EP2234610A2

EP2234610A2 - Use of aminopeptidase inhibitors or azaindole compounds for preventing or treating cancerous metastases from epithelial origin

Info

Publication number: EP2234610A2
Application number: EP09706702A
Authority: EP
Inventors: Pierre Roux; Marion De Toledo; Jean-Paul Leonetti; Christelle. Anguille
Original assignee: Centre National de la Recherche Scientifique CNRS
Current assignee: Centre National de la Recherche Scientifique CNRS
Priority date: 2008-01-22
Filing date: 2009-01-21
Publication date: 2010-10-06
Also published as: WO2009095583A2; WO2009095583A3; JP2011509982A; US20110046154A1; CN101951905A; FR2926463B1; CA2712339A1; FR2926463A1

Abstract

The invention relates to the use of a compound selected from an aminopeptidase-inhibiting compound and an azaindole compound for producing a drug for preventing or treating cancerous metastases in humans or animals.

Description

Use of aminopeptidase inhibitors or azaindole compounds for the prevention or treatment of cancer metastases of epithelial origin

Field of the invention

The present invention relates to the field of therapeutic treatment of cancers, and more particularly to the prevention or treatment of metastases, in the case of cancer in humans or animals.

Prior art

In general, approximately 35% of patients newly diagnosed with cancer are free of metastases. These patients can be cured by local treatments directed against the tumor, such as surgery or exposure to certain radiation. The other part of newly diagnosed patients includes (i) patients already having detectable metastases (30% of patients) and (ii) patients with no detectable metastases at the time of diagnosis (35% of patients). The treatment of this other part of the patients is generally a systemic treatment including, for example, the administration of chemotherapeutic drugs that interfere with the growth of the cancer cells.

Tumor invasion processes are the main cause of death in cancer patients.

It is known that, in some cancers, tumor cells not only have a strong ability to proliferate, but also a strong ability to destroy tissue and infiltrate into nearby tissues, including blood vessels and lymphatics, then to migrate through the bloodstream or lymphatic circulation to tissues located at parts of the body far removed from the primary tumor. Thus, in some cancers, tumor cells have the ability to circulate and then form secondary tumors, also called metastases, in tissues distant from the primary tumor.

The formation of metastases is a multi-stage physiological phenomenon, in which tumor cells break away from the primary tumor, invade the extracellular matrix, enter through the blood vessels, enter the vasculature by intravasation, and then stop migration into the blood or lymphatic circulation at a distant site, extravasate out of the circulation, then attach to a distant tissue and proliferate to form a secondary tumor.

In addition, the epithelium is the most abundant tissue type in the human or animal body. This is probably one of the reasons why more than 90% of human cancers originate from the malignant transformation of epithelial cells. In particular, colorectal epithelial cancer is the second leading cause of cancer death in industrialized countries.

In most cases of cancer, including epithelial cancers, the mortality is not due to the primary tumor, but to metastases that spread systemically in the body, from the primary tumor. This malignant progression that leads to tumor invasion, which is clinically manifested by the generation of metastases, is initially conditioned by the loss of cellular adhesion of certain tumor cells as well as an increase in cell motility, which results in these Invasive tumor cells leave the tissue of the primary tumor and then colonize one or more target tissues, sometimes very distant from the site of the primary tumors, in the human or animal body. In epithelial cancers, the malignant progression is accompanied by a loss of inter-cellular junctions made by E-cadherin, at the level of the primary tumor.

Among epithelial cancers, colorectal cancers represent the second most common cancers in industrialized countries. The evolution of colorectal cancer is a succession of events that begins with an initiation phase due to a genomic dysregulation affecting the proliferative properties of cells. The second phase is a phase of tumor progression that leads to changes in cell morphology and the emergence of transformed cell clones. Finally, the last phase consists of the invasion phase following the epithelio-mesenchymal transition, defined as the ability of an epithelial cell to separate from neighboring cells and migrate to other sites in the body.

In practice, it has been shown that systemic anti-cancer therapeutics have little effect on metastases, and particularly on macro-metastases, which reside in organs distant from the primary tumor site, such as the lung, the liver, bone marrow or brain. Thus, cancer patients often die due to metastatic cancers caused by metastases of cancer cells.

Moreover, until now, the search for anti-invasive molecules has generally been based on (i) the ability of candidate compounds to inhibit the growth of tumor cells, (ii) the ability of the candidate compounds to inhibit cell migration. But, according to the first strategy, the selected compounds have no effect on the spread of invasive cells and therefore no effect on the metastatic process itself. And, in the second case, the compounds may have an activity of inhibiting the ability of cells to migrate into the body and forming metastases, but have no effect on metastatic cells that have already migrated.

There is therefore a need in the state of the art for the identification of active substances having the ability to prevent or treat metastases at patients with cancer, to significantly increase the chances of recovery of these patients.

SUMMARY OF THE INVENTION The present invention relates to the use of a compound selected from an aminopeptidase inhibiting compound and an aza-indole compound for the manufacture of a medicament for the prevention or treatment of cancer metastases in the man or the animal.

It relates in particular to the use, for the manufacture of a medicament for the prevention or treatment of cancerous metastases in humans or animals, of an anti-metastatic compound chosen from compounds of formula ( I), (II), (III), (IV), (V) and (Vl) which are described in detail later in the present description.

The invention relates to the use defined above, for the prevention or treatment of metastases in the case of epithelial cancers, including in the case of colorectal cancer, mammary cancer, liver, pancreas, prostate and uterus.

DESCRIPTION OF THE FIGURES FIG. 1 is a diagram illustrating the anti-metastatic effect of each of the compounds of formulas (I); (II), (III), (IV), (V) and (VI) of the invention. On the ordinate: the invasion capacity of the cells tested, in percentage with respect to the invasion capacity of the cells of the SW620 line (ATCC No. CCL-227) which was arbitrarily set at 100%. On the abscissa, from left to right: (i) cells of the colo-rectal metastatic SW620 line incubated with the culture medium alone, (ii) cells of the non-metastatic Hct1 16 line incubated with the medium alone, and (iii) cells of the metastatic SW620 line incubated with the culture medium supplemented with each of the compounds of formula (I), (II), (III), (IV), (V) and (VI). The results shown in Figure 1 are an average of three experiments. Figure 2 is a diagram illustrating the anti-metastatic effect of each of the compounds of formulas (I); (II), (III), (IV), (V) and (VI) of the invention in other cell lines. On the ordinate: the invasion capacity of the cells tested, in percentage relative to the invasion capacity of the cells of the CoLo205 line (ATCC No. CCL-222) which was arbitrarily set at 100%. On the abscissa, from left to right: (i) cells of the colo-rectal metastatic CoLo205 line (ATCC No. CCL-222) incubated with the culture medium alone or supplemented with each of the compounds of formula (I), (II) , (III), (IV), (V) and (Vl). The results shown in Figure 2 are an average of three experiments.

Figure 3 is a digraph illustrating the anti-metastatic effect of each of the compounds of formulas (I); (II), (III), (IV), (V) and (VI) of the invention. On the ordinate: the invasion capacity of the cells tested, as a percentage of the invasion capacity of the cells of the colo-rectal metastatic SK-CoI line (ATCC No. HTB-77) which was arbitrarily set at 100%. On the abscissa, from left to right: (i) cells of the colo-rectal metastatic SK-CoI line (ATCC No. HTB-77) incubated with the culture medium alone or supplemented with each of the compounds of formula (I), ( II), (III), (IV), (V) and (VI). The results shown in Figure 3 are an average of three experiments.

Figure 4 is a digraph illustrating the anti-metastatic effect of each of the compounds of formulas (I); (II), (III), (IV), (V) and (VI) of the invention. On the ordinate: the invasion capacity of the cells tested, as a percentage relative to the invasion capacity of the cells of the MDA-MB-231 metastatic line of mammary adenocarcinoma (ATCC No. HTB-26) which was arbitrarily fixed 100%. On the abscissa, from left to right: (i) cells of the metastatic SW620 line incubated with the culture medium alone, (ii) cells of the non-metastatic Hct1 16 line incubated with the mileiu alone, and (iii) cells of the line MDA-MB-231 metastatic mammary adenocarcinoma (ATCC No. HTB-26) metastatic incubated with the culture medium alone or supplemented with each of the compounds of formula (I), (II), (III), (IV), (V) and (Vl). The results shown in Figure 4 are an average of three experiments.

Detailed description of the invention

The present invention provides novel means for preventing or treating metastasis in various cancers, particularly in epithelial cancers, including colorectal cancers, breast cancers, liver cancers, pancreatic cancers, Prostate and cancers of the uterus. Surprisingly, it has been shown according to the invention that aminopeptidase inhibiting compounds and compounds of the azaindole family have the ability to inhibit or block invasive properties. of cancer cells initially capable of generating metastases.

More specifically, it is shown according to the invention that aminopeptidase inhibiting compounds and compounds of the azaindole family have the capacity to inhibit or block the migration properties of metastatic cells originating from various epithelial cancers, such as colo-rectal cancers or breast cancers.

The identification of the anti-metastatic properties of these compounds has enabled the applicant to develop pharmaceutical compositions for preventing or treating cancer metastases in humans or animals, comprising as active ingredient at least one selected compound. from an aminopeptidase inhibitor compound and an azaindole compound. This identification of new preventive or therapeutic properties has made possible the use of these compounds for the manufacture of pharmaceutical compositions with anti-metastatic activity. Thus, the present invention relates to a compound selected from an aminopeptidase inhibiting compound and an aza-indole compound for the prevention or treatment of cancer metastasis in humans or animals.

The present invention also relates to the use of a compound selected from an aminopeptidase inhibiting compound and an aza-indole compound for the manufacture of a medicament for the prevention or treatment of cancer metastases in humans or the animal.

As illustrated in the examples, the aminopeptidase inhibiting compounds and the aza-indole compounds identified according to the invention have the capacity to induce a reversion of the invasive phenotype of cancer cells such as metastatic colorectal carcinoma cells and metastatic mammary adenocarcinoma cells.

In addition, the Applicant has shown that the aminopeptidase inhibiting compounds and the azaindole compounds identified according to the invention are not cytotoxic.

Without wishing to be bound by any theory, the applicant believes that the anti-metastatic properties of the aminopeptidase inhibiting compounds and of the aza-indole compounds identified according to the invention are due to the common ability of all these compounds to induce the restoration of cellular junctions associated with the expression of E-cadherin on the surface of metastatic cancer cells.

The ability of the anti-metastatic compounds of the invention to induce the expression of E-cadherin on the surface of the cancerous cells and thus induce the recovery of cellular junctions associated with this expression of E-cadherin can be verified, in particular by performing the test described in the PCT application published under No. WO 2006/134305 (National Center for Scientific Research - ROUX Pierre and TOLEDO Marion).

According to a first embodiment of an anti-metastatic compound which is used according to the invention, said compound consists of an aminopeptidase inhibiting compound of formula (I) below:

[Structural analogs of the compound (I) to be specified by the inventors, please]

The compound of formula (I) is also referred to herein as the compound 2- (4 - [(4-amino-1, 2,5-oxadiazol-3-yl) (hydroxyimino) methyl] -1-piperazinyl) ethanol . The compounds of formula (I) can be synthesized according to any synthesis method known to those skilled in the art. In order to synthesize a compound of formula (I), those skilled in the art may in particular refer to the method described in the following document: Synthesis of secondary and tertiary aminofurazans. Sheremetev, AB; Andrianov, VG; Mantseva, EV; Shatunova, EV; Aleksandrova, NS; Yudin, I. L .;

Dmitriev, D. E .; Averkiev, B. B .; Antipin, M. Yu. N. D. Zelinsky Institute of Organic

Chemistry, Russian Academy of Sciences, Moscow, Russia. Russian Chemical Bulletin

Izvestiya Akademii Nauk, Seriya Khimicheskaya (2004), 53 (3), 596-614.

Publisher: Kluwer Academy / Office Consultants, CODEN: RCBUEY ISSN: 1066-5285. Journal written in English. CAN 142: 21921 1 YEAR 2004: 589877 CAPLUS

According to a second embodiment of an anti-metastatic compound which is used according to the invention, said compound consists of an azaindole compound of formula (II) below:

in which the R 1 group is a linear or branched alkyl having from 1 to 8 carbon atoms

"Alkyl" means a linear or branched aliphatic hydrocarbon group optionally interrupted by a heteroatom, the alkyl may be unsubstituted or substituted on the carbon atoms by one or more identical or different substituents, the substituents being chosen from: aryl, hydroxy, alkoxy, aryloxy, alkyloxy, aralkyloxy. By "branched" alkyl is meant lower alkyl, having

1, 2, 3, 4 or 5 carbon atoms, such as methyl, ethyl or propyl which is linked to an alkyl linear chain. Preferred alkyl groups consist of alkyl groups having 1, 2, 3, 4, 5, 6, 7 or 8 carbon atoms. Examples of alkyl groups are methyl, ethyl, n-propyl, n-butyl, heptyl, decyl or cyclohexylmethyl.

By "alkoxy" is meant an alkyl-O- group in which the alkyl group is as previously defined. Alkoxy groups include methoxy, ethoxy, n-propoxy, n-propoxy, n-butoxy and heptoxy. By "aryloxy" is meant an aryl-O group in which the aryl group is as previously defined. Aryloxy groups include phenoxy and naphthoxy.

By "aralkyl" is meant an alkyl group substituted with one or more aryl groups. By "aralkyloxy" is meant an aralkyl-O- group in which the aralkyl group is as previously defined. Aralkyloxy groups include benzyloxy. A compound of formula (II) which is preferred according to the invention is the compound in which the group R 1 denotes a propyl group, which may also be designated in the present description as the compound methyl 4 - [([3- (4-fluorophenyl) 4-oxo-2-thioxo-1,3-thiazolidin-5-ylidene) methyl) benzoate. The compounds of formula (II) can be synthesized according to any synthesis method known to those skilled in the art. To synthesize a compound of formula (II), those skilled in the art can in particular refer to the process described in the following document: quinazolinone compounds. (Tanabe Seiyaku Co., Ltd., Japan). Jpn. Kokai Tokkyo Koho (1982), 5 pp. CODEN: JKXXAF JP 5701 1970 To 19820121 Showa. Patent written in Japanese. Application: JP 80-86044 19800624. Priority:. CAN 97: 72383 AN 1982: 472383 CAPLUS According to a third embodiment of an antimetastatic compound which is used according to the invention, said compound consists of an azaindole compound of formula (III) below:

in which the groups R ₁ , R ₂ , R ₃ and R ₄ each represent, independently of one another, an alkyl of 1 to 8 carbon atoms, the definition of which is identical to that of the R ₁ group of the compound of formula (II).

A preferred compound of formula (III) is the compound wherein R _{1 is} methyl and R ₂ , R ₃ and R _{4 are} ethyl, said compound may be designated as 3,4 5-triethoxy-N- (2-methyl-4-nitrophenyl) benzamide.

The compounds of formula (III) can be synthesized according to any synthesis method known to those skilled in the art. To synthesize a compound of formula (III), a person skilled in the art can in particular refer to the process described in the following document: Microwave-assisted synthesis of salicylamide via BCI3 mediated coupling. Zhang, Lei; Zhang, John Y. CytRx Laboratories, Inc., Worcester, MA, USA. Journal of Combinatorial Chemistry (2005), 7 (4), 622-626. Publisher: American Chemical Society, CODEN: JCCHFF ISSN: 1520-4766. Journal written in English. CAN 143: 21 1699 YEAR 2005: 538799 CAPLUS According to a fourth embodiment of an antimetastatic compound which is used according to the invention, said compound consists of an azaindole compound of formula (IV) below:

in which the groups R 1, R ₂ and R 3 represent, independently of one another, a halogen selected from a fluorine (F), chlorine (C) I, iodine (I) and bromine atom; (B) r.

A compound of formula (IV) which is preferred according to the invention is the compound in which the groups R 1 and R ³ each signify a chlorine atom and the group R ₂ signifies a bromine atom, which compound can also be designated as the compound 5 - (5-Bromo-3-chloro-2-hydroxybenzylidene) -1- (4-chlorophenyl) -2,4,6 (1H, 3H, 5H) -pyrimidinetrione.

The compounds of formula (IV) can be synthesized according to any synthesis method known to those skilled in the art. In order to synthesize a compound of formula (IV), those skilled in the art may in particular refer to the method described in the following document: Synthesis of pyrimidine derivatives possessing an antioxidative property and their inhibitory effects on picryl chloride-induced contact hypersensitivity reaction. Isobe, Yoshiaki; Hirota, Kosaku. Pharmaceutical and Biotechnology Laboratory, Japan Energy Corporation, Saitama, Japan. Chemical & Pharmaceutical Bulletin (2003), 51 (12), 1451-1454. Publisher: Pharmaceutical Society of Japan, CODEN: CPBTAL ISSN: 0009-2363. Journal written in English. CAN 140: 174446 YEAR 2003: 989349 CAPLUS

According to a fifth embodiment of an anti-metastatic compound which is used according to the invention, said compound consists of an azaindole compound of formula (V) below:

in which :

the group R 1 is a halogen selected from a fluorine (F), chlorine (C) I, iodine (I) and bromine (B) r atom, and the group R ₂ is a linear or branched alkyl having from 1 to 8 carbon atoms, the definition of which is identical to that of the R 1 group of the compound of formula (II).

A preferred compound of formula (V) is the compound in which the group R 1 means a bromine atom and the group R 2 signifies an ethyl group, which compound can also be designated as the 5-bromo-N- (4-butoxyphenyl) compound nicotinamide.

The compounds of formula (V) can be synthesized according to any synthesis method known to those skilled in the art. To synthesize a compound of formula (V), a person skilled in the art can in particular refer to the method described in the following document: Application of organolithium and related reagents in synthesis, 30. Behavior of N-pyridylbenzamides versus benzanilides in the ortho-directed lithiation of masked aromatic carboxylic acids. Jozwiak, Andrzej; Brzezinski, Jacek Z .; Plotka, Mieczyslaw W .; Szczesniak, Aleksandra K.; Malinowski, Zbigniew; Epsztajn, Jan. Department of Organic Chemistry, Institute of Chemistry, University of Lodz, Lodz, PoI. European Journal of Organic Chemistry (2004), (15), 3254-3261. Publisher: Wiley-VCH Verlag GmbH & Co KGaA, CODEN: EJOCFK ISSN: 1434-193X. Journal written in English. CAN 141: 295825 YEAR 2004: 636845 CAPLUS

According to a sixth embodiment of an anti-metastatic compound which is used according to the invention, said compound consists of an azaindole compound of formula (VI) below:

wherein the group R ₁ is a halogen selected from a fluorine (F), chlorine (C) I, iodine (I) and bromine (B) r.

The compound of formula (VI) may also be designated the compound 3-chloro-1-phenyl-4 - [(2-phenylethyl) amino] -1 Hpyrrole-2,5-dione.

The compounds of formula (VI) can be synthesized according to any synthesis method known to those skilled in the art. The compounds of formula (VI) consist of compounds readily available commercially. There may be mentioned in particular the compound of formula VI consisting of 3-chloro-1-phenyl-4 - [(2-phenylethyl) amino] -1H-pyrrole-2,5-dione, which is marketed by the company Chembridge ( San Diego, United States of America) under the reference ID 6137235.

Thus, according to the invention, the preferred active ingredients which are used to manufacture a medicament for the prevention or treatment of cancerous metastases in humans or animals are chosen from: - methyl 4 - [([3- (4-fluorophenyl) -4-oxo-2-thioxo-1,3-thiazolidin-5-ylidene) methyl) benzoate; 3,4,5-triethoxy-N- (2-methyl-4-nitrophenyl) benzamide; 5- (5-bromo-3-chloro-2-hydroxybenzylidene) -1- (4-chlorophenyl) -

2,4,6 (1H, 3H, 5H) -pyrimidinetrione;

5-bromo-N- (4-butoxyphenyl) nicotinamide; and 3-chloro-1-phenyl-4 - [(2-phenylethyl) amino] -1H-pyrrole-2,5-dione.

According to a first preferred aspect, the aminopeptidase inhibiting compounds and the azaindole compounds of interest according to the invention are used for the manufacture of a medicament intended for the prevention or treatment of epithelial cancers. According to a second preferred aspect, the aminopeptidase inhibiting compounds and the azaindole compounds of interest according to the invention are used for the manufacture of a medicament intended for the prevention or treatment of an epithelial carcinoma chosen from a colon cancer. rectal and breast cancer.

Among the pharmaceutical compositions according to the invention, mention may be made more particularly of those which are suitable for oral, parenteral, nasal, percutaneous, transcutaneous, rectal, perlingual or respiratory administration and in particular simple or coated tablets, sublingual tablets, capsules , tablets, suppositories, creams, ointments, dermal gels, and oral or injectable ampoules. The dosage varies according to the sex, the age and the weight of the patient, according to the route of administration, and according to the type of cancer, the state of progression of the cancer, in particular according to whether metastases have been detected or not in the patient. The dosage may also vary depending on the type of the associated anti-cancer treatment (s). In general, an anti-metastatic compound as defined in the present description is used in amounts ranging from 0.01 mg to 1 g per 24 hours for a man or an adult woman with an average weight of 80 kg, in one or more catches.

A pharmaceutical composition according to the invention comprises the anti-metastatic compound in combination with at least one excipient selected from the group consisting of pharmaceutically acceptable excipients.

In general, a pharmaceutical composition according to the invention comprises from 0.01% to 99% by weight, and advantageously from 1% to 90% by weight, of an antimetastatic compound, relative to the total weight of said composition. . In general, a pharmaceutical composition according to the invention comprises from 1% to 99.99% by weight, and preferably from 10% to 99% by weight, of an excipient or a combination of pharmaceutically acceptable excipients.

The pharmaceutical composition of the present invention can be used for parenteral, topical or local administration and prophylactically and / or therapeutically. Thus, the anti-metastatic compound according to the present invention is prepared in a form adapted to the type of administration chosen, for example in liquid form or in freeze-dried form. The pharmaceutical compositions comprising an anti-metastatic compound according to the invention may contain an excipient and / or a pharmaceutically acceptable vehicle, preferably an aqueous vehicle. Many pharmaceutically acceptable excipients and / or carriers may be used, for example, water, buffered water, saline solution, glycine solution and their derivatives as well as agents necessary to reproduce physiological conditions such as buffering agents and pH adjusters, surfactants such as sodium acetate, sodium lactate, sodium chloride, potassium chloride, calcium chloride, this list not being limiting. In addition, the pharmaceutical composition can be sterilized by sterilization techniques well known to those skilled in the art.

For excipients or inert, non-toxic, pharmaceutically acceptable vehicles, mention may also be made, by way of indication and without limitation, of diluents, solvents, preservatives, wetting agents, emulsifiers, dispersing agents, binders, blowing agents, disintegrants, retardants, lubricants, absorbents, suspending agents, dyes, flavoring agents, etc.

When preparing a solid composition in tablet form, the main active ingredient is mixed with a pharmaceutical carrier such as gelatin, starch, lactose, magnesium stearate, talc, gum arabic or the like.

The tablets may be coated with sucrose or other suitable raw materials or may be treated so that they have prolonged or delayed activity and continuously release a predetermined amount of active ingredient.

A preparation in capsules is obtained by mixing the active ingredient with a diluent and pouring the resulting mixture into soft or hard gelatin capsules.

A pharmaceutical composition in the form of a syrup or elixir may contain the active ingredient together with a sweetener, preferably a calorie-free sweetener, methylparaben and propylparaben as antiseptics, as well as a flavoring agent and a suitable colorant.

The water-dispersible powders or granules may contain the active ingredient in admixture with dispersing agents or wetting agents or suspending agents, such as polyvinylpyrrolodinone, as well as with sweeteners or scavengers. taste.

The active ingredient may also be formulated in the form of microcapsules, optionally with one or more supports or additives.

In general, to manufacture a pharmaceutical composition according to the invention, a person skilled in the art can advantageously refer to the latest edition of the European Pharmacopoeia, for example at the 5th edition of the Pharmacopoeia. European published in January 2005, or even the 6th edition of the European Pharmacopoeia, available to the public in June 2007.

Techniques for preparing pharmaceutical compositions according to the invention can easily be found by those skilled in the art, for example in the book Remmingston's Pharmaceutical Sciences, Mid. Publishing Co, Easton, PA, USA.

Physiologically acceptable adjuvants, vehicles and excipients are also described in the book "Handbook of Pharmaceutical Excipients, Second Edition, American Pharmaceutical Association, 1994. To formulate a pharmaceutical composition according to the invention, the skilled person may advantageously refer to the latest edition of the European Pharmacopoeia or the United States Pharmacopoeia (USP).

Those skilled in the art may advantageously refer to the USP 30-NF 25 edition of the American Pharmacopoeia (U.S. Pharmacopeia). Advantageously, a pharmaceutical composition as defined above is suitable for oral, parenteral or intravenous administration.

When the pharmaceutical composition according to the invention comprises at least one pharmaceutical or physiologically acceptable excipient, it is in particular a suitable excipient for administration of the composition orally or of a suitable excipient for administration of the composition parenterally.

The invention also relates to a method for preventing or treating a disorder related to an imbalance of bone metabolism, particularly a disorder associated with loss of bone mass, said method comprising a step in which patients are administered a therapeutically therapeutic amount. effective of an anti-metastatic compound as defined in the present description or a pharmaceutical composition containing said anti-metastatic compound.

A pharmaceutical composition comprising an antimetastatic compound according to the invention is indifferently in a solid form or in a liquid form.

For oral administration, a solid pharmaceutical composition, in the form of tablets, capsules or capsules, will be preferred.

In the liquid form, a pharmaceutical composition in the form of an aqueous suspension or a non-aqueous suspension, or in the form of a water-in-oil or oil-in-water emulsion, is preferred.

Solid pharmaceutical forms may include, as carriers, adjuvants or excipients, at least one diluent, flavor, solubilizer, lubricant, suspending agent, binder, disintegrating agent, and agent. encapsulation. Such compounds are, for example, magnesium carbonate, magnesium stearate, talc, lactose, pectin, dextrin, starch, gelatin, cellulosic materials, cocoa butter, and the like.

The compositions in liquid form may also comprise water, optionally mixed with propylene glycol or polyethylene glycol, and optionally also coloring agents, flavors, stabilizers and thickeners.

The invention is further illustrated, without being limited, by the following example.

Example 1: Anti-invasive properties of the anti-metastatic compounds of the invention.

A. MATERIALS AND METHODS 1. E-cadherin labeling protocol in 96-well plates

1 - Inoculate approximately 10,000 cells per well for colorectal lines (in 96-well Falcon plates, ref: 353072).

2- Make the necessary treatments (control, compound (I), (II) etc. during

48h) and fix 10 minutes to formalin (Formaldehyde 3.7% in PBS)

3- Saturation, 10 minutes PBS-BSA

4- Incubation of the primary antibody, diluted 1: 500 in PBS-BSA, 2 hours to

37 ⁰ C (anti mouse E-cadherin, ZYMED, ref: 13-1700)

5- 2 rapid rinses with PBS - 0.1% Tween

6- Incubation Ac anti-mouse biotinylated, dilution 1: 1000 in PBS-BSA, 30 minutes at 37 ° C, (Anti-Mouse IgG, Heavy and light chain specifies biotin conjugate, CALBIOCHEM, ref: 401213)

7- 2 rapid rinses with PBS - 0.1% Tween

8- Streptavidin-HRP incubation, dilution between 1: 1000 in PBS-BSA, 30 minutes at 37 ° C., (ECL strptavidin-Horseradish peroxidase conjugate, AMERSHAM BIOSCIENCES, ref: RPN1231)

9- 2 rapid rinses with PBS - 0.1% Tween 10- Revelation:

Dissolve one capsule of "Phosphate-citrate buffer containing sodium perborate" (SIGMA, ref: P4922) in 100 ml of deionized water. This buffer must be used within 30 minutes of reconstitution.

Dissolve o-Phenylenediamine (OPD) dihydrochloride tablets (SIGMA, ref: P6787) in this buffer to obtain a final concentration of 0.4 mg / ml (1 OPD 10 mg pellet in 40 ml perborate buffer).

Put 100 μl of OPD / Perborate per well and incubate at room temperature for 5 minutes.

Stop the reaction with 50 μl of 3N HCl.

Read the OD at 490 nm.

2. Invasion test

1 - D-2, defrost the material at 4 ° C all night ...

2- At D-1, pour the material into the nacelles:

Place all necessary equipment in the ice (matrigel, culture medium, cones, eppendorfs, etc.).

Place the fluoroblock inserts (FALCON, ref: 351 152) in the 24-well plates (FALCON, ref: 353504).

Dilute the matrigel to 2 mg / ml in culture medium without cold serum (if the cells must undergo X treatment, include the product in the matrigel, eg: Y27632)

Distribute 100 μl of matrigel 2 mg / ml per insert, avoiding bubbles ...

Leave O / N at 37 ° C in a humid atmosphere

3- On day D, seed the cells on the material:

First, put 700μl of 10% serum medium in a 24-well plate and transfer the inserts into this plate.

Trypsinate the cells (previously treated or not ...) and take them back in 2% serum medium.

Inoculate 50,000 cells per insert in 200 μl of medium 2% serum on the matrigel (optionally put Y27632 in the seeded cells). 4- The migration time is variable depending on the cell type but generally allow to migrate 8 hours by setting a point every 2 hours seems appropriate.

To fix, transfer the inserts into a 24-well plate containing 1 ml of formalin (3.7% formaldehyde in PBS), aspirate the culture medium inside the nacelles and also add formalin (very important, avoid cells to continue to migrate in the matrigel !!!). Incubate 1 0 minutes.

Make 3 quick rinses with PBS

Then, carry out the propidium lodide labeling by incubating the inserts in 1 ml of propidium lodide (SIGMA, ref: P-4864) 1: 500 in PBS, O / N at 4 ° C., protected from light .

B. RESULTS

B.1. The ability of the anti-metastatic compounds of the invention to induce the recovery of inter-cellular junctions

The compounds of formula (I) to (VI) were tested for their ability to induce the restoration of inter-cellular junctions in in vitro cultures of SW620 igneous cells, using the E-cadherin labeling assay described in US Pat. Part Material and Methods. As a negative control, the cells of the SW620 line incubated in the culture medium alone were used in the absence of an anti-metastatic compound. An average O.D. of 0.147 was obtained.

As a positive control, cells of the HCT1 line 16 were used, incubated in the culture medium alone, in the absence of an anti-metastatic compound. An average O.D. of 0.734 was obtained.

The tests were carried out by incubating the cells of the SW620 line with each of the compounds (I) to (VI).

In these tests, the average OD obtained with each of the compounds (I) to (VI) has always been greater than 0.300, which means that the compounds (I) to (VI) are capable of inducing cell spreading, formation of compact cell islands, and to induce the generation of E-cadherin-type inter-cellular neo-junctions.

B.2. Anti-Invasive Properties of the Anti-Metastatic Compounds of the Invention The ability of each of the compounds of formula (I) to (VI) to inhibit or block the invasive properties of metastatic cancer cells was measured with the invasion test described. in the Material and Methods part.

The cells of the SW620 metastatic line incubated in the absence of anti-metastatic compound were used as a negative control, whose control value was arbitrarily set at 100%. Cells of the non-metastatic cancerous line HCT116 were used as a positive control.

The results are shown in FIGS.

In FIG. 1, it is shown that all the compounds of formula (I) to (VI) are capable of blocking the invasive properties of the cells of the metastatic colo-rectal cancer cell line, since the treated cells have a similar invasive activity or lower than that of the HCT1 line 16 of non-metastatic colorectal cancer.

In addition, the results of FIGS. 2, 3 and 4 show that the anti-invasive properties of each of the compounds of formula (I) to (VI) are demonstrated on various types of metastatic human cancer cells, including cells originating from breast cancer (Figure 4).

Claims

1. Use of a compound selected from an aminopeptidase inhibiting compound and an azaindole compound for the manufacture of a medicament for the prevention or treatment of cancer metastasis in humans or animals.

2. Use according to claim 1, characterized in that the aminopeptidase inhibiting compound is the compound of formula (I) below:

3. Use according to claim 1, characterized in that the azaindole compound is the compound of formula (II) below:

in which the group R 1 is a linear or branched alkyl having from 1 to 6 carbon atoms

4. Use according to claim 1, characterized in that the azaindole compound is the compound of formula (III) below:

in which the groups R 1, R 2, R 3 and R 4 each represent, independently of one another, an alkyl of 1 to 6 carbon atoms

5. Use according to claim 2, characterized in that the azaindole compound is the compound of formula (IV) below:

wherein the groups R1, R2 and R3 each independently represent a halogen selected from F, Cl, I and Br.

6. Use according to claim 1, characterized in that the azaindole compound is the compound of formula (V) below:

in which: the group R 1 is a halogen chosen from F, Cl, I and Br, and

the group R2 is a linear or branched alkyl having from 1 to 6 carbon atoms.

7. Use according to claim 1, characterized in that the azaindole compound is the compound of formula (VI) below

wherein the group R 1 is a halogen selected from F, Cl, I and Br.

8. Use according to claim 1, characterized in that the azaindole compound is chosen from the following compounds: methyl 4 - [([3- (4-fluorophenyl) -4-oxo-2-thioxo-1,3-thiazolidine Ylidene) methyl) benzoate; 3,4,5-triethoxy-N- (2-methyl-4-nitrophenyl) benzamide; 5- (5-bromo-3-chloro-2-hydroxybenzylidene) -1- (4-chlorophenyl) -

2,4,6 (1H, 3H, 5H) -pyrimidinetrione;

5-bromo-N- (4-butoxyphenyl) nicotinamide; and

3-chloro-1-phenyl-4 - [(2-phenylethyl) amino] -1H-pyrrole-2,5-dione.

9. Use according to one of claims 1 to 8, characterized in that said medicament is intended for the prevention or treatment of epithelial cancers.

10. Use according to claim 9, characterized in that the epithelial cancer is selected from a colo-rectal cancer and breast cancer.