FR2659333A1 - NOVEL ORGANIC DERIVATIVE OF A PHOSPHOLIPID STRUCTURED METAL, USE OF THE DERIVATIVE AS A BIOMEDIATION AGENT, METHOD OF ISOLATING THE DERIVATIVE, AND PHARMACEUTICAL COMPOSITIONS CONTAINING THE SAME. - Google Patents

Abstract

The invention relates to a new organic derivative of a metal, characterized in that it consists of a derivative of a metal, preferably with a valency of at least 3, with an organic compound comprising a phospholipid which renders the organic derivative of said metal soluble or miscible in water. This phospholipid advantageously contains at least one unsaturated fatty acid of the alpha -linolenic or gamma -linolenic acid type. The metallo-organic derivatives according to the invention are useful biomediators and can be used in this capacity as active principles in pharmaceutical compositions with various activities, in particular an immunomodulatory activity.

Description

New organic derivative of a metal with a phospholipid structure, use of this derivative as a biomediating agent, process for isolating this derivative and pharmaceutical compositions containing it
The invention essentially relates to a novel organic derivative of a metal with a phospholipid structure, the use of this derivative as a biomediating agent, a method of isolating this derivative and pharmaceutical compositions containing it.

 In the present state of the art, various formulations of vanadium have been made to make it bioavailable in the treatment of hyperglycemia. The formulations produced up to now are still very insufficient in that their bioavailability is very low or non-existent.

Moreover, it is known to use various plant extracts as therapeutic agents. For example, in the document
EP-A-0 266 468 (Natura Médita), there is described a therapeutic composition comprising an organic extract of plant of the family Resedaceae useful as an anti-inflammatory agent. Similarly, in document FR-A-2,611,502 (Natura Médica), there is described an extract of umbelliferous root obtained by hydro-alcoholic extraction and its use as a therapeutic agent having a diuretic, cardiovascular, analgesic activity. or anti-inflammatory. Also, it is described in the document
FR-A-2,624,737 (Natura Médica), an extract of black currant leaf obtained by hydro-alcoholic extraction and its use as a therapeutic agent having an analgesic and anti-inflammatory activity.

 These plant extracts described in these documents are very interesting, but they do not make it possible to effectively treat conditions which require the use of metals having a valence of at least 3. However, in the current state of the art, the proposals Formulation of metals having a valency of at least 3 in order to render them bioavailable has failed because their bioavailability is low and their toxicity is high.

 The present invention therefore aims to solve the new technical problem of providing a new formulation of metals, preferably metals having a valence of at least 3, having excellent bioavailability and very low toxicity.

 Preferably, this new formulation must be simple to prepare and be sufficiently stable over time to allow therapeutic use under the best conditions.

 The present invention solves the above-mentioned technical problems in an extremely simple manner, by providing the formulation of a metal, preferably a metal having a valency of at least 3, which has remarkable bioavailability, as well as very low toxicity and excellent stability over time.

 Thus, according to a first aspect, the present invention provides a novel organic derivative of a metal, characterized in that it is constituted by a derivative of a metal, preferably of valence at least equal to 3, with an organic compound comprising at least one phospholipid imparting a water solubility or hydromiscibility to the organic derivative of said metal.

 According to an advantageous variant embodiment, the above-mentioned metal is bonded to the phosphate function of the phospholipid.

 According to another embodiment variant, the aforementioned metal is bonded to a porphyrin group.

 According to an advantageous embodiment, the phospholipid comprises at least one unsaturated fatty acid, preferably in the α or β position with respect to phosphoric acid.

According to an advantageous embodiment, this unsaturated fatty acid is a polyunsaturated fatty acid, preferably without conjugation of the ethylenic bonds. Advantageously, this polyunsaturated fatty acid without conjugation of the ethylenic bonds has a C18 to C20 chain and is chosen from 9, 12-linoleic acid,
9,12,15-linolenic acid, 5,8,11,14-arachidonic acid.

 Still more preferably, it is α-linolenic or γ-linolenic acid.

 According to another particularly advantageous embodiment of the invention, the above-mentioned phopholipid comprises at least one phosphoric acid function, preferably conjugated with an unsaturation of the phospholipidic chain. This acid function can be either in free form or in bound form to an unsubstituted or substituted quaternary ammonium group, oralkylamine in which the alkyl group is a C1-C5, preferably C1-C3, lower alkyl group.

According to another embodiment variant particularly
advantageously, the alkylamine substituent is bound by its function
amine to a phenol or phenolic acid preferably from the series
caffeic acids. As particularly caffeic acid
preferred is caffeic acid, ferulic acid, chlorogenic acid,
salicylic acid and its derivatives such as salicylic aldehyde,
C1-C5 lower alkyl salicylate, in particular
methyl salicylate, monotropitoside, spiréoside,
hyperoside, salicoside or salicin.

According to another embodiment particularly
advantageously, the alkylamine substituent may be bonded to a phenol or
a phenolic acid which is itself bound to a C5 or C6 sugar, in
especially mannitol or mannose.

The phospholipidic chain is preferably at most
of 60 carbon atoms.

The above-mentioned metal, preferably of at least equal valence
at 3 is preferably attached to a phosphate function or to a
porphyrin group. The metals M of valence at least equal to 3 are
preferably selected from Sc, Ti, V, Cr, Mn, Fe, Co, Ni, Cu, Ga,
Ge, Sb, Sn, In, Pd, Rh, Ru, Tc, Mo, Nb, Zr, Y, Hf, Ta, W, Re, Os,
Ir, Pt, Au, Tl, Pb, Bi, Po.

According to another particular embodiment of
The invention, a second acidic function of phosphoric acid can be salified by a valence ion of less than 3 lighter
that the preceding metals and chosen, for example, from Ca, Sr,
Li, Cd, Zn, Mg and Ag.

dans laquelle :
R1 = soit un acide gras en C16 à C20 ayant avantageusement un
faible degré d'insaturation, en particulier une double
liaison),
= soit un noyau porphyrine lié à un métal M1 ayant une valence
supérieure ou égale à 3 telle que précédemment définie,
= soit un groupe X-OH dans lequel X = (H),, n étant égal à 0, 1
ou 2.According to another embodiment particularly
Advantageous of the invention, the organic derivative of a metal is characterized in that it has the following general chemical formula

in which :
R1 = either a C16-C20 fatty acid advantageously having a
low degree of unsaturation, especially double
bond)
= either a porphyrin ring bonded to an M1 metal having a valence
greater than or equal to 3 as previously defined,
= either an X-OH group in which X = (H) n being 0, 1
or 2.

R3 = soit un acide gras insaturé en C à C tel que défini pour 20
R1 ou R2,
= soit un groupe phosphate de formule

à la condition que R2 et R3 ne peuvent pas être simultanément un groupe phosphate. R2 = either unsaturated fatty acid of C16 to C20, preferably having
at least 2 degrees of unsaturation,
= either a phosphate function of formula

R3 = either unsaturated fatty acid C to C as defined for 20
R1 or R2,
= a phosphate group of formula

provided that R2 and R3 can not simultaneously be a phosphate group.

R4 = either H or a quaternary ammonium group (unsubstituted or
substituted, especially with a positive ionic group) -R5,
wherein R5 equals a minimum C5 organic compound of
preferably selected from a sugar, a phenolic acid; is
a C1-C5-alkyl group free or linked to R5; in the preceding formula, a free acid function of the phosphate group of R2 or R3 may be salified by a metaL M3 of valence less than or equal to 2, in particular chosen from Ca, Sr,
Li, Cd, Zn, Mg and Ag.

dans LaqueLLe A1, A2, A3 A4 A5 A6, A7, A8 sont choisis indépendamment parmi les substituants usuels des chlorophylles, en particulier l'hydrogène, un alkyle inférieur en C -C à chaîne droite, ramifiée ou cyclique, un radical vinyle, formyle et ses esters, acétyle et ses esters, propionyle et ses esters, acrylyle et ses esters, et cyclopentanoyles,
M1 étant un métal de valence au moins égale à 3, tel que précédemment défini,
et l'un au moins parmi A1 à A8 étant obligatoirement un substituant phospholipidique tel que précédemment défini.In the case of a substitution in R1 by a porphyrin nucleus, this cycle is characterized rar! j {formula will generate the following:

in which A1, A2, A3 A4, A5, A7, A7, A8 are independently selected from the usual substituents of chlorophylls, especially hydrogen, straight-chain, branched or cyclic lower C -C -alkyl, vinyl, formyl and its esters, acetyl and its esters, propionyl and its esters, acrylyl and its esters, and cyclopentanoyles,
M1 being a metal of valency at least equal to 3, as previously defined,
and at least one of from A1 to A8 being necessarily a phospholipid substituent as previously defined.

The aforementioned customary substituents of chlorophyll are in particular those described by Hendry and Jones in the review
Journal of Medical Genetics, 1980, 17, 1-14, and in particular Table 1, page 5.

 Preferably, the valency of at least one metal M1 or M2 is equal to 3.

According to another advantageous characteristic of
The invention, the novel organic metal derivative according to the invention as defined above is isolated from a plant, preferably from the following families
- legumes, in particular soybeans, broad beans, peas, chickpeas, alfalfa, oats;
- Plantaginés, in particular Plantains,
- Crucifers, in particular rapeseed, cabbage, watercress,
- Salsolaceae, in particular spinach,
- the Urticaceae, especially the nettles dioica, urens and the parietes;
- Umbelliferae, in particular angelica archangelica and angelica sinensis, and foeniculum dulce;
- Résédacées, in particular Réséda, phyteuma, lutera, alba, luteola.

According to a second aspect, the present invention relates to a method of isolating the new organic derivative of a metal, as defined above, characterized in that it comprises
a) extracting a suitable plant known to contain one of the abovementioned metals, by maceration in a hydro-alcoholic solvent for a time sufficient to extract said metal, thus obtaining a crude extract;
b) removing the solvent from the crude extract to obtain a crude powdery product containing said metal;
c) the crude powdery product obtained is redissolved in a minimum of non-ionized water, or distilled, then subjected to precipitation with an organic solvent, preferably chosen from water-miscible solvents, and selected according to the hydrophilicity of the desired metal compound;
d) and optionally, the precipitate obtained containing the organic derivative of the desired metal in the pure state, is subjected to a metal exchange to obtain the salt of a new metal different from that it initially contained in the plant.

 According to a preferred embodiment, for which the phosphoric acid function is linked to a second metal (M3), of valence less than or equal to 2, preferably chosen from the group consisting of calcium, strontium, zinc, lithium, magnesium or silver, the crude powdery product is subjected to precipitation with a saturated solution of cadmium chloride to obtain a precipitate which is separated by centrifugation and then redissolved in a carbonated solution, for example ammonium carbonate and then reprecipitated with a water-miscible organic solvent, preferably an alcohol or a ketone, particularly a lower alcohol such as methanol, methanol or propanol. The purification can be improved in a conventional manner by several steps of precipitation with organic solvent dissolution in water.

 The last precipitate contains the organic derivative of the metal according to the invention, in pure form, optionally mixed with small proportions of other metals.

According to a particular embodiment of this process, when it is desired to change the metal bound to the phospholipid, the precipitated product obtained is redissolved in an aqueous solution containing the metal which it is desired to incorporate, and then a new solvent precipitation is carried out. The above-mentioned water is miscible with organic water, which makes it possible to obtain the organic derivative of the desired metal, according to
The invention.

According to another variant embodiment of the method according to
The invention, the extraction solvent used to obtain the crude extract of the plant is advantageously chosen from water mixtures / lower alcohol C -C, preferably selected from methanol, ethanol, propanol, isopropanol, n-butanol or iso-butanol. In particular, the water / alcohol content is advantageously between 75 and 99% water / 1 to 25% alcohol, by weight.

The identification of the organic derivative of the metal according to
The invention can be carried out by a silica gel thin layer chromatography technique which is well known to those skilled in the art, in particular with the help of the three solvents below:
chloroform / methanol 90/10 by volume
- isopropanol / water 95/5 by volume
- 50/50 ethyl ether / methanol by volume
The chromatogram is then revealed with ninhydrin. There is normally only one task.

According to another variant of identification of the novel organic metal derivative according to the invention, the organic derivative of purified metal is first redissolved as obtained according to the invention.
The process described above, in an aqueous solution which is then added in equal parts with a solution of 2N hydrochloric acid and then in an oven at 80 ° C. for several hours, in particular approximately 8 hours.

 The solution obtained is then subjected to standard techniques for identifying the desired compounds.

For example, fatty acids are identified by gas chromatography
the phenols and phenolic acids are identified by high performance liquid chromatography and ultraviolet detection;
- The sugars and glycerides are identified by HPLC and detection by an apparatus capable of measuring a number of particles after removal of the solvent by vaporization, for example a device marketed by TOUZART and MATIGNON, having the trade name SEDEX 45; ;
Porphyrin is identified by thin layer chromatography according to conventional porphyrin identification techniques,
the metal is identified by atomic absorption spectroscopy and plasma torch, the apparatus being naturally adjusted according to the metal to be identified, for example Applied apparatus
Research Laboratories, USA, Model 3,580 OES.

 The novel organic metal derivative according to the invention described above, can naturally be prepared synthetically from the corresponding constituents which are reacted under operating conditions known to those skilled in the art.

It will be observed that easily accessible sources for the preparation of this new organic metal derivative according to
The invention is constituted not only by the aforementioned plants, but also by biotechnology-derived parts such as the in vitro culture of plants, or so-called callus cultures, ie differentiated cell blocks, multiplied by from complex-rich cells or complex fraction, or parts of animal organs. These various parts of organs or cellular organisms are subjected to specific extraction treatments. As results from the preceding discussion, in the context of the invention, only the extracts made from a solvent having a polarity relative to with the silica of the same order as that of a solvent prepared on the basis of water and methanol in the previously indicated proportions of 75 to 99% of water and 1 to 25% of ethanol, makes it possible to obtain a organic derivative of crude metal which allows to isolate directly or indirectly The new organic metal derivative according to the invention.

 According to a third aspect, the invention also covers a use of the organic metal derivative according to the invention, preferably having a valence of at least 3, as a biomediator or as an active ingredient for the manufacture of a product for therapeutic use.

Indeed, the organic metal derivatives according to
The invention has an immunomodulatory activity and can in this context be usefully used to treat all immune deficiencies, including AIDS, or neuromodulatory activity, or anti-inflammatory activity or anti-rheumatic activity, particularly for combating rheumatoid arthritis.

 Thus, according to a fourth aspect, the present invention still covers a pharmaceutical composition, characterized in that it comprises, as active ingredient, at least one organic derivative of a metal according to the invention as defined above.

 According to a particularly advantageous characteristic, this pharmaceutical composition is characterized in that it is a pharmaceutical composition with immunomodulatory or neuromodulating activity, or anti-inflammatory or antirheumatic, in particular anti-rheumatoid arthritis or hypoglycemic or anti-multiple sclerosis.

 Particularly preferred compounds are an organic derivative of a metal according to the invention in which a gold atom is bonded to a phospholipid having an antirhumatoide activity, in particular against rheumatoid arthritis.

 Another particularly preferred derivative is a metal derivative according to the invention in which the metal is aluminum bound to a phospholipid, whose fatty acid is α-linolenic acid while a ferrulic choline group is bound. at the top of the phosphoric acid or phospholipid chain, this compound being extracted from Reseda phyteuma and having a marked anti-inflammatory activity.

 Another preferred compound according to the invention is a compound whose metal is Sb or a Sb / Sn shot preferably obtained from the Angelica species, in particular Angelica archangelica, and Angelica sinensis having an immunomodulatory activity especially particularly effective against the HIV virus.

 Yet another particularly preferred compound of the invention is a compound whose metal is vanadium, obtained from urtica dioica, having a persistent hypoglycemic activity.

The doses of use of the new organic derivative of a metal according to the invention are from 100 to 300 mg of a final lyophilisate containing at most 5% of the organic metal derivative according to the invention.
The invention for a gastro-resistant capsule. Usually, a dose of 200 mg of purified powdery product, for example in freeze-dried form, has a metal content of less than 1 mg.

 This powdered, purified product, for example in the form of lyophilizate can be dissolved in an aqueous solvent at a dose of 5% so as to produce injectable dosage forms, or instillable or allowing for example an aerosol use.

One of the preferred routes of administration is that of the mucosal pathways (perlingual, pernasal, suppository, eye drops).

Other goals, features and benefits of
The invention will become apparent in the light of the explanatory description which follows, with reference to several examples of the invention given merely by way of illustration and which can not in any way limit the scope of the invention.

In the examples the percentages are given by weight, unless otherwise indicated.

Example 1
Organic derivative of Sb from Angelica archangelica
The plant Angelica archangelica is harvested whole at the maturity of the seeds then dried in the rules of the art and roughly crushed and then stirred mechanically for 4 to 6 days in a mixture at 60 C of ten times its weight of water and once its weight of ethanol at 95% by volume.

 After cooling to room temperature, tangential filtration is carried out in a filter having a pore diameter of 0.2 μm, and then is concentrated on average 10 times by reverse osmosis.

 Finally, the hydroalcoholic solvent is completely removed by lyophilization, thus obtaining a crude extract in the form of lyophilizate.

 The crude extract is redissolved in a minimum of distilled water at the rate of 5 g per 10 ml.

After complete dissolution in water, the resulting aqueous solution is precipitated with methanol or methanol. And we separate the precipitate. It is repeated three times and the last precipitate constitutes the organic derivative of Sb according to
The invention.

This organic derivative comprises in practice a mixture of
Sb / Sn in a substantially equal amount.

 The characteristics of this purified product thus obtained are the following: black oily product, miscible with water in which it is possible to identify after nitric digestion the metal, either by the Rhodamine B reaction well known to the chemist analyst, or by spectroscopy. atomic emission.

Example 2
Organic aluminum derivative from Réséda phyteuma
Starting from the plant Reseda phyteuma, the procedure is as described in Example 1, but the last precipitate is then dissolved in a dilute solution of mineral acid such as 2N hydrochloric acid and then precipitated by the organic solvent that allowed extraction. for example methanol or acetone.

 This precipitate constitutes the organic derivative of the aluminum according to the invention, having mainly aluminum bound by a phosphate bond to a phosphatidylcholine structure, itself linked to ferrulic acid.

 This organic aluminum derivative according to the invention has the following physicochemical characteristics: a green colored fatty product, miscible with water and exhibiting the reactions of choline and aluminum, in which it is possible to demonstrate the phospholipid by thin layer chromatography.

Example 3
Organic vanadium derivative from Urtica dioica
The procedure is as described in Example 1 starting from the plant Urtica dioica until the last precipitate.

This constitutes the organic vanadium derivative
3+
The invention, wherein the V-form vanadium is included in either the porphyrinic ring which is itself substituted by a vanadyl-ionized phospholipid.

 The physico-chemical characteristics of this organic vanadium derivative according to the invention are as follows: brown crystalline powder, soluble in water, the solution of which has a hypoglycemic activity, the powder does not have a melting point but decomposes.

Example 4
Organic gold derivative from Ribes nigrum
The procedure is as described in Example 1 starting from the Ribes nigrum plant to the last precipitate which constitutes the organic gold derivative according to the invention which mainly comprises gold in the form of Au3 + fixed on the phospholipid.

 This organic gold derivative has the following characteristics: Au3 + form, green brown powder, soluble in water, no melting point but decomposes.

Example 5
Organic derivative of a metal having a value of at least 3
included in a porphyrinic cycle
An organic derivative of Sc, Ti, V, Cr, Mn can be obtained.
Fe, Co, Ni, Cu, Ga, Ge, In, Pd, Rh, Ru, Tc, Mo, Nb, Zr, Y, Hf, Ta,
W, Re, Os, Ir, Pt, Tl, Pb, Bi, Po, starting from the organic metal derivative according to the invention as obtained in any one of Examples 1 to 4, proceeding then in the manner next.

 The organic metal derivative according to the purified invention obtained at the end of the procedure of any of Examples 1 to 4 is dissolved in a weakly acid solution of mineral acid such as 2N hydrochloric acid and then precipitated with an organic solvent. having allowed its extraction, for example methanol or methanol.

This precipitate after centrifugation is redissolved in an aqueous solution containing the metal that is to be incorporated into the porphyrin. A final precipitation by said organic solvent makes it possible to obtain the organic metal derivative according to
The invention having at least 50% of the theoretical dose of the desired metal having a valence of at least 3.

Example 6
Preparation of an organic metal derivative according to the invention
having an alkylamine function
The lyophilysate obtained after extraction of Reseda Phyteuma according to the technique described in Example 1 is taken up in distilled water and then reprecipitated with acetone in a volume proportion 1/1.

 The oily precipitate is isolated by centrifugation, washed with pure acetone, taken up with a volume of distilled water and lyophilized in the presence of 10% maltodextrin (weight / volume).

After identification and determination of the metal by atomic absorption according to the conventional atomic absorption technique using the apparatus marketed by the American company
Applied Research Laboratories, model 3,580 OES.

 The resulting lyophilized powder can be used directly for therapeutic purposes, as described later.

Example 7
Preparation of an organic metal derivative according to the invention, comprising an alkylamine function linked to a phenolic acid chosen from ferric, caffeic or chlorogenic acids.

 From the derivative of the above Example 6 having an alkylamine function, the amine function can be bonded to a phenolic acid in the following manner.

 The lyophilizate obtained from Spirea ulmaria is dissolved in 2 parts of distilled water and then reprecipitated with acetone in sufficient quantity.

 The precipitate separated by centrifugation is solubilized in a solution of 2N hydrochloric acid, and then reprecipitated again with a solution of ethanol 95% by volume in sufficient quantity. After centrifugation, a sample of the mass obtained is taken up in distilled water and then cold-titrated with an ethanolic solution of 0.1% ferrulic acid in sufficient quantity until a haze appears, or Using a potentiometer, checking the inflection point.

 The concentration of ferric acid to be added to the rest of the precipitate is deduced from the above assay.

We proceed in a similar way with Caffeic acid, or
Chlorogenic acid.

Example 8
Preparation of an Organic Metal Derivative According to the Invention Linked to an Alkylamine Function Which Is Linked to a Salycilic Acid Derivative
The procedure is as in Example 7, giving an amorphous, yellowish-white, water-soluble product with a very characteristic odor.

Example 9
Example of Biomedicating Agent Formulation According to the Invention
According to a first variant embodiment, a biomedical agent according to the invention is formed by the product obtained directly in the crude state in any one of Examples 1 to 9.

According to another advantageous embodiment,
The crude extract obtained in any one of Examples 1 to 9, in the form of a dry product, is mixed with 5% of its weight of maltodextrin until perfectly homogeneous, and then stored. This mixture constitutes the active principle of a pharmaceutical composition according to
The invention as formulated for example in the following examples.

Example 10
Pharmaceutical composition according to the invention
Capsule specialty gastroresistance
The product obtained according to any one of Examples 1 to 9, is added with its weight of magnesium stearate and ten times its weight of microcrystalline cellulose and intimately homogenized, the mixture obtained is distributed by means of a suitable distribution equipment. in gelatin capsule module 2 at a rate of 240 mg per unit.

 The capsules thus dosed at 25 mg of extract are coated with an apparatus of gum arabic, cellulose ester and titanium oxide. After drying, the capsules obtained are gastroresistant and packaged in blister packs of 15 grouped in a case of 2.

Example 11
Pharmaceutical composition according to the invention
Specialty nasal solution
The product obtained in any of Examples 1 to 9 is dissolved due to 5 mg per ml in a physiological saline nasal spray, diethylene glycol monoethyl ether glycerin. The preparation is packaged in a 30 ml polyethylene bottle equipped with a nasal spray cannula delivering 0.1 ml per pressure.

Example 12
Pharmacological test
A - Investigating an immunomodulatory activity
The organic derivative of an Sb / Sn metal obtained in Example 1 is subjected to tests for the demonstration of immunomodulatory activity, according to the following procedure.

1 - Protocol
The desired effects will be tested in vitro on the cells involved in the immune response (lymphocytes - monocytes and macrophages).

I. Cell (human cells)
Cellular parameters and functions studied - Membrane markers, - Lymphocyte activation and lymphocyte proliferation, - Oxidative metabolism of monocytes.

1. Membrane Markers
The expression of membrane antigens will be studied in a flow cytometer in the presence and absence of each of the products.

Protocol used
From peripheral blood collected on EDTA or heparin, separation of the lymphocyte population.

Markers studied: - CD3 - CD4 - CD8 - B4 for lymphocytes, - M02 for monocytes
These markers are identified by the corresponding monoclonal antibodies which, after fixation, are visualized by a fluochrome-conjugated human anti-Igl antibody (GAM-FITC).

 The study of the distribution of the cells carrying the markers is made with the flow cytometer available to the laboratory (COULTER Epics profile).

- the doses tested: Final dilutions 1/20 - 1/100
* incubation time
short TO-730 min - T60 min
Long TO 6 pm - 24 pm

Materials needed: - flow cytofluometry (COULTER epics profile), - CO2 incubator, - reagents: medium RPMI medium
lymphocyte markers
* monoclonal antibodies against CD3
* anti-CD4 monoclonal antibodies
* monoclonal antibodies against CD8
* monoclonal antibodies anti 84
* anti M02 antibodies
* GAM FITC.

2. Activator and lymphocyte proliferation
Human lymphocytes are collected from peripheral blood by Ficoll gradient centrifugation.

2.1 Activation
Activation is tested under two circumstances - effect of direct activation of the product on the cells, - modulatory effect of the product on activation by
activating substances = PHA - conA - PMA (phorbo myristate
acetate).

 In both cases, activation is assessed by the appearance of membrane markers of lymphocyte activation: CD25 (or II12 receptor) - HLA-DR antigen: I2.

 These two markers are evaluated in flow cytometry.

Protocol
The cells are - on the one hand put in the presence of 2 doses of each product
24 hours and then tested against cells
untreated, - on the other hand, grown in the presence of PHA, conA and PMA with
or without product for 24 hours.

Incubation takes place at 370C in a humid atmosphere with 5% of
C02. The results are appreciated at the flow cytometer.

Material required - flow cytofluorometer, - C02 incubator, - RPMI mid-place, - Ficoll-hypaque mid-site, - Reagents:
* phytohemagglutinin
* LDC
* anti-CD25 monoclonal antibody
* monoclonal antibody anti I2 2.2 P-roliferation:
Activated lymphocytes may or may not proliferate under
The effect of certain drugs.

As the activation, the proliferation will be tested in 2 circumstances - direct proliferation under the effect of the product, - modulation by each product of the proliferation obtained by the
mitogens PHA and conA.

 The protocol is the same as above.

 The proliferation is evaluated on the fourth day of incubation by incorporation into the nucleus of the proliferating cells of radioactive precursor DNA (tritiated thymidine): The large proliferation is indicated by the radioactivity appreciated in the cell extract obtained on day 5 of the incubation.

Material required - C02 incubator, - SKATRON type DNA extractor, - Isotypic liquid scintillation counter, - Reagents
* Mid place RPMI
* Ficoll hypaque
* fetal calf serum
* PHL
* tritiated thymidine 3. Oxidative metabolism of monocytes
The activation of macrophage monocytes stimulates their oxidative metabolism, which can easily be studied by flow cytometry by taking advantage of the oxidation of dichlorofluorescein, of which only the oxidized form is fluorescent, so only the activated cells become fluorescent and are detected by the cytometer. flux.

 By this means, it will be possible to study - direct activation of cells by the product, - the modulatory effect of the product on their activation by PMA.

Protocol
Leukocytes separated from peripheral blood are first contacted with DCF and then - with only the product: at different concentrations of
cells not treated with PMA positive controls, - either with the product and PMA Contacts are made in a water bath
constant agitation and readings are made at
flow cytometer every 5 min and up to 1 h 30, the burst
oxidative reaction not exceeding that duration.

Activation kinetics are then compared
Materials required - flow cytofluorimeter, - continuous stirring water bath, - Reagents:
* Ficoll hypaque * dichlorofluorescein,
* PMA.

2 - Result 2-1 Effect of Sb / Sn Derivative of Example 1 on the Activation Marker
CD25 IL2 receiver:
The product of Example 1 (500 μg), incubated with Ficoll gradient cells and activated by PHA (Phyto hemagglutinin) for 24 h, showed an inhibitory effect on the appearance of IL2 receptors.

Results expressed in% of IL2 receptors (CD25):

<tb><SEP> 1st <SEP> subject <SEP> 2nd <SEP> subject
<tb> cells <SEP> 0.5 <SEP>% <SEP> 4.5 <SEP>%
<tb> cells <SEP> + <SEP> PHA <SEP> 22.5 <SEP> Z <SEP> 14.5 <SE> Z <SEP>
<tb> cells <SEP> + <SEP> PHA <SEP> + <SEP> Sb / Sn <SEP> of <SEP> the example <SEP> 1 <SEP> 2,6 <SEP>% <SEP> 3 , 5 <SEP> X <SEP>
<tb> 2-2 Effect of Sb / Sn of Example 1 on the proliferation of lympho
cats cultured with PHA lectin for 72 h
The product of Example 1, in solution at 5 g per 100 ml, brought into contact with the Ficoll gradient-separated lymphocytes and cultured with PHA lectin for 72 h, shows a significant inhibition of lymphocyte proliferation.

After incorporation of tritiated thymidine, the results are expressed in counts per minute (cpm).

<Tb>

<SEP> 1st <SEP> subject <SEP> 2nd <SEP> subject
<tb> cells <SEP> alone <SEP> 780 <SEP> cpm <SEP> 195 <SEP> cpm
<tb> cells <SEP> + <SEP> PHA <SEP> 47 <SEP> 748 <SEP> cmp <SEP> 50 <SEP> 849 <SEP> cpm
<tb> cells <SEP> + <SEP> PHA <SEP> + <SEP> Sb / Sn <SEP> of <SEP> Example <SEP> 1 <SEP> 11 <SEP> 800 <SEP> cpm <SEP > 795 <SEP> cpm
<tb> 2-3 Effect of Sb / Sn of Example 1 on the Expression of Ag Membra
lymphocytes:
The product of Example 1 (500 μg) incubated for 1 h with Ficoll gradient separated lymphocytes does not give a significant modification in the expression of lymphocyte Ag of the 2 subjects studied.

Results expressed as a percentage:

<tb><SEP> 1st <SEP> subject <SEP> 2nd <SEP> subject
<tb><SEP> cells <SEP> cells <SEP> + <SEP> Sb / Sn <SEP> cells <SEP> cells <SEP> + <SEP> Sb / Sn
<tb><SEP> of <SEP> Example <SEP> 1 <SEP> of <SEP> Example <SEP> 1
<tb> CD2 <SEP> 83.7 <SEP> 79.7 <SEP> 87.3 <SEP> 82.9
<tb> CD19 <SEP> 6.0 <SEP> 7.0 <SEP> 3.2 <SEP> 2.4
<tb> CD4 <SEP> 40.2 <SEP> 45.1 <SEP> 49.4 <SEP> 56.7
<tb> CD8 <SEP> 42 <SEP> 56.5 <SEP> 27.3 <SEP> 20.2
<tb> 2H <SEP> 4 <SEP> 29.5 <SEP> 32 <SEP> 66.1 <SEP> 62.6
<tb> 4B <SEP> 4 <SEP> 72.2 <SEP> 68.1 <SEP> 54.7 <SEP> 56.8
2-4 Effect of Sb / Sn of Example 1 on oxidative metabolism of human leukocytes, studied in flow cytometry
The kinetic study of human leucocytes, in the presence of dichloro fluorescine, marker of intracellular H202, and stimulated by Phorbol myristate acetate, shows a decreased oxidative metabolism over time When the product of Example 1 is added to them, as shown by the curves of FIG. 5. In FIG. 5, the curves t, t-, ... e represent the immune response to the stimulatory action caused by Phorbol myristate acetate alone, respectively for polymorphonuclear leukocytes, Monocytes and lymphocytes, while the curves pub, A. .. represent the immune responses respectively caused by Phorbol myristate acetate in the presence of the product of the Sb / Sn invention of Example 1.

 There is a very significant decrease in the immune response.

B - Anti-inflammatory activity
An anti-inflammatory activity is sought with the organic aluminum derivative according to the invention obtained in Example 2, according to the following procedure based on kaolin edema.

Kaolin edema - modified Winter technique (1)
Introduction
The injection of kaolin under the plantar aponeurosis of the posterior paw of the rat causes an acute inflammatory reaction which can be reduced by the anti-inflammatory substances. This inflammatory reaction results in a rapid and persistent installation edema 48 hours, which makes it possible to obtain a kinetics of anti-inflammatory activity.

Material and methods 1 - Aninals
Male Wistar rats (12 per batch) from the Montpellier breeding center were used in this study.

The animals are kept under observation for 4 days before the start of the tests. At the beginning of the test the animals weigh on average 90 g. During the observation period the animals, distributed in cages of 5, receive food and drinking water ad libitum and are subjected to a temperature between 21 and 23 C and has a day / dark cycle of 12 h.

2 - Products - Organic aluminum derivative of Example 2, - Indometacine (INDOCIDO 250), - Screened kaolin, - Excipients, - Saline.

3 - Protocol
The animals are fasted the day before. On the day of the test the rats are anesthetized with ether. An incision is made in the upper part of the abdomen and the nebulisate injected into the duodenal loop using a syringe. The muscle wall is then saturated and the skin closed with 2 staples. The kaolin suspension is injected 1/2 h later (0.1 ml - 109% solution in physiological saline) into the foot pad of the right hind paw. A batch of control animals serves as control and receives the excipient. For each dose of substance, the concentrations of the suspensions used are adjusted so as to administer exactly 1 ml per 100 g of body weight.

 The plethysmographic measurements (2) are made before 1 h, 2 h, 3 h, 4 h, 5 h, 6 h after the administration of the kaolin suspension.

4 - Doses used
From the organic derivative of aluminum of Example 2,
- 100 mg / kg intraduodenal route.

indomethacin
- 5 mg / kg intraduodenal route.

5 - Expression of results
The percentage of increase in the volume of the paw is obtained for each animal according to the following formula
Measured volume - Initial volume
10% = ~~~~~~~~~~~~~~~~~~~~~~~~~~~~ x 100
Initial volume
The percentages of swelling are analyzed by SNEDECOR F and Student's t tests (3).

The anti-inflammatory activity is expressed as the percentage reduction of edema in treated rats compared to controls according to the following formula
% Swollen Controls -% Swollen Treated xxx 100
Swelling controls
Results
The results are shown in Table II and Figure 2.

Table II
% inhibition compared to controls
1 h 2 h 3 h 4 h 5 h 6 h
INDOMETACIN 0 48 35 35 32 25
5 mg / kg
Aluminum Derivative 2 42 30 28 27 20 of Example 2
100 mg / kg
Conclusion
Examination of the results shows in our experimental conditions that the aluminum derivative of Example 2 at a dose of 100 mg / kg exhibits marked anti-inflammatory activity.

This activity is maximum from the second hour and remains significant for 5 hours. The effectiveness of the aluminum derivative is on the other hand comparable to that obtained with 5 mg / kg of indomethacin.

 Given its activity and its low toxicity the aluminum derivative thus has a real therapeutic interest.

Bibliography
(1) WINTER CA RISLEY. EA and NUSS. GW 1962
Carrageenin induced oedema in hind paw of the rat as an assay for anti-inflammatory drugs. Proc. Soc. Exp. Biol. Med. 1: 1: S44-547
(2) LENCE. P. 1962 A new device for plethysmographic measuring of small objects. Arch. Int. Pharmacodyn. 36: 237-241
(3) SCHARTZ. D. Methods statistics for use by physicians and biologists. Flammarion Edition.

C - Determination of a hypoglycemic effect
With the aid of the organic vanadium derivative obtained in Example 3, the presence of a hypoglycemic activity is determined according to the following procedure.
Male Wistar rats (12 per batch) from the Montpellier breeding center were used in this study. The animals are kept under observation for 4 days before the start of the tests. At the beginning of the test, the animals weigh on average 140 g. During the observation period the animals, distributed in cages of 5, receive food and drinking water ad libitum and are subjected to a temperature between 21 and 230C and a day / dark cycle of 12 h.

2 - Products
Porphyrinometallic derivative (Va) according to Example 3
- Streptozotocin
- Excipients - citrate buffer pH 4.5
- physiological saline 3 - Protocol
20 animals with an average weight of 190 g are anesthetized with ether. An intravenous injection of streptozotocin is performed in the vein of the penis; a control glucose level is performed 48 hours after the administration of streptozotocin.

 Only animals with a blood glucose level greater than or equal to 2 g are selected and subjected to treatment with the test substance. Administration of the derivative according to Example 3 was twice daily (8-18 h) for 3 d, then daily (18 h). Glucose control is performed in the morning at 9:00 am, the time of day when the glycemia of control or treated diabetic rats, which are subject to wide variations during the day, is highest. Diabetic rats receive only the excipient (saline) and serve as diabetic controls.

 Finally, blank control rats of the same weight, having received no streptozotocin injection, are kept for the purpose of comparing consumption of water and food.

Results
The daily control of the blood glucose level did not allow us to demonstrate a hypoglycemic effect during the first 2 days of treatment. A significant and significant drop in blood glucose relative to that of the control rats appears from the third day and is maintained the following days. This is accompanied by a similarly significant reduction in weight gain, polyphagia and polydipsia. The consumption of food and water covering values tends to be similar to that of non-diabetic control rats. / Weight gain of control animals, diabetics and animals treated during the 12 days of treatment (in g).

DAYS J00 J03 J05 J07 J10 J12
WITNESSES 189 215 225 233 255 271
DIABETICS 191 199 218 226 238 246
TREATIES ex. 3 171 194 198 215 220 227 - 2 / Evolution of the glycemia of the control animals, diabetics and animals treated during the 12 days of treatment (mg / 100 ml).

DAYS JOO J03 J05 J07 J10 J12
WITNESSES 104 106 104 98 100 96
DIABETICS 368 460 437 474 560 610
TREATIES 330 366 256 272 267 250 - 3 / Average consumption of food (g).

DAYS J00 J03 J05 J07 J10 J12
WITNESSES 20.8 19.7 20.1 24.1 20 25
DIABETES 21.9 30 34.2 38.2 34.2 32.6
TREATIES ex. 3 18.9 14 25 26 20.3 20.5 - 4 / Average consumption of drinking water (ml).

DAYS JOO J03 J05 J07 J10 J12
WITNESSES 40 40 43.3 46.6 41.7 41.7
DIABETES 88.6 128 141 164 136 150
TREATIES ex. 3 75 55 87.5 75 66 43
Figures 1 to 4 show the weight change (Figure 1), the evolution of blood glucose (Figure 2), food consumption (Figure 3) and water consumption (Figure 4), based on indications. above clearly show the effects indicated below.

 With regard to the four objective parameters above, as well as with regard to the subjective parameters, such as the appearance and the behavior of the animals, it appears that the treatment, under the operating conditions retained, had a hypoglycemic effect, and that the This is major. It has also been possible to observe a residual therapeutic effect even after stopping treatment.

D - Anti-arthritis activity
By Freund's adjuvant test
The presence of an anti-polyarthritis activity is determined with the organic gold derivative obtained in Example 4, as follows
Introduction
The injection of a suspension of mycobacterium butyricum in rats leads to the development of experimental chronic polyarthritis of antigenic origin whose characteristics are close to the pathology of rheumatoid arthritis in humans. It evolves chronically with outbreaks in which is formed destructive articulations generating deformations; joint destruction is the consequence of proliferative synovitis, the site of chronic inflammation of immunological origin.In the absence of treatment, the evolution is irreversible and results in a diffuse, regular swelling, erasing articular reliefs and localized to the hind legs; The involvement of the forelegs is also possible.

 Joint impotence results in a limitation of the active movements, or even a fixation of the joint in its release position.

 Finally, other signs such as the presence of nodules on the tail and ears, erythema located on the swollen areas, particularly the joints, conjunctivitis or genital involvement are all telltale signs of the evolution of arthritis to the chronicity.

Some anti-inflammatory substances can stop
Aggravation of arthritis, or even lead back to a normal joint morphology.

Material and methods 1 - Animals
Male Wistar rats (12 per batch) from the Montpellier breeding center were used in this study. The animals are kept under observation for 4 days before the start of the tests. At the beginning of the test the animals weigh on average 180 g. During the observation period the animals, distributed in cages of 5, receive food and drinking water ad lithium and are subjected to a temperature between 21 and 23 C and has a day / dark cycle of 12 h.

2 - Products - Gold derivative of Example 4, - Indomethacin in capsule form (INDOCID 250), - Suspension of mycobacterium butyric acid DIFCO - Excipients, - Saline, - Gummy water at 3%, - Water for injection , - paraffin oil.

3 - Protocol
Induction of arthritis
A paraffin suspension of bacteria of the mycobacterium tuberculosis strain is injected into the lower third of the rat tail. On the 21st day following the injection, the rats are sorted according to the morphological signs of
The arthritic involvement they present. Only animals with unilateral or bilateral tumefaction of the hind legs are retained.

Constitution of the lots
A measurement of the volumes of the right and left hind legs is carried out with the help of a piethismometer on the previously selected rats. Homogeneous batches by weight and by increasing the volumes of the right and left hind paws are then constituted.

Treatment
The treatment begins on the 21st day after the constitution of the batches and continues for 13 days with a daily administration. The last measurement (J14) is performed 24 hours after the last administration.

Nature of the lots
Five lots of 9 animals each were made according to
The following order:
- a control arthritic lot
- an arthritic lot treated indomethacin 3 mg / kg
- a treated arthritic lot 803 200 mg / kg
- a treated arthritic lot 803 400 mg / kg
- a non-arthritic lot not receiving any treatment.

Evaluation of arthritis
The volume of the hind legs is evaluated before and during the entire course of treatment, in parallel with a morphological examination; this exam helps assess and quantify
Impotence, which indicates a limitation or even the disappearance of the movements of the animal when it is placed on a flat surface, is credited with an index of 0 to 3.

OO no impotence
1 1 average impotence
2 2 strong impotence
3 3 total impotence.

 - The ankylosis reflects the limitation or disappearance of the articular movement of the hind legs is credited with an index of 0 to 2 for each of the legs.

. O no ankylosis
1 1 average ankylosis
2 2 total ankylosis.

 - The edema of the forelegs, which represents an extension of arthritis, is credited with indices O and 1.

OO absence of edema
1 1 presence of edema.

 - Presence of nodules on the ears.

. O absence
1 1 presence
- Presence of nodule on the tail located at the initial point of injection of the adjuvant.

- 0 absence
- 1 presence
- Genital damage.

 - O absence 1 1 present.

 - Eye damage resulting in the presence of unilateral or bilateral conjunctivitis.

OO absence
1 1 presence.

Results
1 - Homogeneity of batches before treatment
Table III
Volume of hind legs
Lots Average Difference
WITNESSES 76.38 + 5.3
INDOMETACIN 76.05 + 4.6 3 then 1.5 mg / kg
Gold - 200 mg / kg 71.66 + 4.4
Gold - 400 mg / kg 78.33 + 5.0 * Gold derivative of Example 4
Table IV
% swelling compared to the absolute control lot
Lots% swelling
WITNESSES 58,33
INDOMETACIN 57.35 3 then 1.5 mg / kg
Gold - 200 mg / kg 48.11
Gold - 400 mg / kg 61.90
An analysis of the variance of the hind paw volume and the weight of the animals of the different batches shows no significant difference before treatment.

2 - Evolution weights the
Table V
Lots OJ J4 J9 J14
WITNESSES 231 + 5.6 240 + 5.5 249 + 9.5 262 + 10
INDOMETACIN 234 + 9.2 240 + 11 261 + 19 296 + 16 3 then 1.5 mg / kg
Gold - 200 mg / kg 234 + 7.3 236 + 7.2 247 + 9.6 259 + 9.7
Gold - 400 mg / kg 243 + 6.5 238 + 5.7 251 + 8.6 271 + 16
NO SIGNIFICANT DIFFERENCE BETWEEN LOTS
Gold derivative of Example 4
3 - Evolution of arthritis during treatment
Table VI
3-1-1 Evolution of the paw volume in mm Hg
Lots OJ J4 J9 J14
WITNESSES 76.38 + 5.3 82.88 + 6.3 85.66 + 7.3 89.94 + 10
INDOMETACIN 76.05 + 4.6 58.00 + 2.7 58.40 + 2.5 56.00 + 2.2 3 then 1.5 mg / kg
However, ex. - 200 mg / kg 71.66 + 4.4 71.83 + 4.2 71.05 + 4.0 73.11 + 5.5
However, ex. - 400 mg / kg 78.33 + 5.0 69.11 + 4.9 64.61 + 4.5 58.77 + 3.4
*
SIGNIFICANT DIFFERENCE FROM WITNESSES
Volumes are expressed in mm of mercury.

3-1-2 Evolution of paw volume in% of base volume.

 The base volume is the volume of the hind legs measured at day 0, before treatment.

Table VII
Lots J4 J9 J14
WITNESSES + 8,5 + 12,14 + 17,75
INDOMETACIN - 23.73 - 23.20 - 26.36 3 then 1.5 mg / kg
Gold - 200 mg / kg + 2,3 - 0,8 + 2,00
Gold - 400 mg / kg - 11.77 - 17.51 - 24.97
* SIGNIFICANT DIFFERENCE WITH RESPECT TO WITNESSES
3-1-3 Percentage of protection.

 The percentage of protection is determined with respect to the volume of the hind legs of the control animals.

Table VIII
Lots J4 J9 J14
INDOMETACIN 30.00 31.82 37.73 3 then 1.5 mg / kg
Gold - 200 mg / kg 13.83 17.05 18.71
Gold - 400 mg / kg 16.61 24.57 34.65
SIGNIFICANT DIFFERENCE FROM WITNESSES 3-1-4 Evaluation of Treatment Efficacy.

 The effectiveness of the treatment is evaluated as a percentage of swelling of the hind legs of the treated groups (treated pathological animals) in relation to the volume of the hind legs of the absolute controls group in which the arthritis has not been conducted (healthy animals) .

Table IX
Lots Volume at 14% swelling
WITNESSES
ABSOLUS 49
WITNESSES
ARTHRITIS 89.94 83.55
INDOMETACIN 3 then 1.5 mg / kg 56.00 14.28
Gold - 200 mg / kg 73.11 49.20
Gold - 400 mg / kg 58.77 19.93 3-2 Evolution of impotence and ankylosis.

Paintings
Treatments Impotence Ankylosis
OJ J18 OJ J18
WITNESSES 2.25 2.40 1.05 1.39
Gold - 200 mg / kg 2.33 2.00 1.35 1.44
Gold - 400 mg / kg 2.13 1.44 * 1.41 0.99 *
INDOMETACIN ** ** 3 then 1.5 mg / kg 2.00 1.30 1.44 0.99
SIGNIFICANT DIFFERENCE AT J18 IN RELATION TO JO 3-3 Evolution of secondary factors
presence of edema on the front paws, nodule on the tail and ears, genital involvement and conjunctivitis.

 With the exception of the presence or absence of edema on the forelegs, the exploitation of the other factors did not give any significant indications as to the effectiveness of the treatments.

Evolution of edema of the front legs during treatment:
Fractions indicate the number of animals with front leg edema over the total number of animals in each lot.

Table XI
OJ processing J18
WITNESSES 5/9 6/9
Gold - 200 mg / k 4/9 4/9
Gold - 400 mg / kg 7/9 4/9
INDOMETACIN 4/9 1/9
Ana lysis of the results - discussion
Arthritic controls: Posterior leg volume increased from 76 to 90 mmHg, which represents an 18% worsening.

On the other hand, the% swelling compared to the absolute control animals changes from 58.33 (OJ) to 83.55 (D14); so there is, in
Lack of treatment, worsening of arthritis. This finding is confirmed by the progression of impotence (2.25 to 2.40), ankylosis (1.05 to 1.39) and extension of arthritis to the forelimb (5 / 9 to 7/9) reflecting overall a chronic phase evolution.

 However, ex 4, - 200 mg / kg - intraperitoneal route: we notice a stabilization of the evolution of the arthritis; The volume of hind legs remains at about 72 mm Hg; the% of swelling compared to absolute control animals also remains stable (48%). Finally, impotence (2.33 to 2.00), ankylosis (1.35 to 1.44) and involvement of the previous limbs (4/9 to 4/9) also confirm this stability.

 At this dose The derivative of the invention of ex. 4 therefore has an efficiency comparable to that of a background treatment.

 However, ex. 4 - 400 mg / kg - intraperitoneal route: we note a significant regression of Edema; the volume of the hind legs increases from 78 to 59 mm Hg (ie - 25% compared to the initial volume); the% swelling compared to the absolute control animals is only 19.93% at the end of treatment. Impotence (2.11 to 1.44), ankylosis (1.41 to 0.99) regressed significantly. The number of animals with anterior leg involvement is reduced (7/9 to 4/9). The X of swelling relative to the absolute control group changes from 61.9 (OJ) to 19.9 (J14). Finally the% of protection compared to the arthritic control lot is 34.65% (J14).

Indomethacin - 3 then 1.5 mg / kg - intragastric way the results obtained are quantitatively similar to those of the previous batch; Significant decrease as of J4 of the volume of the posterior legs (76 to 56 mm of Hg) is - 26%. The% swelling compared to the absolute control animals is 14.28% at the end of treatment. Impotence (2.00 to 1.30), ankylosis (1.44 to 0.99) and
Edema of the forelegs (4/9 to 1/5) decreases significantly. The% of swelling relative to the absolute control group increases from 57.35 (OJ) to 14.28 (J14). The% of protection compared to the arthritic control lot is 37.73% (J14).

 The activity of the gold derivative is therefore, over two weeks of observations, globally similar to that of indomethacin.

If the doses, 3 mg for indomethacin against 400 mg / kg for 803, are uncommon measures, it should be emphasized that indometacin caused death by gastric perforation of 4 animals from the fourth day of treatment. compelling us to reduce the dose to 1.5 mg / kg.

 The gold derivative of the invention, in our experimental conditions, did not show any apparent sign of toxicity except for a slight abdominal bloating at the end of treatment: although the statistical analysis did not reveal no significant difference, the indomethacin lot presents a most favorable weighted evolution. The weighted progression of the animals treated with the gold derivative can be interpreted by the intraperitoneal administration of a large quantity of substance whose resorption may present a problem and explain the bloating noted above.

Conclusion
The analysis of the results shows that, in our experimental conditions, the gold derivative according to the invention has a significant efficacy in the FREUND adjuvant arthritis model. At the doses used, its activity is comparable to that of indomethacin.

Claims (26)

 1. Organic derivative of a metal, characterized in that it consists of a derivative of a metal, preferably of valence at least equal to 3, with an organic compound comprising at least one phospholipid conferring a water solubility or hydromiscibility at organic derivative of said metal.  2. Organic derivative according to claim 1, characterized in that the aforementioned metal is bonded to the phosphate function of the phospholipid.  3. Organic derivative according to claim 2, characterized in that the phospholipid comprises at least one unsaturated fatty acid, preferably at position a or ss relative to the phosphate function.  4. Organic derivative according to claim 3, characterized in that the unsaturated fatty acid mentioned above is a polyunsaturated fatty acid, preferably without conjugation of the ethylenic bonds, more preferably at least two double bonds.  5. Organic derivative according to claim 4, characterized in that the unsaturated fatty acid mentioned above has at least three double bonds, of α-linolenic or γ-linolenic acid type.  6. Organic derivative according to claim 4, characterized in that it is chosen from the group consisting of 9, 12-linoleic acid, 9,12,15-linolenic acid and 5,8,11 acid. , 14-arachidonic.  7. Organic derivative according to one of Claims 2 to 6, characterized in that the phospholipid comprises at least one phosphoric acid function preferably conjugated with unsaturation of the phospholipid chain, this acid function being either free or linked to a group. unsubstituted or substituted ammonium, or alkylamine in which the alkyl group is C1-C5 lower alkyl, preferably C1-C3 lower alkyl.  8. Organic derivative according to one of claims 2 to 7, characterized in that the metal having a valency of at least 3 is selected from the group consisting of Sc, Ti, V, Cr, Mn, Fe, Co, Ni, Cu, Ga, Ge, Sb, In, Pd, Rh, Ru, Tc, Mo, Nb, Zr, Y, Hf, Ta, W, Re, Os, Ir, Pt, Au, Tl, Pb, Bi, Po.  9. Organic derivative according to claim 7 or 8, characterized in that the amine function mentioned above is bonded to a phenol or a phenolic acid, preferably chosen from caffeic acid, ferric acid, chlorogenic acid, salicylic acid or derivatives thereof. Salicylic acid, in particular salicin, a salycoside, a hyperoside, a spyreoside, a mono-tropitoside, salicylic aldehyde or a C1-C5 lower alkyl salycilate, preferably C1-C3, in particular methyl salycilate.  10. Organic derivative according to claim 8 or 9, characterized in that the phenolic acid is itself bonded to a C5 or C6 sugar, in particular mannitol or mannose.  11. Organic derivative according to one of claims 1 to 10, characterized in that the organic compound has the following general chemical formula:

 in which R1 = either a C16-C20 fatty acid advantageously having a  low degree of unsaturation, especially a double bond  = either a porphyrin ring bonded to an M1 metal having a valence  greater than or equal to 3 as previously defined,  = either an X-OH group in which X = (H) n, n being equal to 0, 1  or 2.

 = a phosphate function of formula:  at least 2 degrees of unsaturation,  R2 = either unsaturated fatty acid of C16 to C20, preferably having  provided that R2 and R3 can not simultaneously be a phosphate group.

 = a phosphate group of formula  R1 or R2, R3 = either a C16 to C20 unsaturated fatty acid as defined for Li, Cd, Zn, Mg and Ag.  a C1-C5 alkyl group free or linked to R5; in the preceding formula, a free acid function of the phosphate group of R2 or R3 may be salified by a metal M3 of valence less than or equal to 2, in particular chosen from Ca, Sr,  preferably selected from a sugar, a phenolic acid; is  wherein R5 equals a minimum C5 organic compound of  substituted, especially with a positive ionic group) -R5 R4 = either H or a quaternary ammonium group (unsubstituted or  12. Organic derivative according to one of claims 1 to 11, characterized in that the above phospholipid is bonded to a porphyrin group.  13. Organic derivative according to any one of claims 1 to 12, characterized in that it has the following general chemical formula:

  wherein A1, A2, A3 A4, A5, A7, A7, A8 are independently selected from the usual substituents of chlorophylls, in particular hydrogen, straight-chain, branched or cyclic C 1 -C 6 lower alkyl, radical vinyl, formula and its esters, acetyl and its esters, propionyl and its esters, acrylyl and its esters, and cyclopentanoyl; M1 being a metal of valency at least equal to 3, as previously defined.  And at least one of A1 to A8 is necessarily a phospholipid substituent as defined in any one of claims 1 to 12.  14. Organic derivative according to any one of claims 1 to 13, characterized in that the phosphoric function of the phospholipid is bonded to a cation of a second metal valence less than 3, preferably calcium, strontium, zinc , cadmium, lithium, magnesium and silver.  15. Organic metal derivative, characterized in that it is an organic derivative of gold bound to a phospholipid as defined in any one of claims 1 to 13, in particular obtained from the plant. Rybes nigrum.  16. Organic metal derivative, characterized in that it is the organic derivative of Sb bound to a phospholipid as defined in any one of claims 1 to 13, in particular obtained from the plant Angelica archangelica.  17. Organic metal derivative, characterized in that it is the organic vanadium derivative bound to a phospholipid as defined in any one of claims 1 to 13, in particular obtained from the plant Urtica dioica.  18. Organic metal derivative, characterized in that it is the organic aluminum derivative bound to a phospholipid preferably phosphatidylcholine as defined in any one of claims 1 to 13, in particular obtained from the plant Reseda phyteuma.  19. Biomedical agent, characterized in that it comprises at least one organic metal derivative as defined in any one of claims 1 to 18.  20. Pharmaceutical composition, characterized in that it comprises as active ingredient an organic metal derivative as defined in any one of claims 1 to 18.  21. Pharmaceutical composition with immunomodulatory activity, characterized in that it comprises an organic metal derivative as defined in any one of claims 1 to 18, optionally in a pharmaceutically acceptable excipient, carrier or vehicle.  22. Pharmaceutical composition with immunomodulatory activity, characterized in that it comprises a pharmaceutically effective amount of at least one derivative of Sb or Sb / Sn, preferably extracted from the species Angelica, in particular Angelica archangelica, and Angelica sinensis. optionally in a pharmaceutically acceptable excipient, carrier or vehicle.  23. Pharmaceutical composition with anti-inflammatory activity, characterized in that it comprises as active ingredient an organic metal derivative as defined in any one of claims 1 to 18, in a vehicle, excipient or pharmaceutically acceptable carrier.  24. Pharmaceutical composition with hypoglycemic activity, characterized in that it comprises as active ingredient an organic metal derivative as defined in any one of claims 1 to 18, optionally in a vehicle, excipient or pharmaceutically acceptable carrier.  25. Pharmaceutical composition with antirheumatic activity, in particular with anti-rheumatoid arthritis activity, characterized in that it comprises an organic derivative of a metal as defined in any one of Claims 1 to 18, as active principle optionally in a pharmaceutically acceptable excipient, vehicle or carrier.  26. Process for the preparation of a pharmaceutical composition, characterized in that it comprises the incorporation of at least one organic metal derivative as defined in any one of Claims 1 to 18 into an excipient, vehicle or pharmaceutically acceptable carrier.