ITMI941731A1 - NITRO-INHIBITORS WITH ANTI-INFLAMMATORY ACTIVITIES - Google Patents

Description

Description of the industrial invention

The present invention relates to new products having anti-inflammatory, analgesic and anti-thrombotic activity.

In particular it refers to cyclo-oxygenase (COX) inhibitor products.

It is known that the anti-inflammatory and anti-thrombotic efficacy of NSAIDs (Non steroid antiinflammatory drugs), also known as NSAIDs (non-steroidal anti-inflammatory drugs), but above all their tolerability seem markedly influenced by their cycle inhibitory activity. -oxygenase (COX) both in the inflammatory site and in healthy tissue. See for example FASEB Journal 1, 89, 1987, Bioch. Biophys. Acta 1083, 1, 1991. It is generally believed that the more potent a COX inhibitor a product is, the more effective it is.

The disadvantage of these products is that they are toxic. Furthermore, it is also known that the inhibitory properties of COX seem to depend on some factors linked to the chemical-physical and structural characteristics of the molecules themselves, such as for example the acidic function. See for example J. Pharmacol. Exp. Therap. 196, 226, 1976; Arch. Toxicol. 60, 261, 1987.

The known inhibitors of cyclo-oxygenases are in general acids attributable to general structures, such as:

carboxylic acids, both acetylated, such as for example aspirin and triflusal, and non-acetylated, such as for example salicylates, diflunisal, salsalate;

acetic acids, for example diclofenac, indomethacin, tolmetin, sulindac, etodolac, ketorolac;

propionic acids such as ibuprofen, naproxen, pirprofen, tiaprofenic acid, loxoprofen, indoprofen, oxaprozin, ketoprofen, fenoprofen, fenbufen, flurbiprofen, carprofen, suprofen,

enolic acids such as for example oxifenbutazone, phenylbutazone, piroxicam, sudoxicam, tenoxicam, isoxicam, meloxicam.

See US Patents 3,558,690; USP 3,755,427; USP 3,641,127; FR 2,112,111; USP 4,035,376; USP 3,997,669; USP 3,784,701; USP 3,896,145; USP 3,600,437; USP 3,843,681; USP 3,904,682; USP 3,228,831; USP 4,161,538; USP 4,233,299; USP 3,591,584; DE 2,537,070; USP 3,161,654; USP 4,061,779; USP 4,556,672; USP 4,089,969.

The disadvantage of these products is that they are very effective but have high toxicity.

The importance of the acidic function lies in the fact that the masking of this function in COX inhibitors causes the almost total loss of its properties to inhibit the formation of prostanoids. See Drugs 35, 504, Products are also known which have a high efficacy in inhibiting the cyclo-oxygenase and are endowed with low toxicity even though they do not contain the ad-dic function in their molecule.

These products are known as non-acidic terminated nitric esters. See for example the PCT patents WO 94/04484, which describes a particular group of compounds including the well-known commercial diclofenac product, PCT / EP 93/03193, which describes another specific group of compounds including the commercial products flurbiprofen and indoprofen.

However, these products have a drug-dynamic disadvantage. In fact, in the biochemical assay for evaluating the inhibitory activity of the cyclo-oxygenase, experiments conducted by the Applicant showed a great variability of responses, of the order of 10-40%.

This generally leads to erratic and unpredictable efficacy, making it difficult to establish a correct dosage.

In practice it is necessary to administer higher doses to contain the aforementioned variability. The downside lies in the risks of a higher incidence of side effects.

Another disadvantage is that these products show difficulties from the formulation point of view because the oral or parenteral preparations are prepared with greater difficulty than the conventional preparations based on acidic NSAIDs.

It is well known that the solubility of the molecule is among the most important properties that influence its pharmacokinetic and dynamic processes.

For example, to be administered parenterally, particularly intravenously, the drugs must be formulated in a soluble form.

Likewise, for the oral route, the solubilization process is decisive for the absorption and for the interaction with the effector.

In this regard, the choice of particular solvents and / or excipients, including surfactants, etc., is critical, also from a toxicological point of view. For example, the intravenous formulation must not cause haemolysis or incompatibility with blood components.

However, there is a lot of evidence indicating that non-polar surfactants and solvents can be irritating. See e.g., J. Pharm. Science 72, 1014, 1983.

Experiments conducted by the Applicant in which Tween 800.1% and dimethyl sulfoxide 1% was used to suspend nitroxybutyl flurbiprofen showed that this solvent was irritating towards the gastric mucosa.

Unexpectedly, however, it was found that with a NO-flurbiprofen derivative as described below, and falling within the scope of the present invention, the quantities of Tween 80 and dimethylsulfoxide necessary for the suspension were lower, such as not to cause irritating effects, even with the same results in terms of solubilization.

It has been unexpectedly and surprisingly found after numerous researches that it is possible to prepare anti-inflammatory products, as described below, endowed with high inhibitory activity of the cyclo-oxygenase combined with low toxicity and with good responses in pharmacokinetic terms, and having a variability of the very limited response, with average coefficients of variation of about half compared to known products from the pharmacodynamic point of view, and easier to formulate for oral or parenteral preparations.

This is totally surprising and unexpected as the factors influencing the anti-inflammatory and anti-thrombotic efficacy of NSAIDs depend on various parameters for which it is not possible to predict a priori the pharmacokinetics, for example the fraction of absorbed product, the activity pharmacodynamics, toxicity and inhibitory properties of COX and, above all, no hypothesis can be made to predict or limit the variability of response.

The object of the present invention are compounds, or their compositions, having a general formula:

A - Χλ - NO2

or their salts, for use as medicaments, in particular as anti-inflammatory and antithrombotic drugs, in which:

in which:

Rx is the group 0C0R3; where R3 is methyl, ethyl or linear or branched C3-C5 alkyl, or the residue of a single ring heterocycle having 5 or 6 atoms which can be aromatic, partially or totally hydrogenated, containing one or more hetero-atoms independently selected from 0, N and S;

R2 is hydrogen, hydroxy, halogen, alkyl from 1 to 4 C atoms, linear or branched when possible, alkoxy from 1 to 4 C atoms, linear or branched when possible; a perfluoroalkyl of 1 to 4 C atoms, linear or branched when possible, for example trifluoromethyl, nitro, amino, mono- or di- (Cl-4) alkylamino;

Rx and R2 taken together are the dioxymethylene group, with the condition that when X = NH, then X3 is ethylene and R2 = H; Rx cannot be OCOR-, in position 2 when R3 is methyl, - ni being an integer 0 or 1;

preferably in la) X is equal to 0 or NH, Rx is acetoxy, preferably in position 3 or 4, X1 is ethylene, R2 is hydrogen or halogen, more preferred are the following compounds A Xx N02: 3-acetoxy-N- {2 -nitroxyethyl) -benzamide, 4-acetoxy-N- (2-nitroxyethyl) -benzamide, 3-acetoxy-N- (5-nitroxypentyl) -benzamide 2-acetoxy-N- (5-nitroxypentyl) -benzamide, N-2 - {nitroxyethyl) -2-propionoxy-benzamide, 2-acetoxy-2 -nithroxyethylbenzoate, 2-acetoxy-N- {cis-2-nitroxycyclohexyl) -benzamide, 2-acetoxy-4-chloro-N- (2-nitroxyethyl) -benzamide, N- (2-nitroxyethyl) -2 - ((4-thiazolyldinyl) carbonyloxy) -benzamide hydrochloride, 2-nicotinoyloxy-N- (2-nitroxyethyl) -benzamide, 2-acetoxy-5-nitroxypentylbenzoate,

preferably in Ib) R3 = CH3, ni = 0;

X is equal to 0, X1 is ethylene, in this case Ib) is the residue of acetylsalicylsalicylic acid,

group II) where t = 1

in which :

where the meanings are as follows:

in the compounds of formula (IV), Ketoprofen residue:

<R> im is H, SRIII3 where RIII3 contains 1 to 4 C atoms, linear or branched when possible,

<R> II2 is H, hydroxy;

preferred are the compounds in which RIlt1 and RIII2 are H, R3a is H, and R2e is methyl, X = 0;

in the compounds of formula (XXI), carprofen residue:

Rxxio is H, alkyl from 1 to 6 linear or branched C atoms when possible, C1-C8 alkoxycarbonyl bonded to a C1C6 alkyl, C1-C6 carboxyalkyl, C1C6 alkanoyl, optionally substituted with halogens, benzyl or halobenzyl, benzoyl or halobenzoyl;

Rxxi is H, halogen, hydroxy, CN, C1-C6 alkyl optionally containing OH groups, C1C6 alkoxy, acetyl, benzyloxy, S <R> XXII2 wherein Rxxi is C.-Cg; perfluoroalkyl from 1 to 3 C atoms, carboxyalkyl C-1-C6 optionally containing OH, N02, amino, sulfamoyl, di-alkyl sulfamoyl groups with 1 to 6 C atoms alkyl, or difluoroalkylsulfonyl with 1 alkyl 3 C atoms;

<R> xxyl is halogen, CN, C3-C6 alkyl containing one or more OH groups, C1C6 alkoxy, acetyl, acetamido, benzyloxy, SRIII3 as defined above, 1 to 3 C perfluoroalkyl, hydroxy, 1 to 6 C carboxyalkyl, N02, amino, mono or di-alkyl-amino from 1-6 C; sulfamoyl, di-alkyl sulfamoyl from 1 to 6 C, or di-fluoroalkylsulfamoyl as defined above; or together with Rxx it is an alkylene dioxy from 1 to 6 C, - preferred are the compounds in which Rxxi is H, the connecting bridge is in position 2, Rxxi is H, Rxx is chlorine and is in para position with respect to nitrogen ;

R3B is H, R <2> * is methyl and X is 0;

in the compounds of formula (XXXV) residue of the tiaprofene-CO acid:

Ar is phenyl, hydroxyphenyl optionally mono or polyisubstituted with 1-6 C halogen, alkanoyl and alkoxy, 1-6 C trialalkyl, preferably 1 to 3 C, cyclo-pentyl o-hexyl o-heptyl, heteroaryl, preferably thienyl, furyl optionally containing OH, pyridyl, the preferred compounds of (XXXV) are those in which Ar is phenyl, R3e is H, R <2a> is methyl and X is 0;

in the compounds of formula (II), residue of suprofen,

of which the preferred one has been indicated, its equivalents can also be used as described and obtained in USP 4,035,376 incorporated herein by reference,

in the compounds of formula (VI),

of which the preferred indoprofen have been indicated when R2a and CH- ,;

and indobufen when R2a is equal to H and R3a = C2H5;

its equivalents can also be used as described and obtained in accordance with USP 3,997,669 incorporated herein by reference,

in the compounds of formula (VIII),

the preferred etodolac of which has been shown, its equivalents can also be used as described and obtained according to USP 3,843,681 incorporated herein in full by reference,

in the compounds of formula (VII),

of which the preferred fenoprofen has been shown, its equivalents as described and obtained according to USP 3,600,437 incorporated herein integrally by reference can also be used,

in the compounds of formula (III),

of which the preferred fenbufen has been shown, its equivalents can also be used as described and obtained according to USP 3,784,701 incorporated herein in full by reference,

in the compounds of formula (X) residue of tolmetine,

its equivalents can also be used as described and obtained according to the patent FR 1,574,570 incorporated herein by reference;

in group IV) where t = 1

in which the compounds of group IV) have the following meanings: in the compounds of formula (II):

in group V), the compounds have the following meanings: in the compounds of formula (II)

in the compounds of formula (V),

of which the residue of the known tenidap has been indicated, its equivalents can also be used as described and obtained in USP 4,556,672 incorporated herein by reference;

in these compounds of formula (V) A = R and t - 0,

in the compounds of formula (VII)

of which the residue of the known tenoxicam has been indicated, A is RCO and t = 1 and X is absent or A is R and t = 0;

its equivalents can also be used as described and obtained in DE 2,537,070, incorporated herein by reference;

in the compounds of formula (IX)

where A = R and t = 0, or A = RC0 with t = 1 and X is absent of which the residue of the known piroxicam has been indicated,

its equivalents can also be used as described and obtained in USP 3,591,584 incorporated herein by reference;

in the compounds of formula (III)

where A = RCOO,

of which the residue of the known nabumetone has been indicated, its equivalents can also be used as described and obtained in USP 4,061,779 incorporated herein by reference;

in the compounds of formula (IV)

where A = RCOO,

of which the residue of the known indomethacin has been indicated, its equivalents can also be used as described and obtained in USP 3,161,654 incorporated herein by reference;

The processes for obtaining the compounds containing R of the group I of the type la) are described in the patent W092 / 01668 where the preparation methods are also indicated. This patent is incorporated herein in its entirety by reference. Compounds of type Ib) are for example prepared using the method indicated in The Merck Index, XI ed., 1989, p. 16, n. 95 for the residue of acetylsalicylsalicylic acid. The modifications of the compounds of formula Ib) can be obtained by applying the processes indicated in the patent WO 92/01668.

The products of the present invention of general formula

with connecting bridges X! as defined above, for the compounds of group I), they can be obtained using the methods of the prior art reported above or by modifying the known methods for introducing Xt bridges when these are different from the connecting bridges indicated in the cited patents.

The compounds in which R is of group II) are described in the patents WO94 / 04484 and USP 3,558,690 where the preparation methods are also indicated. These patents are incorporated herein in full by reference.

The connection bridges Xi as defined above, for the compounds of group II), can be obtained using the methods of the prior art reported above or by modifying the known methods for the introduction of the XL bridges when these are different from the connection bridges indicated in the patents cited.

The compounds in which R is of group III) are described and obtained with the processes indicated in the following patents:

patent application PCT / EP / 9303193; for the compounds of formula (IV) see also USP 3,641,127; for the compounds of formula (XXI) see also USP 3,896,145; for the compounds of formula (IX) residue of flurbiprofen see also USP 3,755,427, · for the compounds of formula (II) see also USP 4,035,376; for the compounds of formula (VI) see also USP 3,997,669; for the compounds of formula (VIII) see also USP 3,843,681; for the compounds of formula (VII) see also USP 3,600,437; for the compounds of formula (III) see also USP 3,784,701. All of these cited patents are incorporated herein in full by reference.

The connection bridges X1 as defined above, for the compounds of group III), can be obtained using the methods of the prior art reported above or by modifying the known methods for the introduction of the X ^ bridges when these are different from the connection bridges indicated in the cited patents.

The compounds in which R is of group IV) are described in the English patent application 9320599.5 where the preparation methods are also indicated, this patent is incorporated herein in full by reference.

In group IV) the compounds can also be obtained: for the compounds of formula (II) using USP 3,904,682; the compounds of formula (X) according to USP 4,161,538, the compounds of formula (III) according to USP 3,228,831. These patents indicated herein are reported in full in the present application by reference.

The connection bridges X1 as defined above, for the compounds of group IV), can be obtained using the methods of the prior art reported above or by modifying the known methods for the introduction of the X1 bridges when these are different from the connection bridges indicated in the patents cited.

The compounds in which R is of group V) are described in the Italian patent MI94A 000916 where the preparation methods are also indicated, this patent is incorporated herein by reference.

In group V) the compounds can also be obtained: for compounds of formula (II) using USP 4,089,969 incorporated herein by reference, compounds of formula (V) can be obtained according to USP 4,556,672 incorporated herein in full for reference.

The connection bridges Xx as defined above, for the compounds of group V), can be obtained using the methods of the prior art reported above or by modifying the known methods for the introduction of the bridges Xx when these are different from the connection bridges indicated in the patents cited.

In general, the connection between A and Xt is, as seen, in general, of the ester or amide type (NH or NR1C, as defined in X) when R is of groups I, II, III, IV. For the formation of this connection, all the well known synthesis ways for the formation of such bonds can be used.

In the case of the esters of groups I, III and IV, the most direct synthesis route foresees:

the reaction of the acyl chlorides R-CO-C1 in halogen alcohols of the type HO-Y-Cl, HO-Y-Br, HO-Y-I, under experimental conditions which are part of the known art.

The reaction products of formula R-CO-O-Y-Cl (Br, I) can also be obtained for class II, by reaction of the sodium or potassium salts of said R-CO-OH acids with dihalogen derivatives of general formula YC12, YBr2 or YI2.

The reaction products are transformed into the final products by reaction with AgN03 in acetonitrile, according to what is known in the literature.

In the case of amides, the synthetic sequence foresees the reaction of the same RC0C1 acyl chlorides with amino alcohols of the general formula NH2-Y-OH, NHR1C-Y-0H to give the amides of the general formula:

according to known methods.

The reaction of said amides with halogenating agents such as for example PC15, PBr3, S0C12 etc. leads to the halogen derivatives of the general formula:

The latter by reaction with AgN03 in acetonitrile, according to methods known in literature, lead to the final products A X2 N02.

An alternative way to the formation of the esters is the reaction of the sodium or potassium salts of the acids with the nitric esters of halogen alcohols of general formula:

Synthesis routes analogous to those described above can be used for the products Va and Vb of group V where the dihalogen derivative Br2Y is reacted with the enolates, for example of tenoxicam and piroxicam. The reaction products are then transformed by reaction with AgNO3 into acetonitrile according to the above reaction.

The general scheme is reported here for piroxicam of formula IX of group V.

A further object of the invention is that it has surprisingly been found that the products of the invention containing ON02 groups are also capable of exerting an inhibition effect of the inflammation induced by liposaccharide (LPS) and therefore usable in skeptical shock.

This is surprising given that it is well known that anti-inflammatories in general do not significantly modify the activity of lipopolysaccharide-induced nitrosynthesis in the rat and therefore are not usable in skeptical shock.

The products that can be used for this pharmaceutical use are the products of general formula

A - X1 - N02

specified above, in which the bivalent connection bridge X1 in this case has no limitation, ie known connection bridges are not excluded since for this use nothing was reported in the previous patents.

The following examples are given for illustrative but not limitative purposes of the present invention.

EXAMPLES

To a solution of:

acetylsalicylic acid 5.6 g e

dimethylphormamide 2 0 ml

kept at 0 ° C and under nitrogen flow, it is added in portions:

sodium iodide (80% suspension in vaseline oil) 1.0 g.

The mixture is left under stirring for one hour and is then dropped in 5 hours, in a stirred solution, at 25 ° C of 10.0 g 2,2'-dibromo-diethyl ether and

dimethylformamide 15 ml.

The mixture is left under stirring for 3 days, then it is brought to dryness under reduced pressure. The residue is taken up with:

water 50 ml and

dichloromethane 50 ml.

The phases are separated and the aqueous one is further extracted in dichloromethane 10 ml.

The combined organic phases are washed with water (3 x 25 ml), dried (MgSO4), decoloured with charcoal (1 g), and dried under vacuum.

The residue (11.2 g) is used crude for the next reaction.

Preparation of ASA-NO-DEG:

To a solution of

ASA- (CH2) 2 O (CH2) 2 CI 11.2 g e

acetonitrile 25 ml

kept at room temperature and away from light, it is added:

silver nitrate 8.6 g.

After two days under stirring, 2.2 g of silver nitrate is added.

After another two days under the same conditions, the insoluble salts are filtered, and the filtrate is deprived of the solvent at reduced pressure.

A 7.0 g residue is obtained, which is chromatographed on a silica gel column (500 g of silica) by eluting with a 95/5 v / v toluene / ethyl acetate mixture.

The fractions that are unitary for TLC analysis (Thin Layer Chromatography) are combined and, brought to dryness, provide

Mass spectrometry gave the value of 313 as the molecular weight.

To a solution of

2,2'-dibromo-diethyl ether 12.3 g e

dimethylformamide 15 ml

maintained at room temperature and under nitrogen flow, a solution of

DICLOFENAC sodium salt 13.3 g e

dimethylformamide 25 ml.

The mixture is left to react for two days, then the solvent is eliminated under reduced pressure. The residue is taken up with ethyl acetate (50 ml), washed with a 5% potassium carbonate solution (2 x 10 ml), then with water (20 ml), anhydrified over anhydrous sodium sulphate. The solvent is removed under reduced pressure.

The residue weighs 16 g and is used for the next reaction without any purification.

The mixture is stirred for another two days, then the insoluble salts are filtered, and the filtrate is deprived of the solvent under reduced pressure. The residue is taken up with ethyl acetate (50 ml), then the insoluble salts are filtered and discarded. The filtrate is deprived of the solvent at reduced pressure. A residue of 16.2 g is obtained. Chromatography on a silica gel column (700 g of silica) is carried out on the residue by eluting first with toluene, then with a mixture of toluene / ethyl acetate 99/1 v / v, finally with a mixture of toluene / ethyl acetate 98/2 v / v.

The fractions that are unitary for TLC analysis (thin layer chromatography) are combined and, brought to dryness, provide

Preparation of the 3-nitroxymethyl-phenol intermediate having the formula:

The following reagents are used in the quantities indicated and reacted as indicated below:

The 3-hydroxy-benzyl alcohol in CH2C12 is reacted with the HBr at room temperature for 4 hours.

Then the CH2C12 is evaporated under reduced pressure, at 30 ° C, after being washed with a 5% aqueous solution of NaHC03 and dried on anhydrous Na2SO4.

The oily residue is dissolved in CH3CN (50 ml) then a solution of AgN03 in the remaining quantity of CH3CN is dropped into it. The balloon is kept away from light.

After 8 hours the AgBr precipitate is filtered and the organic phase is evaporated under reduced pressure.

The oily residue obtained is dissolved in toluene (45 ml) and the solution filtered on a silica gel column (400 g). The eluate is brought to dryness under reduced pressure, at 30 ° C to give 20 g of 3-nitroxymethylphenol.

Preparation of the intermediate KETOPROFEN -COC1: 2- (3-benzene) phenyl propionic acid chloride.

They make themselves react:

KETOPROFEN 20 g

Thionyl chloride 50 ml

and the solution is allowed to react under reflux for 45 minutes. The thionyl chloride is evaporated under reduced pressure. A yellow oily residue weighing 21 g is obtained which is used without further purification.

It is left to react for 5 hours at room temperature, then it is diluted with Ha0 (50 ml). The organic phase is washed with 5% by weight NaOH (2 x 10 ml) and evaporated under reduced pressure. The oily residue obtained is chromatographed on silica using a toluene / EtOAc 9.5 / 0.5 v / v mixture as eluent. Evaporation of the eluate gives KETOPROFEN-Ar-NOz with a yield of 85%.

The same procedure was followed as in example la in which R indicated above, residue of IBUPROFEN, was used instead of the residue R of group I indicated in example la.

The same procedure was followed as in example la in which R indicated above, residue of FLURBIPROFEN, was used instead of the residue R of group I indicated in example la.

The same procedure was followed as in example la in which R indicated above, residue of KETOROLAC, was used instead of the residue R of group I indicated in example la.

The same procedure was followed as in example la in which R indicated above was used, residue of TIAPROFENIC ACID instead of residue R of group I indicated in example la.

The same procedure was followed as in example la in which R indicated above was used, residue of NAPROXEN instead of residue R of group I indicated in example la.

EXAMPLE 2: pharmacological examples

The products used above have been characterized from a pharmacological point of view.

Example 2a: ASA-NO-DEG as prepared in example la; Example 2b: DICLOFENAC-NO-DEG as prepared in Example 1b,

Example 2c: KETOPROFEN-NO-DEG as prepared in example le.

Example 2d: IBUPROFEN NO-DEG as prepared in example 1d.

Example 2e: FLURBIPROFEN NO-DEG as prepared in example le.

Example 2f: KETOROLAC NO-DEG as prepared in Example 1f.

Example 2g: TIAPROFENIC ACID NO-DEG as prepared in example ìg.

Example 2h: NAPROXEN NO-DEG as prepared in example ih.

Toxicity

Acute toxicity was evaluated by oral administration of a single dose of 1, 3, 10, 30, 100 mg / kg of product to groups of 10 mice.

The incidence of lethality and the occurrence of toxic symptoms were reported within an observation period of 14 days. Even after administration of a dose of 100 mg / kg the animals showed no signs of apparent toxicity.

Anti-inflammatory activity

The anti-inflammatory activity was determined according to the carrageenan edema method, as described by winter et al. (Proc. Soc. Exp. Biol. Med. III, 544, 1962), in the rat.

Analgesic activity

Analgesic activity was determined in Swiss mice, as reported by Hendershot et al. (J. Pharmacol. Exp. Therap. 125, 237, 1959).

Tolerability

Gastric tolerability was evaluated by oral administration in rats by measuring the severity of induced gastropathy according to the criterion indicated by Wallace et al. {Am. J. Physiol. 259, G642, 1990).

Platelet antiplatelet activity

The anti-aggregating activity of platelets was evaluated in vitro on human platelets stimulated by thrombin according to the method described by Bertele et al. (Science 220, 517, 1983).

Vasodilator activity

Vasodilatory activity was determined in isolated rat aorta by measuring the inhibition of contraction caused by epinephrine with tissue prepared according to the method described by Reynolds et al. (J. Pharmacol. Exp. Therap.

252, 915, 1990).

COX inhibition

The inhibitory activity of cyclo-oxygenases was determined in isolated cells. Bovine aortic endothelial cells were used as a source of COX-1 and macrophage line J774.2 as a source of COX-2. The same conditions were followed for growth and for the verification of cell viability as described by Mitchell et al. (Proc. Nat. Acad. Sci. 90, 11693, 1993).

In summary, the cells were incubated for 30 min with scalar concentrations of test product and the substrate (arachidonic acid) was then added and incubated for another 15 min. The enzymatic activity was determined by measuring the formation of 6-keto-PGF 1 alpha by radioimmunoassay. In the case of the J774.2 lines, the cells were incubated for 12 hours with endotoxin to induce the formation of COX-2.

Inhibition of nitrosynthetase by LPS

The inhibition activity of nitrosynthetase induced by lipopolysaccharide (LPS) was determined in neutrophils and the rat stomach after administration of one of the compounds under examination and compared with that obtained after treatment of the suspending vehicle alone.

In summary, the test product (10 mg / kg) and intravenously (tail vein) LPS (5 mg / kg) are administered to Wistar rats fasted for 24 hours before treatment.

Four hours later, the animals are sacrificed and the blood taken for neutrophil isolation and the stomach.

The enzymatic activity was determined according to the method described by Assreuy et al. (Br. J. Pharmacol. 108, 833, 1993).

Results :

The results obtained are shown below.

As can be seen from the data reported in Tables 1-4, the pharmacodynamic activities (I and II of Tab. 1, - Tab.

2), and the tolerability (Tab. 1 col III) of nitro-derivatives show a better balance than those of native products.

From table 4 it also results that, like diclofenac nitroxybutylester, the nitro-derivative of diclofenac object of the present patent is capable of directly inhibiting the cyclo-oxygenases COX-1 and COX-2, but with a significantly lower variability.

TABLE 1 (Pharmacology col. I and 11; Toxicology col. III) Study of the anti-inflammatory (I), analgesic (II), (Pharmacodynamics) properties; and gastrointestinal (III) tolerability (toxicity) of the compounds under examination after oral administration of doses in the range of 3-30 mg / kg in suspensions based on carboxymethylcellulose and constructing the dose-response curves. The reported results represent the power ratio with respect to the standard-comparison.

The activities were expressed as a power ratio relative to the native product taken as a unit standard. The nitro-derivative is that of the examples indicated, the native reference compound is the one reported as a comparison.

TABLE 2 (Pharmacodynamic activity)

Example of the anti-cyclooxygenase (I), antiplatelet (II) and vasoladalatory (III) properties of the test compounds tested in vitro at concentrations in the range IO <'5> molar -IO <"7> molar of the product in water / alcohol with the addition of small quantities of DMSO (dimethyl sulfoxide) The activities were expressed as a power ratio relative to the native product taken as a unit standard, as stated in Table 1.

TABLE 3 (Biochemistry: action on NOS for the skeptical shock)

Study of the inhibitory properties of nitrosynthetase (NOS) activity induced by lipopolysaccharide (LPS) in rats, using doses ranging from 5-20 mg / kg orally, in suspension based on carboxymethylcellulose.

TABLE 4 (COX inhibitory activity)

Study of anti-cyclooxygenase (COX-1 / COX-2) properties in isolated cells. Response expressed as% of controls with relative variability of the response.

Claims (4)

CLAIMS 1. Compounds, or their compositions, of general formula: in which: R1 is the OCOR3 group; where R3 is methyl, ethyl or linear or branched C3-C5 alkyl, or the residue of a single ring heterocycle having 5 or 6 atoms which can be aromatic, partially or totally hydrogenated, containing one or more hetero-atoms independently selected from 0, N and S; R2 is hydrogen, hydroxy, halogen, alkyl from 1 to 4 C atoms, linear or branched when possible, alkoxy from 1 to 4 C atoms, linear or branched when possible; a peryluoroalkyl from 1 to 4 C atoms, linear or branched when possible, for example trifluoromethyl, nitro, amino, mono- or di-alkylamino, -R3 and R2 taken together are the dioxymethylene group, with the condition that when X = NH , then X3 is ethylene and R, = H; R1 cannot be OCOR3 in position 2 when R3 is methyl; ni being an integer 0 or 1; group II) where t = 1 where is it: in which: R2a and R3a are H, linear or branched alkyl when possible, substituted or not, allyl, with the proviso that if one of the two is allyl the other is H; preferably R2B is H, alkyl from 1 to 4 C, R3a is H; Rla is chosen from where the meanings are as follows: in the compounds of formula (IV), Ketoprofen residue: 1 to 3 C kyl, hydroxy, 1 to 6 C carboxyalkyl, N02, amino, 1-6 C mono- or di-alkyl-amino; sulfamoyl, dialkyl sulfamoyl from 1 to 6 C, or di-fluoroalkylsulfamoyl as defined above; or together with R? cii is an alkylene dioxy from 1 to 6 C; preferred are the compounds in which R * ^ is H, the connecting bridge is in position 2, R ^ is H, Rxiri. it is chlorine and is in a para position with respect to nitrogen; R3a is H, R <2a> is methyl and X is 0; in the compounds of formula (XXXV) residue of tiaprofenic acid: Ar is phenyl, hydroxyphenyl optionally mono or polysubstituted with halogen, alkanoyl and alkoxy of 1-6 C, trialalkyl of 1-6 C, preferably 1 to 3 C, cyclo-pentyl o-hexyl o-heptyl, heteroaryl, preferably thienyl, furyl optionally containing OH, pyridyl; the preferred compounds of (XXXV) are those in which Ar is phenyl, R3a is H, R <2> * is methyl and X is O; in the compounds of formula (II) residue of suprofen, or its equivalents in the compounds of formula (VI) residue of indoprofen, or its equivalents, when R2a is CH3, and residue of indobufen, or its equivalents, when R2a is equal to H and R3a = in the compounds of formula (VIII) residue of etodolac, or its equivalent, in the compounds of formula (VII) residue of fenoprofen, or its equivalents, in the compounds of formula (III) residue of fenbufen, or its equivalents in the compounds of formula (X) residue of tolmetine, or its equivalents, in group IV) where t = 1 ivdi in which : Rrvd and RIVd1 are at least one H and, the other a linear or branched Cx to C6 alkyl whenever possible, preferably C1 and C2, or difluoroalkyl with 1 to 6 C alkyl, preferred C1, or RIvd and RIvd1 together form a methylene group; RIV has the following meaning: in which the compounds of group IV) have the following meanings: in the compounds of formula (II): RIV-II is 1-6 C alkyl, 3 to 7 C cycloalkyl, 1-7 C alkoxymethyl, 1 to 3 C trifluoroalkyl, vinyl, ethynyl, halogen, 1 to 6 C alkoxy, difluoroalkoxy, with 1 to 7 C alkyl, 1 to 7 C alkoxymethyloxy, 1 to 7 C alkylthyomethyloxy with 1 to 7 C alkyl, alkyl methylthio with 1 to 7 C alkyl, cyano, difluoromethylthio, phenyl- or phenylalkyl substituted with alkyl from 1 to 8 C; preferably RIV-I is CH30, Rivd is H and RIvd1 is CH3, and is known as the naproxen residue, X = NH and X1 is equal to (CH2) 4; preferred is also the same compound in which X is equal to 0; in the compounds of formula (X) residue of loxoprofen, or its equivalents in the compounds of formula (III): Rrv-III and a C2-C5 alkyl also branched when possible, C2 and C3 alkyloxy, allyloxy, phenoxy, phenylthio, cycloalkyl from 5 to 7 C atoms, optionally substituted in position 1 by an alkyl in group V), the compounds have the following meanings: in the compounds of formula (II) is i i wherein Y is a linear or branched C1-C20 alkylene when possible, preferably from 2 to 5 carbon atoms excluding this connecting bridge when R is a radical of group I), II), III), IV); or an optionally substituted cycloalkylene of 5 to 7 carbon atoms; wherein R1f = H, CH3 and nf is an integer from 1 to 6; preferably 2-4; excluding this connection bridge when R is a group V). 2. Use of the compounds, or their compositions, according to Claim 1, of general formula: in which: R3 is the group 0C0R3; where R3 is methyl, ethyl or linear or branched C3-C5 alkyl, or the residue of a single ring heterocycle having 5 or 6 atoms which can be aromatic, partially or totally hydrogenated, containing one or more hetero-atoms independently selected from 0, N and S; R2 is hydrogen, hydroxy, halogen, alkyl from 1 to 4 C atoms, linear or branched when possible, alkoxy from 1 to 4 C atoms, linear or branched when possible; a perfluoroalkyl of 1 to 4 C atoms, linear or branched when possible, for example trifluoromethyl, nitro, amino, mono- or di- (C1-4) alkylamino; R3 and R2 taken together are the dioxymethylene group, with the condition that when X = NH, then X3 is ethylene and R2 = H; Ri cannot be 0C0R3 in position 2 when R3 is methyl; ni being an integer 0 or l; group II) where t = 1 RIi6 has the same meaning as RIIS, or when RII5 is H it can be benzyl, RiI is equal to hydrogen, C1-C6 alkyl, or C1 -C6 alkoxy, linear or branched when possible, or Cl, F, Br; RII4 is RIII or bromine; group III) where t = 1 in which: where the meanings are as follows: in the compounds of formula (XXXV) residue of tiaprofenic acid: Ar is phenyl, hydroxyphenyl optionally mono or polysubstituted with halogen, alkanoyl and alkoxy from 1-6 C, triayalkyl from 1-6 C, preferably from 1 to 3 C, cyclo-pentyl o-hexyl o-heptyl, heteroaryl, preferably thienyl, furyl optionally containing OH, pyridyl, the preferred compounds of (XXXV) are those in which in the compounds of formula (II) residue of suprofen, or its equivalents in the compounds of formula (VI) residue of indoprofen, or its equivalents, when R2a is CH3, and residue of indobufen, or its equivalents, when R2a is equal to H and R3e = C2HS; in the compounds of formula (VIII) residue of etodolac, or its equivalent, in the compounds of formula (VII) residue of fenoprofen, or its equivalents, in the compounds of formula (III) residue of fenbufen, or its equivalents in the compounds of formula (X) residue of tolmetine, or its equivalents, in which the compounds of group IV) have the following meanings: in the compounds of formula (II): Riv-I is 1-6 C alkyl, 3 to 7 C cycloalkyl, 1-7 C alkoxymethyl, 1 to 3 C trifluoroalkyl, vinyl, ethynyl, halogen, 1 to 6 C alkoxy, difluoroalkoxy, with 1 to 7 c alkyl, 1 to 7 c alkoxymethyloxy, 1 to 7 c alkylthyomethyloxy with 1 to 7 c alkyl, 1 to 7 c alkyl methylthio, cyano, difluoromethylthio, phenyl- or phenylalkyl substituted with 1 <; > 1 to 8 C alkyl, - preferably RIV-I is CH30, Rivd is H and RIvd1 is CH3, and is known as the naproxen residue; X = NH and X3 is equal to (CH2) 4; preferred is also the same compound in which X is equal to 0; in the compounds of formula (X) residue of loxoprofen, or its equivalents in the compounds of formula (III): Riv-III is a C2-C5 alkyl also branched when possible, C2 and C3 alkyloxy, allyloxy, phenoxy, phenylthio, cycloalkyl from 5 to 7 C atoms, possibly substitutes in group V), the compounds have the following meanings: in the compounds of formula (II) RVII is H or a linear or branched 1-4 C alkyl whenever possible, -RVII! is RVII or alkoxy 1 to 4 C, linear or branched when possible; Cl, F, Br; the position of being o-m-p; preferred is the residue of the known Ketorolac, wherein R ^ and RVII j. are H, and A = R and t = 0, Xt is = (CH2) 4; in the compounds of formula (V), tenidap residue, or its equivalents in these compounds of formula (V) A = R and t = 0, Xi is = (CH2) 4; in the compounds of formula (VII) residue of tenoxicam, or its equivalents, A is RCO and t = 1 and X is absent or A is R and t = 0; in the compounds of formula (IX) where A = R and t = 0, or A = RC0 with t = 1 and X is absent, residue of piroxicam, or its equivalent in the compounds of formula (III) wherein A = RCOO, residue of the known nabumetone, or its equivalent; in the compounds of formula (IV) wherein A = RCOO, residue of indomethacin, or its equivalent; X1 of the formula A-x1-N02, is a bivalent connection bridge chosen from the following: wherein Y is a linear or branched C1-C20 alkylene when possible, preferably from 2 to 5 carbon atoms; or an optionally substituted cycloalkylene of 5 to 7 carbon atoms, rebelliously 2-4; excluding this connection bridge when R is a group V). 3. Compounds according to claims 1 and 2, wherein: in group I, in the compounds of formula la) X is equal to O or NH, R1 is acetoxy, preferably in position 3 or 4, X2 is ethylene, R2 is hydrogen or halogen; in Ib) R3 = CH3, ni = 0; X is equal to 0, X3 is ethylene; in this case Ib) is the residue of acetylsalicylsalicylic acid; in group II where RII3 is H, RII2 and RII3 are chlorine and RII3 is in ortho position with respect to NH; RII5 and RIIS are H, X is equal to O, and X3 is (CH2-CH2-0) 2. 4. Use of the compounds according to claim 1, or their compositions, of general formula: A - X1 - N02 or their salts, as anti-inflammatory and anti-thrombitis, X2 when it is Y0, Y can also be a C1-C20 alkylene, linear or branched when possible, and R can also be a radical of group I.