WO1992022545A1 - New coumarin derivatives - Google Patents

Abstract

Bicoumarin derivatives prepared by the etherification of two 7-hydroxycoumarin radicals with mixed aliphatic or cycloaliphatic bivalent alcohols having formula (I). The invention includes pharmaceutically acceptable salts of these compounds. The bicoumarin compounds of the invention have anti-thrombotic and anti-hypertensive properties and can be used in therapy in connection with vascular pathologies, such as peripheral vascular pathologies, anginal afflictions and cerebral vascular pathologies.

Description

New coumarin derivatives

SUMMARY OF THE INVENTION

The present invention is directed to novel coumarin derivatives, and more precisely bicoumarin

derivatives derived from the etherification of two 7-hydroxycoumarin radicals with bivalent alcohols of an aliphatic or cycloaliphatic nature of a mixed character with regard to this series.

The new derivatives are represented by the

following formula I:

in which each of the substituents R₂-R₅ and R₇-R₁₀ represent hydrogen or a substituent chosen from

the group formed by:

- a halogen,

- a free or esterified carboxylic group,

- a free or esterified or etherified hydroxy group, - a lower alkyl or monocycloarylalkyl or lower

monocycloalkyl-alkyl radical, or a corresponding unsaturated radical, which can be substituted in the aliphatic portion by one or more free or

esterified or etherified hydroxy groups, or by oxo groups or by free or esterified carboxy

groups, and in the aryl portion by one or more lower alkyl groups or halogens or lower hydroxy or alkoxy groups, and in the cycloaliphatic part by one or more lower alkyl groups,

- a monocycloalkyl radical or a corresponding

unsaturated radical, unsubstituted or substituted by one or more lower alkyl groups, a

monocycloaryl radical, unsubstituted or

substituted by one or more lower alkyl groups or halogens or lower hydroxy or alkoxy groups,

- R₁ and R₆ represent an aza-alkyl or

aza-raonocycloalkyl or aza-monocycloalkyl-alkyl or aza-alkyl-monocycloalkyl or

aza-alkyl-monocycloalkyl-alkyl radical, or a corresponding unsaturated radical, with a maximum of 12 carbon atoms, and which may be interrupted in the carbon atom chain by the groups -NH-, -O-, or -S-, and/or may be substituted by free or

esterified or etherified hydroxy groups, or by oxo or by lower alkyl groups, or by free or

esterified carboxy groups. R₄ and R₉ may also represent these moieties, and

- -X- represents a bivalent hydrocarbyl radical

chosen from the group formed by an alkylene, monocycloaryl-alkylene or monocycloalkyl-alkylene radical or a corresponding unsaturated radical, which may be interrupted in the carbon atom chain by heteroatoms chosen from the group formed by -NH-, -O-, and -S-, or by a monocyclo-arylene or monocycloalkylene radical, and may be substituted in the aliphatic or cycloaliphatic part by one or more halogens or free or esterified or etherified hydroxy groups, or by lower amino, alkyl-, or dialkyl-amino groups or

C_5-6 alkylene amino groups, optionally interrupted by -NH-, -O-, or -S- groups, by oxo

groups or by free or esterified carboxy groups,

and in the aromatic moiety by one or more lower

alkyl or halogen groups or lower hydroxy or

alkoxy groups, and a monocycloalkylene radical,

unsubstituted or substituted by one or more lower alkyl groups or free or esterified or etherified hydroxy groups, or by amino, alkyl, or lower

dialkylamino groups, or by oxo groups or by free or esterified carboxy groups.

The novel bicoumarin derivatives of the invention have interesting pharmaceutical properties and can be used in therapy. The invention also encompasses the salts of said compounds, especially those with pharmaceutically acceptable acids or bases.

The present invention also encompasses pharmaceutical preparations containing one or more of the aforesaid bicoumarin derivatives and the

therapeutic use of such compounds. The invention is also directed to preparation methods for the novel compounds and their salts. Due to the close

relationship between the free compounds and salts

with regard to their pharmaceutical properties,

which consititute the basis of the present invention, in the following description whatever is

said of the free substances will be true also of

their salts, where the meaning does not

specifically exclude this possibility.

The new bicoumarin derivatives of formula I and their salts have an anti-thrombotic and anti-hypertensive action and can be used in different

vascular pathologies, for example peripheral vascular pathologies, anginal afflictions and cerebral vascular pathologies.

In the new compounds of formula I, the halogen atoms, both as R₁-R₅ and R₇-R₁₀ substituents and

as substituents of the aromatic radicals optionally present in these substituents or in substituent X, are preferably fluorine, chlorine and bromine

atoms, and of these preferably chlorine.

The R₂-R₅ and R₇-R₁₀ alkyl radicals and, with regard to the aliphatic moiety, the "lower"

monocycloaryl-alkyl and monocycloalkyl-alkyl

groups, or the corresponding unsaturated groups, have a maximum of 7 carbon atoms. Preference is given to alkyl groups having 1 to 4 carbon atoms and

alkenyl groups having 2 to 7 carbon atoms. All these groups can have straight or branched

chains.

The cycloaliphatic radicals R₂-R₅ and R₇-R₁₀ or those contained in such substituents are nionocyclic and have preferably from 3 to 7 carbon atoms in

the ring and more particularly from 5 to 7

carbon atoms.

Of the unsaturated hydrocarbyl radicals, both aliphatic and alicyclic, special mention should be made of those with only one double bond, comprising alkenyl groups and cycloalkenyl groups. In the

monocyclic alicyclic radicals, special mention

should be made of the cyclohexane derivatives,

comprising cyclohexyl and cyclohexenyl radicals.

R₂-R₅ and R₇-R₁₀ aryl groups are monocyclic and

therefore derive from benzene and can be

substituted in the aforesaid manner, that is with lower alkyl, or halogen or lower hydroxy or

alkoxy groups. The term "lower" employed herein and indeed generally in the present description is meant to refer to groups with a maximum of 7 carbon atoms. This is also true of the corresponding alkoxy or alkenyl groups. Such groups have especially a maximum of 4 carbon atoms and are preferably methyl or methoxy groups. The substituents of the aromatic groups are

preferably no more than 3. The aryl groups have

preferably a total of 9 carbon atoms. What is said herein about these aryl groups is true also of the substituting aliphatic groups, for example arylalkyl groups.

In the aza-alkyl, aza-monocycloalkyl, aza-monocycloalkyl-alkyl, aza-alkyl-monocycloalkyl or

aza-alkyl-monocycloalkyl-alkyl radicals, R₁, R₄,

R₆ or R₉ or in the corresponding unsaturated

radicals, any methylene or methyl group can be

substituted by the -NH-group and this group can

therefore be present both in the aliphatic moiety

and in the cyclic parts of the hydrocarbyl^'

radicals. These radicals can be interrupted also by other heteroatoms at other points of the

hydrocarbyl chain or by other -NH-groups. Preferably, oxygen or sulfur atoms can interrupt the chain in the cyclic radicals. Regarding the number of carbon atoms and possible double bonds present, apart from the

condition that such aza-hydrocarbyl groups can have up to 12 carbon atoms, preference is given to those moieties which correspond to the corresponding

hydrocarbyl groups mentioned above for the substituents R₂-R₅ and R₇-R₁₀. In particular, alkyl groups as

substituents of cyclic groups have preferably a maximum of 7 carbon atoms, especially from 1 to 4 carbon

atoms, and they are above all methyl groups.

Cycloalkyl groups have preferably from 5 to 7 carbon atoms and especially 6 carbon atoms. Such

groups can be interrupted by the -NH-group, and are therefore aza-cyclohexyl or aza-cyclopentyl groups, such as the piperidine and pyrrolidine groups and can be interrupted also by other heteroatoms, such as -NH-, -O-, and -S-, and can therefore be

piperazine, morpholine or thiomorpholine residues.

The bivalent -X-radical has, as an alkylene group, preferably from 1 to 8 carbon atoms, and can

however be interrupted or substituted by alicyclic or aromatic carbocyclic radicals. Preferably, in

this case, only one monocyclic radical is present, corresponding preferably to the radicals previously described as preferential forms of R₂-R₅

and R₇-R₁₀ substituents. The total number of carbon atoms of the -X-substituent can therefore be more

than 3, but preferably should not exceed 18 carbon atoms. This is true also in consideration of the

hydrocarbyl functions or groups which can substitute both the aliphatic moiety, and the aromatic or alicyclic moiety. These substituents are preferably those which have in part already been specified for the R₁-R₁₀ substituents and those specified hereafter,

especially with regard to esters and ethers. The -X-group can also represent a cycloalkylene radical, and these radicals preferably correspond to those described above where these are not substituents of the

alkylene group.

The modified functions optionally present in all the aforesaid radicals or present as the R₁-R₁₀

substituents themselves are esterified carboxy

groups, esterified or etherified hydroxy groups and alkyl-, or dialkyl-, or alkylene amino groups.

These functions may, however, be in free form, for example as unsubstituted amino groups.

The carboxy groups in free or esterified form are derived especially from the following acids: formic, acetic, propionic, butyric, trimethylacetic,

n-valerianic, capronic, succinic, phenylacetic,

benzoic, trimethoxybenzoic, chlorobenzoic,

alkylsulfonic acids containing from 1 to 4

carbon atoms, such as methanesulfonic acid or

arylsulfonic acid, especially those containing only one benzene residue, for example p-toluenesulfonic acid, of the inorganic acids, should be mentioned, for example, sulfuric acid or phosphoric acid.

Esterified carboxy groups are preferably those derived from monovalent or bivalent aliphatic

alcohols, saturated or unsaturated, with a maximum of 7 carbon atoms or from monoaryl-aliphatic

alcohols with a maximum of 9 carbon atoms.

Etherified hydroxy groups are preferably those also derived from saturated or unsaturated aliphatic

alcohols with a maximum of 7 carbon atoms or from monoaryl-aliphatic alcohols with a maximum of 9

carbon atoms, while esterified hydroxy groups are derived especially from carboxylic acids of the

aliphatic, araliphatic, aromatic or alicyclic

series having preferably from 1 to 9 carbon

atoms. In the alkyl and dialkylamines the alkyl groups have a maximum of 7 carbon atoms and

preferably between 1 and 4, in the alkyleneamino groups there are 5 or 6 carbon atoms and the

azacyclo-alkyl ring can be interrupted by other heteroatoms, in particular by the -NH-, -O-, and

-S-groups. Substituent amino groups are, for example, those derived from methylamine, ethylamine, propylamine, dimethylamine, diethylamine, pyrrolidine,

piperidine, piperazine or morpholine.

Of the aforesaid alkyl groups, both as substituents of the bicoumarin residue, and of the

aforesaid hydrocarbyl radicals, special mention

should be made of the following groups: methyl,

ethyl, propyl, isopropyl, n-butyl, isobutyl,

tert-butyl; of the alkenyl groups: vinyl, allyl,

propenyl, isobutenyl, 2-butenyl and 2 pentenyl.

Likewise, special examples of cycloalkyl groups

are: cyclopropyl, cyclopentyl, cyclohexyl,

cycloheptyl, and of cylcoalkenyl groups the

cyclopentenyl and cyclohexenyl groups.

Examples of aralkyl groups are benzyl, phenethyl, phenylpropyl and cinnamyl groups. Cycloalkyl-alkyl radicals are, for example, the cyclopentyl, cyclohexylmethyl, cyclopentylethyl and cyclohexylethyl groups and examples of the cycloalkenyl-alkyl groups are

2-cyclohexenyl-methyl or 2-cyclohexenylethyl groups.

Aryl groups, both as substituents of the bicoumarin residue, and of the aforesaid aliphatic

hydrocarbyl radicals are, for example, the phenyl, toluyl, di-and trimethyl-phenyl, ethyl-phenyl,

allyl-phenyl, chlorophenyl, bromophenyl, fluorophenyl, trichlorophenyl, monohydroxy-phenyl,

dihydroxy-phenyl or trimethoxy-phenyl groups.

Particularly valuable are the compounds of formula

I in which -X- represents an alkylene radical having from 1 to 6 carbon atoms, which can be

unsubstituted or substituted by one or two

functions chosen from the groups formed by free or esterified or etherified hydroxy groups, or free or esterified carboxy groups or oxo groups or free

amino groups or lower alkyl or dialkylamino groups or C_5-6-alkyleneamino groups, optionally interrupted by -NH-, -O-, or -S-groups, where the

derivatives of such functions are preferably those mentioned above.

The R₁-R₆ substituents may be identical or

different from the corresponding R₇-R₁₀

substituents. Of particular importance are novel

compounds of formula I wherein the molecule is

symmetrical, the two coumarin residues being mirror images of each other, that is, compounds in which R₁-R₆ are equal to R₇-R₁₀, respectively.

Among these compounds, special mention should be made of those with the formula:

in which

A represents a saturated aza-alkyl, aza-monocycloalkyl-alkyl or aza-alkyl-monocycloalkyl

radical with a maximum of 12 carbon atoms and in

which the cycloalkyl group has 5 or 6 carbon atoms, optionally further interrupted in the carbon atom

chain by one of the groups -NH-, -O-and -S-, and

which can be substituted at the carbon atoms by

alkyl groups with 1 or 2 carbon atoms or by lower hydroxy or alkoxy groups.

B, C and D may represent a hydrogen atom and B may also represent a lower alkyl or alkylene radical or a monocyclic aryl radical or a halogen atom, C

may represent the same aza-hydrocarbyl radical as

defined by the substituent A, or the same hydrocarbyl groups as defined for B, and D may represent the same hydrocarbyl radicals as defined for

B. The moiety -X- represents an alkylene radical having from 1 to 6 carbon atoms and which can be

interrupted in the carbon atom chain by heteroatoms chosen from the group formed by -NH-, -O-,

and -S-, and/or which may be unsubstituted or

substituted by one or two functions chosen from the group formed by halogens, hydroxy groups, free or

esterified with carboxylic acids having from 1 to

9 carbon atoms or etherified with alcohols with

1 to 7 carbon atoms, and carboxy groups,

free or esterified with alcohols having from 1 to

7 carbon atoms and oxo groups and lower alkylamino or dialkylamino groups.

In these selected derivatives the aryl groups may be substituted as in the case of the compounds of

formula I and especially by 1 to 3 methyl or alkoxy groups. In substituent A the cycloalkyl groups not containing nitrogen may be unsubstituted or

substituted by 1 to 3 alkyl groups with a maximum of 3 carbon atoms and likewise those containing the -NH-group. These hydrocarbyl substituents, like the hydroxy groups, are no more than 2. The azacyclic groups are, for

example, those specified above, in particular the

radicals derived from piperidine, piperazine,

morpholine or thiomorpholine. In the hydrocarbylamino groups of substituent -X-, these have preferably a maximum of 4 carbon atoms.

Among the aforesaid bicoumarin derivatives of

formula II according to the present invention, of

special interest are those in which B is one of the aforesaid nitrogen-free hydrocarbyl groups, C and D are hydrogen atoms or one of said nitrogen-free

hydrocarbyl groups or a halogen. In these compounds A is, for example, the diethylamino-ethyl or diisopropylamino-ethyl radical or optionally their

corresponding quaternary groups, obtainable for

example by reaction of the corresponding compounds having a tertiary nitrogen with lower alkyl halogenides, especially with methyl bromide. Moreover, A is especially the morpholino-methyl, piperidinyl-methyl, thiomorpholinyl-methyl, or piperazinyl-methyl radical. B is especially the methyl or

phenyl group, C and D represent hydrogen, allyl or chlorine and X is for example the trimethylene,

3-hydroxy-trimethylene, hexamethylene group or the

2-di-carbethoxy-trimethylene group.

Hereafter is a list of representative compounds of the invention:

1,3-bis[3-(ß-diethylaminoethyl)-4-methylcoumarin-7-yloxy]propane

1,3-bis[3-(ß-diethylaminoethyl)-4-methyl-8-chlorocoumarin-7-yloxy 3 propane

1,3-bis[3-(ß-diethylaminoethyl)-4-phenylcoumarin-7¬

-yloxy] propane

1,3-bis[3-(morpholinomethyl)-4-phenyl-6-chloro-8-methylcoumarin-7-yloxy]propane

1,3-bis[3-(morpholinomethyl)-4-methy1-6,8-diallylcoumarin-7-yloxy]propane

1,3-bis[3-(morpholinomethyl)-4,8-dimethyl-6-allylcoumarin-7-yloxy]propane 1,3-bis[3-(morpholinomethyl)-4-methyl-6-chloro-8¬

-allylcoumarin-7-yloxy]propane

1,3-bis[3-(morpholinomethyl)-4,8-dimethyl-6-chlorocoumarin-7-yloxy]propane

1,6-bis[3-(ß-morpholinoethyl)-4-methylcoumarin-7¬

-yloxy]hexane

bis[3-(ß-diethylaminoethyl)-4-methylcoumarin-7¬

-yloxy] carbethoxy methane

bis[3-(ß-diethylaminoethyl)-4-methyl-8-chlorocoumarin-7-yloxy]carbethoxy methane

bis[3-(ß-diethylaminoethyl)-4-ρhenylcoumarin-7¬

-yloxy] carbethoxy methane

bis[3-(morpholinomethyl)-4-methy1-6,8-diallylcoumarin-7-yloxy]carbethoxy methane

1,3-bis[3-(ß-diethylaminoethyl)-4-methyl-8-chlorocoumarin-7-yloxy]-2-hydroxypropane

1,3-bis[3-(morpholinomethyl)-4-raethyl-6,8-diallylcoumarin-7-yloxy]-2-hydroxypropane

1,3-bis[3-(morpholinomethyl)-4-phenyl-6-chloro-8-allylcoumarin-7-yloxy]-2-hydroxypropane

1,3-bis[3-(ß-morpholinoethyl)-4-methylcoumarin-7¬

-yloxy]-2-hydroxypropane

1,3-bis[3-(ß-morpholinoethyl)-4-methyl-8-chlorocoumarin-7-yloxy]-2-hydroxypropane

and the others listed in the following illustrative Examples.

Other specific compounds include the following:

1,4-bis[3-(ß-diethylaminoethyl)-4-methylcoumarin-7¬

-yloxy]cyclohexane

1,4-bis[3-(ß-diethylaminoethyl)-4-methyl-6,8¬

-diallylcouταarin-7-yloxy]-2,3-dimethylcyclohexane

1,3-bis[3-(ß-diethylaminoethyl)-4-methylcoumarin-7¬

-yloxy]-2-phenylpropane The novel bicoumarin derivatives described above may optionally be salified, if they possess basic or acid functions. It is thus possible to prepare, on the one hand, salts with organic or metal bases and, on the other hand, salts obtained by the addition of acids or alkyl or aryl halogenides or the corresponding sulfonic acids. Particularly important salts are

those which are therapeutically acceptable, and

the new compounds of the invention can be used in this form. The salts can also be derived from bases or from acids which cannot be used for therapeutic purposes and in this case they serve, for example, as intermediate compounds for the purification of the novel products of the invention.

The following are examples of therapeutically acceptable acids: hydrochloric acid, hydrobromic

acid, sulfuric acid, phosphoric acid and

methanesulfonic acid, malic acid, tartaric acid and succinic acid. Picric and picrolinic acid are

particularly suitable for the formation of salts

which may serve for the purification of the

compounds. The amino groups can therefore be

transformed into ammonium salts, in particular the tertiary amino groups can be transformed into

quaternary groups, especially quaternary ammonium groups, such as tetramethylammonium chloride or bromide. Of the metal salts special mention should

be made of alkali metal salts, alkaline earth or

magnesium salts, for example potassium, sodium,

ammonium and calcium salts, but also those with

organic bases, such as primary, secondary or

tertiary amines which are aliphatic or aromatic or heterocyclic, such as methylamine, ethylamine, propylamine, piperidine, morpholine, ephedrine, fur furylamine, choline, ethylenediamine or aminoethanol.

The antithrombotic and antihypertensive activity of a compound in view of a possible application in

vascular pathology, also in hypertensive subjects, can be deduced from in vitro and in vivo experiments according to the following criteria:

1. effects on platelet aggregation in vitro

2. effects on certain leukocyte functions in vitro:

studies on the adhesion and activation of

polymorphonucleate leukocytes

3. effects in vivo in a model of arterial thrombosis in dog

4. effects in vivo in models of arterial hypertension in rat.

The platelet antiaggregating activity of the new products according to the invention can be demonstrated by the following study on human platelets in the presence of various aggregating agents: adenosine diphosphate (ADP), platelet activating factor (PAF) and arachidonic acid (AA).

The study was performed with the following

bicoumarin derivatives.

1) 1,3-bis[3-(ß-diethylaminoethyl)-4-methylcoumarin-7

-yloxy]propane hydrochloride

2) bis[3-(ß-diethylaminoethyl)-4-methyl-coumarin-7-yloxy]dicarbethoxy methane hydrochloride

3) bis[3-(ß-diethylaminoethyl)-4-methyl-coumarin-7-yloxy]carbethoxy methane hydrochloride

4) 1,3-bis[3-(ß-diethylaminoethyl)-4-phenylcoumarin-7

-yloxy]propane hydrochloride

5) bis[3-(ß-diethylaminoethyl)-4-methyl-8-chlorocoumarin-7-yloxy]carbethoxy methane hydrochloride 6) bis[3-(ß-diethylaminoethyl)-4-methyl-8-chlorocoumarin-7-yloxy]dicarbethoxy methane hydrochloride

7) 1,3-bis[3-(morpholinomethyl)-4-methyl-6,8-di- allylcoumarin-7-yloxy]-2-hydroxypropane hydrochloride

8) 1,3-bis[3-(morpholinomethyl)-4,8-di-methyl-6-allylcoumarin-7-yloxy]propane hydrochloride

9) 1,3-bis[3-(morpholinomethyl)-4-methyl-6-allyl-8-chlorocoumarin-7-yloxy3-2-hydroxypropane

hydrochloride

10) 1,3-bis[3-(ß-diethylaminoethyl)-4-methylcoumarin- 7-yloxy3-2-hydroxypropane hydrochloride

11) 1,3-bis[3-(ß-diethylaminoethyl)-4-methyl-8-chlorocoumarin-7-yloxy3-2-hydroxypropane

hydrochloride

The products were εolubilized in saline solution (10μl) and added 1 minute before the aggregating agents. Concentrations ranging between 1-100 μM were tested.

Two specific antagonists for the PAF receptor were also tested, namely: L - 652,731 (Merck Sharp

& Dohme) and SRI 63-675 (Sandoz Res. Inst.). Platelet aggregation

Blood was drawn from healthy volunteers who had taken no drugs for at least two weeks. It was

gathered on 3.8% citrate in a ratio of 9:1. Platelet rich plasma (PRP) was obtained by centrifugation at 190 rpm for 15 minutes at room temperature. Aggregation was induced by Bom's method

(Born G. V. R.: Aggregation of blood platelets by adenosine diphosphate and its reversal. Nature 194: 926-927, 1962), using an Elvi Logos 840 aggregometer.

The platelets were stimulated in the aggregometer with threshold concentrations of aggregating agent (TAC) so as to induce irreversible aggregation.

(The threshold concentration is defined as the lowest concentration at which the biphasic wave has at least 60% optic density variation).

TAC was therefore calculated after incubation (37ºC for 1 minute) of 250 μl of PRP while being

constantly stirred at 1000 rpm and subsequent

stimulation with the aggregating agents (ADP, PAF, AA at concentrations of 1-10 μM, 100-500 nM and

0.7-3 mM respectively).

Aggregation was then monitored for 3 minutes- The compounds were added to the PRP so as to obtain micromolar concentrations. 250 μl of PRP were

incubated with the test products (37ºC for 1 minute while being stirred) and then stimulated with the aggregating agents.

Threshold inhibition concentrations (TIC) were assessed for each compound, that is, the lowest concentration able to inhibit aggregation induced by TAC of the various aggregating agents. Results

The data obtained (Table 1) show that the test products were efficient platelet antiaggregating agents from concentrations of 0.1-2.5 μM. They were active against all the aggregating agents tested (unlike other antiplatelet drugs) . Table 1 :

Effect of the bicoumarin derivatives on aggregation induced by various aggregating agents on human platelets. Values expressed as minimum concentrations (μM) able to inhibit aggregation induced by TAC of the aggregating agents.

Compound ADP PAF AA

1 2.5 2.5 2.5

2 / 0.1 0.1

3 / 0.5 50

4 20 15 50

5 15 1 75

6 35 1 75

7 1000 10 1000

8 / 0.5 50

9 50 0.5 50

10 50 5 500

11 50 2 >400

L-652, 731 50 0.5 50

SRI 63 -675 / 0.5 / (data are means of 3 or 4 replications for each

experiment. The results were comparable to at least two other experi- ments, performed on platelets from two different donors) .

The effects of the bicoumarin derivatives on some leukocyte functions can be studied in vitro by

measuring the adhesion and activation of polymorphonucleate leukocytes. Assessments relative to two specific functions of PMN are reported hereafter:

1. PMN adhesion to endothelial cells (EC)

2. production of superoxide anion by PMN

Materials and Methods

Test substance (solubilization and concentrations) Bicoumarin derivatives 1-4 of those previously

listed were tested.

The products were solublized in saline immediately before the experiment. The cells (PMN) were then

pretreated with increasing concentrations (0.1-500 μM) for 15 minutes at 37ºC.

Indomethacin, nordihydroguaiaretic acid, BN 52021, diltiazem, dexamethazone and Ibuprofen were used

for comparison.

Endothelial cells: preparation of the culture and treatment with interleukin-1 (TL-1)

Endothelial cells (EC) were isolated from umbilical cord and cultivated in culture medium 199 with a

supplement of 20% fetal calf serum, in the presence of 50 μg/ml of specific growth supplement (Sigma Chemical) and 100 μg of pig intestine heparin. The cells were used at confluence (eighth passage). In the experiments to assess adhesion, the growth medium was removed and the cells (1-1.5 × 10⁵ in a 2-cm² well) were washed once with 1 ml of Hank's balanced saline solution

(HBSS) and incubated at 37'C with 600 μl of 199

medium containing 0.25% of bovine serum albumin

(BSA, Sigma) in the presence or absence of IL-1

(purified human natural IL-1, Ultrapure IL-1,

Genzyme), 10 U/ml. After 4 hours at 37ºC the medium was removed, the cells washed twice with 1 ml HBSS

+ 0.25% BSA and used to test adhesion. PMN: preparation and labelling

Immediately before the experiment, PMN were removed from whole, anticoagulated blood taken from healthy volunteers, using dextran sedimentation and

Ficoll-Hypaque gradients (Zimmermann G. A. et al.:

Granulocyte adherence in pulmonary and systemic

arterial blood samples from patients with adult

respiratory distress syndrome. Am. Respir. Pis.

129: 798-804, 1984). The isolated PMN (95-100%)

(10⁷ cells/ml) were suspended in HBSS, in the

absence of Ca/Mg, containing HEPES 20 mM (pH 7.4) and labelled for 15 minutes at room

temperature with 5 μCi/ml 111 Indium-oxine

(Amersham). The cells were radiolabelled and then washed twice with HBSS, in the absence of Ca/Mg, containing 0.25% of BSA and resuspended in the same buffer.

PMN-EC adhesion test

Adhesion was assessed by incubating endothelial cells (in the presence or absence of IL-1) with

known quantities of labelled PMN (500 μl of

suspension of PMN at a final concentration of 1.5 × 10⁶ PMN/dish).

10 μl aliquots of CaCl₂ and MgCl₂ (final

concentrations of 1-1.5 mM, respectively) were added immediately and the cells were incubated for 15

minutes at 37ºC. At the end of incubation the upper phase was carefully removed, the dishes washed

twice with 1 ml of HBSS + 0.25% BSA to remove the non-adhered PMN, incubated for at least 10 minutes with 250 μl of NaOH M + 1% SDS (sodium-dodecylsulfate).

The radioactivity associated with the cells was assessed by means of a gamma counter. Superoxide anion production (O₂-) by stimulated PMN

Experiments were performed in two different conditions of PMN stimulation:

- PMN stimulated by TPA (12-o-tetradecanoyl¬

-phorbol-13-acetate)

- PMN stimulated by FMLP (N-formylmethionyl¬

-leucyl-phenylalanine).

The εuperoxide anion (O₂-) was measured by

cytochrome C reduction, according to the method described by Johnston (Johnston R.: Secretion of superoxide anion. Methods Stud. Mononucl. Phagocytes . : 489-497, 1981) modified by Del Maschio (Del Maschio et al. : Measurement of ionized cytoplasmic calcium mobilization with the photoprotein

aequorin in human polymorphonuclear leukocytes

activated by platelet activating factor (PAF).

Journal of lipid mediators 1, 25-36, 1989) and then assessed by a colorimetric test.

In brief, PMN reεuspended in HBSS (in the presence of Ca²⁺ and Mg²⁺) were preincubated at 22ºC for 5 min in the presence of cytochrome C (10 mg/ml) and then exposed to the agonist for 40 minutes. The cell suspension was centrifuged (1 minute) in

Eppendorf vials and aliquots of the upper phases

(200 μl) were transfered to a 96-well

dish. Absorbance was measured at 550 and 540 nm simultaneously with a Multiskan spectrophotometer (Titertek, Flow Laboratories, Scotland) and the reduction in cytochrome C was calculated in

relationship to the complete reduction in cytochrome C induced by dithionite. The PMNs were stimulated by 0.5 μg/ml of the tumor promotor 12-O-tetradecanoyl-phorbol-13-acetate (TPA Sigma) or by FMLP (10^-7-10^-8M).

Results

Preincubation of the PMNs with the bicoumarin derivatives tested was clearly effective in

reducing adhesion of these cells to the cultured endothelium (both in basal conditions and after exposure to a strong stimulant such as IL-1).

Moreover, the bicoumarin derivatives tested reduce superoxide anion production induced in human PMN's both by the chemotactic peptide FMLP and by the phorbol ester. In particular:

1. PMN adhesion to the endothelial cells (± IL-1) The data (Table 2) on the effect of preincubation of PMN with compounds 1 and 4 indicate that:

compound 1 proved to be active both on control endothelium and that treated with IL-1.

compound 4 induced total inhibition at -500 μM. 2. Superoxide anion production (O₂- ) by stimulated PMN

The data obtained indicate that

O₂- production from TPA-stimulated PMN (Table 3) was markedly inhibited after preincubation with compound 1 and compound 4

Comparison substances such as: indomethacin, nordihydroguaiaretic acid, BN 52021, diltiazem, dexamethasone proved inactive up to 500 μM.

O₂- production from FMLP-stimulated PMN (Table 4) was inhibited after preincubation with

compound 4. Table 2 :

Effect of bicoumarin derivatives on PMN adhesion to endothelial cells treated or not treated with IL-1

Inhibitors + IL-1 - IL-1

% of inhibition compound 1 0.1 μM 26 -7

0.5 14 -10

2.5 15 21

50 28 21

100 45 55

500 61 68 compound 4 500 μM 97 94

The endothelial cells were incubated (4 hrs at 37ºC) with medium containing 10 u/ml of IL-1 or the equivalent volume of saline, then washed and used for the adhesion test. The labelled PMN treated with saline or increasing concentrations of product (15 min at 37ºC) were incubated with the dndothelial cells (+ IL-1). PMN adhesion was assessed after 15 minutes at 37ºC (see Materials and Methods).

Table 3 :

Effect of the bicoumarin derivatives on TPA-induced superoxide production in polymorphonucleate leukocytes .

Inhibitors Conc. (μM) % of inhibition compound 1 1 50 compound 4 1.5 50

Indomethacin 500 0 nordihydroguaiaretic acid 500 0

BN 52021 500 0

Diltiazem 500 0

Dexamethasone 500 0

Ibuprofen 18 0

5555 100

The PMNs were activated with TPA (0.5 μg/ml) and the superoxides measured 40 minutes later (see Materials and Methods)

Data are means of 3 or 4 replications per experiment (as indicated in Table 1)

Table 4 :

Effect of the bicoumarin derivatives on superoxide production induced by FMLP in polymorphonucleate

leukocytes (expressed as % of inhibition) .

Compound μM FMLP 10-⁸M FMLP 10-⁷M

compound 4 1 μM 21% 10%

10 μM 100% 100%

50 μM 100% 100%

100 μM 100% 100%

The PMNs were incubated for 15 minutes at 37 º C with saline or increasing concentrations of the compound. FMLP ( 10 ^-7-10^-8M) was then added and the O₂- production was measured after 40 minutes (see Materials and Methods)

Data are means of 3 or 4 replications per experiment (as indicated in Table 1)

Effects in vivo of the bicoumarin derivatives

in an arterial thrombosis model in dog

Objective

The objective of this experiment was to assess the antithrombotic action of some bicoumarin derivatives in an acute model of peripheral thrombosis in dog. In particular, the experiment focused on the efficacy in inhibiting arterial platelet thrombus formation

(induced by critical stenosis of the femoral

artery) which is the cause of this thrombotic

process and consequent ischemia.

The pharmacological characterization of this model has already been described (Prosdocimi M. et al.:

Stenosis and vascular damage as a cause of

thrombosis in dog femoral artery.

Naunyn-Schmiedeberg's Arch. Pharmacol.: 338:

430-437, 1988). The close causal relationship

between arterial thrombosis and the onset of acute occlusion of various kinds of arteries including the coronaries makes it a very interesting model

(Davies M. J. et al.: Thrombosis and acute

coronary-artery lesions in sudden cardiac ischemic death. N. Engl. J. Med.: 310: 1137-1140, 1984). It is therefore highly predictive of the therapeutic application of new drugs in arterial thrombosis.

Materials and Methods

Test substances (solubilization and concentrations) Bicoumarin derivatives 1, 4, 5 were tested.

The products were solubilised in saline and administered intravenously (i.v.) in εingle doses (0.5 - 3 mg/kg). Comparison drugs were ASA, chlorpromazine,

Ketanserin, dazmegrel, heparin, dipyridamole, and prazosin (versus saline).

In vivo experiments

Male beagle dogs (weighing 9-12 kg) were used in a total of 79 experiments. The animals were

anesthetized with pentobarbital at an initial dose of 35 mg/kg i.v. followed by slow i.v. infusion to ensure constant anesthesia. All the animals were

artificially fanned with air to maintain a constant level of 3.5% CO₂ in the exhalant, which was

constantly monitored with a gas analyzer (LB2,

Beckman Instruments). Catheters were inserted into the femoral veins of both hindlimbs for

administration of anesthetic and drug and

withdrawal of blood samples (Gould USA). ECG was

monitored by subcutaneous electrodes and the

heartbeat with a cardiotachometer. These variables were recorded on a polygraph and visualized on a

screen (Battaglia Rangoni, Italy).

About 2 cm. of the femoral artery were isolated from the surrounding tissues. An electromagnetic

transducer was placed around the artery (2.0-2.5

mm) and the vessel was stenosed with a plastic tube (Lexan). The inner diameter of the tube measured 1.6-1.8 mm. Whenever this procedure failed

to cause the desired cyclic blood flow variations

(CBFV), the vessel was compressed for 10 minutes

with a clamp, as damage to the vessel endothelium favors thrombi formation. The cylinder was placed at the site of compression and in each case CBFV

was subsequently observed. The test products were administered 30 minutes after stabilization of the CBFV. The effects of the products were quantified as scores on a fixed scale of 0 to 4 (see Table 5) as described in the literature (Aiken J. W. et al.: Endogenous prostacyclin contribvites to the efficacy of a thromboxane synthetase inhibitor for

preventing coronary artery thrombosis. J.

Pharmacol. Exp. Ther. 219: 299-308, 1981).

Results

The results obtained (Table 5) indicate that the compounds tested reduce thrombi formation. A marked antithrombotic efficacy was observed in the case of compound 4 (100% responders after 0.5 mg/kg i.v.) and compound 1 (albeit to a slightly lesser degree: 90% responders after 3 mg/kg i.v.).

Table 5:

Antithrombotic effect of the bicoumarin derivatives

Compound Dosage AntithrombResponders No.

mg/kg otic score %

1 3 1.2 ± 0-3 90 10

5 3 0.8 ± 0.3 72 7

4 0.5 2.5 ± 0-3 100 12

ASA 10 2.5 ± 0.6 66 9

CHLORPROMAZINE 0.5 3.3 ± 0.3 100 5

KETANSERIN 0.25 4.0 100 5

DAZMEGREL 0.5 3.4 ± 0.2 100 9

HEPARIN -* 0.6 ± 0.3 28 7

DIPYRIDAMOLE 1.0 0.5 ± 0.2 25 4

PRAZOSIN 0.1 0 0 7

SALINE - 0 0 4

No. indicates the number of experiments

* corresponds to 50 I.U./kg

Score 0 = no effect

Score 1 = slight reduction in occlusion frequency

Score 2 - reduction in occlusion frequency

Score 3 = reduction in occlusion frequency and increase in minimum flow observed

Score 4 = no occlusion Effects in vivo of bicoumarin compound 1 in

models of arterial hypertension in rat

Objective:

The antihypertensive activity of compound 1 in two specific models of arterial hypertension was

assessed to confirm the results of the preliminary screening of the test product.

In particular, the effects of 1 were studied in the following experimental conditions:

1. model of hypertension induced by renal stenosis

(Goldblatt's model)

2. model of hypertension induced by

deoxycorticosterone acetate (DOCA).

Materials and Methods

Test: substance (solubilizations and concentrations)

Compound l was tested in comparison to papaverine. The products were suspended in 0.5% aqueous

tragacanth and administered orally (os) at a dose of 100 mg/kg/die (in a volume of 10ml/kg).

Model of hypertension induced by renal stenosis

(Goldblatt's model)

30 male Wistar rats (120-140 gr) were used, having been rendered hypertensive by means of stenosis according to the method described by Goldblatt et al. (Goldblatt H. et al.: Studies on experimental hypertension. I. The production of persistent

elevation of systolic blood pressure by means of renal ischaemia. J . Exp. Med., 59, 347-379, 1934).

After anesthesia, an incision was made in the

lumbocostal region and the left kidney was

displaced towards the abdomen. The renal pedunculus was exposed, the artery isolated and compressed with a clamp in the vicinity of the abdominal

aorta. The right kidney was removed through a

second incision. The wall was sutured, the skin incisions closed with clips and the animal was left to recover.

All animals were then treated i.m. with 30,000 UI of benzyl penicillin procaine and per os for 4

consecutive days with 100 mg/kg/day of compound 1 in parallel with papaverine.

Systolic blood pressure was measured on the 1st and 4th days, before and after treatment (1st and 4th hrs). (Only those animals with systolic blood

pressure of over 150 mmHg were included in the

study)

Model of hypertension induced by deoxycortisone

acetate (DOCA)

30 male Wistar rats weighing 90-120 g, were

rendered hypertensive by a method similar to that described by Green et al. (Green D. M., Saunders F. J., Wahlgren N., Craig R. L.: Self-sustaining,

post-DCA hypertensive cardiovascular disease. Am.

J. Physiol.: 170, 94-106, 1952). An incision was effected under an anesthetic on the left side of the abdominal wall, and the left kidney was removed. A 50-mg DOCA pellet was implanted s.c.. The abdominal wall was sutured, the skin incision closed with

clips and the animal left to recover. Drinking

water was substituted with a solution containing

0.8% of sodium chloride and 0.1% of potassium

chloride.

The animals then underwent treatment and systolic blood pressure readings, as previously described

(Goldblatt's model). Results

The data obtained show an interesting

anti-hypertensive activity of compound 1, in

particular:

1. Renal stenosis model (Goldblatt)

As reported in Table 6, compound 1 effectively reduces pressure values. This effect was observed on all readings made on the fourth day of the study. Papaverine, on the other hand, had only a modest, not significant, effect on all readings made after treatment.

2. Model of hypertension induced by DOCA

In this model too, treatment with compound 1 proved equally active, with a reduction in pressure values on the 4th day (Table 7).

Table 6:

Effect of repeated doses of compound 1 and papaverine on systolic blood pressure of rats rendered hypertensive according to Goldblatt's method

Group Treatment Mean systolic blood pressure (mmHg)

(10)* (mg/kg) per group on

Day 1 Day 4

Pre-dose 1 h 4 h Pre-dose 1 h 4h

1 Vehicle 183 178 178 174 175 171

2 comp. 1 (100) 196 204 197 152 125 133

3 Papaverine (100) 198 189 178 178 170 176 * in brackets the number of animals per group

Table 7:

Effect of repeated doses of compound 1 and papaverine on systolic blood pressure of rats rendered hypertensive by administration of DOCA/saline

Group Treatment Mean blood pressure (mmHg) per (10)* (mg/kg) group on Day 1 Day 4

Pre-dose 1 h 4 h Pre-dose 1 h 4 h 1 Vehicle 206 218 205 204 209 206

2 comp. 1 (100) 203 197 198 158 179 174

3 Papaverine (100) 201 184 191 186 193 203 * in brackets the number of animals per group

The novel bicoumarin derivatives of formula I of the invention can be prepared in the known way, and more precisely by a procedure which comprises treating a 7-hydroxy-coumarin of the formula:

in which R₁-R₅ are the same substituents as defined for formula I or groups convertible thereto or a metal salt, with a compound of the formula:

Z₁-X-Z₂ (IV) where Z₁ and Z₂ are the same or are different from each other, and each represents a reactive group with

respect to phenol etherification (alkylation) and -X- has the same significance as in formula I or it signifies a substituent convertible in the same, or treating

the aforesaid coumarin compound III with a compound of the formula:

Z ₁-X-Z₃ (V) where Z₁ and -X- have the same significance as in

the previous case and Z3 stands for a non-reactive group with regard to phenol etherification (alkylation) , but is convertible to a reactive group with regard to this reaction, or a hydrogen atom of the unsaturated H-C= structure of the terminal hydrocarbyl group

of -X- , and then converting Z₃ in the obtained

compound of the formula:

to a reactive group with regard to phenol

etherification, and treating the coumarin

compound obtained with a 7-hydroxy-coumarin of the of formula:

or one of its metal salts, in which R₆-R₁₀ have the same significance as the R₁-R₅ substituents of

formula III, but are not necessarily identical to

them, and converting the obtained bicoumarin

compound R₁-R₁₀ and -X- substituents, differing

from those corresponding to their significance in

the compounds of formula I, into substituents of

the same formula, and if desired, converting the

products obtained into their metal or acid addition salts, or into their quaternary ammonium salts.

The procedure which involves treating the compound of formula III with the compound of formula IV in

one single step is preferred when bicoumarin compounds of formula I having a symmetrical structure are to be prepared, in which substituents R₁-R₅ are identical to substituents R₆-R₁₀. In this case

stoichiometric quantities in a ratio of 2 to 1 of

compounds III and IV are used. To obtain compounds with an asymmetric structure, in which at least one of the substituents R₆-R₁₀ differs from the corresponding substituent among R₁-R₅, the

compound of formula III is etherified with the

compound of formula V, in which Z₃ is a

non-reactive group with respect to phenol

etherification, but convertible into a reactive

group. In a second step, when the Z₃ group has been converted into the reactive group, the coumarin

compound, obtained with the 7-hydroxy-coumarin of

formula VII, is etherified. To obtain asymmetrical compounds it is also possible to exploit the

different reactivity of groups Z₁ and Z₂ by

treating compound III with reactive compound IV,

etherifying in a first step in a stoichiometric ratio of about 1 to 1, or in higher ratios, that is, with an excess of compound III, to obtain substantially the compound of the formula:

which can be isolated from the reaction

mixture, and then itself etherified with a 7-hydroxy-coumarin of type VII, different from the compound of formula III. A reactive group with regard to phenol etherification, such as Z₁ and Z₂, may be a reactive functional group and as such it is essentially a

functionally modified hydroxy group, especially a

hydroxy group esterified with appropriate acids, known in the literature, for example with hydracids or with inorganic or organic oxygenated acids, for example sulfonic acids. The esters with the

hydracids are alkylene halogenides, especially

bromides, iodides and chlorides of the Br-X-Br

type, the two reactive groups optionally differing one from the other. Of the oxygenated esters should be mentioned the esters of sulfuric or sulfurous

acid, and especially the esters with alkyl- or

aryl-sulfonic acids, such as methanesulfonic or p-toluenesulfonic acid.

Some terminal epoxide groups present in the

-X-radical between the terminal carbon atom and the one next to it are also reactive groups which may react with the phenol group of 7-hydroxycoumarin to form the ether and a hydroxyl group at the

carbon atom next to the terminal one.

The phenol etherification reaction is effected under known conditions which depend above all on the

nature of the reactive groups Z₁ and Z₂ and on the fact that the 7-hydroxycoumarins themselves or

their metal salts are used as starting compounds, such as in particular the alkaline metal salts, especially sodium, potassium or cesium salts.

Starting with the εalts it is possible to etherify with the compound Z₁-X-Z₂, Z₁ and Z₂ being reactive functional groups, in a neutral organic solvent, such as an alcohol, an ether, for example ethyl or methyl alchol, or dioxane or tetrahydrofuran or a ketone, such as acetone or methyl ethyl ketone, or in amides, such as dimethylformamide, or

sulfoxides, such as dimethylsulfoxide, or

N-methylpyrrolidone, at room temperature or more and especially at a higher temperature, such as between 50º and 150ºC. In this case alkylene halogenides of the Br-X-Br type or the corresponding chlorides or the corresponding compounds having only one reactive group substituting the X are preferable.

Starting with the 7-hydroxycoumarins, the

etherification reaction is performed with said

reactive functional groups in the presence of a

basic compound, such as especially an inorganic

base, for example an alkaline hydroxide or an

alkaline carbonate, or a suitable azotated base, such as pyridine or collidine in one of the

aforesaid solvents. This is especially true when

said alkylene halogenides are used or also the

esters of oxygenated acids, such as alkyl- or

arylsulfonates.

A non-reactive Z₃ group substituting the alkylene -X- radical, but convertible into a reactive

group, is for example a hydroxyl group, free or

esterified with particular acids, such as

mono-esters of carbonic acid, such as the

benzyloxycarbonyl group. Once they have been

introduced into the 7-hydroxycoumarin molecule by the -X- radical, such groups can be converted into said reactive functional groups for example in the known way, e.g., in the case of the benzyloxycarbonyl esters by reduction in the known way and subsequent function conversion at the hydroxy group, for example by conversion into tosylate.

Z₃ may also stand for the hydrogen atom of an unsaturated structure of the terminal

hydrocarbyl group of the -X- radical. This

unsaturated structure may be converted into the

epoxide group in the known way, for example

by reaction with peroxides, such as hydrogen

peroxide or organic peracids, such as perbenzoic

acid or perphthalic acid, thus obtaining a group

capable of etherifying (alkylating or arylating) the phenol group in the 7-position of the coumarin compound. Conversion of said unsaturated compound in the epoxide compound can be effected preferably in a neutral organic solvent such as an ether or in an aromatic hydrocarbon, for example benzene or toluene.

After conversion of Z₃ into a reactive group, etherification of the second stage is; effected with a compound of formula VII under the conditions

described above.

In the bicoumarins obtained according to the

described procedure, R₁-R₁₀ groups and -X- radicals are converted, which are modifications of the groups intended for the desired compounds, for example functional derivatives of substituents

present in such groups, in the intended groups.

Thus, for example, free hydroxy or carboxy groups present in said substituents may be functionally

modified at the end of the procedure, for example esterified or etherified. This is also true for the R₁-R₁₀ substituents, wherever these represent

functional groups, for example carboxy groups.

Vice-versa, modified functional groups can be

converted into free functional groups. Amino groups can be alkylated and also converted into quaternary ammonium salts. All of these reactions can be

performed in the known way.

The bicoumarin compounds obtained by the aforesaid procedure can be converted into their salts. This can be done in the known way, for example by

preparing metal salts of compounds containing carboxy functions by treatment with set quantities of a hydroxide or alkaline carbonate or of an organic base, for example one of those mentioned above.

Compounds which present amino functions in the

alkylene -X- radical can be transformed into salts obtained by acid addition, for example those

mentioned above, or into quaternary ammonium salts. The invention also includes modifications of the aforesaid procedure, in which it is interrupted at any one stage or in which one starts with an

intermediate compound and the remaining steps are carried out, or in which the starting products are formed in situ.

Another object of the present invention is directed to pharmaceutical preparations containing as active substance one or more of the new bicoumarin

derivatives or their salts, and in particular those mentioned above. Such pharmaceutical preparations can be for oral, rectal, parenteral, local or

transdermal use. They can therefore be in solid or semisolid form, for example pills, tablets, gelatin capsules, capsules, suppositories, or soft gelatin capsules. For parenteral use it is possible to use those forms intended for intramuscular,

subcutaneous or transdermal administration, or

suitable for infusions or intravenous injections and can therefore be presented as solutions of the active compounds or as freeze-dried powders of the active compounds to be mixed with one or more

pharmaceutically acceptable excipients or diluents, convenient for the above uses and with an osmolarity compatible with physiological fluids. For local use, preparations in the form of sprays, creams or ointments for topical use may be employed, or suitably treated sticking plasters for transdermal administration.

The preparations of the invention can be administered to humans or animals. They contain preferably about 0.01 to 10% by weight of active component for the solutions, sprays, ointments and creams, and between 100% and preferably between 5 and 50% by weight

of the active compound for the preparations in

solid form. Doses to be administered depend on

individual needs, on the desired effect and on the chosen route of administration. An average daily

dose of 10-30 mg by the intravenous route or 100-400 mg by the oral route is usually advised for humans for the treatment of vascular pathologies (of a thrombotic or hypertensive nature). The following disorders

are particularly indicated in connected with the present invention: peripheral vasculopathies, arteriopathies of the lower limbs, anginal complaints and cerebral

vasculopathies.

The following Examples illustrate the invention:

EXAMPLE 1:

1,3-bis[3-(ß-diethylaminoethyl)-4-methylcoumarin-7¬

-yloxy]propane hydrochloride

70.0 g of AD 112 3-(ß-diethylaminoethyl)-4-methyl¬

-7-hydroxycoumarin are suspended in 500 ml of

ethanol and to this are added 28.5 g of 50% KOH in water. It is left to stand for 1 hour at 50ºC, then concentrated and vacuum-dried.

It is gathered with 500 ml of 2-butanone and 70.3 g of K₂CO₃ are added. It is heated while being

shaken, and then 25.7 g of 1,3-dibromopropane are added and it is left to react for 12 hours. The solvent is evaporated and the residue is

dissolved with 500 ml of ethyl acetate; it is then washed with a 1N solution of NaOH. The solvent is anhydrated and concentrated. The product is purified by chromatography with Prep LC System Waters, using as eluent a mixture of CHCl₃-CH₃OH-NH₄OH (30%) in a

gradient range of 95:5:0.2 to 70:30:0.2. The pure

fractions are concentrated and crystallized from

toluene.

The crystallized product is dissolved in ethyl acetate, anhydrated and treated with HCl in ethanol until a Congo red indicator change. It is filtered and crystallized from isopropyl alcohol. This is filtered and vacuum-dried, obtaining 42.2 g

of a compound to which elementary analysiε (C,H,N) and nuclear magnetic resonance spectroscopy

(protons) both attribute a structure of

1,3-bis[3-(ß-diethylaminoethyl)-4-methylcoumarin-7-yloxy]propane hydrochloride.

EXAMPLE 2:

1,3-bis[3(ß-diethylaminoethyl)-4-methyl-8-chlorocoumarin-7-yloxy3propane hydrochloride

70.0 g of 3-(ß-diethylaminoethyl)-4-methy1-7-hydroxy-8-chlorocoumarin are suspended in 500 ml of

ethanol and to this are added 25.4 g of 50-% KOH in water. It is left to stand for 1 hour at 50ºC and then concentrated and vacuum-dried.

It is gathered with 500 ml of 2-butanone and to

this are added 31.2 g of K₂CO₃. It is heated while

being shaken, and then to this are added 22.8 g of

1,3-dibromopropane and it is left to react for 12

hours. The solvent is evaporated and the residue is

dissolved with 500 ml of ethyl acetate. It is

washed with a 1N solution of NaOH. The solvent is anhydrated and concentrated. It is purified by

chromatography with Prep LC System Waters, using as eluent a mixture of CHCl₃-CH₃OH-NH₄OH (30%) in a gradient range of 95:5:0.2 to 70:30:0.2. The pure fractions are concentrated and crystallized from toluene.

The crystallized product is dissolved in ethyl acetate, anhydrated and treated with HCl in ethanol until a Congo red indicator change. It is filtered and crystallized from isopropyl alcohol. This is

filtered and vacuum-dried, obtaining 36.4 g of a

compound to which elementary analysis (C,H,N) and nuclear magnetic resonance spectroscopy (protons) both attribute a structure of

1,3-bis[3(ß-diethylaminoethyl)-4-methyl-8-chlorocoumarin-7-yloxy]propane hydrochloride. EXAMPLE 3:

1,3-bis[3-(ß-diethylaminoethyl)-4-phenylcoumarin-7¬

-yloxy]propane hydrochloride

63.5 g of 3-(ß-diethylaminoethyl)-4-ρhenyl-7-hydroxycoumarin hydrochloride are suspended in 500 ml of ethanol and to this are added 38.0 g of 50% KOH in water. It is left to stand for 1 hour at 50ºC and then concentrated and vacuum-dried.

It is gathered with 500 ml of 2-butanone and to this are added 23.5 g of K₂CO₃. The resultant is heated while being shaken, and then to this are added 17.1 g of

1,3-dibromopropane and it is left to react for 12 hours.

The solvent is evaporated and the residue is

dissolved with 500 ml of ethyl acetate. It is washed with a 1N solution of NaOH. The solvent is anhydrated and concentrated. It is purified by

chromatography with Prep LC System Waters, using as eluent a mixture of CHCl₃-CH-OH-NH₄OH (30%) in a gradient range of 98:2:0.2 to 85:15:0.5. The pure fractions are concentrated and crystallized from toluene.

The crystallized product is dissolved in ethyl acetate, anhydrated and treated with HCl in ethanol until a Congo red indicator change. It is lyophilized, obtaining 9.2 g of a compound to which elementary analysis (C,H,N) and nuclear magnetic resonance spectroscopy (protons) both attribute a structure of 1,3-bis[3-(ß-diethyl-aminoethyl)-4-phenylcoumarin-7-yloxy]propane hydrochloride.

EXAMPLE 4:

1,3-bis[3-(morpholinomethyl)-4-phenyl-6-chloro-8¬

-methylcoumarin-7-yloxy]propane hydrochloride

30.0 g of 3-morpholinomethyl-4-phenyl-6-chloro-7-hydroxy-8-methylcoumarin) are suspended in 500 ml of ethanol and to this are added 8.7 g of 50%

KOH in water. It is left to stand for 1 hour at

50ºC and then concentrated and vacuum-dried.

It is gathered with 500 ml of 2-butanone and to this are added 13.9 g of K₂CO₃. It is heated while being shaken, and then to this are added 7.8 g of 1,3-dibromoproρane (0.0389 mole) and it is left to react for 12 hours.

The solvent is evaporated and the residue is dissolved with 500 ml of ethyl acetate; it is

washed with a 1N solution of NaOH. The solvent is anhydrated and concentrated, and the product is crystallized from toluene. The crystallized product is dissolved in ethyl acetate, anhydrated and treated with HCl in ethanol until a Congo red indicator change. It is filtered and crystallized from CH₃OH-H₂O 90:10. This is filtered and vacuum-dried, obtaining 4.8 g of a compound to which elementary analysis (C,H,N)

and nuclear magnetic resonance spectroscopy

(protons) both attribute a structure of 1,3-bis[3-(morpholinomethyl)-4-phenyl-6-chloro-8-methylcoumarin-7-yloxy)propane hydrochloride.

EXAMPLE 5:

1,3-bis[3-(morpholinomethyl)-4-methyl-6,8-diallylcoumarin-7-yloxy]propane hydrochloride

20.0 g of 3-morpholinomethyl-4-methyl-6,8-diallyl-7-hydroxycoumarin) are suspended in 200 ml of ethanol and to this are added 6.3 g of 50% KOH in

water. It is left to stand for 1 hour at 50ºC and then concentrated and vacuum-dried. The resultant is gathered with 200 ml of 2-butanone and to this are added 7.7 g of K₂CO₃. It is heated while being shaken, and then to this are added 5.65 g of 1,3-dibromopropane and it is left to react for 12 hours.

The solvent is evaporated and the residue is

dissolved with 200 ml of ethyl acetate; it is

washed with a IN solution of NaOH. The solvent is

anhydrated and concentrated. It is purified by

chromatography with Prep LC system Waters, using as eluent a mixture of CH₂Cl₂-EtAc-CH₃OH in a

gradient range of 85:15:0 to 80:20:0.5. The pure

fractions are concentrated. The residue is gathered in ethyl acetate, anhydrated and treated with HCl in ethanol until a Congo red indicator change. It is filtered, washed and vacuum-dried, giving 12.4 g of a compound to which elementary analysis (C,H,N) and nuclear magnetic resonance spectroscopy

(protons) both attribute a structure of

1,3-bis[3-(morpholinomethyl)-4-methyl-6,8-diallylcoumarin-7-yloxy]propane hydrochloride.

EXAMPLE 6:

1,3-bis[3-(morpholinomethyl)-4,8-dimethyl-6-allylcoumarin-7-yloxy]propane hydrochloride

16.0 g of 3-morpholinomethyl-4,o-dimethyl-6-allyl-7-hydroxycoumarin are suspended in 200 ml of

ethanol and to this are added 5.4 g of 50% KOH in

water. It is left to stand for 1 hour at 50ºC and then concentrated and vacuum-dried. It is gathered with 200 ml of 2-butanone and to this are added 6.6 g of K₂CO₃. It is heated while being shaken, and then 4.85 g of

1,3-dibromopropane are added and the solution is left to react for 12 hours.

The solvent is evaporated and the residue is

dissolved with 200 ml of ethyl acetate; it is

washed with a 1N solution of NaOH. The solvent is

anhydrated and concentrated. It is purified by

chromatography with Prep LC System Waters, using as eluent a mixture of CH₂Cl₂-EtAc-CH₃OH 70:30:5.

The pure fractions are concentrated and crystallized from ethyl acetate.

The crystallized product is dissolved in warm ethyl acetate, anhydrated and treated with HCl in ethanol until a Congo red indicator change. It is filtered and washed in ethyl alcohol. This is filtered and

vacuum-dried, obtaining 5.0 g of a compound to

which elementary analysis (C,H,N) and nuclear magnetic resonance spectroscopy (protons) both attribute a

structure of 1,3-bis[3-(morpho-linomethyl)-4,8-dimethyl-6-allylcoumarin-7-yloxy]-propane hydrochloride. EXAMPLE 7 :

1,3-bis[3-(morpholinomethyl)-4-methyl-6-chloro-8¬

-allylcoumarin-7-yloxy]propane hydrochloride

20-0 g of 3-morpholinomethyl-4-methyl-6-chloro-7-hydroxy-8-allylcoumarin are suspended in 200 ml of ethanol and to this are added 6.4 g of 50% KOH in water. It is left to stand for 1 hour at 50ºC and then concentrated and vacuum-dried. It is gathered with 200 ml of 2-butanone and to this are added 7.9 g of K₂CO₃. It is heated while being shaken, and then to this are added 5.75 g of 1,3-dibromopropane and it is left to react for 12 hours.

The solvent is evaporated and the residue is

dissolved with 200 ml of ethyl acetate; it is

washed with a 1N solution of NaOH. The solvent is anhydrated and concentrated. It is crystallized

from acetone until a clean product is obtained.

The crystallized product is disεolved in ethyl

acetate, anhydrated and treated with HCl in ethanol until a Congo red indicator change. It is filtered and crystallized from ethyl alcohol. This is filtered and vacuum-dried, obtaining 6.7 g of a compound to which elementary analysis (C,H,N) and nuclear magnetic

resonance spectroscopy (protons) both attribute a structure of 1,3-bis[3-(morpho-linomethyl)-4-methyl-6-chloro-8-allylcoumarin-7-yloxy]propane hydrochloride.

EXAMPLE 8:

1,3-bis[3-(morpholinomethyl)-4,8-dimethyl-6-chlorocoumarin-7-yloxy]propane hydrochloride

15.0 g of 3-morpholinomethyl-4,8-dimethyl-6-chloro-7-hydroxycoumarin are suspended in 200 ml of ethanol and to this are added 5.2 g of 50% KOH in water. It is left to stand for l hour at 50ºC and then concentrated and vacuum-dried. It is gathered with 200 ml of 2-butanone and to this are added 6.4 g of

K₂CO₃. It is heated while being shaken, then to this are added 4.8 g of 1,3-dibromopropane and it is left to react for 12 hours.

The solvent is evaporated and the residue is

dissolved with 200 ml of ethyl acetate; it is

washed with a 1N solution of NaOH. The solvent is

anhydrated and concentrated. It is purified by

chromatography with Prep LC System Waters, using as eluent a mixture of CHCl₃-EtAc-CH₃OH 70:30:2.

The pure fractions are concentrated and crystallized from ethyl acetate.

The crystallized product is dissolved in ethyl acetate-chloroform, anhydrated and treated with HCl in ethanol until a Congo red indicator change. It is filtered and crystallized from ethyl alcohol. This is filtered and vacuum-dried, obtaining 4.0 g of a compound to which elementary analysis (C,H,N) and

nuclear magnetic resonance spectroscopy (protons)

both attribute a structure of 1,3-bis[3-(morpholinomethyl)-4,8-dimethyl-6-chlαrocoumarin-7-yloxy]propane hydrochloride. EXAMPLE 9:

1,3-bis[3-(morpholinomethyl)-4-methyl-6-allyl-8-chlorocoumarin-7-yloxy]propane hydrochloride

22.0 g of 3-morpholinomethyl-4-methyl-6-allyl-7-hydroxy-8-chlorocoumarin are suspended in 200 ml

of ethanol and to this are added 7.1 g of 50% KOH

in water. It is left to stand for 1 hour at 50ºC and then concentrated and vacuum-dried. It is gathered with 200 ml of 2-butanone and to this are added 8.7 g of

K₂CO₃. It is heated while being shaken, and then to this are added 6.5 g of 1,3-dibromopropane and it is left to react for 12 hours.

The solvent is evaporated and the residue is

dissolved with 200 ml of ethyl acetate; it is washed with a 1N solution of NaOH. The solvent is

anhydrated and concentrated. It is purified by

crystallization from CH₃OH-EtAc.

The crystallized product is dissolved in ethanol and treated with HCl in ethanol until a Congo red

indicator change. It is filtered and crystallized

from ethyl alcohol. This is filtered and vacuum¬

-dried, obtaining 5.0 g of a compound to which

elementary analysis (C,H,N) and nuclear magnetic

resonance spectroscopy (protons) both attribute a

structure of 1,3-bis[3-(morpholinomethyl)-4¬

-methyl-6-allyl-8-chlorocoumarin-7-yloxy3propane

hydrochloride.

EXAMPLE 10:

1,3-bis[3-(morpholinomethyl)-4-phenyl-6-chloro-8-allylcoumarin-7-yloxy]propane hydrochloride

40.0 g of 3-morpholinomethyl-4-phenyl-6-chloro-7¬

-hydroxy-8-allylcoumarin are suspended in 500 ml of ethanol and to this are added 10.9 g of 50% KOH in water. It is left to stand for 1 hour at 50ºC and then concentrated and vacuum-dried. It is gathered with 500 ml of 2-butanone and to this are added 13.4 g of K₂CO₃. It is heated while being shaken, and then to this are added 10.0 g of 1,3-dibromopropane and it is left to react for 12 hours.

The solvent is evaporated and the residue is

dissolved with 500 ml of ethyl acetate; it is

washed with a 1N solution of NaOH. The solvent is

anhydrated and concentrated. It is purified by chromatography with Prep LC System Waters, using as eluent a mixture of CH₂Cl₂-EtAc 80:20. The pure

fractions are concentrated.

The residue is dissolved in ethyl acetate,

anhydrated and treated with HCl in ethanol until a Congo red indicator change. It is filtered and

washed. It is vacuum-dried, obtaining 4.0 g of a compound to which elementary analysis (C,H,N)

and nuclear magnetic resonance spectroscopy

(protons) both attribute a structure of 1,3-bis[3--(morpholinomethyl)-4-phenyl-6-chloro-8-allylcoumarin-7-yloxy]propane hydrochloride.

EXAMPLE 11:

1,3-bis[3-(morpholinomethyl)-4-phenyl-6-allyl-8-methylcoumarin-7-yloxy]propane hydrochloride

50.0 g of 3-morpholinomethyl-4-phenyl-6-allyl-7-hydroxy-8-methylcoumarin are suspended in 500 ml of ethanol and to this are added 14.3 g of 50% KOH in water. It is left to stand for 1 hour at 50ºC and then concentrated and vacuum-dried. It is gathered with 500 ml of 2-butanone and to this are added 17.6 g ofK₂CO₃. It is heated while being shaken, then to this areadded 12.9 g. of 1,3-dibromopropane and it is left to react for 12 hours.

The solvent is evaporated and the residue is dissolved with 500 ml of ethyl acetate; it is

washed with a IN solution of NaOH. The solvent is anhydrated and concentrated. It is purified by

chromatography with Prep LC System Waters, using as eluent a mixture of CH₂Cl₂-EtAc-CH₃OH 70:30:10.

The pure fractions are concentrated and

crystallized from ethyl acetate/n-hexane 1:2.5. The crystallized product is dissolved in ethyl acetate, anhydrated and treated with HCl in ethanol until a Congo red indicator change. It is filtered and crystallized from ethyl alcohol al 98%. This is

filtered and vacuum-dried, obtaining 14.6 g of a

compound to which elementary analysis (C,H,N) and

nuclear magnetic resonance spectroscopy (protons)

both attribute a structure of 1,3-bis[3-(morpholinomethyl)-4-phenyl-6-allyl-8--methylcoumarin-7-yloxy]propane hydrochloride.

EXAMPLE 12:

1,3-bis[3-(morpholinomethyl)-4-phenyl-6-allyl-8¬

-chlorocoumarin-7-yloxy]propane hydrochloride

30.0 g of 3-morpholinomethyl-4-phenyl-6-allyl-7-hydroxy-8-chlorocoumarin are suspended in 300 ml of ethanol and to this are added 8.2 g of 50% KOH

in water. It is left to stand for 1 hour at 50°C and then concentrated and vacuum-dried. It is gathered with 300 ml of 2-butanone and to this are added 10.1 g ofK₂CO₃. It is heated while being shaken, then to this are added 7.4 g of 1,3-dibromopropane and it is left to react for 12 hours.

The solvent is evaporated and the residue is

dissolved with 300 ml of ethyl acetate; it is

washed with a 1N solution of NaOH. The solvent is

anhydrated and concentrated. It is purified by

chromatography with Prep LC System Waters, using as eluent a mixture of CH₂Cl₂-EtAc 90:10. The pure

fractions are concentrated and crystallized from

95% ethanol.

The crystallized product is dissolved in ethyl acetate, anhydrated and treated with HCl in ethanol until a Congo red indicator change. It is filtered and crystallized from ethanol/ chloroform 4 : 3 . This is filtered and vacuum-dried, obtaining 11.1 g of a

compound to which elementary analysis (C, H, N) and

nuclear magnetic resonance spectroscopy (protons) both attribute a structure of

1 , 3 -bis [3 - (morpholinomethyl) -4-phenyl-6-allyl-8-chlorocoumarin-7-yloxy]propane hydrochloride.

EXAMPLE 13 :

1,3-bis[3-(morpholinomethyl)-4-phenyl-6,8-diallylcoumarin-7-yloxy]propane hydrochloride

30.0 g of 3-morpholinomethyl-4-phenyl-6,8-diallyl-7-hydroxycoumarin are suspended in 300 ml of

ethanol and to this are added 7-8 g of 50% KOH in

water. It is left to stand for 1 hour at 50ºC and then concentrated and vacuum-dried. It is gathered with 300 ml of 2-butanone and to this are added 9.9 g of K₂CO₃. It is heated while being shaken, and then to this are added 7.2 g of 1,3-dibromopropane and it is left to react for 12 hours.

The solvent is evaporated and the residue is

dissolved with 300 ml of ethyl acetate; it is

washed with a 1N solution of NaOH. The solvent is

anhydrated and concentrated. It is purified by

chromatography with Prep LC System Waters, using as eluent a mixture of CH₂Cl₂-EtAc 80:20. The pure

fractions are concentrated.

The residue is dissolved in ethyl acetate,

anhydrated and treated with HCl in ethanol until a Congo red indicator change. It is filtered and crystallized from isopropyl alcohol. This is filtered and

vacuum-dried, obtaining 10.0 g of a compound to which elementary analysis (C,H,N) and nuclear magnetic

resonance spectroscopy (protons) both attribute a structure of 1,3-bis[3-(morpholinomethyl)-4-phenyl6,8-diallylcoumarin-7-yloxy]-propane hydrochloride.

EXAMPLE 14:

1,3-bis[3-(ß-morpholinoethyl)-4-methylcoumarin-7-yloxy]propane hydrochloride

19.0 g of 3-(ß-morpholinoethyl)-4-methyl-7-hydroxycoumarin are suspended in 300 ml of ethanol and to this are added 5.5 g of 50% KOH in water. It is left to stand for 1 hour at 50ºC and then

concentrated and vacuum-dried. It is gathered with 300 ml of 2-butanone and to this are added 9.0 g of K₂CO₃. It is heated while being shaken, and then to this are added 6.6 g of 1,3-dibromopropane (0-0328 mole) and it is left to react for 12 hours.

The solvent is evaporated and the residue is

dissolved with 300 ml of ethyl acetate; it is

washed with a 1N solution of NaOH. The solvent is

anhydrated and concentrated. It is crystallized

from methanol.

The crystallized product is dissolved in ethyl acetate, anhydrated and treated with HCl in ethanol until a Congo red indicator change. It is filtered and washed. It is further filtered and vacuum-dried,

obtaining 5.8 g of a compound to

which elementary analysis (C,H,N) and nuclear

magnetic resonance spectroscopy (protons) both

attribute a structure of 1,3-bis[3-(ß-roorpholinoethyl)-4-methylcoumarin-7-yloxy]propane hydrochloride.

EXAMPLE 15:

1,3-bis[3-(ß-morpholinoethyl)-4-methyl-8-chlorocoumarin-7-yloxy]propane hydrochloride 30.0 g of 3-(ß-morpholinoethyl)-4-methyl-7-hydroxy-8-chlorocoumarin hydrochloride are

suspended in 300 ml of ethanol and to this are

added 18.7 g of 50% KOH in water. It is left to

stand for 1 hour at 50ºC and then concentrated and vacuum-dried. It is gathered with 300 ml of 2-butanone and to this are added 11.5 g of K₂CO₃. It is heated while being shaken, and then to this are added 8.4 g of

1,3-dibromopropane and it is left to react for 12 hours.

The solvent is evaporated and the residue is

dissolved with 300 ml of ethyl acetate; it is

washed with a 1N solution of NaOH. The solvent is anhydrated and concentrated. It is washed with warm CH₃OH.

The product is dissolved in chloroform/ ethyl acetate, anhydrated and treated with HCl in ethanol

until a Congo red indicator change. It is filtered and washed. This is filtered and vacuum-dried,

obtaining 22.0 g of a compound to which elementary analysis (C, H,N) and nuclear magnetic resonance

spectroscopy (protons) both attribute a structure of 1 , 3-bis [ 3- (ß-morpholinoethyl) -4-methyl-8-chlorocoumarin-7-yloxy] propane hydrochloride.

EXAMPLE 16 :

1,6-bis[3-(ß-morpholinoethyl)-4-methylcoumarin-7¬

-yloxyjhexane hydrochloride

25.0 g of 3-(ß-morpholinoethyl)-4-methy1-7-hydroxycoumarin sulfate are suspended in 500 ml of ethanol and to this are added 22 g of 50% KOH in

H₂O. It is left to stand at 50ºC for 1 hour and then concentrated and vacuum-dried. It is gathered in

2-butanone and to this are added 9.0 g of K₂CO₃. It is heated while being shaken. To this are then added 8.0 g of 1.6 dibromohexane and it is left to react for 20 hours. The solvent is evaporated and the residue is dissolved with 500 ml of CHCl₃ and washed with H₂O.

The solvent is concentrated and the product

precipitated by adding methanol. It is gathered with a few ml of CHCl₃ and it is reprecipitated

with ethyl acetate (EtAc). It is dissolved in a mixture of chloroform/methanol and HCl in ethanol is added until a Congo red indicator change.

The product is concentrated slightly and precipitated by adding ethyl acetate. This is filtered and vacuum-dried, obtaining 10.7 g of a compound to which

elementary analysis (C,H,N) and nuclear magnetic

resonance spectroscopy (protons) both attribute a

structure of 1,6-bis [3-(ß-morpholinoethyl)-4-methylcoumarin-7-yloxy]hexane hydrochloride.

EXAMPLE 17:

Bis (3-(ß-diethylaminoethyl)-4-methylcoumarin-7-yloxy] dicarbethoxy methane hydrochloride

50.0 g of 3-(ß-diethylaminoethyl)-4-methyl-7-hydroxy-coumarin are placed in a 500-ml reactor

equipped with a shaking and separating apparatus. 300 ml of toluene and 49.7 g of K₂CO₃ are added. It is heated, while separating and eliminating the

reaction water. The toluene is then concentrated to about 50 ml, and it is cooled to room temperature and to this are added 100 ml of DMSO. One hour later 31.7 g of the diethyl ester of dibromomalonic acid are added, and the mixture is left to react for 12 hours. It is gathered with toluene and washed with H₂O and a 1N solution of NaOH. It is anhydrated and the solvent is concentrated. It is purified by chromatography with Prep LC System Waters using as eluent a mixture of

CH₂Cl₂-CH₃OH-NH₄OH (30%) 95:7:0.2.

The pure fractions are concentrated, gathered with ethyl acetate, anhydrated and the hydrochloride is precipitated with HCl in ethanol. 9.0 g of a compound are obtained to which elementary analysis (C,H,N) and

nuclear magnetic resonance spectroscopy (protons)

both attribute a structure of bis(3-(ß-diethylaminoethyl)-4-methylcoumarin-7-yloxy]dicarbethoxy methane hydrochloride.

EXAMPLE 18:

Bis[3-(ß-diethylaminoethyl)-4-methyl-8-chlorocoumarin-7-yloxy]dicarbethoxy methane hydrochloride

57.0 g of 3-(ß-diethylaminoethyl)-4-methyl-7-hydroxy-8-chlorocoumarin are placed in a 500-ml

reactor equipped with a shaking and separating

apparatus. 300 ml of toluene and 49.7 g of K₂CO₃

are added. It is heated, separating and eliminating the reaction water. The toluene is concentrated to about 50 ml, cooled to room temperature and to this are adde'd 100 ml of DMSO. One hour later to this are added 31.7 g of the diethyl ester of dibromomalonic acid and it is left to react for 12 hours. The resultant is gathered with toluene and washed with H₂O and a 1N solution of NaOH. The solvent is anhydrated and

concentrated. It is purified by chromatography with Prep LC System Waters using as eluent a mixture of

CH₂Cl₂-CH₃OH-NH₄OH (30%) 98:2:0.2.

The pure fractions are concentrated, gathered with ethyl acetate, anhydrated, and the hydrochloride is precipitated with HCl in ethanol. 13.0 g of a compound are obtained to which elementary analysis (C,H,N) and nuclear magnetic resonance spectroscopy (protons) both attribute a structure of bis [3-(ß-diethylaminoethyl)-4-methyl-8-chlorocoumarin-7-yloxy]dicarbethoxy methane hydrochloride. EXAMPLE 19:

Bis[3-(ß-diethylaminoethyl)-4-phenylcoumarin-7-yloxy]dicarbethoxy methane hydrochloride

25.0 g of 3-(ß-diethylaminoethyl)-4-phenyl-7-hydroxy-coumarin are placed in a 500-ml reactor

equipped with a shaking and separating apparatus. 300 ml of toluene and 27.7 g of K₂CO₃ are added. It is

heated, separating and eliminating the reaction

water. The toluene is then concentrated to about 50 ml, cooled to room temperature and to this are

added 100 ml of DMSO. One hour later 12.7 g of the diethyl ester of dibromomalonic acid are added, and it is left to react for 12 hours. It is gathered with toluene and waεhed with H₂O and a 1N solution of NaOH. The solvent is anhydrated and concentrated. It is

purified by chromatography with Prep LC System

Waters using as eluent a mixture of

CH₂Cl₂-CH₃OH-NH₄OH (30%) 95:5:0.2.

The pure fractions are concentrated, gathered with ethyl acetate, anhydrated, and the hydrochloride is precipitated with HCl in ethanol. 12.5 g of a

compound are obtained to which elementary analysis

(C,H,N) and nuclear magnetic resonance spectroscopy

(protons) both attribute a structure of

bis [3-(ß-diethylaminoethyl)-4-phenylcoumarin-7-yloxy]dicarbethoxy methane hydrochloride.

EXAMPLE 20:

Bis [3-(ß-diethylaminoethyl)-4-methyl-coumarin-7¬

-yloxy]carbethoxy methane hydrochloride 81.0 g of 3-(ß-diethylaminoethyl)-4-methyl-7-hydroxy-coumarin are placed in a 500-ml reactor equipped

with a shaking and separating apparatus. 300 ml of toluene and 48.4 g of K₂CO₃ are added. It is

heated, separating and eliminating the reaction water. The toluene is concentrated to 50 ml, cooled to

room temperature and to this are added 100 ml of

DMSO. One hour later 23.55 g of ethyl dichloroacetate are added and it is left to react for 48

hours.

The reaction mixture is gathered with ethyl acetate and washed with H₂O and a 1N solution of NaOH. The solvent is anhydrated and concentrated. It is purified by chromatography with Prep LC System Waters using a

mixture of CHCl₃-CH₃OH 80:20 as eluent. The pure

fractions are evaporated. This is gathered in ethyl acetate, anhydrated, and the hydrochloride is

precipitated with HCl in ethanol. It is filtered,

washed with ethyl acetate and vacuum-dried,

obtaining 28.0 g of a compound to which elementary analysis (C,H,N) and nuclear magnetic resonance

spectroscopy (protons) both attribute a structure

of bis [3-(ß-diethylaminoethyl)-4-methyl-coumarin-7-yloxy]carbethoxy methane hydrochloride.

EXAMPLE 21:

Bis[3- (ß-diethylaminoethyl) -4-methyl-8-chlorocoumarin-7-yloxy] carbethoxy methane hydrochloride

50 .0 g of 3-(ß-diethylaminoethyl) -4-methyl-7-hydroxy-8-chlorocoumarin are placed in a 500-ml

reactor equipped with a shaking and separating apparatus .

300 ml of toluene and 26.7 g of K₂CO₃

are added. It is heated, separating and eliminating the reaction watfer. The toluene is concentrated to 50 ml, cooled to room temperature and to this are added 100 ml of DMSO. One hour later 12.6 g of ethyl

dichloroacetate are added, and it is left to react for 48 hours.

The reaction mixture is gathered with ethyl acetate and washed with H₂O and a 1N solution of NaOH. The

solvent is anhydrated and concentrated. It is purified by chromatography with Prep LC System Waters using a

mixture of CHCl₃-CH₃OH-NH₄OH (30%) 95:5:0.2 as

eluent. The pure fractions are evaporated. It is

gathered in ethyl acetate, anhydrated, and the

hydrochloride is precipitated with HCl in ethanol.

It is filtered, washed with ethyl acetate and

vacuum-dried, obtaining 17.0 g of a compound to

which elementary analysis (C,H,N) and nuclear

magnetic resonance spectroscopy (protons) both

attribute a structure of bis [3-(ß-diethylaminoethyl)-4-methyl-8-chlorocoumarin-7-yloxy)carbethoxy methane hydrochloride.

EXAMPLE 22:

Bis [3-(ß-diethylaminoethyl)-4-phenylcoumarin¬

-7-yloxy]carbethoxy methane hydrochloride

25.0 g of 3-(ß-diethylaminoethyl)-4-phenyl-7-hydroxycoumarin hydrochloride are placed in a

500-ml reactor equipped with a shaking and separating apparatus. 300 ml of toluene and 27.7 g of K₂CO₃

are added. It is heated, separating and eliminating the reaction water. The toluene is concentrated to 50 ml, cooled to room temperature and to this are added 100 ml of DMSO. One hour later 6.26 g of ethyl

dichloroacetate are added, and it is left to react for 48 hours. The reaction mixture is gathered with ethyl acetate and washed with H₂O and a 1N solution of NaOH. The solvent is anhydrated and concentrated. It is purified by chromatography with Prep LC System Waters using a

mixture of CHCl₃-CH₃OH-NH₄OH (30%) 95:5:0.2 as

eluent. The pure fractions are evaporated. It is

gathered in ethyl acetate, anhydrated, and the

hydrochloride is precipitated with HCl in ethanol, obtaining 5.5 g of a compound to which elementary analysis (C,H,N) and nuclear magnetic resonance

spectroscopy (protons) both attribute a structure

of bis [3-(ß-diethylaminoethyl)-4-phenylcoumarin-7-yloxy]carbethoxy methane hydrcchloride. EXAMPLE 23:

Bis[3-(morpholinomethyl)-4-methyl-6,8-diallylcoumarin-7-yloxy]carbethoxy methane hydrochloride

20.0 g of 3-(morpholinomethyl)-4-methyl-6,8-diallyl-7-hydroxycoumarin are placed in a 500-ml

reactor equipped with a shaking and separating

apparatus. 300 ml of toluene and 11.6 g of K₂CO₃

are added. It is heated, separating and eliminating the reaction water. The toluene is concentrated to 50 ml, cooled to room temperature and to this are added 100 ml of DMSO. One hour later to this are added 4.39 g of ethyl dichloroacetate and it is left to react for

43 hours.

The reaction mixture is gathered with ethyl acetate and washed with H₂O and a 1N solution of NaOH. The

solvent is anhydrated and concentrated. It is purified by chromatography with Prep LC System Waters using a

mixture of CHCl₃-CH₃OH-NH₄OH (30%) 95:5:0.2 as

eluent. The pure fractions are evaporated. This is gathered in ethyl acetate, anhydrated, and the hydrochloride is precipitated with HCl in ethanol.

5.6 g of a compound are obtained to which

elementary analysis (C,H,N) and nuclear magnetic

resonance spectroscopy (protons) both attribute a

structure of bis[3-(morpholinomethyl)-4-methyl-6,8-diallylcoumarin-7-yloxy]carbethoxy methane

hydrochloride.

EXAMPLE 24:

Bis [3-(morpholinomethyl)-4-methyl-6-chloro-8-allylcoumarin-7-yloxy]carbethoxy methane hydrochloride

30.0 g of 3-(morpholinomethyl)-4-methyl-6-chloro-7-hydroxy-8-allylcoumarin are placed in a 500-ml

reactor equipped with a shaking and separating

apparatuε. 300 ml of toluene and 16.5 g of K₂CO₃

are added. It is heated, separating and eliminating the reaction water. The toluene is concentrated to 50 ml, cooled to room temperature and to this are added 100 ml of DMSO. One hour later to this are added 6.75 g of ethyl dichloroacetate and it is left to react for

48 hours.

The reaction mixture is gathered with ethyl acetate and washed with H₂O and a 1N solution of NaOH. The solvent is anhydrated and concentrated. It is purified by chromatography with Prep LC System Waters using a

mixture of CHCl₃-CH₃OH-NH₄OH (30%) 95:5:0.2 as

eluent. The pure fractions are evaporated. This is gathered in ethyl acetate, anhydrated, and the

hydrochloride is precipitated with HCl in ethanol.

It is filtered, washed with ethyl acetate and

vacuum-dried, obtaining 10.0 g of a compound to

which elementary analysis (C,H,N) and nuclear

magnetic resonance spectroscopy (protons) both

attribute a structure of bis[3-(morpholinomethyl) -4-methyl-6-chloro-8-allylcoumairin-7-yloxy]

carbethoxy methane hydrochloride.

EXAMPLE 25:

Bis[3-(morpholinomethyl)-4,8-dimethyl-6-allylcoumarin-7-yloxy]carbossimetano hydrochloride

30.0 g of 3-(morpholinomethyl)-4,8-dimethyl-6-allyl-7-hydroxycoumarin are placed in a 500-ml

reactor equipped with a shaking and separating

apparatus. 300 ml of toluene and 25.2 g of K₂CO₃

are added thereto. It is heated, separating and

eliminating the reaction water. The toluene is

concentrated to 50 ml, cooled to room temperature and to this are added 100 ml of DMSO. One hour later to this are added 7.2 g of ethyl dichloroacetate and it is left to react for 48 hours.

The reaction mixture is gathered with ethyl acetate and washed with H₂O and a 1N solution of NaOH. The solvent is anhydrated and concentrated. It is purified by chromatography with Prep LC System Waters using a

mixture of CH₂Cl₂-EtAc-CH₃OH 70:30:2 as eluent.

The pure fractions are evaporated and gathered in ethyl acetate, anhydrated, and the hydrochloride is

precipitated with HCl in ethanol.

It is filtered, washed with ethyl acetate and

vacuum-dried, obtaining 9.1 g of a compound to

which elementary analysis (C,H,N) and nuclear

magnetic resonance spectroscopy (protons) both

attribute a structure of bis[3-(morpholinomethyl)-4,8-dimethyl-6-allylcoumarin-7-yloxy]carboxymethane hydrochloride. EXAMPLE 26:

Bis[3-(morpholinomethyl)-4-methyl-6-allyl-8-chlorocoumarin-7-yloxy]carbethoxy methane hydrochloride

30.0 g of 3-(morpholinomethyl)-4-methyl-6-allyl-7-hydroxy-8-chlorocoumarin are placed in a 500-ml reactor equipped with a shaking and separating

apparatus. 300 ml of toluene and 17.8 g of K₂CO₃

are added. It is heated, separating and eliminating the reaction water. The toluene is concentrated to 50 ml, cooled to room temperature and to this are added 100 ml of DMSO. One hour later 6.75 g of ethyl dichloroacetate are added thereto, and it is left to react for

48 hours.

The reaction mixture is gathered with ethyl acetate and washed with H₂O and a 1N solution of NaOH. The solvent is anhydrated and concentrated. It is purified by chromatography with Prep LC System Waters using a

mixture of CH₂Cl₂-EtAc-CH₃OH 70:30:1 as eluent.

The pure fractions are evaporated, gathered in

ethyl acetate, anhydrated and the hydrochloride is precipitated with HCl in ethanol. It is filtered,

washed with ethyl acetate and vacuum-dried,

obtaining 9.9 g of a compound to which elementary

analysis (C,H,N) and nuclear magnetic resonance

spectroscopy (protons) both attribute a structure

of bis[3-(morpholinomethyl)-4-methyl-6-allyl-8-chlorocoumarin-7-yloxy]carbethoxy methane

hydrochloride. EXAMPLE 27:

1,3-bis[3-(ß-diethylaminoethyl)-4-methyl-8-chlorocoumarin-7-yloxy]-2-hydroxypropane hydrochloride

50.0 g of 3-(ß-diethylaminoethyl)-4-methyl-7-hydroxy-8-chlorocoumarin are suspended in 500 ml of ethanol and to this are added 18.1 g of 50% KOH in H₂O. It is left to stand at 50ºC while shaking for one hour and then concentrated and vacuum-dried. It is gathered with 500 ml of 2-butanone and to this are added 22.0 g of K₂CO₃. It is heated while being shaken, then to this are added 9.9 g of

epibromohydrin and it is left to react for 3 days.

The solvent is evaporated and the residue is

dissolved with 500 ml of ethyl acetate. It is

washed with a 1N solution of NaOH. The solvent is anhydrated and concentrated. It is purified by

chromatography with Prep LC System Waters using as eluent a mixture of CH₂Cl₂-CH₃OH-NH₄OH (30%) 95:5:0.2.

The pure fractions are evaporated, crystallized from ethanol, gathered in ethyl acetate,

anhydrated, and the hydrochloride is precipitated with HCl in ethanol. The hydrochloride is

crystallized from ethanol. This is filtered and

vacuum-dried, obtaining 6.8 g of a compound to

which elementary analysis (C,H,N) and nuclear

magnetic resonance spectroscopy (protons) both

attribute a structure of 1,3-bis[3-(ß-diethylaminoethyl)-4-methyl-8-chlorocouraarin-7-yloxy]-2-hydroxypropane hydrochloride.

EXAMPLE 28:

1,3-bis[3-(morpholinomethyl)-4-methyl-6,8-diallylcoumarin-7-yloxy]-2-hydroxypropane hydrochloride

50.0 g of

3-(morpholinomethyl)-4-methyl-6,8-diallyl-7¬

-hydroxycoumarin are suspended in 500 ml of ethanol and to this are added 15.0 g of 50% KOH in H₂O. It is left to stand at 50ºC while shaking for one hour and then concentrated and vacuum-dried. It is gathered with 500 ml of 2-butanone and to this are added 24.8 g of K₂CO₃. It is heated while being shaken, then to this are added 9.5 g of epibromohydrin and it is left to react for 3 days. The solvent is evaporated and the residue is dissolved with 500 ml of ethyl acetate. It is

washed with a 1N solution of NaOH. The solvent is

anhydrated and concentrated. It is purified by

chromatography with Prep LC System Waters using as eluent a mixture of CH₂Cl₂-EtAc-CH₃OH 70:30:1.

The pure fractions are evaporated, gathered in ethyl acetate, anhydrated, and the hydrochloride is precipitated with HCl in ethanol. The hydrochloride is crystallized from a mixture of ethanol-ethyl acetate or acetone. This is filtered and

vacuum-dried, obtaining 16.3 g of a compound to

which elementary analysis (C,H,N) and nuclear

magnetic resonance spectroscopy (protons) both

attribute a structure of 1,3-bis[3-(morpholinomethyl)-4-methyl-6,8-diallylcoumarin-7-yloxy3-2-hydroxypropane hydrochloride.

EXAMPLE 29:

1,3-bis[3-(morpholinomethyl)-4-methyl-6-chloro-8-allylcoumarin-7-yloxy]-2-hydroxypropane hydrochloride

20.0 g of 3-(morpholinomethyl)-4-methyl-6-chloro-7-hydroxy-8-allylcoumarin are suspended in 200 ml of ethanol and to this are added 6.4 g of 50% KOH

in H₂θ. It is left to stand at 50ºC while being

stirred for one hour, and then concentrated and

vacuum-dried. It is gathered with 200 ml of 2-butanone and 7.9 g of K₂CO₃ are added. It. is heated while being shaken, then to this are added 3.8 g of epibromohydrin and it is left to react for 3 days.

The solvent is evaporated and the residue is

dissolved with 500 ml of ethyl acetate. It is

washed with a 1N solution of NaOH. The solvent is anhydrated and concentrated.

The reaction product is crystallized from ethyl ether/ethyl acetate, 10:1. It is gathered in ethyl acetate, anhydrated, and the hydrochloride is

precipitated with HCl in ethanol. The hydrochloride is crystallized from ethanol. This is filtered and

vacuum-dried, obtaining 6.0 g of a compound to which elementary analysis (C,H,N) and nuclear magnetic

resonance spectroscopy (protons) both attribute a structure of 1,3-bis[3-(morpholinomethyl)4-methy1-6-chloro-8-allylcoumarin-7-yloxy]-2-hydroxypropane hydrochloride.

EXAMPLE 30:

1,3-bis[3-(morpholinomethyl)-4-phenyl-6-allyl-8-methylcoumarin-7-yloxy]-2-hydroxypropane hydrochloride

50.0 g of 3-(morpholinomethyl)-4-phenyl-6-allyl-7-hydroxy-8-methylcoumarin are suspended in 500 ml of ethanol and to this are added 14.3 g of 50% KOH in H₂O. It is left to stand at 50ºC while being

shaken for one hour, and then concentrated and

vacuum-dried. It is gathered with 500 ml of 2-butanone and to this are added 26.5 g of K₂CO₃. It is heated while being shaken, then to this are added 8.7 g of

epibromohydrin and it is left to react for 3 days.

The solvent is evaporated and the residue is

dissolved with 500 ml of ethyl acetate. It is

washed with a 1N solution of NaOH. The solvent is anhydrated and concentrated. It is purified by chromatography with Prep LC System Waters using as eluent a mixture of CH₂Cl₂-EtAc-CH₃OH 70:30:1.

The pure fractions are evaporated, gathered in ethyl acetate, anhydrated, and the hydrochloride is precipitated with HCl in ethanol. It is washed, filtered and vacuum-dried, obtaining 5.0 g of a compound to which elementary analysis (C,H,N) and nuclear magnetic resonance spectroscopy (protons) both attribute a structure of 1,3-bis[3-(morpholinomethyl)-4-phenyl-6-allyl-8-methylcoumarin-7-yloxy]-2-hydroxypropane hydrochloride.

EXAMPLE 31:

1,3-bis[3-(morpholinomethyl)-4,8-dimethyl-6-allylcoumarin-7-yloxy3-2-hydroxypropane hydrochloride

30.0 g of 3-(morpholinomethyl)-4,8-dimethyl-6-allyl-7-hydroxycoumarin are suspended in 500 ml of ethanol and to this are added 10.2 g of 50% KOH in H₂O. It is left to stand at 50ºC while being shaken for one hour, and then concentrated and vacuum-dried. It is gathered with 500 ml of 2-butanone and to this are added 12.6 g of K₂CO₃. It is heated while being shaken, then to this are added 6.2 g of

epibromohydrin. and it is left to react for 3 days. The solvent is evaporated and the residue is

dissolved with 500 ml of ethyl acetate. It is

washed with a 1N solution of NaOH. The solvent iε anhydrated and concentrated. It is purified by chromatography with Prep LC System Waters using as eluent a mixture of CH₂Cl₂-EtAc-CH₃OH 70:30:3.

The pure fractions are evaporated, gathered in ethyl acetate , anhydrated, and the hydrochloride is precipitated with HCl in ethanol . The

hydrochloride is crystallized from a mixture of ethanol-ethyl acetate . This is filtered and

vacuum-dried, obtaining 9. 2 g of a compound to

which elementary analysis (C, H,N) and nuclear

magnetic resonance spectroscopy (protons) both

attribute a structure of 1, 3-bis [3- (morpholinomethyl) -4 , 8-dimethyl-6-allylcoumarin-7-yloxy3 -2-hydroxypropane hydrochloride.

EXAMPLE 32 :

1,3-bis[3-(morpholinomethyl)-4-methyl-6-allyl-8-chlorocoumarin-7-yloxy]-2-hydroxyproρane hydrochloride

20.0 g of 3-(morpholinomethyl)-4-methyl-6-allyl-7-hydroxy-8-chlorocoumarin are suspended in 200 ml of ethanol and to this are added 5.8 g of 50% KOH in H₂O. It is left to stand at 50ºC while being

shaken for one hour, then concentrated and

vacuum-dried. It is gathered with 300 ml of 2-butanone and to this are added 7.9 g of K₂CO₃. It is heated while being shaken, and then to this are added 3.8 g of epibromohydrin and it is left to react for 3 days.

The solvent is evaporated and the residue is

dissolved with 500 ml of ethyl acetate. It is

washed with a 1N solution of NaOH. The solvent is anhydrated and concentrated. It is purified by

chromatography with Prep LC System Waters using as eluent a mixture of CH₂Cl₂-CH₃OH-NH₄OH (30%) 98:2:0.2.

The pure fractions are evaporated, gathered in ethyl acetate, anhydrated and the hydrochloride is precipitated with HCl in ethanol. The hydrochloride iε crystallized from ethanol. This is filtered and vacuum-dried, obtaining 3.3 g of a compound to

which elementary analysis (C,H,N) and nuclear

magnetic resonance spectroscopy (protons) both attribute a structure of 1,3-bis[3-(morpholinomethyl)-4-methyl-6-allyl-8-chlorocoumarin-7-yloxy]-2-hydroxypropane hydrochloride. EXAMPLE 33:

1,3-bis[3-morpholinomethyl)-4-phenyl-6-chloro-8-allylcoumarin-7-yloxy]-2-hydroxypropane hydrochloride

50.0 g of 3-(morpholinomethyl)-4-phenyl-6-chloro-7-hydroxy-8-allylcoumarin are suspended in 500 ml of ethanol and to this are added 13.5 g of 50% KOH in H₂O. It is left to stand at 50ºC while being

shaken for one hour and is then concentrated and

vacuum-dried. It is gathered with 500 ml of 2-butanone and to this are added 16.6 g of K₂CO₃. It is heated while being shaken, and then to this are added 8.2 g of epibromohydrin and it is left to react for 3

days. The solvent is evaporated and the residue is dissolved with 500 ml of ethyl acetate. It is

washed with a 1N solution of NaOH. The εolvent is anhydrated and concentrated. It is purified by

chromatography with Prep LC System Waters using as eluent a mixture of CH₂Cl₂-EtAc 95:5.

The pure fractions are evaporated, gathered in ethyl acetate, anhydrated, and the hydrochloride is precipitated with HCl in ethanol-ethyl acetate.

This is filtered and vacuum-dried, obtaining 7.7 g of a compound to which elementary analysis (C,H,N) and nuclear magnetic resonance spectroscopy

(protons) both attribute a structure of 1,3-bis[3¬

-morpholinomethyl) -4-phenyl-6-chloro-8-allylcoumarin7-yloxy ] -2-hydroxypropane hydrochloride. EXAMPLE 34 :

1,3-bis[3-(morpholinomethyl)-4-phenyl-6,8-diallylcoumarin-7-yloxy]-2-hydroxypropane hydrochloride

30.0 g of 3-(morpholinomethyl)-4-phenyl-6,8-diallyl-7-hydroxycoumarin are suspended in 300 ml of ethanol and to this are added 7.8 g of 50% KOH in H₂O. It is left to stand at 50ºC while being

shaken for one hour and is then concentrated and vacuum-dried. It is gathered with 300 ml of 2-butanone and to this are added 9.9 g of K₂CO₃. It is heated while being shaken, and then to this are added 4.9 g of epibromohydrin and it is left to react for 3 days.

The solvent is evaporated and the residue is

dissolved with 500 ml of ethyl acetate. It is

washed with a 1N solution of NaOH. The solvent is anhydrated and concentrated. It is purified by

chromatography with Prep LC System Waters using as eluent a mixture of CH₂Cl₂-EtAc 80:20.

The pure fractions are evaporated, gathered in ethyl acetate, anhydrated, and the hydrochloride iε precipitated with HCl in ethanol. The hydrochloride is freeze-dried, obtaining 5.0 g of a

compound to which elementary analysis (C,H,N) and nuclear magnetic resonance spectroscopy (protons) both attribute a structure of 1,3-bis[3-(morpholinomethyl)-4-phenyl-6,8-diallylcoumarin-7-yloxy]-2-hydroxypropane hydrochloride.

EXAMPLE 35:

1,3-bis[3-(morpholinomethyl)-4-phenyl-6-allyl-8-chlorocoumarin-7-yloxy]-2-hydroxypropane hydrochloride

33.0 g of 3-(morpholinomethyl)-4-phenyl-6-allyl -7-hydroxy-S-chlorocoumarin are suspended in 300 ml of ethanol and to this are added 8.8 g of 50%

KOH in H₂O. It is left to stand at 50ºC while being shaken for one hour and then concentrated and

vacuum-dried. It is gathered with 300 ml of 2-butanone and to this are added 11.1 g of K₂CO₃. It is heated while being shaken, then to this are added 5.5 g of

epibromohydrin and it is left to react for 3 days.

The solvent is evaporated and the residue is

dissolved with 300 ml of ethyl acetate. It is

washed with a 1N solution of NaOH. The solvent is anhydrated and concentrated. It is purified by

chromatography with Prep LC System Waters using as eluent a mixture of CH₂Cl₂-EtAc 80:20.

The pure fractions are evaporated, gathered in ethyl acetate, anhydrated, and the hydrochloride is precipitated with HCl in ethanol. The hydrochloride is crystallized from ethanol. This is

filtered and vacuum-dried, obtaining 5.4 g of a

compound to which elementary analysis (C,H,N) and nuclear magnetic resonance spectroscopy (protons) both attribute a structure of 1,3-bis[3-(morpholinomethyl)-4-phenyl-6-allyl-8-chlorocoumarin-7-yloxy]-2-hydroxypropane hydrochloride.

EXAMPLE 36:

1,3-bis[3-(ß-morpholinoethyl)-4-methylcoumarin-7¬

-yloxy]-2-hydroxypropane hydrochloride

25.0 g of 3-(ß-morpholinoethyl)-4-methyl-7-hydroxycoumarin sulfate are suspended in 300 ml of ethanol and to this are added 22.0 g of 50% KOH in

H₂O. It is left to stand at 50ºC while being shaken for one hour and then concentrated and vacuum-dried. It is gathered with 300 ml of 2-butanone and to this are added 9.0 g of K₂CO₃. It is heated while

being shaken, and then to this are added 4.5 g of epibromohydrin and it is left to react for 3 days.

The solvent is evaporated and the residue is

dissolved with 300 ml of ethyl acetate. It is

washed with a 1N solution of NaOH. The solvent is anhydrated and concentrated. It is crystallized

from CH₃OH, filtered, gathered in ethyl acetate,

anhydrated, and the hydrochloride is precipitated

with HCl in ethanol. It is washed, filtered and

vacuum-dried, obtaining 9.0 g of a compound to

which elementary analysis (C,H,N) and nuclear

magnetic resonance spectroscopy (protons) both

attribute a structure of 1,3-bis[3-(ß-morpholinoethyl)-4-methylcoumarin-7-yloxy]-2-hydroxypropane

hydrochloride.

EXAMPLE 37:

1,3-bis[3-(ß-morpholinoethyl)-4-methyl-8-chlorocoumarin-7-yloxy]-2-hydroxypropane hydrochloride

46.0 g of 3-(ß-morpholinoethyl)-4-methyl-7-hydroxy-8-chlσrocoumarin hydrochloride are

suspended in 500 ml of ethanol and to this are

added 28.6 g of -50% KOH in H₂O. It is left to stand at 50°C while being shaken for one hour and then

concentrated and vacuum-dried. It is gathered with 500 ml of 2-butanone and to this are added 17.6 g of K₂CO₃. It is heated while being shaken, then to this are added 8.9 g of epibromohydrin and it is left to react for 3 days. The solvent is evaporated and the residue is

dissolved with 300 ml of ethyl acetate. It is

washed with a 1N solution of NaOH. The solvent is

anhydrated and concentrated. It is purified by

chromatography with Prep LC System Waters using as eluent a mixture of CH₂Cl₂-EtAc-CH₃OH 70:30:10.

The pure fractions are evaporated, gathered in

CHCl₃-CH₃OH 2 :1 and the hydrochloride is

precipitated with HCl in ethanol. This is filtered and vacuum-dried, obtaining 12.0 g of a compound to which elementary analysis (C,H,N) and nuclear

magnetic resonance spectroscopy (protons) both attribute a structure of 1,3-bis[3-(ß-morpholinoethyl)-4-methyl-8-chlorocoumarin-7-yloxy3-2-hydroxypropane hydrochloride.

EXAMPLE 38:

1,3-bis[3-(ß-diisopropylaminoethyl)-4-methylcoumarin-7-yloxy)-2-hydroxypropane hydrochloride

25.0 g of 3-(ß-diisopropylaminoethyl)-4-methyl-7-hydroxycoumarin are suspended in 300 ml of ethanol and to this are added 8.2 g of 50% KOH in H₂O. It

is left to stand at 50ºC while being shaken for one hour and then concentrated and vacuum-dried.

It is gathered with 300 ml of 2-butanone and to

this are added 10.0 g of K₂CO₃. It is heated while

being shaken, 5.0 g of epibromohydrin are added and it is left to react for 3 days. The solvent is

evaporated and the residue is dissolved with 300 ml of ethyl acetate. It is washed with a 1N solution of NaOH. The solvent is anhydrated and concentrated. It is

purified by chromatography with Prep LC System Waters using as eluent a mixture of CH₂Cl₂-CH₃OH-NH₄OH (30%) 95:5:0.2.

The pure fractions are evaporated, gathered in ethyl acetate, anhydrated, and the hydrochloride is precipitated with HCl in ethanol. The hydrochloride is crystallized from isopropyl alcohol. This is filtered and vacuum-dried, obtaining 5.5 g of a compound to which elementary analysis (C,H,N) and nuclear magnetic resonance spectroscopy (protons) both attribute a structure of 1,3-bis[3-(ß-diisopropylaminoethyl)-4-methylcoumarin-7-yloxy)-2-hydroxypropane hydrochloride.

EXAMPLE 39:

1,3-bis[3-(ß-diethylaminoethyl)-4-methylcoumarin¬

-7-yloxy]-2-hydroxypropane hydrochloride

35.0 g of 3-(ß-diethylaminoethyl)-4-methyl-7¬

-hydroxycoumarin are suspended in 500 ml of ethanol and to this are added 12.6 g of 50% KOH in H₂O. It is left to stand at 50ºC while being shaken for one hour, and then it is concentrated and vacuum-dried. It is gathered with 500 ml of 2-butanone and to this are added 15.5 g of K₂CO₃. It is heated while being shaken, then to this are added 6.9 g of epibromohydrin and it is left to react for 3 days. The solvent is evaporated and the residue dissolved with 300 ml of ethyl acetate. It is washed with a 1N solution of NaOH. The solvent is anhydrated and concentrated. It is purified by chromatography with Prep LC System Waters using as eluent a mixture of CH₂Cl₂-CH₃OH-NH₄OH (30%) 90:10:0.4.

The pure fractions are evaporated, gathered in ethyl acetate, anhydrated, and the hydrochloride is precipitated with HCl in ethanol. The hydrochloride is crystallized from ethanol. This is filtered and vacuum-dried, obtaining 18.0 g of a compound to which elementary analysis (C,H,N) and nuclear magnetic resonance spectroscopy (protons) both attribute a structure of 1,3-bis[3-(ß-diethylaminoethyl)-4-methylcσumarin-7-yloxy]-2-hydroxypropane hydrochloride. EXAMPLE 40 :

1-[3-(ß-diethylaminoethyl)-4-methylcoumarin-7-yloxy]-3-[3-(ß-diethylaminoethyl)-4-methyl-8-chlorocoumarin-7-yloxy]propane hydrochloride

35.0 g of 3-(ß-diethylaminoethyl)-4-methyl-7-hydroxy-coumarin are suspended in 500 ml of ethanol

and to this are added 14.2 g of 50% KOH in water.

It is left to stand for 1 hour at 50ºC and then

concentrated and vacuum-dried. It is gathered with 500 ml of 2-butanone and to this are added 35.1 g of K₂CO₃. It is heated while being shaken, then to this are added 25.7 g of 1.3 dibromopropane and it is left to react for 12 hours. The solvent is evaporated and the residue is dissolved with 500 ml of ethyl acetate. It is washed with a 1 N solution of NaOH. The solvent is

anhydrated, concentrated and crystallized

from isopropyl alcohol. It is filtered and the

1-[3-(ß-diethylaminoethyl)-4-methylcoumarin-7-yloxy]-3-bromopropane crystals are dried. At the same

time, the potassium salt of 3-(ß-diethylaminoethyl)-4-methyl-7-hydroxy-8-chlorocoumarin is

prepared by treating 39.0 g of this compound with 14.2 g of 50% KOH in water in 500 ml of ethanol. This mixture is left to stand for l hour at 50ºC and then it is concentrated and vacuum-dried.

The potassium salt is gathered with 500 ml of

2-butanone and to this are added 35.1 g of K₂CO₃,

It is heated while being shaken, and then to this are added the 1-[3-(ß-diethylaminoethyl)-4-methylcoumarin-7-yloxy]-3-bromopropane crystals and it is left to react for 12 hours.

The solvent is evaporated and the residue is

dissolved with 500 ml of ethyl acetate. It is washed with a 1N solution of NaOH. The solvent is anhydrated and concentrated. The reaction product is purified by chromatography with Prep LC System Waters, using as eluent a mixture of CHCl₃-CH₃OH-NH₄OH (30%) at a gradient of 95:5:0.2 to 70:30:0.2.

The pure fractions are concentrated and crystallized from toluene. The crystallized product is dissolved in ethyl acetate, anhydrated and treated with HCl in ethanol until a Congo red indicator change is noted. It is filtered and crystallized from isopropyl alcohol. This is filtered and vacuum-dried, obtaining 36.4 g of a

compound to which elementary analysis (C,H,N) and

nuclear magnetic resonance spectroscopy (protons)

both attribute a structure of

1-[3-(ß-diethylaminoethyl)-4-methylcoumarin-7-yl-oxy]-3-[3-(ß-diethylaminoethyl)-4-methyl-8-chlorocoumarin-7-yloxy]propane hydrochloride.

EXAMPLE 41:

1-[3-(ß-diethylaminoethyl)-4-methylcoumarin-7-yl-oxy]-6-[3-(ß-diethylaminoethyl)-4-methyl-8-chlorocoumarin-7-yloxy]hexane hydrochloride

35.0 g of 3-(ß-diethylaminoethyl)-4-methyl-7-hydroxycoumarin are suspended in 500 ml of ethanol and to this are added 14.2 g of 50% KOH in water.

It is left to stand for 1 hour at 50ºC and then

concentrated and vacuum-dried. It is gathered with 500 ml of 2-butanone and to this are added 35.1 g of K₂CO₃. It is heated while being shaken, then to this are added 31.0 g of 1.6 dibromohexane and it is left to react for 12 hours. The solvent is evaporated and the residue is dissolved with 500 ml of ethyl acetate. It is washed with a 1 N solution of NaOH. The solvent is

anhydrated, concentrated and crystallized by

ethyl alcohol. It is filtered and the 1-[3-(ß-di ethylaminoethyl)-4-methylcoumarin-7-yloxy]-6-bromohexane crystals are dried.

At the same time, the potassium salt of 3-(ß-diethylaminoethyl)-4-methyl-7-hydroxy-8-chloro

coumarin is prepared by treating 39.0 g of this compound with 14.2 g of 50% KOH in water in 500 ml of ethanol. It is left to stand for 1 hour at 50ºC and then concentrated and vacuum-dried. The potassium salt is gathered with 500 ml of 2-butanone and to this are added 35.1 g of K₂CO₃. It is heated while being shaken, and then to this are added the 1-[3-(ß-diethylaminoethyl)-4-methylcoumarin-7-yloxy3-6-bromohexane crystals

and it is left to react for 12 hours.

The solvent is evaporated and the residue is

dissolved with 500 ml of ethyl acetate; it is

washed with a 1N solution of NaOH. Th solvent is

anhydrated and concentrated. It is purified by

chromatography with Prep LC System Waters, using as eluent a mixture of CHCl₃-CH₃OH-NH₄OH (30%) at a

gradient of 95:5:0.2 to 70:30:0.2.

The pure fractions are concentrated and precipitated by methahol. The precipitate is dissolved in a mixture of chloroforra/methanol and treated with HCl in ethanol until a Congo red indicator change is observed. It is slightly concentrated and then precipitated by

the addition of ethyl acetate. This is filtered and vacuum-dried, obtaining 35.7 g of a compound to

which elementary analysis (C,H,N) and nuclear

magnetic resonance spectroscopy (protons) both

attribute a structure of 1-[3-(ß-diethyl-aminoethyl)-4-methylcoumarin-7-yloxy3-6-[3-(ß-diethyl-aminoethyl)-4-methyl-8-chlorocoum£irin-7-yloxy]hexane hydrochloride. EXAMPLE 42:

1-[3-(ß-diethylaminoethyl)-4-methylcoumarin-7-yloxy]-1-[3-(ß-diethylaminoethyl)-4-methy1-8-chlorocoumarin-7-yloxy]dicarbethoxy methane hydrochloride

25.0 g of 3-(ß-diethylaminoethyl)-4-methyl-7-hydroxycoumarin are placed in a 500-ml reactor

having shaking and separating equipment. 300 ml of toluene and 24.8 g of K₂CO₃ are added. It is

heated, separating and eliminating the reaction

water; the toluene is concentrated to about 50 ml, cooled to room temperature and to this are added

100 ml of DMSO. One hour later 31.7 g of dibromomalonic ester are added thereto, and it is left to react for 12 hours. The potassium salt of 3-(ß-diethylaminoethyl)-4-methyl-7-hydroxy-8-chlorocoumarin is

added, having been previously prepared by treating

28:5 g of the 8-chlorocoumarin compound with 10-1 g of 50% KOH in water for 1 hour in 250 ml of ethanol at 50ºC and evaporating to dryness under high vacuum.

The reaction mixture is left to react for 24 hours, and then gathered in toluene and washed with water and a 1N solution of NaOH. The solvent is anhydrated and concentrated. It is purified by chromatography with Prep LC System Waters using as eluent a mixture of

CH₂Cl₂-CH-OH-NH₄OH (30%) 96:4:0.2.

The pure fractions are concentrated, gathered with ethyl acetate, anhydrated, and the hydrochloride is precipitated with HCl in ethanol. A pulp is obtained which is freeze-dried, obtaining 11.2 g of a compound to which elementary analysis (C,H,N) and nuclear

magnetic resonance spectroscopy (protons) both

attribute a structure of 1-[3-(ß-diethylaminoethyl)-4-methylcoumarin-7-yloxy]-1 -[3-(ß-diethylaminoethyl)-4-methyl-8-chlorocoumarin-7-yloxy)dicarbethoxy methane hydrochloride.

EXAMPLE 43:

1-[3-(ß-diethylaminoethyl)-4-methylcoumarin-7-yl-oxy]-1-[3-(ß-diethylaminoethyl)-4-methyl-8-chlorocoumarin-7-yloxy)carbethoxy methane hydrochloride

25.0 g of 3-(ß-diethylaminoethyl)-4-methyl-7-hydroxy-coumarin are placed in a 500-ml reactor

equipped with a shaking and separating apparatus. 300 ml of toluene and 24.8 g of K₂CO₃ are added. It is

heated, separating and eliminating the reaction

water; the toluene is concentrated to about 50 ml, cooled to room temperature and to this are added

100 ml of DMSO. One hour later 15.7 g of ethyl

dichloroacetate are added, and it is left to react for 12 hours. The potassium salt of 3-(ß-diethylaminoethyl)-4-methyl-7-hydroxy-8-chlorocoumarin is

added, having been previously prepared by treating

23.5 g of the 8-chlorocoumarin compound with 10.1 g of

50% KOH in water for 1 hour in 250 ml of ethanol at 50ºC and evaporating to dryness under high vacuum.

The reaction mixture is left to react for 24 hours, and then gathered in toluene and washed with water and a IN solution of NaOH. The solvent is anhydrated and concentrated. It is purified by chromatography with Prep LC System Waters using as eluent a mixture of

CH₂Cl₂-CH₃OH-NH₄OH (30%) 95:5:0.2.

The pure fractions are concentrated, gathered with ethyl acetate, anhydrated, and the hydrochloride is precipitated with HCl in ethanol. It is filtered, washed with ethyl acetate and vacuum-dried, obtaining 15.4 g of a compound to which elementary analysis (C,H,N) and nuclear magnetic resonance spectroscopy (protons) both attribute a structure of

1-[3-(ß-diethylaminoethyl)-4-methylcoumarin-7-yloxy)-1-[3-(ß-diethylaminoethyl)-4-methyl-8-chloroc oumarin-7-yloxy]carbethoxy methane hydrochloride.

EXAMPLE 44:

Examples of pharmaceutical compositions for

inj ection : Preparation No. 1: one 2-ml ampoule contains:

- active principle 30 mg

- mannitol 100 mg

- water for injection q.s. ad 2 ml Preparation No. 2: one 2-ml ampoule contains:

- active principle 40 mg

- mannitol 100 mg

- water for injection q.s. ad 2 ml Preparation No. 3: one 30-mg freeze-dried vial plus a 5-ml ampoule of solvent in which:

a) one freeze-dried vial contains:

- active principle 30 mg

- mannitol 30 mg b) one 5-ml ampoule of solvent contains:

- sodium chloride 45 mg

- water for injection q.s. ad 5 ml

Preparation No. 4: one 40-mg freeze-dried vial plus one 5-ml ampoule of solvent in which:

a) one freeze-dried vial contains:

- active principle 40 mg

- mannitol 25 mg b) one 5-ml ampoule of solvent contains:

- sodium chloride 45 mg

- water for injection q.s. ad 5 ml EXAMPLE 45:

Examples of pharmaceutical compositions for oral use:

Preparation No. 1: one 100-mg capsule contains:

- active principle 100.00 mg - saccharose 92.77 mg

- corn starch 30.93 mg

- magnesium stearate 34.60 mg

- povidone 25.48 mg

- monobasic potassium phosphate 20.80 mg - trimellitate cellulose acetate 95.42 mg

- gelatin capsule 77.00 mg

Preparation No. 2: one 200-mg capsule contains:

- active principle 200-00 mg - saccharose 92.77 mg

- corn starch 30.93 mg

- magnesium stearate 34.60 mg

- povidone 25.48 mg

- monobasic potassium phosphate 20.80 mg - trimellitate cellulose acetate 95.42 mg

- gelatin capsule 77.00 mg

Preparation No. 3: one 100-mg capsule contains:

- active principle 100.00 mg - vegetable oil 187.00 mg

- hydrogenated soybean oil 2-40 mg

- hydrogenated vegetable oils 0.60 mg

- soybean lecitin 1.00 mg

- yellow wax 0.60 mg - ethyl vanillin 0.30 mg

- gelatin 79-00 mg

- glycerol 30.00 mg

- titanium bioxide E 171 1.10 mg

- cupric chlorophyll E 141 0.10 mg

- yellow iron oxide E 172 0.60 mg

- brown iron oxide E 172 0.03 mg

- sodium ethyl p-hydroxybenzoate 0.30 mg

- sodium propyl p-hydroxybenzoate 0.20 mg

Claims

The following is claimed:

1. Bicoumarin derivatives of the formula:

wherein each of the substituents R₂-R₅ and R₇-R₁₀ represents hydrogen or a substituent chosen from the group formed by:

- a halogen,

- a free or esterified carboxy group,

- a free or esterified or etherified hydroxy

group,

- a lower alkyl or lower monocycloaryl-alkyl or lower monocycloalkyl-alkyl radical, or a corresponding unsaturated radical, which can be substituted in the aliphatic moiety by one or more free or esterified or etherified hydroxy groups or by oxo groups or by free or esterified carboxy groups, and in the aryl part by one or more lower alkyl groups or halogens or lower alkoxy or hydroxy groups, and in the cycloaliphatic part by one or more lower alkyl groups, or

- a monocycloalkyl radical or a corresponding unsaturated radical, unsubstituted or substituted by one or more lower alkyl groups, a monocycloaryl radical, unsubstituted or substituted by one or more lower alkyl groups or halogens or lower alkoxy or hydroxy groups,

- R₁ and R₆ represent an aza-alkyl or aza-monocycloalkyl or aza-monocycloalkyl-alkyl or aza-alkylmonocyclo-alkyl or

aza-alkyl-monocycloalkyl-alkyl radical, or a corresponding unsaturated radical, with a

maximum of 12 carbon atoms, and which may be interrupted in the carbon atom chain by the

-NH-, -O-, or -S-groups, and/or may be

substituted by free or esterified or etherified hydroxy groups, or by oxo groups or by lower alkyl groups, or by free or esterified carboxy groups, and wherein R₄ and R₉ may have the same meanings as R₁ and R₆, and

- -X- stands for a bivalent hydrocarbyl radical chosen from the group formed by an alkylene radical, monocycloaryl-alkylene radical or monocycloalkyl-alkylene radical or a corresponding unsaturated radical, which may be interrupted in the carbon atom chain by

heteroatoms chosen from the group formed by

-NH-, -O-, and -S-, or by a monocyclo-arylene or monocyclo-alkylene radical, and may be

substituted in the aliphatic or cycloaliphatic part by one or more halogens or free or

esterified or etherified hydroxy groups, or by lower amino, alkyl, or dialkylamino groups, or C_5-6 alkyleneamino groups, optionally

interrupted by -NH-, -O-, or -S-groups, by oxo groups or by free or esterified carboxy groups, and in the aromatic part by one or more lower alkyl groups or halogens or lower hydroxy or alkoxy groups, and a monocycloalkylene

radical, unsubstituted or substituted by one or more lower alkyl groups or free or

esterified or etherified hydroxy groups, or by lower amino, alkyl, or dialkylamino groups, or by oxo groups, or by free or esterified carboxy groups and their basic or acid salts.

2. Bicoumarin derivatives according to claim 1, wherein the halogen atom is chlorine, bromine or fluorine.

3. Bicoumarin derivatives according to claim 1, wherein the R₂-R₅ and R₇-R₁₀ alkyl radicals have a maximum of 7 carbon atoms.

4. Bicoumarin derivatives according to claim 1, wherein the R₂-R₅ and R₇-R₁₀ monocyclo-aryl-alkyl and monocycloalkyl-alkyl

radicals have a maximum of 7 carbon atoms in the aliphatic moiety.

5. Bicoumarin derivatives according to any one of claims 3-4 , wherein the R₂-R₅ and R₇-R₁₀

cycloaliphatic radicals or those contained in such substituents are monocyclic and have from 3 to 7 carbon atoms in the ring.

6. Bicoumarin derivatives according to claim 1,

wherein unsaturated R₂-R₅ and R₇-R₁₀

hydrocarbyl radicals corresponding to the lower alkyl, lower monocycloaryl-alkyl, lower

monocycloalkyl-alkyl radicals have a double

bond in the aliphatic and/or cycloaliphatic

part.

7. Bicoumarin derivatives according to any one of

claims 5 and 6, wherein the R₂-R₅ and R₇-R₁₀

cycloaliphatic radicals or those contained in such substituents are monocyclic and have

from 5 to 7 carbon atoms.

8. Bicoumarin derivatives according to any one of

claims 1, 4 and 6, wherein the R₂-R₅ and R₇-R₁₀ aryl substituents or aryl groups contained in εuch substituents are phenyl groups,

unsubstituted or substituted by 1 to 3 lower

alkyl groups or lower alkoxy groups, or hydroxy groups or halogens.

9. Bicoumarin derivatives according to any one of

claims 1, 6 and 8, wherein the lower alkyl or alkoxy groups have a maximum of 7 carbon atoms.

10. Bicoumarin derivatives according to claim 9, wherein said lower alkyl or alkoxy groups have a maximum of 4 carbon atoms.

11. Bicoumarin derivatives according to claim 1,

wherein the R₁ and R₆ and the R₄ and R₉ radicals are aza-alkyl, aza-monocycloalkyl, aza-monocycloalkyl-alkyl or aza-alkylmonocyclo -alkyl groups which have no more than 7 carbon atoms in the aliphatic parts and between 3 and

7 carbon atoms in the cyclic parts thereof.

12. Bicoumarin derivatives according to claim 11,

wherein the R₁, R₄, R₆ and R, radicals have no

more than 4 carbon atoms in the aliphatic parts and 5 or 6 carbon atoms in the cyclic parts thereof.

13. Bicoumarin derivativeε according to

claim 11 or 12, wherein the aza group interrupts the aliphatic chain of carbon atoms.

14. Bicoumarin derivatives according to

claim 12 or 13, wherein the aza group

interrupts one of the cyclic parts of the

carbon atom chain.

15. Bicoumarin derivatives according to claim 14,

wherein the cyclic group is a radical derived from piperidine, piperazine or morpholine.

16. Bicoumarin derivatives according to claim 11,

wherein the aza-alkyl group is derived from an alkyl with a maximum of 7 carbon atoms.

17. Bicoumarin derivatives according to claim 16,

wherein the aza-alkyl group is a 3-aza-3-ethyl-pentyl group.

18. Bicoumarin derivatives according to any one of

claims 11, 14 and 15, wherein the cyclic parts are substituted by free, esterified or

etherified organic groups.

19. Bicoumarin derivatives according to any one of claims 11, 14 and 15, wherein the cyclic parts are substituted by free or esterified carboxy groups.

20. Bicoumarin derivatives according to any one of

claims 11, 14 and 15, wherein the cyclic parts are substituted by alkyl groups with a maximum of 4 carbon atoms.

21. Bicoumarin derivatives according to any one of

claims 1-20, wherein the -X- moiety represents an alkylene radical or a corresponding unsaturated radical with only one double bond and having from 1 to 8 carbon atoms.

22. Bicoumarin derivatives according to any one of

claims 1-20, wherein the -X- moiety represents a monocyclo-alkylene radical with 5 or 6 carbon atoms.

23. Bicoumarin derivatives according to any one of

claims 1-20, wherein the -X- moiety represents a monoaryl or monocycloalkyl-alkylene radical with a maximum of 8 carbon atoms in the aliphatic

part, the cyclic part optionally being substituted by 1 to 3 alkyl groups with a maximum of

4 carbon atoms.

24. Bicoumarin derivatives according to any one of

claims 20-23, wherein the alkylene or cycloalkylene radical is substituted by one or more functions chosen from the group formed by a

hydroxyl group, free or esterified or etherified, a free or esterified carboxy group, an oxo group and an amino group which is free or alkylated with alkyl groups having a maximum of 4 carbon atoms.

25. Bicoumarin derivatives according to any one of

claims 20-24, wherein the alkylene or cycloalkylene radical is interrupted in the

aliphatic or cycloaliphatic part or in both parts by a heteroatom chosen from the group formed by

-NH-, -O-and -S-.

26. Bicoumarin derivatives according to any one of

claims 1-25, wherein etherified or esterified hydroxyl groups as substituents R₂-R₅ and

R₇-R₁₀ or present in such substituents or in

substituents R₁, R₄, R₆ and R₉ or in the

-X-radical are derived from saturated or unsaturated aliphatic alcohols having a maximum of 7 carbon atoms or from monoarylaliphatic alcohols having a maximum of 9 carbon atoms or, respectively,

from acids of the aliphatic, aromatic,

araliphatic or heterocyclic series with a

maximum of 9 carbon atoms.

27. Bicoumarin derivatives according to any one of

claims 1-25, wherein esterified carboxylic

groups as substituents R₂-R₅ and R₇-R₁₀ or

present in such substituents or in the

substituents R₁ and R₄ and R₆ and R₉ or in the

-X-radical, are derived from monovalent or bivalent aliphatic alcohols, saturated or unsaturated, with a maximum of 7 carbon atoms or from monoaryl-aliphatic alcohols with a maximum of 9 carbon atoms.

28. Bicoumarin derivatives according to any one of claims 1-25, wherein -alkylamino, dialkylamino or alkyleneamino groups optionally present in the -X-radical, are derived from alkyl groups having a maximum of 4 carbon atoms or from aza-cyclo- alkyl groups having 4 to 6 carbon

atoms in the ring, optionally interrupted in the hydrocarbyl chain by heteroatoms chosen from the group formed by -NH-, -O- and -S-.

29. Bicoumarin derivatives according to any one of

claims 1-28, wherein -X-represents an alkylene radical having from 1 to 6 carbon atoms,

unsubstituted or substituted by one or two

functions chosen from the group formed by free or esterified hydroxy groups with lower

aliphatic acids or oxo groups and free amino groups, lower alkyl and dialkylamino groups and C_5-6 alkyleneamino and alkyleneamino groups

interrupted in the carbon atom chain by a

heterocyclic group or atom chosen from the

group formed by -NH-, -O- and -S- .

30. Bicoumarin derivatives according to any one of

claims 1-29, wherein the substituents R₁-R₆ are identical to substituents R₇-R₁₀.

31. Bicoumarin derivatives according to claim 1 of

the formula

wherein

A represents a saturated aza-alkyl, aza-monocycloalkyl-alkyl or aza-alkyl-monocycloalkyl radical with a maximum of 12 carbon atoms and wherein the cycloalkyl group has 5 or 6 carbon atoms, optionally being further interrupted in the carbon atom chain by one of the -NH-,

-O-and -S-groups, and which may be substituted at the carbon atoms by alkyl groups with

1 or 2 carbon atoms or by lower hydroxy or alkoxy groups.

B, C and D may represent a hydrogen atom and B may represent also a lower alkyl or alkenyl radical or a monocyclic aryl radical or a halogen atom, C may represent the same aza-hydrocarbyl radical as defined for substituent A, or the same hydrocarbyl groups as defined for B, and D can represent the same hydrocarbyl radicals as defined for B, and -X-, represents an alkylene radical having from 1 to 6 carbon atoms and which may be interrupted in the carbon atom chain by heteroatoms chosen from the group formed by -NH-, -O-, and -S-, and/or which can be unsubstituted or

substituted by one or two functions chosen from the group formed by halogens, hydroxy groups, free or esterified with carboxylic acids having from 1 to 9 carbon atoms or etherified with alcohols having from 1 to 7 carbon atoms, and free or esterified carboxy groups with alcohols having from 1 to 7 carbon atoms and oxo groups and lower alkylamino or dialkylamino groups.

32. Bicoumarin derivatives according to claim 31, wherein in formula II, B represents a lower alkyl or alkenyl radical or a monocyclic aryl radical, C and D represent a hydrogen atom, a halogen or a lower alkyl or alkenyl radical.

33. Bicoumarin derivatives according to claim 32, wherein in formula II of claim 31, A represents the diethylamino-ethyl or di-isopropyl-amino-ethyl radical or their corresponding quaternary ammonium groups with lower alkyl groups or the morpholinyl-methyl, piperidinyl-methyl, thiomorpholinyl-methyl or piperazinyl-methyl radical.

34. Bicoumarin derivatives according to claim 33, wherein B represents the methyl or phenyl

group, C and D are a hydrogen citom, the allyl group or chlorine.

35. Bicoumarin derivatives according to any one of claims 3.3 and 34, wherein -X- is the trimethylene, 3-hydroxy-trimethylene, hexamethylene or 2-di-carbethoxy-trimethylene radical.

36. A bicoumarin derivative according to claim 1

selected from the group consisting of:

1,3-bis[3-(ß-diethylaminoethyl)-4-methylcoumarin- 7-iloxy]propane hydrochloride, bis[3-(ß-diethylaminoethyl)-4-methylcoumarin-7-yloxy3dicarbethoxymethane hydrochloride, bis[3-(ß-diethylaminoethyl)-4-methylcoumarin-7-yloxy]carbethoxymethane hydrochloride,

1,3-bis[3-(ß-diethylaminoethyl)-4-phenylcoumarin- 7-yloxy]propane hydrochloride, bis[3-(ß-diethylaminoethyl)-4-roethyl-8-chlorocoumarin-7-yloxy]carbethoxymethane hydrochloride, bis[3-(ß-diethylaminoethyl)-4-methyl-8-chlorocoumarin-7-yloxy]dicarbethoxymethane hydrochloride,

1,3-bis[3-(ß-diethylaminoethyl)-4-methylcoumarin- 7-yloxy]-2-hydroxypropa.ne hydrochloride, and

1,3-bis[3-(ß-diethylaminoethyl)-4-methyl-8- chlorocoumarin-7-yloxy]-2-hydroxypropane

hydrochloride.

37. Metal or base salts of any one of the

bicoumarins of claims 1-36 containing acid groups.

38. Therapeutically acceptable salts according to claim 37.

39. Salts obtained by acid addition of any one of the bicoumarins of claims 1-36 containing basic groups.

40. Therapeutically acceptable salts according to claim 39.

41. A process for the preparation of the

bicoumarin products of claim 1 characterized by the steps of treating:

7-hydroxy-coumarin of the formula

wherein R₁-R₅ are the same substituents as

defined for formula I or groups convertible

into the same, or one of its metal salts, with a compound of the formula

Z₁-X-Z₂ (IV) wherein Z, and Z₂ are the same or different from each other, and each represents a reactive

group with regard to phenolic etherification

and -X- has the same significance as in

formula I or it signifies a substituent

convertible into the same, or treating said

coumarin compound III with a compound of the

formula

Z₁-X-Z₃ (V) wherein Z₁ and -X- have the same significance as recited above and Z₃ stands for a

non-reactive group with regard to phenolic

etherification, but is convertible into a reactive group with regard to said reaction or a

hydrogen atom of the unsaturated H-C= structure of the terminal hydrocarbyl group of -X-, and then converting Z₃ of the compound obtained of

the formula:

into a reactive group with regard to phenolic

etherification , and then treating the coumarin compound obtained with a 7 -hydroxy-coumarin of the formula

or one of its metal salts, wherein R₆-R₁₀ has

the same significance as for substituents R₁-R₅ of formula III, but if R₆-R₁₀ are not identical to R₁-R₅, converting, in the bicoumarin compound obtained, substituents R₁-R₁₀ and -X- which differ from those corresponding to the significance which they have in the compounds of formula I, into substituents of said formula, and if desired, converting the products obtained into their metal or acid addition salts, or into their quaternary

ammonium salts.

42. The process according to claim 41, wherein

bicoumarin derivatives with a symmetrical

structure are prepared by reacting the compound of formula III with the compound of formula

Z₁-X-Z₂ in a stoichiometric ratio of 2:1.

43. The process according to claim 41, wherein

bicoumarin derivatives with a symmetrical

structure are prepared by reacting the compound of formula III with a compound of formula

Z₁-X-Z₂ in a stoichimetric ratio of about 1:1 or with an excess of coumarin reagent and the product obtained is reacted with a coumarin of formula VII.

44. The process according to claim 41, wherein

bicoumarin derivatives with an asymmetrical stucture are prepared by reacting a compound of formula III with a compound of formula V

wherein Z₃ represents a free or esterified hydroxy group with esters of carbonic acid or of lower carboxylic aliphatic acids or a

hydrogen atom of the unsaturated structure H-C= of a terminal hydrocarbyl group of the

-X-radical.

45. The process according to any one of claims 41-44, wherein the reactive group Z₁ or Z₂ is a hydroxy group esterified with hydracids, with inorganic oxygenated acids or with organic sulfonic acids.

46. The process according to claim 45, wherein alkylene halogenides are used.

47. The process according to claim 45, wherein the methanesulfonic or p-toluenesulfonic esters of an -OH-X-OH alcohol are used.

48. The process according to any one of claims 41-44, wherein the reactive group Z₁ or Z₂ represents an epoxide group.

49. The process according to any one of claims 41-44, wherein the 7-hydroxycoumarin of formula III or formula VII is used in the form of one of its metal salts.

50. The process according to claim 49, wherein

sodium, potassium or cesium salts are used.

51. The process according to any one of claims 41-44, wherein the 7-hydroxy-coumarin of formula III or VII are used in their free form and

etherification is performed with the compounds of formula IV and V, respectively, in the

presence of a basic agent.

52. The process according to any one of claims 41 and 48, wherein the 7-hydroxy-couma.rin of formula III or VII is etherified with the compound Z₁-X-Z₂ wherein Z₁ and Z₂ are epoxide groups.

53. The process according to any one of claims 41 and 44, wherein in the compound of formula VI an esterified hydroxy group is converted into a free hydroxy group in a known way and the

hydroxy group is converted into a reactive

ester in a known way.

54. The process according to any one of claims 41 and 44, wherein in the compound of formula VI an unsaturated structure -X- is converted into an epoxide group by hydrogen peroxide or an

organic peroxide in a known way.

55. A pharmaceutical preparation containing a

bicoumarin of formula I of claim 1 together

with a pharmaceutically acceptable excipient.

56. A pharmaceutical preparation according to claim

55, containing any one of the compounds of

claims 1-40.

57. The use of a bicoumarin derivative of formula I

of claim 1 in therapy for the treatment of vascular pathologies.

58. The use according to claim 57 for the treatment of peripheral vascular pathologies, anginal ailments and cerebral vascular pathologies.

59. The use according to claim 57 as an antithrombotic agent.

60. The use according to claim 57 as an antihypertensive agent.