LU83307A1 - ADENOSINE DERIVATIVES WITH ANTI-INFLAMMATORY AND ANALGESIC ACTIVITY, AS WELL AS THERAPEUTIC COMPOSITIONS CONTAINING THESE DERIVATIVES AS ACTIVE INGREDIENT - Google Patents

Description

"I" 1 'The present invention relates to ade- \ nosine derivatives with anti-inflammatory, analgesic and: {antipyretic) activity, as well as therapeutic compositions !! ques containing these derivatives as active ingredient.

! i jî S The compounds with therapeutic activity according to jj the present invention correspond to the general formula: j NH-R, i '' '10 L 1 / (°> »; CH0-SR (I) i. d', 15 0R2 or2 4 i in which | R represents a linear or branched chain alkyl group containing 1 to 18 carbon atoms, or a phenylalkylene group in which the alkylene chain contains f 20 1 to 6 carbon atoms, | R - represents a hydrogen atom, an aliphatic acyl group containing 1 to 6 carbon atoms or an "aromatic acyl group, | R2 represents a hydrogen atom, an aliphatic acyl group containing 1 to 6 carbon atoms or a aromatic acyl group, or alternatively, the radicals R0 together form an isopropylidene chain, n is equal to 0 or 1.

In addition, when R ^ represents a hydrogen atom, the invention also relates to the acid addition salts of the compounds of formula (I).

? Preferably, R represents a methyl group, an ethyl group, a propyl group, an isopropyl group. Pyle, butyl group, isobutyl group, sec-butyl group, pentyl group, hexyl group, heptyl group, octyl group, decyl group, dodecyl group, hexadecyl group, octa group - "2: î decyl or a benzyl group.

Preferably, R ^ represents a hydrogen atom, an acetyl group, a propionyl group, a butyryl group, a benzoyl group or a tosyl group, preferably Ro represents a hydrogen gene, an acetyl group, a propionyl group, a butyryl group, a benzoyl group, or a tosyle group.

Preferably, the acid addition salts of the (compounds of formula (i) are chlorides, sulfa-10 tes, phosphates, formates, acetates, citrates, tartrates, methane sulfonates or the | p-toluene-sulfonates.

To | 'The compounds of formula (I) are, in part, ί new.

The compounds of formula (I) are prepared by different methods, depending on the meaning of the different radicals.

j To prepare the group of compounds corresponding to the formula: 20 NH2 i f yf ^

; kX / CH * -E-R

i 0 25 f X (la)

Vr

OH OH

i in which R has the meanings indicated above, the method of Legraverand is followed (Legraverand M. Ibanez S., et al. (1977) "Eut. J. Med. Chem.” 12, 105-108) in which is converted from adenosine to 5'-chloro ~ 5'-deoxy- I adenosine by reaction with thionyl chloride i in hexamethylphosphoramide.

I · j 35 Then, we transform 51-chloro-51-deoxy-! i adenosine to thioether required by reaction with the corresponding mer-captan in a solution of 3N 2N sodium hydroxide at 80 ° C.

The thioethers obtained are purified by recrystallization from water or from lower aliphatic alcohols.

Then, the compounds of formula (la) can be subjected to salification with the stoichiometric amount of the acid required.

The compounds of general formula: NH-R were prepared.

1 ° 1 L X (Ib) ch2-s-r

P

0R20Ro in which R, R ^ and R ,, have the meanings defined above, provided that R. and R ,, are different from a hydrogen atom or an isopropyl chain lidene, by the method of Satom and Makino (Satom K, Makino K (1951) "Rature" 167, 23S) by reacting · the corresponding compounds of formula (la) with the required acyl chloride in anhydrous pyridine, Preferably, the products are recrystallized from a 1: 1 mixture of chloroform and petroleum ether.

The compounds of formula: NH-R1 ïV> are prepared

V ^ / o I

35, ^ Vr - £> S '\ - <CtÇ CH3 I · ji 4 i in which R and have the meanings defined above, by reacting the corresponding compounds of formula (i) in which represents a hydrogen atom , with acetone in the presence of 5 ZnCl 2, again by the process of Satom and Makino Aâ (mentioned above) · Preferably, the products obtained are purified by crystallization from a 111 ml mixture of chloroform and ether of petroleum · j The compounds of formula: j 10 NH-Rj: are prepared. | Y%? (Id)

kA / o CH * -S-R

15 P

i 0R.? 0Ro, i * in which R, R ^^ and R., have the meanings defined above, by oxidation of the corresponding thioethers! dants obtained by the methods indicated above, in j; using bromine or hydrogen peroxide in solution! aqueous (Green Stein J.P., Winitz M. (1961) - Chemistry j, of the amino acids - John Wiley & Sons Inc. 2146), The products obtained are purified by recrystallization j in water.

Among all the products prepared, the one which has proved to be particularly advantageous for the subject of the present invention is 51-deoxy-54-methyl-30 io thioadenosine of formula: k [! I "5 ψο; i I>

I ^ Λ / CH -S-CH

Τί N -5 i 5 1 ^ ° Χ (II)

TT

OH OH

'10 which is a physiological compound already present in living organisms.

To prepare this product, a particularly simple and economical process was found from the industrial point of view. This new process essentially consists in carrying out the hydrolysis of S-adenosylmethionine under critical conditions strictly regulated in order to achieve practically total hydrolysis and complete crystallization of 5 ′ -deoxy-51-methylthioadenosine NH ,,

20 I

It> + CHo-S-CHo-CH “-CH-C00H - iftl“

OH OH

NH9 6c> CH.-S-CH3 0 J **

^ T -T HO-CH „-CH0-CH-COOH

h-v + "+‘ 35 Tn "6

The controlled hydrolysis process can be applied to S-adenosylmethionine prepared in any way.

However, the process for preparing the solution of S-adenosylmethionine is also an influencing factor when the new process is implemented in an economically advantageous manner.

The following operating steps constitute the most economical embodiment of the process: a) Normal baker's yeast is enriched in S-adenosylmethionine by treatment with methionine in accordance with the Schlenk process * (Schlenk F. (1965) Enzymologie 29, 283).

b) The yeast cells suspended in * 15 water are subjected to lysis by treatment with! ethyl acetate or methyl acetate at room temperature (patent application of the Federal Republic of Germany DT-OS P 23 36 401 * 4) ·

By adjusting the pH to a value between 20 4 and 6 and by carrying out a filtration, an aqueous solution is obtained containing practically all of the i! S-adenosylmethionine present in the initial yeast.

I £ c) The solution is concentrated in vacuo at 35-40 ° C

to about 1/10 of its original volume.

D) The concentrate is boiled under reflux for about 30 minutes, then the pH is adjusted to 7; with soda.

e) The solution is left to stand at 0-5 ° C., while the precited 5′-deoxy-5methylthioadenosine is collected almost completely and with good purity.

Steps c, d and e which, as indicated, are critical and necessary for · ensuring hydro- | complete selective lysis of S-adenosylmethionine in 135 i 5’-deoxy-5‘-methylthioadenosine without any formation of by-products, are new.

üî f i.

Γ 7 "

The preparation of certain I ‘f products used according to the present invention will be described below, (EXAMPLE 1

Preparation of 5 * -deoxy-5'-methylthioadenosine.

5 11 liters of ethyl acetate and 11 liters of water at room temperature are added to 90 kg of baker's yeast which has been enriched in S-adenosyl-methionine by adding methionine until that the content of S-adenosylmethionine is 6.88 g / kg, 10 After having vigorously stirred for 30 minutes, the pH is adjusted to 4 * 5 with dilute H 2 SO 4, the mixture is filtered and the residue is washed with water 1 to obtain 140 liters of a solution having an S-adenosylmethionine content of 4 * 4 g / 1 * or 99 * 5% of the (15 S-adenosylmethionine present in the initial material.

The lysate thus obtained is concentrated under vacuum (30 mm Hg 5 35-40 ° C) to a volume of about 14 liters. The solution is boiled under reflux; centered under normal pressure for 30 minutes. It is cooled to 20 ° C, the pH is adjusted to / with; 40% sodium hydroxide and left to stand overnight in a refrigeration cell (+ 3 ° C),

A white precipitate is formed which is filtered, which is dissolved in 10 liters of distilled boiling water, then crystallized by cooling this solution.

410 g of crystalline 5'-deoxy-5 * -methylthio-adenosine of high purity are obtained, ie a yield of 90% with respect to S-adenosylmethionine subjected to hydrolysis, "The characteristics of the product thus obtained tt, agree with those of pure 5'-deoxy-5'-methylthio-adenosine obtained by other means, EXAMPLE 2 Preparation of 51-deoxy-51-ethylthioadenosine.

j Under a nitrogen atmosphere, 1 kg is dissolved! - ^ b? · Adenosine in 10 liters of hexamethvlphosphoramide

J

i · i f i ”! i i ί {and, while cooling, 7> 5 liters of thionyl chloride are added.

The mixture is left to react at room temperature for 20 hours. 10 liters of water are added and the mixture is neutralized with 2N NaOH. The S'-deoxy-S'-chloradenosine thus formed is left to crystallize overnight at 3 ° C., then it is separated by filtration. We obtain 0.950 kg of 5 * ~ 5-deoxy-1-chlor a denos ine (yield: 89%) · We dissolve 0.950 kg of 5 * -deoxy ~ 5'-10 chloradenosine in 10 liters of 2N NaOH and 200 ml are added ethane-thiol. The mixture is heated to 80 ° C and allowed to react for one hour. We neutralize it; read with glacial acetic acid. The i-deoxy-S’-ethylthioadenosine thus formed is allowed to precipitate overnight at 3 ° C. Then, it is separated by filtration and recrystallized from water. 0.830 kg of product is obtained (yield: 80% vis-à-vis the previous step).

EXAMPLE 3 f i 20 Preparation of other compounds of formula la.

I The process described in Example 2 is carried out, but using respectively propane-thiol, butane-thiol, isobutane-thiol, pentane-thiol,! Hexane-thiol and benzylthiol in place of ethane-thiol.

! EXAMPLE 4 ί „Preparation of NO, 2’, 3’-triacetyl-51 -deoxy-5 * -thioadé-I 'nosine.

1 kg of 5'-deoxv-5 '-methylthioadenosine 30 is suspended in 10 liters of anhydrous pyridine and 3 liters of acetic anhydride are added. The mixture is left to react for 4 hours. 20 liters of water are added and the mixture is concentrated under vacuum to obtain an oily mass free of pyridine. This mass is dissolved in 10 liters of a hot L1: 1 mixture of petroleum ether and chloroform and allowed to crystallize. The product is recrystallized from a 1: 1 mixture of petroleum ether and chloroform.

1.140 kg of product are obtained (yield: 8θ> "). EXAMPLE 5

Preparation of other compounds of formula Ib.

5 The process described in Example 4S is carried out

but using other thioethers or propionic anhydride, butyric anhydride, benzoyl chloride or tosyl chloride instead of 5 * -deoxy-5 * -methylthioadenosine.

10 EXAMPLE 6

Preparation of 5 ’“ deoxy - ^ ', 3’-isopropylidene-S’-methylthioadenosine.

1 kg of 5 * -deoxy-51-methylthioadenosine is suspended in 25 liters of anhydrous acetone and 2.5 kg of molten ZnC 4 are added. The reaction is carried out at reflux for 5 hours. Then, the mixture is concentrated under vacuum to 1/3 of its initial volume, then 7.5 kg of barium hydroxide is added to 8 molecules of water in aqueous suspension. Then, carbon dioxide is bubbled through until neutral. The mixture is filtered and the residue is washed with acetone.

The filtrate is concentrated under vacuum to obtain a syrupy residue. The latter is taken up in 10 liters of a hot 1: 1 mixture of chloroform and petroleum ether, filtered and allowed to crystallize. The product obtained is recrystallized from a 1: 1 mixture of chloroform and petroleum ether to obtain 0.795 kg of product (yield: 70%).

EXAMPLE 7 Preparation of other compounds of formula Ie.

The process described in Example 6 is carried out, but starting from the corresponding derivatives of adenosine in place of 5'-deoxy-5'-methylthioadenosine. EXAMPLE S

35,] Preparation of 5 1 -deoxy-5 * sulfoxide -methylthioade-

J

nosxne.

"10 I 1 kg of 5'-deoxy-5'-methylthioadenosine is suspended in 10 liters of water and added; bromine while cooling, j | The aqueous solution containing bromine is de-!, 5 colored immediately by the oxidation of 5 * ~ deoxy— | 5’-methylthioadenosine sulfoxide,! Continue adding bromine until i i no longer discolour the solution,

The solution becomes discolored when small amounts of 51-deoxy-5 '-methylthioadenosine are added further; The aqueous solution is treated with the resin "Amberlite IRA 93" (registered trademark of "Rohm and Haas" for a weakly basic ion exchange resin with a polystyrene matrix) until that the reaction of bromide ions ceases, | The mixture is filtered and the residue ji washed with water. The aqueous solution is concentrated to 10! liters, it is treated with 100 g of activated carbon and it is lyophilized, ‘20 0.950 kg of product is obtained (yield: 90%).

; EXAMPLE 9

Preparation of other compounds of formula Id,: The process described in Example S is followed, but i ‘starting from the corresponding derivatives of adenosine in place of 51-deoxy-5'-methylthioadenosine.

As noted at the outset, the compounds of formula I have been found to exert potent anti-inflammatory activity in conjunction with analgesic and antipyretic activity; 30 The anti-inflammatory activity was initially demonstrated for certain compounds by practicing the test of experimental edema caused in rats by! ; carrageenan by determining the percentage of protection provided by the Winter method ("J, Pharm.

35 "exper. Therap. "141, 369, 1963). The results are obtained in the table below,

i ~ K

* 11

BOARD

Compound of formula X Dose Pourcen— administration of protected nistrea 5 by oral route calculated (mg / kg) on - development of 10 __________11 edema n = 0 R = -CH2, Rx = Rz = H 37 50 n = 0 R = -CH, Ri = R2 = H 23 (a) 50 n = 0 R = -CH2-C6H5, R1 = R2 = H 47 10 ^ CH3 15 n = 0 R - -CH2-CH, Rx = R2 = H 85 62 NVch3 n = 0 R = - (CH2) 6-CH3, Rx - R2 = H 95 20! n = 0 R = - (CH2) 11-CH3J Rx = R2 = H 112 10 n = 0 R = - (CH0) —CH, R, = R9 = H 90 25 20 n = 0 R = - CH0-CH3, Rj = R2 = H 50 44 n = 0 R = - (CH0) 0-CH, R = R0 = H 8θ 53 / CH3 n = 0 R - -CH, Rx = R2 = H 80 45

^ CH

; 25 n = 0 R = - (CH2) 3-CH3J = R £ = H 85 39 'n = 0 R = CH-CH.-CH-, R, = R0 = H 85 35. CH3 • n = 0 R = - (CH2) 7-CH3, Rj_ = R2 = H 100 47 n = 0 R = - (CH0) n-CH0 3 R, = R „= H 106 33 2931 u 30 η = 1 R = -CHOJ R. = R9 = H 150 50 O * ^ η = 1 R = -CH, Rj = R, - KS, 6 (a) 50 n = 0 R = -CH-, R = R0 = -C0 -CH. 114 47

O ** O

n - 0 R = -CH3j Ra = R9 = tosyle 204 15 n = 0 R = -CH ^, R1 = R2 = -CO-C6H5 104 10 35; n = 0 R = -CH3, R1 = H, R0-R? = isopropylidene 91 20 ώΪ Indomethacin 9 50 \ / »12 (a) means that the product has been administered by the intramuscular route.

: i _ - _ "

As can be seen from this table, 5 the effective dose at 50% (DE ^ _q) 5 1 -deoxy-5 1 -methylthioadenosine is 37 mg / kg and, therefore, this va-; their is the weakest of those of the subject compounds! '' under test when administered orally.

| ! In the same test, the ED, -q value of indo-methacin is 9 mg / kg. At these doses, we see appa; have serious gastric lesions while | ED, -q, 5deoxy — 5 ’—methylthioadenosine has no side effects on the gastrointestinal system. It should also be noted that the lethal dose! 15 to 505® (LD50) of indomethacin in rats is 12 mg / kg (Martelli A, in "Aspetti di farmacologia dell * j infiammazione”, page 73j published by Tamburini - Milan [1973) t while the value LD ^ of 5 '* - deoxy-5' - methylthioadenosine in rats is greater than 200 to 20 mg / kg / orally.

i Consequently, the following therapeutic indices are obtained:

11indomethacin therapeutic index = 1.3

5'-deoxy ~ 5'-25 methylthioadenosine therapeutic index => 54d05

The compounds according to the invention are also subjected to a series of pharmacological tests in order to confirm their anti-inflammatory activity and in order to demonstrate their analgesic and antipyretic activity.

The results obtained in some of these trials will be given below with 5'-deoxy-5'-methylthioadenosine, which is a product which, in any case, appears to be most active when administered by Orally and which, without a doubt, is the safest product, since as well, as already mentioned, it is physiologically present in the body.

13 d:

Likewise, from the industrial production point of view, the process for the preparation of 51-deoxy-5 * -methylthioadé-nosine from S-adenosylmethionine according to the invention is by far the simplest and most economical, {5 while '' it can be carried out on an industrial scale at a particularly low cost.

As can be seen from the data. From the above table, methylthio-adenosine sulfoxide is particularly active when administered intramuscularly. The strongest activity exerted by this compound during intramuscular administration has been confirmed in all the trials which have been carried out. Some significant data relating to the sulfoxide are also given. -deoxy-5'-methylthioadenosine, however, it should be noted that, in all cases, all of the compounds tested have been found to be active, albeit at different levels.

A - Anti-inflammatory activity The products were subjected to provo tests

as for pleuritis in rats by carrageenan according to the Velo method (Velo G.P., DUNN

C.J. et al. (1973) "J. Path." 111, 149).

In a dose of 75 mg / kg orally, 5deoxy-5'-methylthioadenosine provided protection of 42.4 ^, calculated on the volume of exudate, and 48.8 ^, calculated on the number total cells present in the exudate.

^ Comparable protection was obtained with 10 13 mg / kg of indomethacin, ie a dose much closer to the DL ^ q value. In the same 1 test, the sulfoxide of 51-deoxy-51-methylthioadé nosine provides a protection of 75.8%, calculated on i the volume of the exudate, and 76.4 / ^ * calculated on P 35 i the total number of cells present in the ex-I Jt dat when administered intramuscularly at a dose of 80 mg / kg.

V

t 11!) 14 j f s B - Anti-inflammatory activity.

ί i When testing the granuloma caused in cotton rats by cotton pellets (Winter CA, Rise- ley EA, et al, (1963) 5 "J * Pharm, Exper, Ther.", j 5 141? 369), this trial being significant for chronic inflammation, 5'-deoxy-5'-methylthio-j adenosine provided protection of 30%> in an oral dose of 9 mg / kg with a therapeutic index of 222.

• 10 C - The analgesic activity of the products was the subject of two tests considered to be very significant.

'- In the heating plate test performed on the mouse following Roberts (Roberts E, Simonsen DG (i960) "Biochem. Pharmac.” 15y 1875) j S 15 51-deoxy-5'-methylthioadenosine provides protec 50% in an oral dose of 37 µg / kg. Approximately equivalent protection of 58% is obtained with 100 mg / kg of amidopyrin administered orally.

20 In the same test, 5 * -deoxy— 5’ — methylthioadenosine sulfoxide provides 50% protection in a dose of 20 mg / kg when administered

Itration intramuscularly and in a dose * of 100 mg / kg when administered orally.

- During the phenylquinone extension test (Siegmund E., Cadmus R., G0-LU (1957) "Proc. Soc. Exp. Biol. Med." 95, 729) 5 '5-deoxy -5 '-methyl-I thioadenosine provides protection in an oral dose of 37 mg / kg.

I In the same test, 5’ — deoxv— I 51-methylthioadenosine sulfoxide has a DEr „value of 10 I 50? mg / kg law’s for intramuscular administration.

I 35 D - Antipyretic activity I f This activity is measured for new products by causing fever in rats using

Y

"" 15 I of brewer's yeast (Winter CV et al. (L96l), | "J. Pharmacol. Exp. Ther.” 133, 117) · The antipyretic effect evaluated for one hour | after the oral administration of 5'-deoxy-51-methyl- | thioadenosine in a dose of 300 mg / kg, gave a temperature reduction of 459% with respect to the controls which were treated only with yeast. corresponds to a drop in temperature from 38.8 ° C to 37.4 ° C · 10 By comparison, amidopyrine administered orally in a dose of 200 mg / kg gives a reduction in temperature of 4 * 69 $ , while the intramuscular administration of 51-deoxy-5 '- J: methylthioadenosine in a dose of 80 mg / kg provides Π' 15 a reduction in temperature of 2.35 ^ · 1 E - Inhibitory activity of Platelet Agglomeration The compounds of the invention have also been evaluated for their ability to inhibit platelet agglomeration.

It is known that the agglomeration of platelets is a complex phenomenon which can be divided into a primary stage due to the direct action of a stimulus (for example, adenosine diphosphate or epi-! Nephrine ) and in a secondary stage due to agglomeration

H

25 adenosine diphosphate induced meration | * released by platelets. In this regard, when |: the platelets come into contact with the colla- I: subendothelial gene, the collagen triggers a whole series of reactions leading to the release of adenosine diphosphate by the platelets.

It is precisely this adenosine diphosphate that is the cause of the second wave of platelet agglomeration. The tests described below were carried out in order to evaluate the new compounds 35 j concerning their 1k agglomeration inhibiting effect. platelets: a 16 1) "In vitro" tests on the agglomeration of platelets caused by adenosine diphosphate and: | collagen, in the presence of new products; 2) "in vitro" tests on the agglomeration of platelets caused by arachidonic acid; | 3) "in vivo" tests on the agglomeration of pla—! quetbes caused by adenosine diphosphate! and collagen in people treated with | new products.

j, 10 In this case, the most significant results are again obtained with 5’— deoxy-5’— ^. methylthioadenosine; therefore, the results obtained using this product are given to indicate the behavior of the entire class of compounds.

1) "In vitro" tests.

We take blood without stasis and add one; j anticoagulant (sodium citrate at $ 3.8 S) to · obtain i! a ratio of Ç: 1 between blood and citrate. Platelet-rich plasma and platelet-poor plasma are obtained by temperature centrifugation! ambient ture. The agglomeration of platelets- 'I tes is evaluated by adopting the Born & Cross (G.V.R.

J Born and M.J. Cross, "J. Physiol.", London 168, 17S, 25 1963) on the fraction of platelet-rich plasma.

| i We use the agents causing the agglomeration of j! . platelets in the following concentrations: diphos-jl adenosine phage (Sigma) 1 µm; collagen (Hornj '1 5 ^ ig / ntl; arachidonic acid 4 x 10 ^ M.

30 We used adenosine in one concentration. * t ~ 'of 1 x 10 ^ M as a reference substance exerting an activity I inhibiting platelet agglomeration.

I The results obtained with adenosine diphosphate are illustrated by the graphs of r; 35 figure I where the time (in minutes) is taken in abs-! j- cisse, while the agglomeration percentage is f Ji '• 1 plotted on the ordinate.

: •: \ i.

»17

Curve 1 relates to the controls, curve 2, to the samples treated with adenosine in a concentration -5 tration of 1 x 10 M and curve 3j to the samples [treated with 5 * -deoxy-5 * -methylthioadenosine in a concentration of 5 x 10 ~ ^ M.

From the shape of the curves, it can be seen that 5'-deoxy-51-methylthioadenosine strongly reduces the primary agglomeration of platelets due to the adenosine diphos-phate and that, consequently, it inhibits the second stage of 'agglomeration.

Same tests with collagen give negative results, i.e. 5'-deoxy-51-methylthioadenosine has no inhibitory effect on platelet agglomeration caused by collagen, which is worthwhile to be noted, 2) FIG. 2 illustrates the effects exerted by different concentrations of 51-deoxy-51-methylthioadeno-sine on the agglomeration of platelets caused by arachidonic acid at a concentration of 4 × 10 ^ M, i 20 Curve 1 relates to the controls, curve 2, to 5'-deoxy-51-methylthioadenosine in a concentration of 2.5 × 10 -4 M, curve 3} to 5 * -deoxy-5 ' -methylthioadenosine in a concentration of 5 x 10 * ”^ M and the curve 4j with 5'-deoxy — 5 * -methylthioadenosine in a concentration —3 25 centration, As can be seen, the effect exerted by 51-deoxy-5'-methylthioadenosine on the inhibition of platelet agglomeration is proportional to its concentration. The ability of 51-deoxy-51-methylthioadeno-30 sine to enhance the inhibitory effect of prostacyclin (PGI0) on the agglomeration caused by ara-chidonic acid has also been studied. In FIG. 3j, curve 1 relates to the controls, curve 2, to 51-deoxy-5’-methylthioadé-nosine in a concentration of 5 × 10, curve 3,

.Q

35 to PGI2 in a concentration of 5 x 10 'M and the curve 4 * to a mixture consisting of 51-deoxy-5'-methyl-thioadenosine in a concentration of 5 x 10 ~ ^ M and "! | i8 _û 1 * GI9 in a concentration of 5 x 10 M. According to j ^ I Figure 3, we see that by using the mixture, we | strongly increases the agglomeration inhibiting effect at concentrations which, in themselves, are ineffective * 3) "In vivo" tests.

Three apparently healthy * volunteers, aged 35, 42 and 48, respectively, and having taken no medication for at least 15 days, 10 were subjected to platelet agglomeration tests before and after consuming the new produced in a dose of 100 mg every eight hours! for 3 days ; then the assessments were done.

\ The blood sample for the determination of! The agglomeration of platelets was taken two hours before consumption of the last dose of the product under test. FIG. 4 gives the results I obtained with 5 * -deoxy-5’-methylthioadenosine.

More specifically, the solid lines 20 relate to the values obtained with blood samples taken from untreated patients,! while the dashed lines refer to the values obtained in the same patients treated with 5’-deoxy-5’-methylthioadenosine.

;; It is found that 5 ′ -deoxy-5 ′ —methylthioadé— - * nosine strongly inhibits the agglomeration of t-platelets caused by adenosine diphosphate (1 ^ iM) "in I vivo".

By repeating the same tests, but adding 30 5 yag / ml of collagen to the blood, it was found that 5'-deoxy-5'-methylthioadenosine was not effective in inhibiting the agglomeration of platelets caused by collagen, but that it simply prolonged the latency time of the phenomenon.

The fact that 5'-deoxy-5'-methylthioadenosine t (and, in a more or less comparable way, the other products of the same class) strongly inhibits the agglomeration of platelets caused by the adenosine diphosphate, while it has practically no effect on the agglomeration caused by collagen,; indicates that 5'-deoxy-5'-methylthioadenosine inhibits the first stage of agglomeration, while it has a negligible direct effect on the second stage of agglomeration.

fi | As a result, it is particularly interesting; resent of using it in combination with other known drugs which inhibit platelet agglomeration; quets which are generally active on the second stage of agglomeration, whereas they have only a mediocre effectiveness compared to the first stage.

The activity thus demonstrated suggests the use of 5 * -deoxy-5 * -methylthioadenosine (and the other compounds of the series, even if they are less effective) 5 not only as a drug for inhibiting agglomeration tion of platelets, but also as an antithrombotic and anti-atherosclerotic drug I; indeed, in addition to the fact that it constitutes the basis of the mechanisms of thrombogenesis, the modified relationship between the platelets and the walls of the vessels also plays a role; an essential role in atherosclerotic diseases.

f 25 F - Soporific activity.

j | ; We did the Morris test (Morris R.W.

f (1966) "Int. Arch. Pharmacodyn" 16l, No. 2, 380).

K "During this trial, 5’-deoxy-5’-methyl- ΐ thioadenosine increased by S7a> sleep duration? 30 caused by pentobarbital in mice at an intramuscular dose of 20 mg / kg.

G - Acute toxicity.

The compounds of the present invention are practically free from acute toxicity when administered orally. They are practically free of toxicity at therapeutic doses for any method of administration.

-'rv-! 20 *! , The va] eui, ¾ below are obtained for 5 * ~ jr deoxy-5 * —methy] thioadenosine and the sulfoxide of 5 '-' 1 deoxy— 5 1 - methy] thioadenosine: t I 5 "- deoxy-5 * -methylthioadenosine - LDrn in mice J 5 orally> 2,000 mg / kg I intravenously 360 mg / kg I 5 1 -deoxy — 5 * sulfoxide —methylthioadenosine - LDj-q | , in mice orally> 2,000 mg / kg

Il 0 intravenously 400 mg / kg

The adenosine derivatives of formula (i) can be administered, in dilution with appropriate and pharmacologically acceptable excipients, in any form therapeutically useful, orally, parenterally, orally. venous or by: i rectally. They can also be used in

NOT

f | products for external use by topical application.

Below are given for illustrative purposes only certain examples of specific pharmaceutical compositions containing certain 51-deoxy-5> -methyl thioadenosine, "100 mg capsules 51-deoxy -5 * -methylthioadenosine 100.2 mg if 1 Mannitol 195.0 mg d 25 Magnesium stearate 5.0 mg | ; - \) 300.2 mg 11 - 50 mg capsules jl 51 -deoxy-51 -methylthioadenosine 50.1 mg 30 s Mannitol 100.0 mg | Magnesium stearate 30 mg _:. -t 153.0 mg! * * I! ! \

V

"21 - 100 mg tablets 5 * -deoxy-5 * -methylthioadenosine 100.2 mg

Starch 100.0 mg

Magnesium stearate 15 * 0 mg 5 Lactose 85.0 mg! _ I 300.2 mg!} Ji ‘- 50 mg tablets I 51-deoxy-5'-methylthioadenosine 50.1 mg | 10 Starch 120.0 mg * Magnesium stearate 15> 0 mg | Lactose 115 »0 mg | 300.1 mg * 15 - Suppositories at 100 mg d 5'-deoxy-51-methylthioadenosine 100.2 mg | Mass for suppositories 1,700.0 mg ï | 1,800.2 mg 20 -50 mg suppositories 5 * -deoxy-51-methylthioadenosine 50.1 mg

Mass for suppositories 1,450.0 mg 1,500.1 mg * 25 - Vial for injection at 50 mg i '5'-deoxy-51-methylthioadenosine.

j, HCl (56.15 mg basically I equivalent) 50 mg? Lidocaine.HCl 25 mg 30 Water, to make up to 3 ml j “Vial for injection at 25 mg î I 5'-deoxy-5'-methylthioadeno! sine. HCl (28.07 mg basically equivalent.) 25 mg I 35) Lidocaine. HCl 20 mg 1. -4. Water, to make up to 2 ml V ^ * ϊ ο Ο; s ί '- Oral dose at ÎOQ mg 5 * -deoxy-51-methylthioadeno-sine. HCl (112.3 mg basically equivalent) 100 mg 5 Lemon flavor 0.025 mg | Sugar 1 g! Fermentation inhibitor 50 mg

Water, to make up to 5 ml î - Oral dose at 50 mg ‘10 51-deoxy-51-methylthioadenosine. HCl (56.15 mg basically equivalent) 50 mg, Lemon flavor 0.015 mg

Sugar 0.5 g 15 Fermentation inhibitor 30 mg

Water, to make up to 5 ml - Ointment at 100 g 51-deoxy-51-methylthioadenosine 5 g

Base for water-soluble ointment, 20 to complete 100 g I Antioxidant 0.1 g.

Jh ‘F? Γ ^ /: /! »» \! I, jj;

; I

Claims (14)

1. Therapeutic composition with anti-inflammatory, analgesic and antipyretic activity, characterized in that it comprises, as active principle, '5 at least one compound corresponding to the general formula: j NH-RX 11 / (0) ni 10 ÇH9-SR :. $! OR2OR2 I 1 15 in which I R represents a straight or branched chain alkyl group containing 1 to 18 carbon atoms, or a group! phenylalkylene in which the alkylene chain contains; 1 to 6 carbon atoms; R 1 is a hydrogen atom, an aliphatic acyl group containing 1 to 6 carbon atoms or an aromatic acyl group; R2 represents a hydrogen atom, an acyl group; aliphatic containing 1 to 6 carbon atoms or one! 25 aromatic acyl group, or alternatively, the i j "radicals R ^ together form an isopropylidene chain, n is equal to 0 or 1. 2. Therapeutic composition according to claim 1, characterized in that it comprises, as active principle, a compound of formula (Ij in which R, = R ,, = H, R = CH, n - 0. * · - J 3 · therapeutic composition according to claim 1, characterized in that it comprises, as I active principle, a compound of formula (i) in which 35. Rj = R0 = H, R = CH3, n = 1.; ü * 24> L · 4 · Therapeutic composition according to re- | vendication 1, characterized in that it comprises, "as active ingredient, a compound of formula (X) in which R. = R„ = H, R = straight-chain or branched chain alkyl group containing 1 with 12 carbon atoms and n - 0. 5. Therapeutic composition according to re vendication 1, characterized in that it comprises, as active principle, a compound of formula (l) in i 10 which = H, R = benzyl, n = 0. 6. Therapeutic composition according to the re- || vendication 1, characterized in that it comprises, jf "as active ingredient, a compound of formula (i) in it I which R ^ = R ^ = benzoyl, R = methyl, n = 0. 'f 15 7 · Therapeutic composition according to re- i | vendication 1, characterized in that it comprises,. "! ! as active principle, a compound of formula (i) in which R ^ = R9 = t.osyle, R = methyl, n = 0. • 'SI _: | i 8. Therapeutic composition according to re- 'Ji! 20 vendication 1, characterized in that it includes,: | as active ingredient, a compound of formula (i) in; | which R? - R2 = isopropylidene, R = methyl, n = O. I 9 »Process for the preparation of 51-deoxy-51-I methylthioadenosine, characterized in that I 25 hydrolyses S-adenosyl-methionine, in the form of a solution; . | > concentrated aqueous, by heating at reflux, then jf Jf A * · J § separates the 5 * —deoxy-5’-methylthioadenosine formed by: 'i cooling after neutralization of the reaction mixture. 10. The method of claim 9, characterized in that the aqueous solution i of S-adenosyl-methionine is concentrated by heating under vacuum to;! a temperature of 35 to 40 ° C. : *! 11 * A method according to claim 9, characterized in that precipitates 51-deoxy-5 '-methvl-; J i ..., 1' thioadenosine formed by cooling to a temperature • J * rature 0-5 ° C. y \% 25 m r 12. Use of a compound corresponding to the general formula: f NH-R ism I 'S Λ, / <ï> »1" w ch9-sr (I)! Ol!: 10 II OR OR I * in which U * R represents a straight or branched chain alkyl group containing 1 to 18 carbon atoms, or a phenylalkylene group in which the alkylene chain contains 1 to 6 carbon atoms; II A ^ p R ^ represents a hydrogen atom, an aliphatic acyl group containing 1 to 6 carbon atoms or an aromatic acyl group I represents a hydrogen atom, an aliphatic acyl group containing 1 to 6 carbon atoms or a aromatic acyl group 1, or alternatively the radicals together form an isopropylidene chain for the preparation of pharmaceuticals with anti-inflammatory, analgesic and antipyretic activity. 13. Use of the compound of formula (i) in which R ^ = R £ = H, R = CH ^ j n = 0, for the preparation of pharmaceutical products with anti-inflammatory, analgesic and antipyretic activity. 14. Use of the compound of formula (Ij in which R ^ = R £ = H, R = CH ^, n = 1> for · the preparation of pharmaceutical products with anti-inflammatory, analgesic and antipyretic activity. _________ i ! ίι