Method of thieno(3-2-c)pyridine's new derivatives production

(54) A process for the preparation of new thieno [3,2-c] pyridine derivatives

The invention relates to a process for the preparation of novel thieno (3,2-c) pyridine derivatives which can be used for therapeutic purposes.

The present invention provides a process for the preparation of novel thieno [3,2-c] pyridine derivatives of the formula I by condensing 4,5,6,7-tetrahydrothieno [3,2-c] pyridine of the formula II

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ÍIIJ

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with α-chlorophenylacetate of formula III

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(III) where R is where

R is a straight or branched lower alkyl radical having 1 to 4 carbon atoms, and

X represents a hydrogen atom, a chlorine atom or a fluorine atom or a methyl group, and addition salts of these compounds with pharmaceutically acceptable inorganic or organic acids, as well as the two enantiomers or mixtures thereof, and X is as defined above; optionally converting the resulting compound into an acid addition salt by treatment with an acid acceptable from a pharmaceutical standpoint, or optionally separating the racemate into enantiomers.

The esters of formula (III) may be obtained in a manner known per se, according to EL Eliel, T. Fisk and T. Prosser, Organic Syntheses, Coll. IV, J. Wiley and Sons, Inc., New York, 1963, p.

The condensation of tetrahydrothienopyridine with an ester of formula III is preferably carried out in the presence of an alkali metal carbonate such as potassium carbonate in an inert solvent such as dimethylformamide, tetrahydrofuran or 1,2-dimethoxyethane. The reaction is carried out at 60 ° C to the boiling point of the solvent.

The invention will be illustrated by the following examples:

Example 1 α- (4,5,6,7-Tetrahydrothieno [3,2-c] -5-pyridyl) -o-chlorophenylmethyl acetate (R = —CN, X = 2-C1], derivative No. 1

To a solution of 20 g (0.144 mol) of 4,5,6,7-tetrahyrothieno [3,2-c] pyridine in 200 ml of dimethylformamide was added 31.47 g (0.144 mol) of C-chloro-o-chlorophenylmethyl acetylate and 19.82 g (0.144 mol) of potassium carbonate and the mixture was heated at 90 for 4 hours ^CC. The reaction mixture was cooled to room temperature, the inorganic salts were collected by filtration and the solvent was evaporated. The residue is treated with water and extracted with ethyl ether. The ether extracts were washed with water, dried over sodium sulfate and evaporated to a yellow oil which was purified over hydrochloride to give the title product as white crystals, m.p. 130-140 ° C, recrystallized from ethyl acetate / isopropanol. The yield is 45%.

Example 2 α (4,5,6,7-tetrahydro-thieno [3,2-c] -5-pyridyl) -phenylmethyl acetylate (R = - N, X = Ή), derivative No. 2

The title compound is obtained as described above. in Example 1 by alkylation

4,5,6,7-Totrahydrothione O '[3,2-c] pyridine' by treatment with 2-chlorophonylmethyl acetate.

The hydrochloride of the resulting product is white crystals, m.p. 200 DEG C. after recrystallization from ethanol. The yield is 50%.

Example 3 α- (4,5,6,7-Teteahydrothieno [3,2-c] -5-pyridyl) -o-fluorophenyl methyl acetate (R = -OH 3, X = 2-F), Derivative No. 3

This resulting compound can also be obtained by the method of Example 1 by alkylation of 4,

5.6.7-Tetra-α-thieno [3,3-c] pyridine by treatment with C-chlorofluorophenyl methyl methylacotate.

The hydrochloride of this material is white crystals, which at 100 ° C becomes a pasty mass. Yield 75.5%.

Example 4 α- (4,5,6,7-tetrahydro-thiothio [3,2-c] -5-pyridyl] -o-methylphenyl-ethyl-acylate (R = —CH 2 —CH 3, X = 2-N 3), derivative No. 4

The title compound was obtained as described in Example 1 by alkylation of 4,5, [3,2-c] pyridine with C-chloro-o-methylphenylethyl acetate.

The hemisulphate is in the form of white crystals, m.p. 188-190 ° C after recrystallization from isopropanol. Yield 54%.

The pharmacological and toxicological results obtained with the compounds according to the invention give data on the toxicity, tolerability and action of the compounds of the formula I, in particular with regard to the inhibition of platelet aggregation and the autotrombotic activity.

The compounds of formula (I) may be formulated as a medicament having an inhibitory effect on platelet aggregation and an antithrombotic effect. The pharmaceutical composition will contain as an active ingredient a compound of formula I or a pharmaceutically acceptable acid addition salt thereof with an inorganic or organic acid.

Toxicological tests

The compounds of formula I are well tolerated and have low toxicity.

Tests were conducted for acute, chronic, subchronic and delayed toxicity in various animal species, with no local or general adverse reaction, no disturbances or anomalies of a macroscopic, microscopic or biochemical nature during any of the experiments.

Pharmacological tests.

1. Inhibition of platelet aggregation

The experiments were performed on rats given the active ingredients for 3 days, at 4848 hours, 2424 hours, and hodinách2 hours in gum arabic suspension. At time 0, 4 ml of blood was drawn from the jugular vein according to the Renaud method in an auesthetized animal. This citrate blood was used for clustering tests.

a) Agglutinated by ADP ml of citrate blood is quickly poured into a small vessel equipped with a magnetic stirrer. After stirring for a few seconds, 0.4 ml containing 0.66 µg ade246062 nosindiphosphate (ADP) per ml is poured into the vial. After 90 seconds of mixing, add two 0.5 ml blood samples:

- the first sample is mixed with 0,5 ml of the solution of ethylenediaminetetraacetic acid (EDTA) in formol, - the second sample is mixed with 0,5 ml of the EDTA solution.

The addition of EDTA and formol is intended to stabilize the platelet count in EDTA-formol._nn —-------- χ 100 - = number of plates in

EDTA

The test substance causes greater inhibition of platelet aggregation, the closer the ratio is to 100.

The results of an experiment in which the blood blood count and fix the clusters, the addition of EDTA alone should dissolve the formed clusters.

After 10 minutes, the two mixtures are centrifuged for 5 minutes at low speed to separate red blood cells and the platelet-rich plasma supernatant is removed, diluted and the plates counted.

Aggregation intensity can be determined by the relationship of non-agglutinated platelets of platelets is the average of groups of five rats, either experimental or control, as shown in Table I below.

TABLE I

<td colspan="2">Group</td><td>Dose mg / kg</td><td>Method of administration</td><td>Result</td>

<td>control</td><td></td><td></td><td>AFTER</td><td> 16 ± 4</td>

<td>derivative no.</td><td> 1</td><td>3 x 25</td><td> —</td><td> 94 - ± 3</td>

<td>control</td><td></td><td></td><td> —</td><td> 20 ± 11</td>

<td>derivative no.</td><td> 1</td><td>3x5</td><td> —</td><td> 82 - ± 11</td>

<td>control</td><td></td><td></td><td> —</td><td> 23 ± 15</td>

<td>derivative no.</td><td> 1</td><td>3 x 2.5</td><td> —</td><td> 56 - ± 17</td>

<td>control</td><td></td><td></td><td> —</td><td> 13 ± 3</td>

<td>derivative no.</td><td> 4</td><td>3 x 100</td><td> —</td><td> 89 - ± 1</td>

<td>control</td><td></td><td></td><td> !-------------------</td><td> 3 ± 1</td>

<td>derivative no.</td><td> 4</td><td>3 x 100</td><td> —</td><td> 89 - ± 4</td>

<td>control</td><td></td><td></td><td> —</td><td> 4 ± 0</td>

<td>derivative no.</td><td> 2</td><td>3 x 100</td><td> ..-------------------------------------</td><td> 72 ± 12</td>

b) Measurement of platelet aggregation in collagen

1.5 ml of citrate blood is added to 0.10 ml of a solution containing 10 µg of collagen per ml. The mixture was stirred and the plate counting was performed without interruption.

The reduction in the number of free platelets is monitored as a function of time continuously and makes it possible to create a curve from which the initial clumping rate can be read as a function of its slope.

Results are the average of groups of five rats, both experimental and control, and are shown in Table II below.

TABLE II

Collagen test

<td>Group</td><td>Dose mg / kg</td><td>Method of administration</td><td>Results</td>

<td>control</td><td></td><td>AFTER</td><td> 3,12 ± 0,47</td>

<td>derivative No. 1</td><td>3 x 25</td><td> —</td><td> 0,14 ± 0,03</td>

<td>control</td><td></td><td> —</td><td> 2,17 ± 0,64</td>

<td>derivative No. 1</td><td>3x5</td><td> —</td><td> 0,19 + 0,04</td>

<td>control</td><td></td><td> —</td><td> 5,00 - ± 1,02</td>

<td>derivative No. 1</td><td>3 x 2.5</td><td> —</td><td> 0,60 ± 0,20</td>

<td>control</td><td></td><td> —</td><td> 3,41 '± -0,55</td>

<td>derivative No. 4</td><td>3 x 100</td><td> —</td><td> 0,12 ± 0,02</td>

<td>control</td><td></td><td> —</td><td> 4,73 ± 0,55</td>

<td>derivative No. 4</td><td>3 x 100</td><td> —</td><td> 0,30 ± 0,02</td>

<td>control</td><td></td><td> —</td><td> 5,00 - ± - 1,06</td>

<td>derivative No. 2</td><td>3 x 100</td><td> —</td><td> 0,51 - ± 0,18</td>

c) Measurement of ido-bleeding

Assessment of anti-platelet aggregation activity also had to be supplemented by determining bleeding time.

For this purpose was. a modification of the method described in L; Stella, Μ. B. Donati · and G. de Gaetano, Thromb. Res., 1975, 7, 709-716.

The experiments were performed on rats given 65 hours, 41 hours and 17 hours prior to the experiment with the test substance orally in suspension in 10 ml / kg of 5% aqueous gum arabic solution. After introduction into pentobarbital-anesthesia. The tail of the rat is separated from the end of the blood by allowing the blood to flow for seconds. At that time, blood is initially aspirated so that the aspirating material does not touch the pailey.

The effect is achieved if the blood stops spontaneously within one minute.

The results indicate the average bleeding time in seconds for a group of five rats in Table III and it is clear that the compounds prolonged the bleeding time with time data longer than 1,200 seconds (20 minutes) were no longer counted.

TABLE · · III

Bleeding time

<td>Group</td><td colspan="3">Dose mg / kg</td><td>Method of administration</td><td>Result</td>

<td>control</td><td></td><td></td><td></td><td>AFTER</td><td> 600</td>

<td>derivative No. 2</td><td> 3</td><td>x</td><td> 200</td><td> —</td><td> &gt;1200</td>

<td>control</td><td></td><td></td><td></td><td> —</td><td> 420</td>

<td>derivative No. 1</td><td> 3</td><td>x</td><td> 25</td><td> —</td><td> &gt;1 200</td>

<td>derivative No. 1</td><td> 3</td><td>x</td><td> 5</td><td> —</td><td> 1080</td>

<td>control</td><td></td><td></td><td></td><td> —</td><td> 435</td>

<td>derivative · No. 1</td><td> 3</td><td>x</td><td rowspan="2"> 12,5</td><td> —</td><td> &gt;1 200</td>

<td>control</td><td></td><td></td><td> —</td><td> 780</td>

<td>derivative · No. 3</td><td> 3</td><td>x</td><td> 200</td><td> —</td><td> &gt;1 200?</td>

2. Antithrombotic efficacy

This efficacy was measured in inducing experimental thrombosis with a silk thread.

The principle of this procedure is the adaptation of the method, experimental thrombosis caused by extracorporeal circulation. . as described in Tor.uhiko^! Umetsu and Hazuko. Sansi (Thromb. Kaemost., 39, 1, 1978).

On . Narcotized rats (intraperitoneal ... injection of pentobarbital with the left, jugular vein and right. A. Carotis externa are dissected.

An arteriovenous shunt is formed by one central catheter and two lateral catheters. In the central part, the thread is introduced from natural white silk and the circulation is renewed after 20 minutes. After the staple arrest, the thread is carefully removed and weighed immediately. The wet weight of the silk is determined in advance so that the weight of the precipitate can be determined from the difference.

The test substance is administered 48 hours, 24 hours and 2 hours before the start of blood flow, by short circuit. .per orally, v. suspension in 10 ml / / kg arabic gum · concentration 5 ·%, control. receive only · 5% · solution of Arabian gum,

The results showing the weight of thrombus in mg are shown in Table IV below.

..... ............ TABLE IV

Antithrombotic efficacy

<td>Group</td><td>Dose mg / kg</td><td>Method of administration</td><td>Thrombus weight (mg)</td><td>% Change</td>

<td>control</td><td></td><td> —</td><td> 42,65 ± 3,30</td><td></td>

<td>derivative · · No. 3</td><td>3 x 200</td><td> —,</td><td> 2,60 ± 0,24</td><td> —94</td>

<td>control</td><td></td><td> —</td><td> 36,24 ± 2,05</td><td></td>

<td>derivative · -no. 1</td><td>3 x 25</td><td> —</td><td> 6,56 ± 0,51</td><td> —82</td>

<td>derivative · No. 1</td><td>3 x 12.5</td><td> —</td><td> 15,98 ± 1,81</td><td> —56</td>

<td>control</td><td></td><td> —</td><td> 40,86 ± 2,02</td><td></td>

<td>derivative · No. 1</td><td>3x5</td><td> —</td><td> 27,7 ± 2,82</td><td> —32</td>

<td>control.</td><td></td><td> — </td><td> 40,68 ± 1,74</td><td></td>

<td>derivatives. 2</td><td>3 x 200</td><td> —</td><td> 8,42 ± 3,28</td><td> —79</td>

246092

Toxicological and pharmacological tests confirm the low toxicity of the compounds of formula (I), which are also well tolerated and have a strong inhibition of platelet aggregation, moreover having antithrombotic activity, so that they can be used in human and veterinary medicine.

The pharmaceutical compositions containing these agents can be for oral administration and are in the form of tablets, dragees, capsules, drops, granules or syrups. They can also be administered rectally in the form of suppositories or parenterally in the form of injectable solutions.

Each unit dose preferably contains from 0.005 to 0.250 g of the compound produced by the process of the invention, the daily dose may be in the range of 0.005 to 1.00 g, depending on the age of the patient and the severity of the disease.

The following are examples of useful pharmaceutical compositions.

1. Tablets

Tablets may be prepared from the following mixture: The active compound in an amount of 0.050 g is mixed with excipients, for example lactose, sugar, rice starch, alginic acid or magnesium stearate.

2. Dragee

Dragee can be obtained by mixing 0.100 g of the active ingredient with excipients such as magnesium stearate, corn starch, gum arabic, shellac, sugar, glucose, white wax, carbonic wax, paraffin and the like.

3. Capsule

Capsules can be obtained by mixing 0.100 g of active ingredient with excipients such as magnesium stearate, corn starch or lactose.

4. Solution for injection

An injectable solution may be obtained by mixing 0.075 g of the active compound with the isotonic solution to a final volume of 3 ml.

5. Cípky

Suppositories can be obtained by mixing 0.100 g of active ingredient with semisynthetic triglycerides in an amount sufficient to produce 1 suppository.