FR2642973A1 - Medicinal composition containing as active principle a dioxymethyl carbohydrate derivative of strophanthidin - Google Patents

Abstract

A composition containing 2',3'-dioxymethyl-6beta-methyl-3beta-D-glucosylstrophanthidin I or Mansonin, which is a medicament for the treatment and prevention of cardiac insufficiency, is prepared with the object of improving cardiotonicity in order to improve cardiac rhythm and to combat fibrillation and arterial hypertension originating from a calcium excess, to combat arteriosclerosis.

Description

soit, dans ea représentation spatiale

The present invention relates to the dioxymethylated carbohydrate derivative of strophanthidine, which is known as Mansonin, namely 2 ', 3' O dioxymethyl-6-methyl-3ssD-glucose-strophanthidine of the formula
C31H46O10, of digital-mimetic structure, and corresponding to the following formula (I):

either, in the spatial representation

 The present invention relates, on the one hand, to a novel process for the preparation of the compound of formula (I), and, on the other hand, to the use of this compound, or its pharmaceutically acceptable salts thereof, in pharmaceutical preparations for the preparation thereof. treatment and prevention of heart failure and cardiovascular disease.

The invention also relates to a fraction containing the compound of formula (i) and its applications also for the treatment and prevention of heart failure and cardiovascular diseases.

 The preparation of the compound of formula (I) has been described for the first time by Allgeier et al., In "Hely Chim Acta 50, 1967, p 455", and consisted of a purification of seed extracts. Mansonia altissime var altissima a sterculiacea of the great West African forests (Ivory Coast, Ghana, Nigeria and Cameroon) This tree was described by Chevalier A. in "Rev. Bot.

Agr. Too much. 27, 1947. p. 422 In West Africa, bark preparations of M. altissima are traditionally used as arrow poisons, and as decoctions in anticedemic external baths.

The present invention therefore firstly relates to a process for preparing the compound of formula (I), this process being characterized by the fact that
in a first step, an extraction is carried out
hot water bark of Mansonia altissima, for
obtain an extract P1;
- in a second step, an extraction of
the P1 extract with an organic solvent capable of
solubilize P1 to obtain an extract P2;
- In a third step, we conduct a split
of the P2 extract by a succession of chromatographies
of exchange with anionic resins and
cationic agents capable of eliminating
chromogens contained in the extract P2, while retaining A
each chromatography that the fraction capable
to inhibit the ATPase Na, K, to obtain a
fraction P3 ;;
in a fourth step, a fractionation is carried out
of 1 P3 fraction by elution chromatography or
sharing, in order to collect the fraction P4 that is
capable of inhibiting the Na +, K +, and
compound of formula (I> sought, and
if necessary, converting the compound of formula (I)
in one of its pharmaceutically acceptable salts.

 As a starting material, the bark of Mansonia altissima var. altissima. The first step of the process is therefore an aqueous extraction, hot, especially boiling, at atmospheric pressure or, if desired, under pressure.

 At the second stage. which can be conducted cold, is used as extraction solvent, a chlorinated organic solvent, such as chloroform.

 Preferably, between the first and the second stage, delipidation of the extract P1 is carried out, in particular using a conventional delipidating agent, such as petroleum ether.

 In the third step, anionic resins, in particular resins of the dextran or polysaccharide type, modified by the introduction of anionic functions, such as the methyl carbamate function, are used as the anionic resins.

 At this same stage, cationic resins, especially dextran or polysaccharide-type resins, modified by the introduction of cationic functions, such as the dimethylaminoethyl function, and resins obtained by condensation of formaldehyde with phenol acids and phenyl-sulphonic.

 In the fourth step, fractionation of the P3 fraction can be carried out by elution chromatography, for example on a silica gel column, or by thin layer partition chromatography.

 The compound of formula (I) is obtained in particular in the form of fine crystals, in the highly concentrated state in a water-ethanol mixture (1: 1). This compound can therefore be presented in the form of very dilute solutions in the bidistilled water or saline.

 Moreover, the compound of formula (I) may be salified at its hydrophobic sites.

 The present invention also relates to the P3 fraction as obtained by the process which has just been defined.

 The present inventors have now moreover demonstrated that the compound of formula (I) has at the same time tonicardial, vasodilatory activities. anticalcic and antiarrhythmic.

 Certainly. It had been previously shown that aqueous extracts of bark of Mensonia altissima var.

aitissima inhibit the activity of ATPase Na +, K of rabbit brain microsomes (C.R. Acad Sc, Paris, t 26-4, dated June 6, 1977). However, this total extract has no definite properties, being a complex mixture of many substances. The work carried out by the present inventors has now made it possible to demonstrate that the compound of formula (I) possesses tonicardial properties as well as antihypertensives, whereas most products of this type, if they are tonicardial, are rather, hypertensives.

This dual complementary potentiality is to be emphasized.

 Furthermore, it has been confirmed that the compound of formula (I) has anti-calcium properties, thus avoiding in hypertensives the accumulation and absorption of calcium by blood platelets, which can clog the blood vessels. It has also been confirmed that the compound of formula (I) has antiarrhythmic activity, which avoids the use of electric shock to combat fibrillation.This unique combination of properties makes the compound of formula (I) very interesting as drug for the treatment and prevention of cardiac insufficiency and cardiovascular pathology, as a substitute for classical digitalis.

It has thus been shown that the compound of formula (I) presents, in the anesthetized dog
a) a positive inotropic action, with an increase of
the contraction is 70% above normal s the dose
10 nM plasma, and the tone of the contraction of the
200% transversus at 50 nM dose in sheep
b> a peripheral vasodilator action with reduction
total peripheral resistance of 30% at the dose
10 nM plasma
c> an anti-calcium effect with reduction of a current
Calcium returning from 30% to 70% at doses of 5 to 50 nM
on the frog-auricular fiber of the frog.

 Moreover, the compound of formula (I) has antiarrhythmic effects at picomolar doses on the rat arrhythmic heart and anti-fibrotic effects a.

50 nM, by an extension of the diastolic delay of the monkey ventricular trabeculae.

This drug may therefore be prescribed in the treatment and prevention of medical conditions such as
- cardiac insufficiencies for which the
digitalis are prescribed, with the aim of improving the
cardiotonicity, to improve the heart rate, and
to fight against fibrillation
- hypercalcic hypertension, for
to fight arteriosclerosis
The present invention therefore relates to a medicinal composition for the treatment and prevention of heart failure and cardiovascular diseases in humans and animals, characterized in that it contains, as active principle, the fraction P as defined above, or 2'.K'-omegamymethyl-6-methyl-36D-glucose-strophanthidine of formula (I), or a pharmaceutically acceptable salt thereof, in association with a vehicle pharmaceutically acceptable for the type of administration considered.

 The subject of the present invention is also the use of the P3 fraction as defined above, the derivative of formula (I) or a pharmaceutically acceptable salt thereof, for the manufacture of a medicinal composition having the above-mentioned purpose. .

 The present invention also relates to a method for preparing a medicinal composition for the treatment and prevention of cardiac insufficiency and cardiovascular diseases in humans and animals, characterized in that a mixture of pharmaceutically effective fraction P3 as defined above, or 2 ', 3'-dioxymethyl-6ssmethyl3ssD-glucose-strophanthidine, or one of the p: arreueutiquement acceptable of the latter, with at least one excipient pharmaceutically acceptable compatible.

 The pharmaceutical composition according to the invention is suitable for oral administration, in particular in the form of capsules or tablets, or for administration by intravenous or intramuscular injection, or for intraperitoneal administration. In particular, it is suitable for the treatment of heart failure and cardiovascular diseases, in particular hypertensive and hypercalcic type arteriosclerosis, providing an effective amount of the compound of formula (I) or of one of its pharmaceutically acceptable salts, up to 300 ng per kg of body weight, or preferably up to 250 ng per kg of body weight.

 We will now describe in detail, firstly in (A) obtaining the compound of formula (I) in the pure state according to the process of the present invention, and its identification, then in (B), the character of the pharmacodynamic properties of this compound.

(A) PREFERING THE FORMULA COMPOUND (I>
1 - Extraction
the compound of formula (I) was extracted from Mansonis barks altissima var. altissillies
in the primary tropical forest of Ivory Coast.

500 g of these barks freed of their cuticle
are chopped, brought to a boil, for 2 hours, in
2 liters of bidistilled water. The brown-and-syrupy decoction
obtained is filtered on medical gauze, to eliminate
the residue, then on Whatmann n 3MM filter paper doubled
of hydrophilic cotton. The filtrate is then evaporated at
Dry in Rotavapor BUchi rotary evaporator at 30 ° C
and at reduced pressure.

The 100 g of the resulting P1 extract is
presenting in the form of a brown, viscous paste, are
delipidated with petroleum ether (2 ml / g P1), with
periodic agitation for 48 hours. The product
insoluble delipidation is redissolved in 300 ml of water
double distilled and chloroform extract (final report
CHCl3 / H2O 80:20, v / v) in a separatory funnel
for 24 hours. It is stirred by periodic reversals.

The chloroform phase, evaporated to dryness at 30.degree.
reduced pressure gives 10 g of a powdery extract P2,
yellow, bitter and very hygroscopic.

2) Purification
The 10 g of the extract P2 dissolved in
chloroform <1 ml / g> are purified by chromatography on
a series of three columns
the first (1 x 10 cm) is a carbamate column of
Methyl (CM -) - Anionic Sephadex C-25 (Pharmacia Fine
Chemicals);
- the second column (2.5 x 20 cm), a column
of Amberlite 20-50 type IRA-68 cationic lot 40280
(Reagenzen); and
- the third column (1.5 x 15 cm), a column of
DEAE + -Sephadex A25 (Pharmacia Fine Chemicals)
lot 4994.

 The substances are eluted through the column, pre-equilibrated with CHCl 3 / H 2 O (50:50, v: v), using CHCl 3 / H 2 O (50:50, v / v) as eluent for 6 hours. At an elution rate of about 62.5 ml / hour, 500 ml of colorless, clear eluates are collected and lyophilized. Na +, K + pork kidney ATPase, according to the method of Schooner et al described in Eur J. Biochem 1 (1967), p 334-343, to give 1 g of a P3 fraction, present in the form of a dirty white powder.

Fraction P3, redissolved in 10 ml of
CHCl3 / H2O (90:10, v / v). is chromatographed on a column (2.5 x 20 cm) of Silica gel (60 F 254, Merck Darmstad), equilibrated in a mobile phase CHCl3 / H20 (90:10, v / v) as described by Allgeier et al. in Helv Chim.

Acta. 50, 1967, pp. 456-460. The elution rate is 62.5 ml / h and the desired fraction P4, selected by the aforementioned Schöner method, is eluted in the first liter of the collected mobile phase. These 1000 ml of eluate are lyophilized and give 400 mg of semi-crystallized P4, which are redissolved in 20 ml of ethanol and crystallize in this solvent at room temperature.

3) Analvse fractions P3 and P4
Fractions P3 and P4 are thin-layer chromatographed plates (20 × 20 cm 3) of cellulose coated with fluorescent polyethylene imide (lot 5579-1166005, Merck
Darmstadt). 10 mg of each fraction dissolved in CHCl 3 / H 2 O (50:50 v / v) were deposited in comparison with the same amounts of Ouabain (lot 7887-8545885, Merck
Darmstad) and reference Mansonin (0.5 ml 10-3 M, lot 67). provided by Dr. Allgeier of the Institute of
Organic Ball Chemistry.

 The chromatography is up-mobile phase, composed of butanol / water / acetic acid (5: 3: 2, v / v: v> in a chamber saturated with butanol vapors elution lasts 4 hours.

The UV fluorescence reading of this chromatogram gives
- For P3: four spots, one with a Rf of 0.700
- For P4: a spot with an Rf of 0.700
- For the reference Mansonine: a spot at a Rf
0, 700
- For Ouabain: a spot with a Rf of 0.650.

 The P3 fraction was therefore resolved into four spots, which were removed by scraping. All fractions were dissolved in 3 ml of water each and then centrifuged for 20 minutes at 10,000 x g.

UV spectra (from 0 to 350 nm), made on 100 g / ml solutions of CHCl3 / H20 (50:50, v / v>, make it possible to evaluate the degree of purity of chromato graphite fractions.
P3: 4 peaks at 160, 217, 218 and 230 nm
- P4: 1 single peak at 217 nm
- Mansonin reference: 1 single peak at 217 nm
Further silica gel thin-layer plate chromatography, performed simultaneously and comparatively with ouabain and P4 fraction, was revealed with KEDDE reagent <Pharmacol. Weekblad, 82, 1947, p. 741>, H2SO4 2N and Cl3Sb.

These tracings make it possible to identify an unsaturated lactone and the steroidal and glycoside nature of the P4 fraction, as in ouabain and reference Mansonin.
The characterization of P4, thus identified with
Mansonine, consisted of the determination of the gross formula of molecular weight and structure by ion mass spectroscopy (Model CH4 Ionic T04 Einlassystem). The results are confirmed by a Brucker model nuclear magnetic resonance spectrum (FIG. 1).

The UV spectrum (FIG. 2) is also confirmed by high performance liquid phase chromatography (HPLC> (Model Reverse System Model 440), in isocratic condition <FIG. 3).

Legends of Figures 1, 2 and 3:
Figure 1: Resonance NMR of H + in the form [3H].

Solvent: pure ethanol
Figure 2: UV Optical density according to the
wavelength in nm.

Solvent: alccol-water <50:50)
Concentration: 100 μg / ml
Figure 3: HPLC Optical Density vs. Time
elution in minutes (at 254 nm).

Eluent: methanol / water (30:70, v / v)
Elution rate: 1 ml / minute
Recording speed = 1 cm / minute
5> Results
It results from the treatment of P4 by mass spectroscopy and by nuclear magnetic resonance (FIG. 1) that the substance is a carbohydrate steroid (C31H46O10). who reacts positively to the tests of
Libermann, antimony trichloride (Cl3Sb) and Kedde's reaction. These reactions respectively confirm the glycosidic and steroidal nature carrying an unsaturated lactone. The HPLC spectrum determines the degree of purity with a single peak (Fig 3), and the UV spectrum gives a maximum abscrption at 217 nm (Fig 2> .The melting temperature is 172 + 2'C, the rotational power [ &alpha;] D20 = +6.7C in methanol and molecular weight (MW) = 578.71.

(B) CHARACTERIZATION OF THE PHARMACODYNAMIC PROPERTIES OF
FORMULA COMPOUND (I)
The studies first focused on (1) the toxicity of compound (I), then <2) on its biochemical effects, and (3) on its hemodynamic and physiological effects.

(1) Limits of toxicity of the compound <I>
The lethal dose toxicity tests (DL>, the lethal dose 50 (LD50) and the maximum tolerated dose (MTD) were performed in rats, guinea pigs and rabbits <Table I>.

Table I: Terminal dose limits for toxicity of the compound (I)

<tb> Parameters <SEP> Rat <SEP> Guinea pig <SEP> Rabbit
<tb> DMT <SEP> (g / kg) <SEP> 120 <SEP>#<SEP> 5 <SEP> 130 <SEP><SEP>#<SEP><SEP> 3 <SEP> 12 <SEP>#<SEP><SEP> 0.02
<tb> LD50 <SEP> (g / kg) <SEP> 150 <SEP>#<SEP><SEP> 5 <SEP> 110 <SEP>#<SEP><SEP> 5 <SEP> 20 <SEP>#<SEP><SEP> 0.05
<tb><SEP> DL100 <SEP> (g / kg) <SEP> 200 <SEP>#<SEP> 5 <SEP> 130 <SEP>#<SEP><SEP> 7 <SEP> 100 <SEP>#<SEP><SEP> 0.5
<Tb>
The toxicity of the compound (I) is twice as low in the rat, reputedly resistant to digitalis, as in the guinea pig recognized quite sensitive In contrast, in the rabbit, this toxicity is 5 times stronger than in cobeye.

 In addition, tests on the contracility <inotropic effect), heart rate (chronotropic effect), conductivity (dromotropic effect> and excitability <bathmotropic effect) were performed on the isolated rabbit and monkey heart (Table II). ).

[Table II: Limits of toxicity of the compound (I) on the
myocardium (monkey and rabbit)

<tb> I <SEP> Dose <SEP> therapeutic <SEP> Dose <SEP> toxic
<tb> Effects <SEP>#<SEP><SEP> 100 <SEP> nM <SEP>#<SEP> 100 <SEP> nM
<tb><SEP>(<SEP> low <SEP> strong
<tb><SEP> (5x10-8M <x <10-7M) <SEP>(#<SEP> 10-7M)
<tb> Inotropic <SEP> (+) <SEP> (++) <SEP> * <SEP> (+++)
<tb> Chronotropic <SEP> (-) <SEP> (-) <SEP> * <SEP> (++)
<tb><SEP> Dromotrope <SEP> (-) <SEP> (-) <SEP> * <SEP> (---)
<tb> Sathmotrope <SEP> (-) <SEP> (+) <SEP> * <SEP> (++)
<Tb>
t Effects increasing in magnitude with concentrations in
compound of formula (I).

(2) Biochemical effects of the compound (I)
The compound (i) exhibits inotropic effects on the hemodynamics of the anesthetized dog. This inotropic effect is corroborated by inhibitory effects on Na + K + ATPase. The compound (I), by inhibiting ATPase Na K, binds to sites specific to digitalis with an affinity of the order of Ki = 142 nM. Compound (I) recognizes, in this action, all isoforms a (high affinity sites KD 10 nM) and a (low affinity sites KD 10 M).

 This differential recognition is well marked in rats, dogs, guinea pigs and monkeys by participation in well-defined percentages of populations of high affinity and low affinity sites.

The sensitivities compared to the compound (I) of the Na K ATPase isoforms of rat, guinea pig, dog and monkey heart sarcolemomas are reported in the literature.
Table III.

Table III: Comparative Table of Sensitivity to + +
compound I) isoforms of ATPase Na, K
sarcolemmas of the heart of the rat, guinea pig, dog
and monkey

<tb><SEP> Species <SEP> CI @@ (nm) <SEP><SEP> Kn <SEP>% <SEP> contribution <SEP> SR / Activity
<tb><SEP> e <SEP> fixation
<tb><SEP>&alpha; + (nM) <SEP><SEP>&alpha; (vM) <SEP><SEP> HA <SEP> BA <SEP> D &alpha; / kD &alpha;; +
<tb><SEP> at <SEP> nonmal
<tb><SEP> years old <SEP> Ca2 + <SEP> (n = 8) <SEP>! <SEP> 20 + 1 <SEP> I <SEP> 9 + 1 <SEP> 9 # 1 <SEP><SEP>!<SEP> 54 # 5 <SEP><SEP> 4656 <SEP> 1000
<tb> with <SEP> Ca2 + <SEP> nM + <SEP> (n = 7) <SEP> 1000 # 12 <SEP> 9 # 1 <SEP><SEP> 6 # 3 <SEP><SEP> 38 # 2 <SEP><SEP> 62 # 2 <SEP><SEP> 670
<tb> Ret <SEP> in <SEP> SA
<tb> shock <SEP> C + Ca2 + mM <SEP> (n = 5) <SEP> 1200 # 20 <SEP> 60 # 7 <SEP><SEP> 50 # 6 <SEP><SEP> 30 # 2 <SEP><SEP> 30 # 2 <SEP><SEP> 830
<tb> Hypertrophy <SEP> (n = 5)
<tb><SEP> + <SEP> Ca2 + <SEP> 2 <SEP> mM <SEP> 500 # 10 <SEP> 20 # 2 <SEP> 50 # 2 <SEP> 66 # 6 <SEP> 34 # 6 <SEP> 2500
<tb> Guinea pig
<tb> normal <SEP> (n = 8) <SEP> 100 # 10 <SEP><SEP> 10 # 1 <SEP><SEP> 3 # 1 <SEP><SEP> 57 # 4 <SEP><SEP> 43 # 4 <SEP><SEP> 300
<tb> Ineffective <SEP> (n = 2) <SEP> 100 # 10 <SEP><SEP> 10 + 1 <SEP> 15 + 5 <SEP> i <SEP> 48 + 5 <SEP> 5255 <SEP> 1500
<tb> well <SEP> normal <SEP> 30 # 5 <SEP><SEP> 0.5 # 0.1 <SEP><SEP> 0.1 # 0.00 <SEP><SEP> 30 # 5 <SEP><SEP> 70 # 5 <SEP><SEP> 167
<tb> normal inge <SEP>
<tb> fried <SEP> frozen <SEP> 600 # 50 <SEP><SEP> 75 # 10 <SEP><SEP> 55 # 10 <SEP><SEP> 68 # 2 <SEP><SEP> 32 # 2 <SEP><SEP> 167
<Tb>
shock: same operating conditions as hypertrophy.

 In this Table, it can be seen that the percentage ratio of the populations of the low affinity (BA) sites to those of the high affinity site (HA) populations, which marks the selectivity in the binding of the compound (I) to the isoforms. of the Na + K + ATPese, can be compared to the notion of therapeutic index IT which is determined in vivo by the ratio of the toxic dose to the therapeutic dose. In this table, the compound (i) presents, for the rat, unanimously recognized as resistant to digitalis (selectivity in the fixation of the order of 250), an index four times higher than digitalis (selectivity in the setting equal to 1000>. In pathological cases, this ratio increases from 1000 to 2500 in the hypertrophied rat heart, from 300 to 1500 in the guinea pig heart.

 In the monkey, for which this ratio is 750, the sensitivity of the isoforms to the compound (I) is perfectly represented by the curves of Figure 4.

Legend of Figure 4
ATPase Na K of the heart of the Mangabe + EGTA monkey sensitivity to ouabain (@) and compound (I) (@).

The curve gives the percentage of residual activity as a function of the logarithm of the oubain or compound (I) concentration.
(S) Hemodymnal and physiological effects of the compound (I)
These effects will be described with reference to
Figures 5 to 9 below
Figure SA: Inotropic effect of the compound (I)
3.3 nmol / kg / min and for 20 minutes on the pressure
systolic anesthetized dog (Jareau method and
al (1984) LND 623. Eur. Heart J., supplement F),
309-314. )
The curve shown is the variation curve (#%) of the pressure over time.

Figure 5B: Evolution of the potential action torque
intracellular (FA) / contraction
right ventricular transversus of sheep
(I) in the presence of the compound (I) 500 nM (toxic dose)
in a time ranging from 0 to 20 minutes
PA variations
N + 1 o to 6 minutes: 5%
2 10 mn: 9%
3 15 min: 15%
4 minutes: 26% (with OAP appearance>
Variations in the tone of the contraction
N 100%
1 120%
2 140%
3 175%
4,200% <with appearance of OAC)
PAO: oscillation after potential
OAC: oscillation after contraction
CII) as a function of concentrations of the compound <I) of O
at 500 nM at 20 minutes.

PA variations
N + 1: 5% <0-5. 10-8M)
2: 10% (10-7M)
3: 15% (2.10%)
4: 28% (5.10-7M) (with appearance of PAO)
Variations in the tone of the contraction
N 100%
1 120%
2,140
3 175%
4,200% <with appearance of OAC)
The technique used is that of the microelectrode <iE) (Ling et al., 1949V, J. Cell Comp.

Physiol. 34, 383-396; Coraboeuf et al (1949), C.R. Soc.

Biol., Paris, 143, 1329-1331). The infusion solution is tyrode + sucrose.

Figure 6: Vasodilator effect of the compound <I)
3.3 nmol / kg / min and for 20 minutes on several
parameters of the anesthetic dog hemodynamics.

The curves shown describe the variation of a parameter a to d below as a function of time
to Average Blood Pressure: WFP
b Kotr index: PAM / FC (HR = heart rate)
c Total Peripheral Resistance (RPT): PAM / DC
(DC = cardiac output)
of Oxygen Conscmmation: PAM x FC
The solution of the injected compound is diluted with NaCl
Control is attached to NaCl and mark #% = 0
The method is the aforementioned method of Jarreau et al.

Figure 7: Calcium current of depolarization of the fiber
sino-atrial frog by the method of
voltage-clamp (50 mV) (Deck et al <1964), Pflugers
Arch., 280, 50-62). The perfusion medium is
Physiological Ringer + Tetrodontoxin (TTX> + Mannitol
+ 2.5 mM Ca2t
without Compound (i) 0 M (N)
in the presence of compound (I) 5 x 10 9 M <1) #
Figure e # Effect of the <I) picomolar compound <20 μM on
heart made arrhythmic by hypothermia <heart
isolated from rat)
A: Normal functioning of the arrhythmic heart
B: Addition of the compound (I) to 20 μM <concentration
final)
C: Modification of the start-stop operation by the
compound <I)
D: C suite after 3 minutes Vertical scale: i cm - Horizontal: 10s
Figure 9: Effect of the compound (I) on time. of the
diastolic depolarization of trabeculae
ventricular monkey
Microelectrode technique.

9A Evolution of the diastolic phase of responses
electric trabeculae in the presence of Ca
and compound (I)
1. 2mM K +
2. 2mM K + and compound (I) 5 x 10-8M
3. 2mM K ++ and 0.5mM Ca ++
4. 2mM K +, 0.5 mM Ca ++ and compound (I) 5 x 10 -M
9B Comparative action of adrenaline and compound (I)
on the diastolic phase of the electrical response
transmembrane of monkey trabecula <same technique
only for 9A>.

1. 0.5 mM Ca
2. 0.5 mM Ca and Adrenaline 10 5 mM
3. 0.5 mM Ca and compound <I) 5 x 10 8 M
Results
1> Hemodynamic results in dogs
The anesthetized dogs are perfused for 20 minutes with the compound (I) or ouabain, at a dose of 3.3 nanomol / kg per minute, at a rate of 1 ml / min iv (saphenous). The controls are perfused under the same conditions with a solution of NaCl 0.9% (w / v). This protocol makes it possible to observe the evolution up to 180 minutes of the following parameters, in the presence of the compound (I) and ouabain in equimolar doses and under the same experimental conditions. The values are expressed in #% of the action of the drug compared to the control =
The final concentration of the drugs, subjected to the diffusion being estimated at 60 nM, this inotropic dose exerts a global evolution from 0 min beginning of observation) to i80 min (end of observation) of the following parameters, recorded in Table IV.

a) at the level of the heart
- Systolic pressure (P'max) from 0 to + 70% <Fig. 5A)
b) at the ship level
- Average arterial pressure (MAP) ranges from 0 to -30%
(Fig. 6a>
- Oxygen consumption (PAMxFC) varies from o to 27%
<Fig. 6d>
- The index Katz CPAM / FC) varies from 0 to -33% (Fig 6b)
- The total peripheral resistance (RPT) varies from O to
-34% (Fig. 6c)
It is found that the 80 mM compound (I) varies the parameters of intravascular pressure # below zero. There is thus vasodilator action of the compound (I).

Table IV
Variation of haemodynamic parameters in the anesthetized dog in the presence of the compound (I) (MSN) and ouabain (OAU): 3.3 nanomoles / kg / min x 20 min.

<Tb>

<SEP> Parameters <SEP> Drugs <SEP>% # <SEP> to <SEP> 0 <SEP> min <SEP>% # <SEP><SEP> to <SEP> 20 <SEP> min <SEP>% # <SEP> max <SEP> (min) <SEP><SEP>%#<SEP> to <SEP> 90 <SEP> min
<tb>P'max<SEP> MSN <SEP> 0 <SEP> 70 # 1 <SEP><SEP> 75 # 1.5 <SEP> (45 ') <SEP> 60 # 2
<tb><SEP> OAU <SEP> o <SEP> 45 <SEP># 2 <SEP><SEP> 90 <SEP># 1,5 (45 ') <SEP><SEP> 75 <SEP> t2 <September>
<tb>P'max / P40 <SEP> MSN <SEP> 0 <SEP> 65 # 1.2 <SEP><SEP> 65 # 1.2 (20 ') <SEP><SEP> 40 # 1.5
<tb><SEP> OUA <SEP> 0 <SEP> 30 # 2 <SEP><SEP> 55 # 2 (40 ') <SEP><SEP> 45 # 2
<tb>P'min<SEP> MSN <SEP> 0 <SEP> 18 # 2 <SEP><SEP> 18 # 2 (20 ') <SEP><SEP> -40 # 0.8
<tb><SEP> OAU <SEP> 0 <SEP> 50 # 2 <SEP><SEP> 60 # 2 (30 ') <SEP><SEP> 35
<tb> Index <SEP> relaxation <SEP> MSN <SEP> 0 <SEP> -1 # 0.6 <SEP><SEP> 0.50 (90 ') <SEP> 0.5
<tb> PTVG <SEP> OAU <SEP> 0 <SEP> -1 # 0.6 <SEP><SEP> 0.50 (5 ') <SEP> -1
<tb> Frequency <SEP> MSN <SEP> 0 <SEP> 4 # 0.5 <SEP> 5 # 1 (15 ') <SEP><SEP> 4 # 0.5
<tb> cardiac <SEP> FC <SEP> OAU <SEP> 0 <SEP> 0 <SEP> 7 # 2 (60 ') <SEP><SEP> 7 # 2
<tb> Flow <SEP> ejection <SEP> MSN <SEP> 0 <SEP> 12 # 1 <SEP><SEP> 13 # 1 (60 ') <SEP><SEP> 11 # 0.5
<tb> systolic <SEP> OF <SEP> OAU <SEP> 0 <SEP> 10 # 2 <SEP><SEP> 15 # 2 (25 ') <SEP><SEP> 2 # 0.5
<tb> Index <SEP> cardiac <SEP> MSN <SEP> 0 <SEP> 5 # 2 <SEP><SEP> 7 # 1 (40 ') <SEP><SEP> 5 # 1
<tb><SEP> OAU <SEP> 0 <SEP> 10 # 2 <SEP><SEP> 15 # 1 (35 ') <SEP><SEP> 5 # 2
<tb><SEP> Consumption <SEP> O2 <SEP> MSN <SEP> 0 <SEP> 20 # 1 <SEP><SEP> 20 # 1 (20 ') <SEP><SEP> -27 # 3
<tb> PAM <SEP> x <SEP> FC <SEP> OAU <SEP> 0 <SEP> 45 # 2 <SEP><SEP> 45 # 2 (20 ') <SEP><SEP> 25 # 2
<tb> Index <SEP> Katz <SEP> MSN <SEP> 0 <SEP> 10 # 0.6 <SEP><SEP> 12 # 1 (10 ') <SEP><SEP> -33 # 2
<tb> PAM / FC <SEP> OAU <SEP> 0 <SEP> 45 # 3 <SEP><SEP> 45 # 3 (20 ') <SEP><SEP> 2 # 0.6
<tb> Work <SEP> (TWg) <SEP> MSN <SEP> 0 <SEP> 33 # 2 <SEP><SEP> 33 # 2 (20 ') <SEP><SEP> -26 # 1
<tb> PAM <SEP> x <SEP> DC <SEP> OAU <SEP> 0 <SEP> 60 # 3 <SEP><SEP> 65 # 2 (25 ') <SEP><SEP> 15 # 2
<tb> Resist. <SEP> MSN <SEP> 0 <SEP> -0.13 <SEP> 7 # 1 (10 ') <SEP><SEP> -33 # 1
<tb> perif. <SEP> tot. <SEP> OAU <SEP> 0 <SEP> 25 # 2 <SEP><SEP> 25 # 2 (20 ') <SEP><SEP> 5 # 1
<tb> RPT <SEP> = <SEP> PAM / DC
<tb> Press. <SEP> art. <SEP> MSN <SEP> 0 <SEP> 19 # 1 <SEP><SEP> 19 # 1 (20 ') <SEP><SEP> -14 # 1
<tb> systol. <SEP> NOT <SEP> OUA <SEP> 0 <SEP> 40 # 1 <SEP><SEP> 45 # 2 (30 ') <SEP><SEP> 15 # 2
<tb> Press. <SEP> art. <SEP> MSN <SEP> 0 <SEP> 18 # 0.5 <SEP><SEP> 18 # 0.5 (20 ') <SEP><SEP> -40 # 1
<tb> diast. <SEP> OUA <SEP> 0 <SEP> 40 # 2 <SEP><SEP> 40 # 2 (20 ') <SEP><SEP> 5 # 1
<tb> TWg / batt. <SEP> MSN <SEP> 0 <SEP> 28 # 1 <SEP><SEP> 28 # 1 (20 ') <SEP><SEP> -29 # 1
<tb><SEP> OUA <SEP> 0 <SEP> 65 # 3 <SEP><SEP> 70 # 5 (25 ') <SEP><SEP> 20 # 2
<Tb>
physiological results
These results characterize the action of the compound (I) on the electrical and mechanical activities of the right ventricular sheep transversus, on the calcium current and depolarization of the left frog atrium and on the carotid blood pressure of the rat anesthetized with Nembutal.

a) Electrical and mechanical activities of the transversus
The electrical activity is represented by PA of amplitude equal to 115 millivolts in absolute value, and a duration of 450 milliseconds. The amplitude of this AP is reduced by 5, 10, 20 and 30% of its normal value from 0 to 20 minutes with the compound (I) 5 x 10 7 M (Figure 5B <I)). On the other hand, the variationaln of the concentration of the compound (I) from x 10-8 to 5 x 10-7 M induces a reduction of up to 25% in 20 minutes of the amplitude of the AP. As for the duration of the AP, it is reduced to 50 to 60% of its normal value by a concentration of the compound (I) of 5 × 10 7 M in 20 minutes (Fig. 5B (II)).

 The mechanical activity is represented by the plots (Fig. 5S (I) and <II>) of a normal contraction of 200 mg of tone and 400 milliseconds of duration. These parameters which evolve up to 200% of the normal tone by 5 × 10 -7 M of compound (I), vary with the concentrations of compound (I) (5 × 10 5 x 10 7 M) and with time up to 20 min. The dose of compound (I> <5 x 10 7 M) gives the first signs of toxicity by the appearance of extra-potential oscillation Oscillation after potential <O.A.P. >, as well as oscillating extra-contraction oscillation after contraction (O.A.C.) on the contraction (Fig. 5B (I> and <II>).

b) The calcium current of depolarization
The calcium current was made with a voltage clamp of 50 mV in the presence of TTX which eliminates the fast sodium current. The presence of compound (I) 5 × 10 9 M gives a reduction of 30%, worsened to 70% of the normal calcium current by the compound (I) 5 × 10 8 M of final concentration, in a Ringer + Ca 2 + 2 medium, 5 mM (Fig. 7).

Claims (7)

 wherein one of its pharmaceutically acceptable salts, in association with a pharmaceutically acceptable vehicle suitable for the type of administration considered.

 i. A medicinal composition for the treatment and prevention of cardiac insufficiency and cardiovascular disease in humans and animals, characterized in that it contains, as an active ingredient, 2 ', 3'- dioxymethyl-6-methyl-3-beta-D-glucose-strophanthinine, also known as Mansonin, represented by the formula (  2. Drug composition according to claim 1, characterized in that it is intended for the treatment and prevention of pathological conditions such as:  - the cardiac insufficiencies for which digital ics are prescribed, in order to improve the carotonicity, to improve carding rhythm and to fight against fibrillation;  Hypertension of hypercalcic origin. to fight against arteriosclerosis.  3. Drug composition according to one of the re vendicat ions I and 2, characterized in that it is suitable for oral administration, especially in the form of capsules or tablets, or for administration by intravenous or intramuscular injection , or for intraperitoneal administration.  4. Drug composition according to one of claims 1 to 3 for the treatment of heart failure and cardiovascular diseases, in particular hypertension and hypercalciarteriosclerosis, providing an effective amount of the compound of formula (I), or a pharmaceutically acceptable salt thereof, up to 300 ng per kg of body weight, or preferably up to 250 ng per kg of body weight.  5. Medicinal product containing mansonine.  6. Use of 2 ', 3'-dioxymethyl-6-betamethyl-1-3-beta-D-glucose-strophanthin, represented by the formula I) or a pharmaceutically acceptable salt thereof to make a drug composition for the treatment of heart failure and cardiovascular disease in humans and animals.  7. Process for the preparation of a medicinal composition for the treatment and prevention of cardiac insufficiency and cardiovascular diseases in humans and animals, characterized in that a pharmaceutically effective amount of 2 is mixed 3 'dioxymethyl-6-beta-D-glucose-strophanthidine, or one of the pharmaceutically acceptable salts with at least one pharmaceutically acceptable compatible excipient.