DD158397A5 - PROCESS FOR PREPARING ISOPHTHALSAEUREPICOLYLAMID MONOHYDRATE - Google Patents

Abstract

N, N'-bis (3-picolyl) -4-methoxy-isophthalamide monohydrate in crystalline form is a stable chemical compound and has high pharmaceutical activity for the treatment of various thromboembolic disorders.

Description

Berlin, 31. 8. 81 AP C 07 D / 229 409/5

Process for the preparation of isophthalic acid picolylamide monohydrate _ _i

Field of application of the invention

The invention relates to a process for the preparation of N, N'-bis (3-picolyl) -4-methoxyisophthalamide monohydrate and to pharmaceutical compositions (preparations) containing it

According to the invention, a novel organic molecule is prepared in the monohydrate crystal form which has a pharmaceutical activity which counteracts the aggregation of the platelets, combats thromboembolic disorders in the blood and delays the clumping of the blood,

Charat eri stic hang the known technically en Solu

It is known that N, N'-bis- (3-picolyl) -4-methoxyisophthalamide (hereinafter referred to as "picotamide" according to the international name) is a compound having a high fibrinolytic and anticoagulant activity (see FR-PS 2 100, 850, "Chimie Therapeutique", (5, 203-207, 1971) and also has good platelet anti-aggregation activity (see, U.S. Patent 3,973,026, Age and Aging, 7, 246, 1978).

Picotamide, as described in the above publications and patents, is known in the anhydrous form, and its melting point is as indicated in FR-2 100 850, in the Kofler block at 124 ⁰ C.

This compound is used in the abovementioned

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th patent procedures specified by

Crystallization of the crude product / containing an amount of impurities in anhydrous and apolar organic solvents.

The crystallization of the crude picotamide, as it is obtained in the synthesis reaction of anhydrous and apolar organic solvents such as benzene, results in a powdery product, which under the microscope the characteristics of a fibrous substance, such as. B. voluminous down, shows. This powder easily absorbs an electrostatic charge and is relatively unstable. These properties are particularly disadvantageous when the compound is prepared for pharmaceutical use. The electrified particles repel each other and volatilize as they are weighed and introduced into the device for producing the various pharmaceutical preparations, resulting in a change in particle weight, thereby making titer stabilization difficult.

aim the invention

The aim of the invention is to provide a stable N, N'-bis- (3-picolyl) -4-methoxyisophthalamide, which is easy to handle and precisely metered.

Explanation of the essence of the invention

The invention has for its object to produce a crystalline monohydrate of this compound.

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It has now been found that N , U'-bis (3-picolyl) -4-methoxy-isophthalamide monohydrate is obtained by reacting a functional derivative of 4-tiethoxy-isophthalic acid with 3-picolylamine and crystallizing the crude product in an aqueous solution Having such characteristics of chemical-physical structure and stability that it is clearly superior to the already known anhydrous picotamide. "

It has also surprisingly been found that the

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picotamide monohydrate is more active and more effective than anhydrous picotamide, both from a pharmacodynamic standpoint and clinical pharmacology,

The subject of the present invention is therefore N, N'-bis (3-picolyl) -4-methoxy-isophthalamide monohydrate [alpha] n ^ A ^ .H ₂ O having a molecular weight of 394.4.

The conventional chemical formula can be given as follows:

CO-NH-CH ₂ -,

The picotamide monohydrate according to the invention is a white, odorless, bitter-tasting crystalline powder which is stable in the air and readily crystallizable in water and has a melting point on the Kofler block of 95 to 97 ° C.

For the sake of simplicity, this compound will be referred to in the following description as "picotamide monohydrate ¹¹ .

The compound of the invention differs from the physico-chemical point of view of the already known compound by an unpredictable and clear improvement in its stability. This improvement is due to the fact ρ that the Molekülhydratationswasser on the molecular ^">: participates in structure ^I of the Invention according en connection, since it is arranged in the crystal lattice in a precisely defined position, wherein the oxygen atom of the water of hydration well identifiable hydrogen bonds with specific atoms as indicated below, to give a single crystal of the compound having well-defined properties, these properties affect, in an unpredictable manner, the pharmaceutical behavior of the novel compound and its bioavailability in mammalian organisms at a faster rate Absorption when administered to this class of animals including humans.

Not only does the picotamide monohydrate according to the invention in the novel crystalline form not only surprisingly avoid the abovementioned disadvantages of the anhydrous picotamide, but in a still more surprising manner yields a compound which is more active and effective in terms of its pharmacological usefulness, in view of the beneficial effects of which are found on administration to animal and human organisms.

Although a well-founded theoretical explanation for

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This test result is not yet available, it can be assumed that a solution of the crystalline compound of the invention in water follows a different mechanism than a solution of the known anhydrous compound in amorphous form in water.

Another object of the invention is a process for the preparation of the novel hydrate compound (hydrate molecule),

The invention also provides a pharmaceutical composition containing the novel monohydrate compound (the monohydrate molecule) in various suitable pharmaceutical forms as an active ingredient for the clinical treatment of thromboembolic disorders of the blood.

As indicated above, the melting point of picotamide monohydrate is 95 to 97 C,

In this connection, it should be noted that, in contrast, anhydrous picotamide has a melting point of 124 C. This difference between the melting points of the two compounds is already an indication of the different molecular structure, which brings about the essential difference between anhydrous picotamide and picotamide monohydrate »

Exporting run c \ s at play

The invention will be explained in more detail below with reference to the accompanying drawings, in which:

1 shows a three-dimensional representation of the molecule of picotamidone nonohydrate obtained from an X-ray spectrum;

FIG. 2 is a three-dimensional representation of the monocrystal of picotamide monohydrate; FIG.

Fig. 3 is the same single crystal as shown in Fig. 2, in more detail; and

Fig. 4 is a graph showing a direct comparison between the platelet anti-aggregation activity of picotamide monohydrate and anhydrous picotamide.

The structure of a crystal of picotamide monohydrate, apart from the physical and chemical analysis, is also characterized by the X-ray spectrum of its monocrystal. '

The data of the X-ray diffraction spectrum showing the spatial arrangement of the atomic centers in the new molecule in its entirety as a dense, stable and non-hygroscopic crystal are listed in the following Table I, in which the various atoms of the molecule are represented by their chemical symbols and an identification number subsequently specified is displayed. The specific arrangement and identification number of these atoms can be seen from Fig. 1, which shows the geometric three-dimensional position of the atomic centers, as shown in Table I.

Table I

Space Coordinates of Picotamid Monohydrate (X-Radiation) Maximum = 92.52 Minimum »-86.27 Multiplied by 167.8829

Projection atom height X / A

0.1895 0.1967 0.2670 Oi2047 0.2299 0.2944 0.2164 0.1627 0.2795 0.135 0.2847 0.1665 0.2547 0.3600 0.3326 0.1082 0.3771 0 , 4250 -0.0616 0.5422 0.5202 0.0384 0.0004 0.4606 -0.0875 0.0127 0.4483 -0.0519 0.5071 96.0 0.0376 92.0Q5364 84 _f OQ2173 84, OQ3392 83,0Q3204

number 08 1 C38 2 Cl 3 C3 4 C5 5 C6   6 C7 7 09 8th ClO 9 Nile 10 012 11 C13 12 C14 13 C15 14 N19 15 C22 16 027 17 C221 18 N222 19 C223 20 N224 21 C227 22 C228 23 C229 24 C231 25 C232 26 C234 27 C235 28 C244 29 Q 1 30 Q 2 31 Q 3 32 Q 4 33 Q 5 34

Y / B

0,6286 0,7283 0,5074 0,4937 0,4539 0,4562 0,5904 0,3432 0,6035 0,4642 0,3716 0,4277 0,6442 0,4594 0,5077 0,3996 0, 7056 0.4199 0.3513 0.3476 0.3510 0.3880 0.3668 0.3864 0.3574 0.3932 0.414 0.3732 0.3758 0.3935 0.3978 0.3847 0.3922 0 , 6259

Mo- Er-Z / C S.OeF. le- _hökül hung

0.8386, 1.0000 0.8919 1.0000 0.3663, 1.0000 0.6312 1.0000 0.4720 0.2040 0.6832

0.6819 0.4219 0.8824 0.1698 0.7347 0.5815

-0.0529 0.1101 1.0097 0.1461 0.0573 0.9680 0.2266 0.0056 0.9075 1.0498

-0.0822 0.7563 0.6853

χ * \ ν · / ν ** \ s

1,0000 1,0000 1,0000 1,0000 1,0000 1,0000 1,0000 1,0000 1,0000 1,0000 1,0000 1,0000 1,0000 1,0000 1,0000 1,0000 1, 0000 1,0000 1,0000 1,0000 1,0000 1,0000

vu ^^ jl. * ν \ _ / »_ / vx

0,2819 1,0000

0,6042 1,0000

0.3636 1.0000

0.5361 1.0000

0.3476 1.0000

0.4186 1.0000

-0.0552 1.0000 0.3130

1.0000

1 1.26 1 0.27 1 0.82 1 1.60 1 "L, 44 1 0.75 1 1.11 1 2.75 1 0.35 1 2.43 1 1.24 1 2.31 1 0.51 1 0.11 1 0.21 1 3.28 0 0.00 1 1.34 1 2.34 1 3.43 1 2.34 1 2.47 1 2.96 1 1.28 1 1.23 1 1.33 1 2.47 1 0.72 1 3.53 1 0.88 1 3.48 1 1.70 1 0.51 1 0.00

Bonds (comprising the symmetrically arranged atoms)

1-2 1 , 44 3-5 1.40 4-5 1.39 3-6 1 50 6-11 1 , 22 4-12 1.51 8-12 1.23 10-12 1 35 14-15 1 , 49 10-16 1.48 14-18 1.54 20-21 1 , 37 18-24 1 40 21-24 1.39 19-25 1.33 22-26 1 , 38 20-29 1 , 36 27-29 1.39 26-30 1.21 20-31 1 , 08 18-33 1 , 89 9-34 1.31

In particular, it can be stated that the atom no. 17, which is indicated as 027, which represents hydration water, while the atom No. 8, indicated as 09, is an oxygen atom of the methoxy group, the atom No. 15, indicated by N 19, is a nitrogen atom of the amide group, and the atom No. 21, designated N224, is a nitrogen atom of the pyridine group.

In Table I, the symbols X / A, Y / B and Z / C represent the space coordinates of the various atoms.

As can be seen from Fig. 1 showing the structure of picotamide monohydrate, the oxygen atom of the hydration water in the crystal lattice is located in a well-defined position in relation to the picotamide molecule.

From FIG. 2 it can be seen that the oxygen atom of the water of hydration inside the monocrystal, represented by a rectangle, is bound by hydrogen bonds.

- ίο -

shown by a dashed line) is bound to well-defined atoms of different picotamide molecules, wherein the molecules are arranged in an ordered three-dimensional lattice and interconnected by the hydrogen bonds with the hydration oxygen atom.

This binding is shown in Fig. 3 more in _detail? from which it can be seen that the oxygen atom 027 of the water of hydration is bound via hydrogen bonds in each case to:

(1) The oxygen atom of = C0 of the methoxy

group (Ο ") of a first picotamide holeküls (bond length 2.81 Ä) J

(2) the nitrogen atom of the amide group = NH (N, ^) of a second picotamide molecule arranged in the same plane as the above-mentioned molecule (bond length 2.96 X);

(3) the nitrogen atom of the pyridine ring ( ^N ₂ 24 ^ of a third picotamide molecule, which is located in the underlying or in the overlying plane, based on the other two bound molecules ^ (bond length 2.80 S).

The presence of these three bonds explains both the firmness with which the water of crystallization is trapped between different picotamide molecules forming the crystal and the compactness (strength) of the crystal itself. This compactness, evoked by the molecule water of crystallization, is explained as the reason

for the improvement of the bioavailability of the monohydrate form compared to the hitherto known anhydrous form, which, as will be shown in more detail below, from a pharmacological point of view results from a comparison between the absorption time, the blood content and the activity of both. ^: Medicines «

On the basis of previous knowledge of the Techv technology on the anhydrous Picotaraid molecule namely achieved by the introduction of a water of crystallization improvement effect in terms of compactness (density) of the spatial structure of picotamide and the improvement in terms of a significantly better bioavailability by no means -vorhersehbar.

The chemical elemental analysis also gave results which correspond to the structure explained above:

J Elemental analysis for C ₉₁ H ₉ N, 0 ~ .H ^ O (molecular weight 394.4)

C 63.94 H 5.62 N 14.20 0 Difference

gef. 63,87 5,72 14,18 % difference

The process for the preparation of the novel picotamido-anohydrate molecule follows, as far as the synthesis of the compound is concerned, the already known process for the preparation of anhydrous picotamide.

However, when the r.ohe picotamide has been obtained, so

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is surprisingly found to be obtained by recrystallization of the crude product from an aqueous mixture as opposed to an anhydrous mixture _v as in the already known process, the crystalline monohydrate product as defined above »

The following examples illustrate the process for preparing the inventive picotamide monohydrate from 4-methoxy-isophthaloyl dichloride or another functional derivative of 4-methoxy-isophthalic acid in the presence of proton acceptors.

Preparation example

4-methoxy-isophthaloyl dichloride (molecular weight 233) 100 g (0.43

3-picolylamine (molecular weight 108) 130 g (1.2

moles)

Triethylamine 120 ml

Tetrahydrofuran (anhydrous) 120 + 200 ml

The 3-picolylamine, the triethylamine and 120 ml of anhydrous tetrahydrofuran were introduced into a 3 liter flask equipped with a reflux condenser, a dropping funnel and a mechanical stirrer.

The 4-methoxyisophthaloyl dichloride was dissolved separately in ml of anhydrous tetrahydrofuran. The solution was slowly introduced through the dropping funnel into the reaction mixture contained in the flask with stirring. The

should be done within one and a half to two hours, with the following exothermic reaction takes place;

COCI

+2

COCI

OCH,

II

III

After this addition of dichloride, the reaction mixture was refluxed for about 2 hours, diluted slowly with water to 2 liters and stirred until a crystalline slurry consisting of crude picotamide passed away.

This slurry was separated on a suction filter and crystallized in a wet state into 700 to 800 ml of an acetone / water mixture (6 parts by volume of acetone + 11 parts by volume of water). The product thus obtained was recrystallized from water to obtain picotamidine nonohydrate having a melting point on the Kofler block of 95 to 97 ° C.

The above-described process of the present invention is characterized in that the recrystallization of crude picotamide as obtained in the synthesis reaction is carried out by using an aqueous solvent unlike the prior art in which

anhydrous and apolar organic solvents, such as benzene, which yield an anhydrous product. ,

Pharmacological tests

It has been found that ₉ Picotamidmonohydrat has a high activity as platelet anti-aggregation agent, and a fibrinolytic agent. It is therefore useful in clinical pharmacology and human therapy.

These activities were tested in vivo by spectrophotometric determination of Born's platelet anti-aggregation activity and Fearnley's blood clot dissolution test to determine fibrinolytic activity.

example 1

Platelet anti-aggregation activity in vivo in rabbits'

New Zealand rabbits, which had been fasted for 12 hours on water intake, were anesthetized with a 20% ethanol solution of urethane at a dose of 0.6 ml / 100 g intraperitoneally. Blood was withdrawn from the carotid artery before (control) and 1 1/2 hours after intraperitoneal injection of picotamide monohydrate at a dose of 25-50-100 mg / kg. The blood samples were made coagulatable by a 3.8% solution of sodium citrate in a 9: 1 volume ratio and then centrifuged for 15 minutes at 1000 rpm,

to obtain platelet-rich plasma (PRP). A part of this plasma was then centrifuged at 8000 rpm for 10 minutes to obtain a platelet-poor plasma (PPP).

The PPP was used to set the zero point of a Born aggregometer and 1 ml of PRP was introduced into the reservoir of the measuring apparatus. Platelet aggregation was induced by disodium adenosine diphosphate (ADP) at concentrations that varied with platelet responsiveness. Platelet anti-aggregation activity was calculated as a 50% inhibition of the aggregation curve after treatment, relative to that of control (IDc _n ). The results obtained are shown below.

Example 2

Time effect on platelet aggregation and levels of the drug in the blood in dogs

Picotamide monohydrate was administered orally at a dose of 100 mg / kg to Beagel dogs 0 which were not fed any amount of water intake for 18 hours.

Blood was withdrawn before (control) and 2-4-6-8-10 hours after the treatment and, depending on the time, the platelet anti-aggregating activity (using the method described above) and the levels of the drug in the Blood determined ..

Blood levels were determined using UV spectrophotometry according to the following procedure: 5 ml of plasma obtained by centrifuging at 100 rpm for 10 minutes was added with 2 ml of concentrated HCl and placed on a 100 ° C water bath for one hour C hydrolyzed. It was cooled, taken up in 2 ml H ₂ O and filtered. The filtrate was made strongly alkaline by NH, OH and extracted with 30 mL CHCl 2. The chloroform layer, after drying, was extracted over anhydrous Na ₂ SO 4, with O, In H ₂ SO 4, to which was added 0,900 In H ₂ SO ₂ to 100 ml, and the spectrophotometer became 228 nm read. The results are given below.

Example 3

Fibrinolytic activity in vivo in guinea pigs

Fibrinolytic activity was determined in Italian guinea pigs after oral administration of picotamide monohydrate at a dose of 100 mg / kg. To carry out this test, the Fearnley method was used, which was modified as follows: In tubes kept at 0 C, 1.7 ml of phosphate buffer (pH 7.4) and 0.1 ml of a 50 NIH / well throbmin solution were introduced. After the addition of 0.2 ml guinea pig whole blood. The tubes were kept at 0 C for 3 minutes to cause agglomeration, then held on a 37 C water bath for 30 minutes to achieve dissolution. Then the blood clot weight was determined.

The fibrinolytic activity was calculated as a percentage of the decrease in the blood lump weight of the treated animals, based on the blood lump weight of the control animals.

Example 4

'Platelet anti-aggregation activity and fibrinolytic activity in humans (volunteers)

J A confirmation of the two activities in vivo of Pico tamidmonohydrat resulted in healthy people (volunteers) of both sexes in the age from 35 to 65 years who were treated orally with a single dose of 12 mg / kg.

The inhibitory effect (inhibitory action) of platelet aggregation was tested by using disodium ADP as an antagonist by the same method as in Example 1. The fibrinolytic activity was determined by determining the dissolution time of the euglobins. The results obtained are shown below.

Results

From the results obtained in the assay of platelet anti-aggregation activity in vivo in guinea pigs (Example 1) by probit analysis, the dose capable of inhibiting (inhibiting) platelet aggregation by 50% was calculated ) (ID '), and this dose was 54.10-l * 1.43 mg / kg.

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2Λ A / A H Γ

L. J Hr U%? "J

- 18 -

A maximum effect (53.82 %) in dogs (Example 2) was found in the fourth hour. The corresponding blood level was 22.6 yen / ml of plasma and corresponded to a maximum value, as indicated in the diagram of FIG.

The fibrinolytic activity in guinea pigs (Example 3) tested at a dose of 100 rag / kg orally was found to be 27.83 %. "

In humans (Example 4), at a single dose of 12 mg / kg orally, the maximum platelet anti-aggregation effect occurred 4 hours after treatment and was 81.4 %; the maximum fibrinolytic activity was determined as 54.2% decrease in the dissolution time of the euglobins.

Verqleichsbetrachtunqen

From the comparison of the activity and toxicity results for picotamide monohydrate with those for anhydrous picotamide, given in US Pat. No. 3,973,026, it can be seen that there are substantial differences in activity and that these differences are for the monohydrate form are favorable, which was certainly not foreseeable due to the fact that only one crystallization water molecule characterizing the new compound had been introduced,

For comparison purposes, a test was carried out with respect to the time

40

effect and blood levels in dogs when administered orally under the same experimental conditions using the already known anhydrous picotamide to determine its bioavailability after administration.

The results of this test are shown in the graph of FIG. 4 which shows the time effect on platelet aggregation and on the blood levels in dogs at a dose of 100 mg / kg p.o. shows.

The axis of ordinate indicates the time in hours and the axis of abscissa indicates the blood levels, measured in γ / ml of plasma, and the platelet anti-aggregation activity, measured in%. Lines 1 and 1 * represent the anti-aggregation activity of picotamide monohydrate and anhydrous picotamide, respectively, and solid lines 2 and 2 'represent the blood levels of picotamide monohydrate and anhydrous picotamide, respectively.

A review of the results presented in the graph shows that anhydrous picotamide gives a maximum platelet anti-aggregation effect in the eighth hour, which corresponds to 49 % , while the maximum blood content is 19.75 g / ml occurring in the sixth hour (timed is shifted).

By comparison, in picotamide monohydrate, the maximum platelet anti-aggregation effect occurs in the fourth hour and, moreover, the maximum activity decreases.

taktpeak coincidentally with the maximum peak of blood levels, indicating faster bioavailability of picotamide monohydrate compared to anhydrous picotamide.

The results of the pharmacological tests carried out with picotamide monohydrate are listed in the following Table II, which also gives the data of the same test with anhydrous picotamide.

Table II

test

Platelet aggregation in vivo (rabbit), intraperitoneally

Bone platelet aggregation in vivo (dog) 100 mg / kg p.o.

parameter

Platelet aggregation in humans after administration of a single dose

Picotamidmonohydrat

ID ₅₀ (mg / kg) 54.1

Maximum inhibition of aggregation {%) 53.83

maximum activity time found 4. hrs.

maximum blood content in the plasma (γ / ml) 22,6

found time of maximum blood content 4. Std.

maximum incarnation of aggregation 00 81.4

anhydrous · it picotamide

108.2

48.12

8.Std.

19.75

6th hour *

70.11

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~ 2i -

maximum activity time found 4 hours 8 hours

Acute toxicity in rats and

Dogs po _# DL ₅₀ (mg / kg)> 3 000> 3 000

A comparison between the test results are given in Table II for the two molecules, shows that Picotaraidmonohydrat improved fect pharmacological ef- '"\ over anhydrous picotamide has. Such unexpected improvement seems to improved bioavailability of the new Arnzeimittels in the form of the monohydrate crystal with a stable structure.

Therapeutic use

Picotamide monohydrate, because of its low toxicity, high tolerability, and lack of adverse side effects in human therapy for.) Can treat various thromboembolic disorders, especially cerebral vascular disorders, myocardial infarctions, arterial and flebothromboses, pulmonary embolism, general arteriosclerotic conditions, and in general cardiac surgery be used.

For the applications mentioned various pharmaceutical preparations can be used which contain 10 to 500 mg of the active substance (active agent), for which a few examples are given below:

(a) Oral: capsules, tablets, pills containing IO to 500 mg for a daily dosage of 50 to 3000mg / day;

(b) parenteral; sterilized intravenously injectable vials containing 10 to 50 mg for a daily dosage of 10 to 200 mg / day.

It can also be administered rectally in the form of suppositories.

The pharmaceutical compositions (preparations) according to the invention may, of course, in addition to the active ingredient (active ingredient) still contain conventional pharmaceutically acceptable excipients and adjuvants, as well known in the pharmaceutical field. It is also clear that the administration forms of picotamide monohydrate and the respective dosage regimens may be varied according to the clinical circumstances and experience of the physicians.

Claims (2)

31. 8. 81 AP C 07 D / 229 409/5 59 131 11 - 23 - Invention claim A process for the preparation of the N, N'-bis (3-picolyl) -4-methoxy-isophthalamide monohydrate in crystalline form of the formula CO-NH-CH ₂ -I CO-NH-CH OCH with a melting point, on the Kofler block, of 95 to 97 C and an X-ray diffraction spectrum of its monocrystal, as indicated in Table I in the preceding description, characterized in that a functional derivative of 4-methoxy-isophthalic acid in an anhydrous organic solvent is reacted with 3-picolylamine, the reaction product is precipitated by water and the crude product is recrystallized from an aqueous solution and then from water. 2. The method according to item 1, characterized in that the reaction product is crystallized after precipitation with water in an acetone / water solution and recrystallized from water.