et al., Science 208: 11555-56 (1980)). 30 to 40 g, which were dissected on ice and then homogenized in 10 volumes of 50 mM tCris-HCl buffer (pH 7.4 at 0-4 ° C). The resulting suspension was centrifuged, the supernatant was removed and the obtained mass was washed by preparing a new suspension in Iris-HCl buffer, and then centrifuged again. The final mass obtained was resuspended in 20 volumes of 10 mM Hepes buffer (pH 7.2 at 23 ° C) containing 130 nM HaCl, 4.7 nM KOI, 5 nM MgCl2 ÂμM EDTA, 5 mg / ml bovine albumin and bacotracin (0.25 mg / ml). The saturation studies were incubated with cerebral cortical membranes at 23 ° C for 120 minutes in a final volume of 50 micro -literols of Hepes buffer (pH 7.2) along with tritiated penta-gastrin at a concentration of 0.2 to 20 nM (Amersham International, England). the displacement experiments these membranes were incubated with a single concentration (2 nM) of ligand together with increasing concentrations (10 â † '10 â †') of competitive assay compound. In each case the non-binding binding was defined as the binding that persists in the presence of the unlabeled octapeptide CCK2gβ (10 ÂμM). - 19 -

After incubation the membrane bound radioactivity was separated from the free radioactivity in solution by rapid filtration through Wiiatman GrP / B filters and washed three times with ice-cold Iris-HCl buffer (4 ml). Filters of the samples incubated with tritiated pentagastrin were placed in polyethylene vials containing 4 ml of scintillation mixture and the radioactivity was assessed by liquid scintillation spectrophotometry (efficiency ranging from 47 to 52%). Specific binding to the sites of QOK reactants was defined as the total amount of tritiated pentagastrin bound less to amounts of tritiated pentagastrin bound in the presence of the β2K2 β2 octapeptide at the concentration of 10 ÂμM.

The saturation curves of the specific tritiated pentagastrin which binds to the cortical membranes of the mice were analyzed using the methods of Scatchard (Ann.Lew York Acad.So 51: 660-72 (1949), and Hill J, Physiol 40: IY-VIIX (1910) in order to provide an evaluation of the maximum number of binding sites (Bm_ ") and the equilibrium dissociation constant (Ka).

The displacement experiments the inhibition curves were analyzed alternately by the swallowed " logit-lot " or by the iterative curve fitted by the computer program (DeLean, Munson and Redbard, 1978) in order to provide an assessment of the values CI ^ q and nH (Hill apparent coefficient).

By definition the IC50 values correspond to the concentration of the test compound required to produce 50% inhibition of the specific binding).

The inhibition constant (Kj) of the compounds tested in accordance with the above-

Cheng-Prusoff equation 1 + ZX7 / Ka where the symbol L L-7 represents the concentration of the radioactive identifier and the symbol K "represents the equilibrium dissociation constant cl.

In the following Table II, several K 2 / M values are grouped for some representative compounds of the present invention.

Table II. connection to the central CCCE

Compound K ± m (n) (D 7.16 + 1.95 9 (2) 5.44 (4.23-7.10) 9 (3) (4) 7.80 (5.56-12.9) 3 (5) 9.20 (6.86-11.8) 30.40 (26.6-35.5) 3 (7) (n) = Number of tests

Data Electrofisioldgioos It is possible to find explanatory texts and protocols for experiments similar to these in J. Hughes, Proo. lati. Acade. Know. 87: 6728-6732 (1990) and βεΐί3,

songs cited there. Figure 1 depicts the dose / response type eye for C0K8s before and during exposure to compound (2) and also a curve for compound (2) alone. These curves represent the total number of action potentials that occur during response to CCK8s to compound (2) and are plotted in terms of counting unit (y-axis) as a function of dose (X-axis). Curves for compound (2) indicate that this compound has factitious properties of a partial agonist in this experiment. The curve drawn with the symbol -I- is for CCK8s, the curve drawn with the symbol == == is for the compound (2) and the curve drawn with the symbol .... A .... is for the CCK-8s + compound (2) (1μM) combination. Figure 2 depicts dose / response curves for the BH-CCK and CCK4 analogs in the same neuromondance in the VM. The total number of action potentials occurring during the response is marked by counting units (Y-axis) as a function of the ligand dose (X-axis).

The curves for the function (X * A) / (X + B) were adjusted to these points using the computer program ESI (A = maximum possible response, B = CE) of the compound calculated from the curve. These curves indicate that both compound (4) and compound (5) have agonist-like properties in this experiment. The curve plotted with the dash == o == is for compound (5). the curve drawn with the trace ..., J [... is for the compound BH-CCK, the curve drawn with the trace ---- J ---- is for the compound (4), the curve drawn with the dash - - - - is (1078, 2924441) / (5.395304e-03 + X), the curve drawn with the dash - is for 1149.663878X *) / (0.010686 + X), the curve drawn with the trait ........ is for (1100 * X) / (X + 0.47953), the curve plotted with the trace -------- is for (IlOOseX) / (X + 0.014358), the curve drawn with the trace ........ is smoothed (0.5) for compound CGK and the curve drawn with. is for compound GCK only. - 22

Figures 3 and 4 show the dose / response curves for CCK and for compounds (4), (5) and (6) both before and after exposure to the potent CCK-8 PD 134308 antagonist. The total number of action potentials occurring during the response is plotted as counting units (Y-axis) as a function of dose (X-axis). The curves for the function (X * A) / (X + B) were adjusted to these points using the ESI software. Such curves indicate that compounds (4), (5) and (6) have properties similar to those of a CCK antagonist which are antagonized by CCK-8 antagonists.

In Figure 3 the curve plotted with the -.- trait is for CCK. the curve drawn with the dash == J; = is for compound (8), the plotted curve -------------- 4 for the combination of compound (6), + PD 134308, the curve drawn with the ------ - is for (X & 1126.907666) /) 1 + 9.879552), the curve drawn with the dash is for (1123 * Χ) / (X + 0.140626) and the curve plotted with the dash ======== is for (1127 * X) / (X + 0.031476).

Ia Piggra 4 the curve drawn with the trace. - is for CCK, the curve plotted with the dash and for the compound (5), the curve drawn with the dash is for the compound (4), the curve drawn with the dash- --- $ ---- is for the combination of compound (5) + PD 134308 and the trace-plotted curve -4 is for the combination of compound (4) + PD 134308.

Tables III and IV summarize the four figures. - 23

TABLE III

(2) partial agonist 1 - (4) Agonist 2 14.0 (5) weak agonist 2 479.5 (6) Agonist 3 524.0 CCK8s Agonist 1-4. 5.4

Such compounds have also been shown to be agonists in accordance with experiments in which there was stimulation with the potent and selective CCK-B antagonist described above, the reference compound PD 134308 designated [(R, R 1, R 2) -4-Z " Z-2-Z " 3- (1H-indol-3-yl) -2-methyl-1-oxo-2 - [(trichloro-2-yloxy) ) carbonyl] amino] -Z-propyl] amino] -1-phenethyl] amino] -4-oxobutanoic acid. TABLE I?

Compound Graphic Ke (PD 134308) / nM (4) reference compound 4 4.67 (5) + reference compound 4 3.02 (6) + reference compound 3 14.4 CCK8s + reference compound - 5.4 -

The results demonstrate that compound (4) is equipotent with the compound CCK8s whereas compound (6) has a slightly lower electrophysiological activity and compound (5) possesses a relatively weak agonist-activity. Compound (2) is a partial agonist.

The compounds of the present invention are useful as an appetite suppressant as it is concluded by taking the tests described below as the basis.

The assay designated by " Tasty Diet Food " the male and adult mice of the Hooded Lister strain with a variable mass between 200 and 400 g are housed individually and trained to eat a tasty diet. This diet consists of condensed and sugary milk from the lestlé, powdered food for rats and water, all of these components being mixed together until a product of a firm consistency is obtained, a quantity between 20 and 30 g of tasty diet for 30 minutes per day during the light period of the daily light / scurry cycle during a training period corresponding to 5 days. The ingestion of a tasty diet is measured by weighing the food container before and after the 30 minutes of the access period (with 0.1 g precision limits). It is necessary to take the necessary precautions to collect any spilled diet and make the appropriate corrections. The rats have free access to a food granule and water except for the 30 minute period of the test.

After the training period the dose / response curves were constructed for 0CK8 compound and for various representative compounds of the present invention (n = 8-10 mice per dosage level). EMP40 values (confidence limits at + 95%) were obtained for the anorectic effects of these compounds. - 25 -

In their therapeutic use as appetite suppressing agents the compounds of the present invention are administered to a patient at dosage levels of about 200-2800 mg per day.

As agonists the compounds of the present invention have therapeutic utility as appetite suppressants and as antagonists have utility as agents in the treatment of gastrointestinal disorders, in the treatment of central nervous system disorders such as when applied as suppressors to the CNS, these compounds exhibit their effects as antipsychotics, neuroleptics, anxiolytics and anticonvulsants.

As antagonists it is to be understood that the compounds of the present invention have utility for reducing gastric acid secretion, for reducing anxiety, for treating gastrointestinal ulcers, for treating psychotic behavior, for blocking the reaction caused by suppression of a drug, alcohol, cocaine, benzodiazepine, diazepam or nicotine and also to enhance the effect of morphine in cases of pain treatment.

CCE analogs containing? -Methyl amino acids were prepared by three methods. Method A consists of a procedure in which there is a solution phase in which the peptides are pooled using standard protocols in which there is a solution phase to provide fully protected intermediates or a standard salt coupling which provides aspartic acid side chain unprotected intermediates. Examples 1 and 2 described below illustrate Method A. - 26

Said peptides may also be prepared by method B which is a method in which there is a solid phase wherein the peptides are constructed on solid phase resins designed to provide C-terminal amides either by treatment of the resin with ammonia in methanol or by cleavage directly from a suitably substituted resin using trifluoroacetic acid, in the presence of the necessary purifiers / obtaining the amides directly.

This latter protocol was used with DuPont RapidAmide® or Nova Biochem Novasyn KR® resins in a simple bubbler (DuPont resin) or an automated synthesizer (Nova Biochem resin).

This method is illustrated in example 3 below. The third method is Method C and is a solvent-free method where the amino component and the pentafluorophenyl ester are heated together without any solvent at a temperature comprised between 50 ° C and 100 ° C for a variable period of time between 10 and 120 minutes or possibly higher.

This method is illustrated in Example 4 below.

Individual male lodges of Hooded Lister strain (175-250 g) were fasted overnight and fasted overnight (with water-free agar). They were then anesthetized with urethane (1.5 g / kg IP) and a cannula was inserted through the trachea to aid spontaneous breathing. An infusion was performed continuously in the stomach using a modification of the original method described by Ghosh & Schild in " Continuous recordis of acid secretion in the rat ". (Br. D. Pharmac, 13: 54-51, 1956 in co-signi fi cance as described by Parsons in the publication " Quantitative studies of drug-induced gastric acid secretion " (Ph.D. Thesis, University of London, 1969). The infusion in 27

cavity was performed at a flow rate of 3 ml / min with a 5.4% w / v glucose solution via esophageal and body cannula. The fluid was driven by a centrifugal bonnet (Gilson, Minipuls 2) through a heating coil to raise its temperature to 37 ± 1 ° C. The infusion fluid was collected by a funnel collecting funnel and passed through a pH sensing electrode connected to a Jenway pH meter (PHML6). The signal obtained at the output of that pH reader was transmitted to a Rika-denki chart recorder in order to allow the real-time recording of the pH value of the gastric perfusate.

A frozen aliquot of pentagastrin was stored and diluted to the desired concentration values using a sterile 0.9% w / v NaCl solution. These new compounds were dissolved in a sterile 0.9% w / v NaCl solution on the day of the experiment. The drugs were administered via a cannula applied to the jugular vein from a large ampoule at a volume dose corresponding to 1 ml / g with 0.15 ml dilution of a 0.9 w / v NaCl solution. The basal pH value was allowed to stabilize prior to commencing administration of said compounds. Usually 30 minutes elapse between surgical intervention and administration of the first compound.

The compounds of the present invention may also antagonize the stimulation of gastric acid secretion which is produced by a standard dose of pentagastrin at a dose of 1 nmole. One of the compounds also attenuates the amount of gastric acid secreted in response to a dose of pentagastrin at 1 nmole / kg (the initial response to pentagastrin was 254 μmol / 1 H) and after administration of the compound (cumulative dose of 1.1 μmol / g) was 128 μmol / 1 H). With both compounds the antagonism is reversible with complete recovery of pentagastrin response. 28 -

The compounds of the present invention are also useful as anti-ulcer agents as hereinafter described.

The gastric lesions induced by aspirin were evaluated in 10-rater groups. All animals were fasted 24 hours prior to treatment and during the experiment. Drug or vehicle was given 10 minutes before an oral dose of 1 ml of an aspirin suspension in 0.5% carboxymethylcellulose (CMC) at a concentration corresponding to 45 mg / ml was given.

The animals were sacrificed 5 hours after administration of aspirin and the stomachs were removed and the guai were opened for examination.

The gastric lesions were classified and ranked as follows: classification 1 small hemorrhage 2 large hemorrhage 3 small ulcer 4 large ulcer 5 perforated ulcer The mean value for the classification of ulcers in the control group and the saline solution applied was of 12.1 + 6.85 (+ DP). Treatment with ranitidine (15 mg / kg PO) inhibits the formation of ulcers by 74% providing a score value for ulcers corresponding to 3. 2 + 2.35 (p < 0.001 as compared to control animals). Treatment with a compound of the present invention (10 mg / kg PO) provided a paramount classification value.

ulcers corresponding to 6.3 + 4.14 (p < 0.05 compared to control animals) which means a 48% reduction in ulcer formation.

However, the specific dosage levels may vary according to the patient, according to the state of the condition to be treated and according to the activity of the compound used. Determination of optimal doses is within the skill of the art.

The compounds of the present invention are also useful as the anxiolytic agents as described above and described.

Anxiolytic activity was evaluated in the lux / dark holding test performed with mice (B. u. Jones, et al., Br. J. Pharmacol 93: 985-993, (1988).) The number of mice used was 5 and the pretreatment time was 40 minutes The test compound was administered po in doses corresponding to 0.1, 1 and 10 mg / kg The apparatus used was an open cap box with 45 cm of tablet, 27 cm of width and 27 cm height divided into a small area (2 x 5) and a large area (3 x 5) by a partition wall extending 20 cm above the side walls. The floor of each compartment is divided into squares with 9 cm of side. The white compartment has been illuminated with a tungsten lamp with the power of loo watt placed 17 cm above the box and the black compartment was i illuminated in the same way but by a red lamp with the power of 60 watts. The entire laboratory was illuminated with red light. 30

All tests were performed between 1 pm and 6 pm. Each mouse submitted to the experiment was placed in the center of the white area and allowed to explore the surrounding environment for 5 minutes. The behavior of the mice was recorded on tape for tape deck and from that record the behavior was subsequently analyzed. Five parameters were measured: the latency time until the entry into the dark compartment # the time consumed in each area. the number of transitions between compartments, the number of lines traversed in each compartment and the number of indents in each compartment.

In this test an increase in the time taken to light is a significant, ie, directly correlated, measure of the anxiolytic effects of various standard anxiolytic drugs. The drugs are dissolved in water or physiological saline and then administered subcutaneously, intraperitoneally or through the mouth (PO) using a stomach needle.

Such compounds are subcutaneously active. In the case of control animals, 3% of crossings were observed in the dark area during measurement periods of 5 minutes. In the case of. mice treated with a compound at the dose of 1 mg / kg (SC) we observed 85 crossings into the illuminated area and only 24 crossings into the dark area, which represents the existence of a significant difference (p · & lt 0.01) relative to the anxious control mouse. The compound diazepam (0.25 mg / kg IP) provides an effect similar to that of one compound in the same experiment.

The compounds of the present invention are useful as antipsychotic agents. These compounds were tested for their ability to reduce the effects of intra-acumbene amphetamines in rats, as described below. 31

Male Sprague Dawley (Cd) Bradford rats were used. The rats were housed in groups of 5 at 21 ± 2 ° C and subjected to a daily cycle of 12 hours of lus / 12 hours of darkness with the lights on between 07.00 and 20.00 hours. The rats were fed a CRM diet (Labsure) and allowed to consume water ad libitum.

The rats were anesthetized with chloral hydrate (40 mg / kg SC) and placed in a stereotactic Kopf structure. Chronically resident guide cannulas (made of 0.65 mm diameter stainless steel tube supported bilaterally on supports *, PerspexH) were implanted using standardized stereotactic techniques to terminate 3.5 mm above the center of acumbene nucleus (Ant (Fig. 9.4, Vert. 0.0, Lat. 1.6) or to end 5.0 mm above the central nucleus of the amygdala (Ant. 5.8, Vert. -1.8, Lat. + 4.5) (De Groot atlas, 1959). These guides were maintained during a 14-day recovery period using 0.3 mm diameter stainless steel stilettos which extended 0.5 mm beyond the tips of these guides.

The rats were manually contained and the stylets were removed. Intracerebral injection cannula (s) 0.3 mm in diameter was injected and drugs were dispensed in an amount corresponding to a volume of 0.5æl for 5 seconds (it was expected that an additional 55 seconds would elapse to allow deposition) using Hamilton-type syringes connected through a polyethylene tube to the injection units. The animals were only used on a single occasion.

Experiments were performed on behavior between 07:00 and 30 minutes and at 21 and 30 minutes in a calm environment at 22 ± 2 ° C. However,

were relocated to their housing and for an hour they were allowed to adapt to the new environment. The locomotor activity was evaluated in individual net cages (Perspex " (25 x 15 x 15 cm (height)) (in groups of 30) each equipped with a photoelectric cell unit disposed along the major axis and spaced 3.5 cm It has been found that this position minimized counting units over the residual activity caused, for example, by head movements and body-adjusting movements when the animal is stationary. At this time animals are also observed with the aim of investigating the presence of any non-specific modification in the locomotor activity, for example, sedation, prostation, stereotyped movements and others that may interfere with the recording of the locomotor activity.

The ability of the compounds of the present invention to inhibit the hyperactivity caused by the injection of amphetamine within the acumbene nucleus was also measured.

Is there an increase in locomotor activity after bilateral injection of amphetamine (20æg) in the acumbeno nucleus? the peak hyperactivity (50 to 60 counting units every 5 minutes) occurs after 20 to 40 minutes after injection. Intraperitoneal injection of the mice with the compound at doses of 20 mg / kg or 30 mg / kg or 10 mg / kg reduces the hyperactivity caused by intra-acumbeno amphetamine injection. This test is known to predict antipsychotic activity (Costall, Domeney & Naylor & Tyers, Brit J Pharmac 92: 881-894). 33 -

Example 1 N-1- (1,1-Dimethylethoxy) carbonyl 7a-methyl-DL-tryptophyl-L-methionyl-L-aspartyl-L-phenylalaninamide

Boc-D-Me-Trp-Met-Asp-Phe-NH 2 (1)

A solution of Asp-Phe-NH 2 (0.6 g, 2.02 mmol) in DMF (10 mL) was treated and treated with diisopropyl-ethyl amine (261 mg, 2.02 mmol) followed by followed by treatment with the pentafluorophenyl ester of Fmoc-methionine (Fmoc-Met-OPFP - 1.10 g, 2.05 mmol) and the mixture was stirred overnight. After removal of DMF, the residue was triturated with a mixture of diethyl ether: ethyl acetate (5: 1), and the resulting solid was filtered. A crude product, Fmoc-Met-Asp-Phe-NH / / was obtained which was used without further purification (1.22 g, 96%). The procedure was repeated to give 1.29 g of product. The above prepared solid (2.00 g, 3.16 mmol) was dissolved in DMF containing piperidine (20% to 50% v / v, 16 mL) and then stirred for 20 minutes. After removal of DMF and piperidine, the residue was triturated with ether to give a white, solid product, Met-Asp-Fhe-NH4, in quantitative yield. It was also possible to use this product without further purification.

With the free tripeptide (205 0.5 mmol) a suspension was prepared in DMF (5 mL) and diisopropylethylamine (129 mg, 1.0 mmol) was added. Boc-DL-MeTrp-OPFP (242 mg, 0.5 mmol) was added and the mixture was then stirred until clean (approximately 2 days). Removal of the volatiles afforded a light tan residue which was triturated with a mixture of diethyl ether and ethyl acetate (1: 1) affording an off-white solid. Chromatography of this solid product (silica gel, 5% MeOH in dichloromethane with 1% AcOM) afforded an off-white solid product in a total of 34%

of 279 mg and in 78% yield. (DMSO-d 6) δ 1.21 and 1.28 (3H, 2S), 1.42 and 1.43 (9H,

(2H, m), 2.81 (1H, dd), 3.11 (2H, dm), 4.18-4.4 (3H, m), 4.5 (1H, (1H, mbr), 8.06 (1H, brs), 7.98 (1H, brs), 6.78 (1H, brs), 6.86-7.34 (11H, t br), 8.24 (1H, br d). (M + H) +, 739 (M + Na), 711 (M + H)

Microanalysis; Calc. Calc'd for C16 H39 N3 O: C, 58.03; H, 6.54; N, 11.60: S, 4.42. found: C, 58.13; H, 6.48; N, 11.35; S, 4.40. In an analogous manner, the compound N- [1,1-dimethylethyl] carbonyl] -L-methylethyl-L-methionyl-L-aspartyl-L -phenylalanineamide.

Boc-L-Me-Met-Asp-Phe-KH "/ (2), and the N-N- (11-dimethylethoxy) carbonyl] -Î ± -methyl-D-tryptophan- L-methionnyl-L-aspartyl-L-phenylalaninamide.

Boc-D-a-MeTrp-Met-Asp-Phe-ISIB3 (3), NMR (2: DMSO-d6): 1.25 (3H, s), 1.44 (9H, s), 1.33

(2H, m), 4.20 (1H, m), 4.33 (1H, m), 4.50 (1H, (1H), 6.92-7.47 (12H, m), 7.82 (2H, m), 8.11 (1H, br d), 10.91 (1H, s). (3H, s), 1.44 (9H, s), 1.92 (2H, m), 2.02 (3H, s), 2.42 (2H, t), 2.60 (1H, bs) ), 2.84 (1H, dd), 3.13 (2H), 3.31 (2H + water), 4.19 (1H, m), 4.36 (1H, m), 4.48 (1H, m), 6.94-7. 24 (12H, m), 7.32 (1H, d),

7.47 (1H, d), 7.84 (1H, d br), 8.14 (1H, d br), 8.22 (1H, br.s), 10.92 (q, s). 35 -

Example 2

By this method the α-methyl-L-tryptophyl-L-methionyl-L-α-aspartyl-L-phenylalaninamide compound was also prepared. L-c-MeTrp-Met-Asp-Pbe-NH4, (4), (4-methyl-D-tryptophyl-N-methionyl-L-cystectyl-L-phenyl-alanine-mide. D-a-MeTrp-Met-Asp-Phe-NH2 / (5) and α-methyl-DL-tryptophyl-L-methionyl-L-α-aspartyl-l-phenylalanineamide. DL-ph-MeTrp-Met-Asp-Phe-NH2 / (6).

In these last three cases the final residue used was the appropriate Fmoc-cc-MeTrp activated ester which affords a protected tetrapeptide which was treated with a solution of piperidine in DMF to remove the protecting group. This treatment afforded the tetrapeptides (4-6) completely deprotected. Purification was performed by high pressure liquid chromatography (MeGN-H₂O / 0.1% TFA) affording the trifluoroacetate salt of each peptide. NMR (δ, D₂O): 1.73 (3H, s), 1.85 (2H, q), 2.04 (3H, s), 2.34 (2H, t), 2.66 (2H, qd), 3.07 (2H, qd), 3.41 (2H, s), 4.34 (1H, t), 4.54 (2H, m), 7.14 (1H, t), 7.21-7.37 (9H, m), 7.48-7.56 (2H, dd). RM (5); (2H, m), 1.97 (3H, s), 2.54 (2H, m), 2.93-3.28 (6H, m), 4.13 (1H, t), 4.42 (1H, t), 4.55 (1H, m), 7.13-7.36 (8H, m), 7.51 (1H, d), 8.18 (1H, d). NMR (6); (3H, s), 1.85 (2H, m), 1.94 and 2.03 (3H, 2b), 2.34 (1H, t), 2.74 (2H, , 4.01 and 4.34 (1H, 2t), 4.60 (2H, m), 7.11-7.48 (10H, m), 7.53 (2H, t). 36 -

The following compounds were prepared by the procedures described above in Method A.

Glycyl-Î ± -methyl-D-tryptophyl-L-methionyl-β-Î ± -aspartyl-1β-phenylalaninamide (Gly-DMePrp-Met-Asp-Phe-NH2),

Glycyl-Î ± -methyl-DL-tryptophyl-L-methionyl-L-β-aspartyl-L-phenyl-alanamide (Gly-DL-MeTrp-Met-Asp-Hie-NH2),

Glycyl-α-methyl-L-tryptophyl-L-methionyl-L-C-aspartyl-L-phenylalaninamide. (Gly-L-MeTrp-Met-Asp-Phe-NHg) /

Trp-Met-Asp-DL-MePhe-NH2.CP3COH, and Trp-Met-DL-MeAsp-Phe-NH2CF3CO2H2. can also be prepared according to the following example. S-3-L-tryptophen-3-methyl-DL-methionyl-L-α-aspartyl-L-phenylalaninamide Ιι-Trp-DL-g-MeMet-Asp-Phe-NH 2 (7)

Using an automated peptide sys- temizer model Nova Biochem 4170 and the Bioplus (r) toxicology application the peptide was constructed from the C-terminus using DHBt or pentafluorophenyl esters of the Fmoc amino acids and using the catalysis with HOBt. Each residue was present in an amount five times in excess to ensure rapid and complete acylation. On a scale corresponding to 0.095 mmol the peptide (4) was isolated followed by cleavage with TFA (94% TFA, 5% anisole, 1% ethanedithiol) from resin (2 hours at room temperature) with a 45% yield (32 mg isolated). High pressure liquid chromatography (CLEP) (C18, 20-80% MeCN-E2 O + 0.1% TFA) demonstrated that the purity was greater than 90% (detection at 254 nm) and that the product obtained was a mixture of diastereomers in the ratio of 1: 1. NMR (d 20) & (3H, s), 2.59 (2H, m), 3.09 (2H, m), 3.38 (3H, m), 4.31 (1H, , m), 4.51 (2H), 7.17-7.37 (10¾ m), 7.52 (1H), 7.68 (1H), 10.94 (1s). The product obtained was the same.

In a similar manner, the product (a) in the same manner as the product prepared above was also prepared.

Example 4

Trp-Met-Asp-DL-MeEh and -NH 2. CF 3 COOH (8)

Fmoc-Asp (OBu ") - DL-MePhe-

A mixture of MePhe-NH 2 (90 mg, 0.5 mmol) and Fmoc-Asp- (OBu1 ') pP (290 mg, 0.5 mmol) was prepared and dissolved in the required minimum amount of ethyl acetate and then The solvent was distilled off immediately. The resulting viscous oil was heated in boiling barium for 2 hours and then allowed to cool. The resulting resin was triturated with diethyl ether to give a precipitated product. A total amount of 159 mg was obtained in 56% yield. It would have been possible to obtain another crop of product by chromatography of the ether washing products.

(900 mg, 1.58 mmol) was N-deprotected using a 20% solution of N-

piperidine in DMF for 20 minutes and then the solvent was completely removed. The residue was triturated repeatedly with n-hexane to provide the amino group free dipeptide (293 mg, 0.84 mmol). To this was added Fmoc-Met-Qpfp (450 mg, 0.84 mmol) and then the required minimum amount of ethyl acetate was further added to provide a clear solution. The solvent was removed immediately and the resulting viscous oil was heated in a water bath at 65 ° C for 45 minutes. Trituration of the resulting resin with diethyl ether gave the protected tripeptide (418 mg, 78%).

(300mg, 0.43mmol) was N-deprotected using a 20% solution of piperidine in DMF for 20 minutes. Removal of the solvent followed by trituration of the residue with n-hexane afforded the free tripeptide (198 mg, 0.41 mmol). To this was added Fmoc-Trp-OPfp (243 mg, 0.41 mmol) and a sufficient amount of ethyl acetate to provide a clear solution. The solvent was removed immediately and the resulting mixture was heated at 65 ° C for 15 minutes. Trituration of the residue with diethyl ether gave a white solid solid in a total of 283 mg in 78% yield.

(8) The above tetrapeptide was N-deprotected using a 20% solution of piperidine in DMF for 20 minutes and then the solvent was removed and the residue was triturated. residue with n-hexane to afford the free amino group tetrapeptide. This product was treated with thianisole and ethanedithiol (100-250 æl / mmol and 50-100 æl / mmol) and then treated with an aqueous solution of 95% TFA ((3-5 mL / mmol ) for 5 minutes.All volatiles were removed and the residue was triturated with diethyl ether to give a white product as a white solid which was purified by high pressure liquid chromatography (HPLC ) preparatory.

Trp-Met-DL-MeAsp-Phe-NEU. CF 3 CO (9)

Fmoc-DL-MeAsp (OBu1) -Phe-NH2

An amount of Pmoc-DL-MeAsp (OBu ') OPfp (1.46 g, 2.47 mmol) and H-Phe-NH 2 (0.41 g, 2.5 mmol) was dissolved in the minimum amount of ethyl acetate required to provide a solution clear and then the solvent was removed immediately. The mixture was then heated at 100 ° C for 1 hour and allowed to cool. Trituration of the residue with diethyl ether gave the dipeptide. of the protected product in a total of 993 mg in 71% yield.

Fmoc-Met-DL-MeAsp (OBr) -Phe-NH The above dipeptide (571 mg, 1.0 mmol) was N-deprotected as previously described. After trituration with n-hexane the residue was mixed with Fmoc-Met-OPfp (350 mg, 0.65 mmol) and with sufficient ethyl acetate to allow dissolution. After immediate removal of the solvent the residue was heated at 65 ° C for 4 hours. Trituration of the residue with diethyl ether provided the protected tripeptide in a total of 210 mg and in 46% yield. It would have been possible to isolate another crop from the ether washing products using the chromatography technique.

Fmoc-Trp-Met-PL-MeAsp (OBuO -Eh-NHg

The above tripeptide (400 mg, 0.57 mmol) was N-deprotected as previously described. After trituration of the residue with n-hexane it was possible to isolate an amount of 258 mg of product. An amount of 200 mg (0.42 mmol) of this product and Fmoc-Trp-OPfp (246 mg, 0.42 mmol) was mixed together with a sufficient amount of ethyl acetate which allowed dissolution. The solvent was removed immediately and the residue was heated to 65 ° C.

Claims (2)

for 20 minutes. Trituration of the resulting mixture with diethyl ether provided the protected tetrapeptide in a total of 283 mg and in a yield of 77%. Trp-Met-DL-MeAsp-Phe-NH2. The above tetrapeptide was treated by the same procedure to provide compound (8) after final trituration with ether. CLAIMS or a pharmaceutically acceptable salt thereof wherein R3 is hydrogen, or a BOC group, 1-Adoc, 12 13 14 2-AdOC, Z # FMOC, R 2 R 2 Si (CH 2) O CO- wherein R 1 and R 2 are independently and independently a lower alkyl group having 1 to 3 ring atoms, carbon and m is an integer from 0 to 3 ## STR7 ## wherein R 15 is a straight or branched chain alkyl group having 1 to 6 carbon atoms or a group 1 or 2-adamantylo group A represents a bond or represents a group Gly, ρ-ALA-GABA, DAVA, Aib, NH (CH2) p wherein p is an integer from X to 6, or NH3, Met-Gly, Tyr-Met-Gly or Asp-Tyr-Met-Gly, Asp-Tyr (OSO3 H) -Met-Gly? R 2, R 3 and R 4 are individually or independently hydrogen, with the proviso that not all are simultaneously hydrogen, lower alkyl, -CH = CH 2, (CH2) q N (R) 2 wherein q is an integer from 0 to 0, wherein R1, R2, R3, and 3, R is hydrogen or lower alkyl, Ar is an unsubstituted or substituted carbo- or heteroaromatic or mono- or polycyclic hydroaromatic radical; R 2 is 2- or 3-indolyl, 1- or 2-naphthyl, 2- or 3-benzothienyl, 2- or 3-benzofuranyl, 2- or 3-indazoyl or phenyl with the possibility of being 0 and 3 substituents independently selected from hydrogen, fluorine, chlorine, bromine, iodine, or between methyl, methoxy, trifluoromethyl, nitro, hydroxy, nitro and trifluoromethyl groups; R 2 is a straight or branched chain lower alkyl group having 1 to 6 carbon atoms or represents a (CH 2) n CH 2 group in which R 2 represents an integer of from 1 to 6 and s is an integer from 0 to 6; n is an integer from 1 to 3; R 7 is -OH, -COOH, tetrazole, triasol, -COOR 16, COOR 10 R 1: L, 1 wherein R is lower alkyl and R 10 and R 11 are as defined below; R is selected from the groups 3 defined above for R '  € ƒâ € ƒâ € ƒR1 is -CH2 NR1 R2 or CH3 OR is hydrogen, wherein R1 and R2 are independently and independently hydrogen or lower alkyl; alkoxycarbonyl, carboxyalkyl or the radicals R 2 and R 3 together with the nitrogen atom to which they are attached form a ring having from 3 to 10 atoms, said ring having atoms selected from carbon, nitrogen , oxygen and sulfur; R 2 has the same meanings as defined for R1 or is CHCHOCH3 or -CH2 CH2 OH; or CONTAIN; The radicals represented by the symbols R 1 and R 2, R 2, R 3 and R 4 are independently selected from the group consisting of: carbon, nitrogen, oxygen and sulfur. A compound according to claim 1 wherein R 3 is Pmoc, Z, Boc, or Me 2 Si (CH 2 >20002H; A is a bond or a Gly, a group Asp-Tyr-Met-Gly: R, R, R and R are each independently and independently an alkyl group having from 1 to 3 carbon atoms, a carboxyalkyl group having 1 or 2 atoms of carbon or a CH20H? R 3 is 3-indazolyl, 3-benzothienyl, 2-benzofuranyl, 3-benofuranyl or 2-bromo-3-benzofuranyl; R 5 is an isobutyl, sec-butyl-ethyl, n-propyl ortho-propyl group; R 2 is a CONH 2, CONMe, or OH group; R 2 is selected from the groups defined for R 3; R 2 is a group in which R 10 and R 11 are individually and independently an alkyl group having 1 to 3 carbon atoms or together the groups represented by the symbols and form a ring having from 4 to 8 atoms including the nitrogen atom to which they are attached. A compound according to claim 1 wherein R 1 is hydrogen or BOC? the symbol A represents a bond or the group p-ALA? R 2, R 3, R 4 and R 5 are independently and independently a C 1-4 -alkyl group; R4 is a 3-indolyl, 1-naphthyl or 2-naphthyl group; R 2 is n-butyl # n-pentyl # (ch 2) 2sch 3 or (ch 2) 2sch 2ch? R4 is tetrazole, triazol or COOH; R 9 is a phenyl, 1-naphthyl or 2-naphthyl group; R 2 represents a group R 3 and R 11 is independently and independently hydrogen or CH 2, or together these R 10 and R 11 radicals form a ring having 5 or 6 atoms including the nitrogen atom to which they are attached. A compound designated as α-methyl-DL-trip-tolyl-L-methionyl-L-α-aspartyl-L-phenyl-alaninaraamide. A compound designated as α-methyl-D-trypto-phthal-1-methionyl-L-α-aspartyl-L-phenylalaninamide. A compound designated as α-methyl-L-tryptophyl-β-raetionyl-L-α-aspartyl-L-phenyl-alaninamide. (1,1-dimethyl-ethoxy) -carbonyl-7-α-methyl-L-tryptophoryl-Irmethionyl-L-α-aspartyl-L-phenylalaninamide. A compound designated as N- [1- (1-dimethyl-ethoxy) carbonyl] -7-methyl-7-DL-tryptophyl] -L-methionyl-L- A compound designated L-tryptophyl-2-methyl-3-ethionyl-1α-aspartyl-L-phenylalaninamide. A compound designated as (1,1-dimethyl-ethoxy) -carbonyl-α-methyl-β-D-tryptophenyl-L-methionyl-L-α-aspartyl-L-phenylalaninamide. Compound designated by glycyl-α-methyl-D-tryptophyl-1-methionyl-L-α-aspartyl-β-phenylalaninamide. A compound designated as glycyl-Î ± -methyl-DL-tryptophyl-L-methionyl-L-Î ± -aspartyl-L-phenyl-alaninamide. A compound designated as glycyl-Î ± -methyl-β-tryptophyl-L-methionyl-L-Î ± -aspartyl-L-phenylalaninamide. 46 - Pharmaceutical coapposition characterized in that an amount of a compound according to claim 1 is effective to suppress the appetite of a mammal together with a pharmaceutically acceptable carrier. The compound selected from the group consisting of Fmoc-MePhe-NK2, Fmoc-Asp (OBu **) -MePhe-NH2, Fmoc-Met-Asp (OBu3) ) -MePhe-KH3 / Asp (OBu **) -MePhe-NH3 / Asp-MeFhe-NH2, Fmo c-Met-Asp-MeFhe-NH2, Met-Asp (OBut) -MePhe-KH2-Mst (OBu) -MePhe-NH2, Fmoc-Trp-Met-Asp-MePhe-NKg / 4 * Fmoc-MeAsp (OBu) -Phe-MHg Fc c- MeAsp-Phe-KH 2 # Fmoc-Ket-MeAsp (OBu) -Phe-NH 2, Fmoc-Trp-Met-MeAsp (OBu **) -Phe-NK 4 # 4 MeAsp (OBu) MeAsp-Phe-NH 2 # Met-MeAsp (OBu **) - Fhe-NH 4 47 - Metho-Me-Asp-Phe-NH 2 MeMet-Asp (OBut) -Phe-NH 2 Fmoc-MeMet-Asp (OBu) Phe-NH and Fmoc-Metho-Asp-Phe-NH2 MeMet-Asp-Ehe-NH-Fmoc-Trp-MeMet-Asp (OBi) -Fe-NK2 Fmo c-Trp-MeMet-Asp-Phe-NH2 Fmoc-MeTrp -Met-Asp (OBu) -Phe-NH2 Fraoc-MeTrp-Met-Asp-Phe-NH2 Frnoc-Gly-Trp-Met-Asp-MePhe-NH2 Fmoc-Gly-Trp-Met-Asp (OBu) MePhe (Glu-Gly-Trp-Met-MeAsp-Phe-NH 2, Fmoc-Gly-Trp-Met-MeAsp (OBu)) - Phe- Fmoc-Gly-Trp-MeMet-Asp-Fhe-NH2, t Fmoc-Gly-Trp-MeMet-Asp (OBu) -Phe- NH-Gly-Trp-MeMet-Asp (QBu2) -Phe-NH2, Fmoc-Gly-MeTrp-Met-Asp-Phe-NH2, Fmoc-Gly-MeTrp-Met-Asp (OBi3) -Phe- , MeOH (MeOH), Fmoc-MeAsp (OBtt) (OH), and MeOH (MeOH), and MeOH (MeOH) The applicant claims the priority of the U.S. application filed on April 24, 1991, under Serial No. 690,754, Lisbon, April 24, 1992 49 -