ITTO950126A1 - NEW POLYAMIDE DERIVATIVES OF ORNITHINE, LYSINE AND ANALOGUES WITH CCK-B AND GASTRIN ACTIVITIES, PROCEDURE FOR THEIR PREPARATION - Google Patents

Abstract

Compounds which can be represented by the general formula (I) indicated below and in which: (FORMULA I) r1 is a simple or mono or disubstituted phenyl group with chlorine; R2 is H or CH3; R3 is a spiro heterocyclic group represented by: (FORMULA II) in which Y is chosen independently between CH2 and O (oxygen) or 1: s is an integer between 1 and 4; m, r and t are integers chosen independently and included between O (zero) or and 2; z is equal to O (zero) 1; R4 is independently selected from a simple or mono or disubstituted phenyl group with methyl or chlorine, a 1 (or2) -naphtyl group, a 2 (or3) -indolyl group, a 2 (or3) -kinolinyl group. The configuration of the chiral center marked with * in the general formula (I) can be independently R or racemic, while that of the chiral center indicated with ** can be independently R, racemic or S.

Description

Description of the industrial invention entitled:

"NEW POLYAMIDE DERIVATIVES OF LYSINE ORNITHINE AND ANALOGS TO ACTIVITIES" CCK-B AND GASTRINE? ANTAGONIST, PROCEDURE FOR THEIR

PREPARATION AND THEIR PHARMACEUTICAL OS

DESCRIPTION

The present invention relates to new polyamide derivatives which can be represented by the general formula (I) indicated below and in which:

where Y? independently chosen between CH2 and O (oxygen) and n? 0 (zero) or 1;

s? an integer between 1 and 4;

m, r and t are integers chosen independently and included between 0 (zero) and 2;

z? equal to 0 (zero) or 1;

? independently selected from a simple or mono or disubstituted phenyl group with methyl or with chlorine, a 1 (or 2) -naphthyl group, a 2 (or 3) -indoleIe group, a 2 (or 3) -quinolinyl group.

The configuration of the chiral center marked with * in the general formula (I) can? be, independently, R (Rectus), or racemic (RS) while the configuration of the chiral center marked with ** in the general formula (I) can? be independently R (Rectus), racemic (RS) or S (Sinister). Preferably R1? a phenyl substituted with chlorine in positions 3 and 5, R3? the 8-azaspiro group [4.5] decan-8-yl, s? 2, R *? the group 1-naf? ile, and if z? 0 (zero) at the same time m and r are both 1 while t? 0 (zero) or 1, while when z? 1, m and t are both 0 (zero) and r? 2.

The compounds of the present invention prove to be potent peripheral gastrin receptor antagonists, i.e. at the level of the gastrointestinal system, and the powerful cholecystokinin receptor antagonists (CCK) at the level of the central nervous system (CCK-B-antagonists). Can you? therefore to believe that they can be used with advantage in the therapy of various human diseases linked to imbalances in the physiological levels of gastrin and CCK or other bioactive polypeptides related to them, both at the level of the gastrointestinal system and at the level of the nervous system central (CNS) or other organs or systems in which these bioactive peptides play a physiological or pathological role. So, for example, you can? advocate an advantageous use of these compounds for the treatment, at the gastrointestinal level, of diseases related to motility disorders? and mucous trophism such as gastritis, peptic ulcer, colitis or certain gastro-intestinal tumors caused by gastrin or related polypeptide hormones, and at the CNS level, for the treatment of mental disorders such as anxiety, panic, psychosis, such as schizophrenia, anorexia, etc. Another use could be that of the treatment and prevention of some eye diseases such as myosis induced in the course of surgical treatments of cataracts or by chronic ocular inflammations or other affections of the sense organs.

Pharmaceutical forms of the compounds object of the invention can be prepared according to conventional techniques such as for example tablets, capsules, suspensions, solutions and suppositories and can be administered orally, parenteral, rectal, ocular or transdermal or other suitable forms to obtain the effect therapeutic.

The active ingredient is typically administered to the patient with a reference dose ranging from 0.01 to 10 mg / kg of body weight per dose. For parenteral and ocular administration? it is preferable to use a water-soluble salt of the compounds in question such as the sodium salt or another non-toxic and pharmaceutically acceptable salt. As inactive ingredients, substances commonly used in pharmaceutical technique such as excipients, binders, flavoring, disaggregating agents, dyes, humectants, etc. can be used. The process for the preparation of the polyamide derivatives according to the invention consists of an amidation reaction of the basic derivatives of formula (II):

Alternatively, instead of the anhydrides of formula (III) and specifically if m and r are both 0 (zero), the corresponding monoacid chlorides are preferably used.

The basic starting derivatives of formula (II) having the chiral center marked with (*) in the racemic configuration (R, S) were prepared with conventional methods starting respectively from diaminopropionic acid, diaminobutyric acid, omithine and lysine, all in the configuration (R, S) to give the respective bases of formula (I) in which "s" assumes the value of 1, 2, 3 and 4 respectively (Compounds H-1 - II-4 of table 2).

The basic starting derivatives of formula (?) Having the chiral center marked with (*) in the (R) configuration can be conveniently prepared from the corresponding acids, as for example in the case of (R) -4- (3,5 -dichlorobenzoylamino) -5- (8-azaspiro [4.5] dec-8-yl) -5-oxopentanoic, the reference compound among the CCK-B-antagonists derived from 2-aminopentanedioic acid [CR 2194, Makovec et al. , J. Med. Chem., 1992 (35), 28-38], This compound? was converted into the corresponding acylazide, which by means of a Curtius transposition, gave the corresponding isocyanate, which after hydrolysis, formed the desired basic compound? -5 (see table 2). In the same way, yes? obtained the corresponding enantiomer (S) II6 synthesized for comparative purposes. Some physico-chemical data of the basic intermediates of formula used in the synthesis of the compounds of formula (I) object of the invention are shown in table 2. The anhydrides of formula (IH) are available on the market, or are described in the literature or are been prepared with conventional methods. The following examples are reported to better illustrate the invention.

Example. 1

l- (8-azaspiro [4.5] dec-8-yl) -l-oxo-2- (RS) - (3,5-dichlorobenzoylamino) -5 aminopentane [Compound? -3 - Table 2]

7 g of (RS) -N <a> -Z-N <e> -BOC-omithin (0.0172 mol) prepared according to Scott J. W. et al. [Synth. Comm., 1981 (11). 303-314] are dissolved in 100 mL of tetrahydrofuran (THF); to this solution, 2.5 mL of triethylamine (0.0181 moles) are added and the whole is cooled to -5 ° C. At this temperature 1.7 mL of ethyl chloroformate (0.0181 moles) dissolved in 20 mL of THF are dropped. The reaction ? kept under agitation at -5 ° C for 20 minutes and after this period, always keeping the temperature at -5 ° C, we drop 2.9 g (0.0189 moles) of 8-azaspiro [4.5] decane prepared according to Najer H. et al. [Bull. Soc. Chim. 1964 (IQ), 2572-2581], dissolved in 30 mL of THF. The reaction is then left for 30 minutes always at -5 ° C and overnight at room temperature. The precipitate (triethylammonium chloride) is filtered and the THF is evaporated; the oily residue? dissolved in ethyl acetate and? washed first with HC12N and then with 2N NaOH; after neutral washing, the solvent is anhydrified and evaporated recovering 7 g of oil (0.0144 moles) which are dissolved in 150 mL of methyl alcohol and hydrogenated at room temperature and pressure, in the presence of 0.2 g of palladium on 10% carbon. After 5 hours the catalyst? filtered and the solvent evaporated; 4.1 g of oil (0.0116 moles) are obtained which are dissolved in 100 mL of THF. To this solution are added 6.1 mL of 2N NaOH (0.0122 moles) and 40 mL of water; at 20 ° C, 2.5 g of 3,5-dichlorobenzoyl chloride (0.0116 moles) and 6.1 mL of 2N NaOH (0.0122 moles) are dropped simultaneously. After one night at room temperature the solution? acidified and the separated oil is extracted with ethyl acetate. The organic phase? washed with HC12N and neutral water. The solvent? dried and evaporated, recovering 6.1 g of oil (0.0115 moU) which are dissolved in 100 mL of ethyl acetate; the solution is cooled to 0 ° C and saturated with gaseous HCl. After an hour at 0? C the solution? treated with excess isopropyl ether. The precipitating solid is filtered, washed with isopropyl ether and dried in vacuo, obtaining 4.4 g (0.0096 moles) of compound II-3 hydrochloride. Yield 56% (C21H29C12N302-HC1).

Melting point 206-210 ° C; TLC (ButOH-Ac. Acetic-H20 5: 2: 2): Rf 0.75. Rotating power [aD] = 0 (C = 1% in chloroform)

Compounds? -1, 11-2 and II-4 of table 2 were prepared with the same procedure.

Example 2

l- (8-azaspiro [4.5] dec-8-yl) -l-oxo-2- (R) - (3,5-dichlorobenzoylamino) -4 -aminobutane [Compound II-5 - Table 2]

5 g (0.0113 moles) of (R) -4- (3,5-dichlorobenzoylamino) -5- (8-azaspiro [4.5] dec-8-yl) -5-oxo-pentanoic acid (CR2194), are dissolved in 100 mL of dioxane; 1.58 mL of triethylamine (0.0113 moles) are added to this solution and the whole is cooled to -5? C. 1.08 mL of ethyl chloroformate dissolved in 25 mL of THF is added dropwise and then, after about 15 minutes, 1.8 mL of trimethylsilylazide dissolved in 25 mL of THF. At the end, the temperature is allowed to rise to the ambient one and one proceeds by similitude with what described by Altman Y. et al. [Tetrahedron: Asymmetry 1994 (5), p. 891, compound 9].

With the same procedure? Compound II-6 of Table 2 was prepared, synthesized for comparative purposes.

Example 3

Acid-4- (RS) - (l-naphthoylamino) -5N - [[3- (R) - (3,5-dichlorobenzoylamino) -4- (8-azaspiro [4.5] dec-8-yl) -4- oxo] -l-aminobutyl] -5-oxo-pentanoic [Compound 10 - Table 1]

7 g of the compound II-5 of Table 2 (0.017 moles) are dissolved in 200 mL of acetonitrile and 6 mL of triethylamine (0.0373 moles) are added to this solution; the mixture is cooled to -5 ° C and 6 g (0.020 moles) of (RS) -N- (1-naphthoyl) -glutamic anhydride (compound III-5 of Table 3) are added. After 20 minutes cold, it is left at room temperature for one night. The reaction mixture is diluted with water and acidified with HC12N. The oil that is obtained? extracted with ethyl acetate which is subsequently washed in neutral with water, anhydrified and evaporated. The residue obtained is precipitated by treatment with ligroin and subsequently purified by treatment with isopropyl ether.

According to this procedure, all the derivatives of table 1 were prepared with the exception of derivatives 3, 4 and 8 which were synthesized using respectively phenyl or benzylmalonic acid as described in the following example.

Example 4

Acid-2- (RS) -benzyl-3N - [[5- (R, S) - (3,5-dichlorobenzoylamino) -6- (8-azaspiro [4.5] dec-8-yl) -6-oxo] -l-aminohexyl] -3-oxo-propionic [Compound 8-Table I] 8 g (0.04 moles) of benzylmalonic acid were converted into the corresponding monochloride by treatment with donyl chloride in analogy with what described by Rumar G.N. et al. [Drug Des. and Discovery, 1993 (10V p. 13]. The chloride dissolved in THF is then dropped at about 10 ° C in a solution containing 18 g (0.04 moles) of compound II-4 and 10.1 ml (0.08 moles) of triethylamine dissolved in 400 ml of THF. After 12 h of reaction the solution is evaporated under vacuum and the oily residue is taken up with ethyl acetate and H20, the organic phase is washed with diluted HCl and H20 and dried under vacuum. The residue is precipitated by repeated washing with petroleum ether and subsequently purified by treatment with isopropyl ether.

In the following table 1 some derivatives of formula (I) cos? obtained with some chemical-physical characteristics that identify them. Table 2 shows some chemical-physical data concerning the basic intermediates of formula (II) used in the preparation of the derivatives described in table 1, while in table 3 there are some literature data concerning the anhydrides of formula (III) gi? previously described as well? some chemical-physical characteristics of the anhydrides (III) not previously known.

DESCRIPTION OF THE ACTIVITY PHARMACOLOGICAL

1) Activities anticolecystokinin infants CCK-B 1 in vitro

To evaluate the ability? to interact by the compounds object of the invention with the central CCK-B receptor, yes? used [3-H] [N-methyl-N-leucine] CCK-8. This ligand has been shown to be selective for CCK-B receptors, having an affinity for it. about 4000 times pi? elevated for receptors of the cortex (CCK-B) compared to those of the pancreas (CCK-A) in the guinea pig [Knapp et. to the; J. Pharmacol. and Exp. Therap. 255 (3) (1990), 1278-1286], Cerebral cortices of male albino guinea pigs were therefore used, following the aforementioned method, slightly modified [Makovec et al .; Bioorgani c & Med. Chem. Letters 3 (5) (1993), 861-866] and so as to obtain a membrane content corresponding to about 300 mcg of proteins / ml. The results obtained are shown in table 4, which shows the IC50, that is? the concentration (in micromoles / liter) of the antagonist capable of displacing [3-H] [N-methyl-N-leucine] CCK-8 from the receptor by 50%.

From the data reported in table 4 it can be seen that many of the compounds object of the invention, such as compounds 2 and 10, are potent inhibitors of the binding of [N-methyl-N-leucine] CCK-8 to the receptors of the cortical membranes of guinea pig, being only 4-8 times less active than the specific pentagastrin antagonist and 100-200 times more? potent compounds of the reference compound CR 2194. The activity? unsettling? strongly conditioned by the stereochemistry of the carbon atom indicated with (*) in the general formula (I). In fact, for example, compound 2? about 30 times pi? active component of its S diastereomer, compound 17.

2) Activities antigastrinic (peripheral) in rabbit gastric mucosal cells in vitro

The parietal cells of the gastric mucosa are responsible for the secretion of HC1. They have specific membrane receptors which can be activated by gastrin and which have been defined as gastrin or cholecystokinin type B (CCK-B) receptors.

Since? ? it has been observed that the activation of CCK-B receptors by gastrin leads to an increase in the levels of cytosolic calcium ion,? a technique was used to measure the increase in intracellular calcium induced by gastrin, in the presence and absence of the compounds object of the invention, as an index of the activity? antigastrinic of the compounds themselves.

Suspensions (0.8 x 106 / ml) of rabbit gastric mucosa cells were prepared by conventional techniques using collagenase and pronase as digestive enzymes; the estimate of the values of [Ca2 *] ;, basal or reached after stimulation of the cellular system,? was conducted in accordance with Grynkiewicz et al [J. Biol. Chem. 260 (1985), 3440], In the control samples the cells were stimulated with gastrin 5x10-8, while in the samples in which? The effect of the compounds in question was evaluated, the cells were incubated with them before stimulation with gastrin. The results were expressed as percentages of [Ca <2 +>] i compared to the control value. The activity? antigastrinic of compounds? been expressed as IC ?, that is? the concentration (in micormoles / liter) at which the response to the stimulus induced by gastrin? reduced by 50%. The results cos? obtained for some compounds object of the invention are shown in table 5, which also shows an index obtained from the ratio between the activity? peripheral antigastrinica now described and the activity? disturbing obtained from the study of binding to the cortical receptors of guinea pig described previously.

From the data reported in table 5 it can be seen that many of the compounds object of the invention are potent inhibitors of the cytosolic calcium increase induced by gastrin in the cells of the rabbit gastric mucosa.

Basically the activity? peripheral antigastrinic? in good agreement with the activity? antigastrinica obtained centrally with the binding studies illustrated previously in table 4. In fact, compounds 2 and 10 are also in this case the most? potent among the compounds described, exhibiting ICS0 of the nanomolar order of magnitude (5 and 3 nM respectively). Generally the compounds in question, carry out the activity? antigastrinic on this model at concentrations 2-8 times lower than those obtained centrally.

3) Activities anticolecystokinin infants CCK-A1

In order to verify the hypothesis that the compounds in question were specific CCK-B antagonists, yes? tested for all the compounds illustrated in tables 4 and 5 also a possible activity? CCK-A antagonist. As an experimental model yes? used the guinea pig gallbladder stimulated in vitro by CCK-8 according to the method described by Makovec et al. [(Arzneim. Forsch. / Drug. Res. 35 (7), 1048-1051 (1985)].

None of the compounds tested was shown to have an activity? CCK-A antagonist pi? powerful of 10 x 10 <-6> M.

Comparing these activities? with the CCK-B antagonist previously illustrated in table 5 you can? conclude that the compounds in question are specific antagonists for the CCK-B receptor, exhibiting the compounds pi? as powerful as compounds 2 and 10, an affinity? at least 1000 times more? elevated for the gastrin receptor (CCK-B) than for the cholecystokinin (CCK-A).

4) Activities anxiolytic

Among the possible activities? of the compounds in question on the Central Nervous System, linked to imbalances in the neuronal physiological levels of gastrin or other peptides related to it, their potential activity appears particularly interesting? anxiolytic.

In fact, an important role for the central CCK-B receptor in anxiety has recently been postulated. This is in agreement with studies also conducted on humans which have shown that central CCK-B mechanisms play an important role in mediating panic attacks [Bradwejn, J. et al; J. Psychopharmacology 6 (1992), 345], In order to confirm this hypothesis, yes? evaluated the activity? anxiolytic of some of the pi? powerful CCK-B antagonists object of the invention by challenging the "elevated plus-maze" method in rats, carried out in accordance with Pellow et al. [J. ofNeurosc. Meth. 14 (1985), 149-167], Yes? used a labyrinth whose length of the cross arms was 45 cm and placed at a height of 70 cm from the ground. A compound with activity? anxiolytic produces on this experimental model a% increase in the time spent in open arms and a% increase in the number of entries in open arms.

The results obtained are shown in the following table 6, where the activities are reported. obtained with different doses of compound 10 administered intraperitoneally (IP) in comparison with a group of animals treated with physiological solution by the same route.

From the examination of table 6 it can be seen that compound 10 exerts a powerful anxiolytic effect.

In fact, it can be seen that at doses 0.1 and 1.0 mg / kg IP the compound increases by about 50-60% and significantly against the controls the% of entries in the open arms on the total number of entries.

Compound 10 also increases at all doses the% of the residence time in the open arms; such increase? significant for the doses 0.1 and 1 mg / kg IP compared to the group of control animals, treated with physiological solution alone.

Claims (11)

CLAIMS 1) Compounds that can be represented by the general formula (I) indicated below and in which: where Y? independently chosen between CH2 and 0 (oxygen) and n? 0 (zero) or 1; s? an integer between 1 and 4; m, r and t are integers chosen independently and included between 0 (zero) and 2; z? equal to 0 (zero) or 1; R4? independently selected from a simple or mono or disubstituted phenyl group with methyl or with chlorine, a 1 (or 2) -naphthyl group, a 2 (or 3 bindolyl) group, a 2 (or 3) -quinolinyl group; the configuration of the chiral center marked with * in the general formula (I) pu? be, independently, R (Rectus), or racemic (RS) while the configuration of the chiral center marked with ** in the general formula (I) can? be, independently, R (Rectus), racemic (RS) or S (Sinister) j preferably R1? a phenyl substituted with chlorq in positions 3 and 5, R3? the 8-azaspiro group [4.5] decan-8-yl, s? 2, R4? the l-naphthyl group and if z? 0 (zero) at the same time m and r are both 1 while t? 0 (zero) or 1, while if z? 1, m and t are both 0 (zero) and r? 2. 2) Compounds according to claim 1 of general formula (I) wherein Rj? the 3,5-dichlorophenyl group, R2? H or CH3, R3? the 8-azaspiro group [4.5] decan-8-yl, R4? the l-naphthyl group, s? Are 2 or 3, m, r, t independently 0 (zero) or 1, z? 0 (zero) and the stereochemistry of the chiral center marked with (*) in (I)? R (Rectus) or (RS). 3) Compounds according to claim 1 of general formula (I) wherein Rj? the 3,5-dichlorophenyl group, R2? H or CH3, R3? the 8-azaspiro group [4.5] decan-8-yl, R4? the I-naphthyl group, s? 2 or 3, m and t are 0 (zero), r? 2, z? 1 and the stereochemistry of the chiral center marked with (*) in (I)? R (Rectus) or (RS). 4) Pharmaceutical preparation comprising as active substance at least one of the compounds according to claim 1 or a pharmaceutically acceptable salt thereof. 5) Pharmaceutical preparation according to claim 4 for use in therapy for the purpose of its activity? antiulcer. 6) Pharmaceutical preparation according to claim 4 for use in the therapy of tumor affections caused by gastrin and other bioactive polypeptides related thereto. 7) Pharmaceutical preparation according to claim 4 for use in the therapy of affections of the gastrointestinal apparatus such as non-ulcerative dyspepsia and irritable colon. 8) Pharmaceutical preparation according to claim 4 for the treatment of pathological situations of the CNS linked to imbalances in the neuronal physiological levels of gastrin or other bioactive polypeptides related thereto, such as for example anxiety, panic attacks, psychosis, anorexia etc. or to other causes related to the mechanism of action of the compounds according to claim 1. 9) Pharmaceutical preparation according to claim 4 for use in the therapy and prevention of eye diseases such as myosis induced by surgical treatments of cataracts or chronic ocular inflammations or for the prevention of the pathology of other sense organs connected to the mechanism action of the compounds according to claim 1. 10. Pharmaceutical preparation according to claim 4 further comprising pharmaceutically acceptable inactive ingredients, selected from the group consisting of carriers, binders, disintegrating flavoring agents, preservatives, humectants, and mixtures thereof, or ingredients which facilitate transdermal absorption. 11) Process for the preparation of a derivative of general formula (I), in which R1, R2, R3 R4, s, m, z, r, t have the meaning reported in claim 1 and in which the substituents on the asterisk chiral center with (*) have the configuration R or (RS), and the substituents on the chiral center asterisk with (**) have the configuration R, (RS) or S, which consists of an amidation reaction of the basic derivatives of formula (II ) in which R *, m, r, t, z and (**) have the meaning previously indicated, in molar ratio from 1 to 3, in the presence or absence of a tertiary base, at a temperature between -15 ° C and 20? C, or with other equivalent conventional synthetic methods, using for example the monochlorides of the appropriate dicarboxylic acids with the same modalities. previously indicated for the corresponding anhydrides, in recovering the compounds (I) from the reaction mass as such or as pharmaceutically acceptable salts and purifying them with conventional methods.