WO2013045826A1

WO2013045826A1 - Compositions containing spermine together with cadaverine, putrescine and/or spermidine

Info

Publication number: WO2013045826A1
Application number: PCT/FR2012/052162
Authority: WO
Inventors: Jacques-Philippe Moulinoux
Original assignee: Nutrialys Medical Nutrition Sa
Priority date: 2011-09-29
Filing date: 2012-09-26
Publication date: 2013-04-04
Also published as: FR2980709B1; EP2760442A1; FR2980709A1

Abstract

The present invention relates to novel compositions including a mixture of cadaverine, putrescine, and spermidine at a concentration of 0.3 to 0.6 nanomoles per gram of composition, said composition also including spermine at a concentration of 150 to 17,000 nanomoles per gram of composition, as well as to the uses thereof for treating a patient for diseases associated with cell hyperproliferation.

Description

COMPOSITIONS CONTAINING SPERMINE WITH CADAVERINE, PUTRESCINE AND / OR

spermidine

The present invention relates to novel compositions containing spermine, pharmaceutical compositions containing them, and their uses in the preparation of medicaments for the treatment in a patient of pathologies related to cellular hyperproliferation.

Polyamines are polycationic aliphatic amines which include putrescine, spermidine, spermine in eukaryotes. Cadaverine, other polyamine, is present only in prokaryotes.

Polyamines are synthesized from L-arginine or L-methionine. Their pathway of biosynthesis and catabolism is based on a limited number of enzymes. The synthesis of putrescine from ornithine (arginine degradation product) involves Ornithine Decarboxylase (ODC). Spermidine synthetase (SpdS) allows the synthesis of spermidine from putrescine, which in turn will be supported by spermine synthetase (SpmS) to form spermine. Spermidine / spermine Nl-acetyltransferase (S S AT) is the key enzyme for catabolism of polyamines that allows the conversion of spermine to spermidine and spermidine to putrescine. SSAT allows fine regulation of the intracellular levels of the different polyamines which may be toxic to the cell.

Spermidine and spermine can also be synthesized from methionine by the action of S-adenosylmethionine decarboxylase (SAMdc).

The data in the literature show that a daily food intake of 2000 kcal (kilo calories) provides a dose of logically "active" organic polyamines which varies from 200,000 to 700,000 nanomoles (nmol). This means that a daily food ration provides a supply of 100 to 350 nmol of logically active organic polyamines per kilocalorie (Bardocz et al., 1995, Br J Nutr, 73 (6): 819-828). This difference depends on the nature of the foods consumed that contain varying amounts of polyamines.

In order to be able to convert into grams (g) quantities of logically active bioamines expressed in nanomoles, it is necessary to consider an average molecular weight for all the logically active bio-polyamines. This average molecular weight of logically active organic polyamines is an approximation and is estimated at 145.24 g / mol. Thus, a daily ration of 2000 kcal brings from 14.5 to 51 μg of logically active organic polyamines / kcal. This corresponds to a contribution of 29 to 102 mg of logically active organic polyamines / day or to a contribution of 29 to 102 mg of logically active organic polyamines for 2000 kcal.

It may be necessary to consider, either the pool of logically active polyamines as a whole but the level of each of the logically active polyamines considered alone. Foods can be divided into three classes according to their polyamine content (www.guerir.org/magazine/nutrition-guide- nutrialys.pdf). This distribution makes it possible to roughly estimate the levels of each of the polyamines (cadaverine, putrescine, spermidine and spermine) for each category of food. Thus, a "medium" daily food intake provides respectively about 10.8 mg of putrescine, about 4.4 mg of cadaverine, about 15.7 mg of spermidine and about 6.7 mg of spermine for 2000 kcal. This corresponds to an overall polyamine level (cadaverine, putrescine, spermine and spermidine) of approximately 37.6 mg for a daily "average" food ration of 2000 kcal. Consequently, putrescine represents 28.7%, cadaverine represents 11.7%, spermidine represents 41.8% and spermine represents 17.8% of the overall level of logically active organic polyamines of the aforementioned food ration.

Logically "active" organic polyamines can be identified in particular according to at least one of the following methods:

1) In culture:

A logically "active" organic polyamine or one of its derivatives must be able to participate in the physiological cellular metabolism of polyamines, or even be able to interfere with the latter or to dysregulate.

at. A logically "active" organic polyamine must therefore be able to associate or even be recognized by the transport system or systems intended to internalize it in a living cell. The addition of a logically "active" bio-labeled polyamine radiolabeled in the culture medium makes it possible to verify its internalization.

b. A logically "active" organic polyamine must be able to suppress the inhibition of cell proliferation caused by the inhibition of the endogenous anabolism of polyamines (eg by Γα-DFMO). vs. A logically "active" organic polyamine, including synthesis, by dysregulating the natural metabolism of polyamines must be able to modulate the level of cell proliferation. 2) In vivo:

In the grafted tumor bearing animal, the exogenous supply (gastrointestinal tract) of "active" polyamines must suppress the beneficial anti-cancer effects caused by the "active" polyamine deficiency induced by the decrease of endogenous and exogenous sources of "active" polyamines ", This exogenous intake being coupled or not with anticancer drugs.

Polyamines are essential compounds for cell growth and differentiation. The inventors have shown that a diet depleted of polyamines makes it possible to potentiate the effects of inhibitors of ODC on tumor growth (EP 0 703 731 B1).

In addition, studies have shown that cancer cells contain higher levels of polyamines than those observed in healthy cells (WO 1997/11691). Depletion of polyamines has been used in anticancer strategy. This depletion is achieved either by inducing the expression of SSAT or stabilization of its messenger RNA by an analogue of spermine, in particular by N ^J N ¹² - diethylnorspermine (DENSPM) (Parry L et al, 1995, Biochem J 305 451-458), either by inhibiting ODC with reversible or irreversible inhibitors such as α-DiFluoroMethylOrnithine (α-DFMO) (WO 2002/15895), or by blocking the transport of polyamines at cell membranes .

Although no particular gene has been identified as the polyamine transporter, three distinct polyamine transport systems have been demonstrated in mammals (Igarashi et al., 2010, Plant Physiology and Biochemistry, 48, 506). 512). The antizyme, a protein involved in the degradation of ODC, is capable of modulating the entry or exit of polyamines from the cell according to a mechanism that has not yet been elucidated. A mechanism of endocytosis of polyamines mediated by glypican-1 and caveolin-1 has also been reported. More recently, studies have shown the presence of a protein complex consisting of SLC3A2 and LAT proteins capable of excreting putrescine from the cell and returning arginine in exchange.

The development of inhibitors of polyamine transport is the subject of much research. Various classes of molecules have been developed, including spermine analogues (Burns MR, 2009, J Med Chem, 52, 1983-1993) or polyamine dimers (US 2005/0267220 A1), optionally linked to an anthracene nucleus ( WO 2010/148390).

Inhibitors of polyamine anabolic enzymes and inhibitors of polyamine transport play an important role in the endogenous synthesis of polyamines by the cells of the body.

The bacteria of the intestinal flora constitute the second source of endogenous synthesis of polyamines. The use of antibiotics targeting these bacteria allows a reduction of this other endogenous source of polyamines.

Spermine is the final product of the metabolism of polyamines. Its ability to convert spermidine by two different enzymatic pathways (Spermine oxidase or SSAT and polyamine oxidase) gives it an essential role in maintaining the homeostasis of intracellular polyamines. This polyamine is involved in many cellular functions. The prior art shows that spermine has rather deleterious effects on cellular metabolism.

Indeed, spermine participates in the metastatic process in prostate cancer where it has been shown that the treatment of HT-29 cells with spermine leads to a decrease in the expression of molecules involved in cell adhesion, promoting thus metastatic invasion (Tsujinaka et al., 2011, Int J Oncology, 38: 305-312).

In addition, the treatment of macrophages by spermine inhibits the cytotoxic activity of these cells, preventing them from exerting their antitumor auxiliary function (WO 2000/006546).

A recent study of 375 patients with colorectal cancer treated for three years with a combination of α-DFMO and sulindac (anti-inflammatory compound) found that patients with basal spermine / spermidine ratio less than 0.30 are more sensitive. to the chemoprotective effects of this drug combination that patients with basal spermine / spermidine greater than 0.30 (Thompson et al., 2010, Gastroenterology, 139: 797-805).

In the context of cancer treatment, increasing the survival or lifespan of patients remains a major concern. There is therefore a real need for the development of new compositions capable of participating in the treatment of pathologies related to cellular hyperproliferation, and in particular to the treatment of cancer.

Surprisingly, the inventors have demonstrated a beneficial effect of spermine on the metabolism of cancer cells which makes it possible to distinguish it from other polyamines participating in cellular homeostasis.

One of the aims of the invention is to provide compositions for their uses in the treatment of pathologies related to cellular hyperproliferation, in particular in the treatment of cancer.

Another object of the invention is to provide novel compositions spermine enriched more effective than the compositions described in the prior art in the context of the treatment of pathologies related to cellular hyperproliferation, particularly in the treatment of cancer.

Another object of the invention is to propose compositions that can be administered to patients together with an anti-cancer treatment.

(chemotherapeutic) leading to a synergistic effect in terms of cancer treatment.

The present invention relates to compositions comprising a mixture of cadaverine, putrescine and spermidine at a concentration ranging from 0.3 to 0.6 nmol per gram of composition, said composition also comprising spermine at a concentration ranging from 150 to 17000 nmol by gram of composition.

Thus, spermine represents from 99.60% to 100% of the logically active organic polyamines of the compositions of the present invention. The distribution of the logically active organic polyamines of the compositions of the present invention is very different from that of foods constituting a "medium" food ration.

Surprisingly, the inventors have shown the existence of a synergistic effect in the treatment of pathologies related to cellular hyperproliferation, based on the combination of compositions comprising on the one hand an impoverished mixture cadaverine, putrescine and spermidine and spermine at a concentration higher than that provided by a standard food diet.

The compositions of the present invention may be ingested by a patient who is a mammal. By "mammal" means a human being or an animal, said animal may be a pet or a farm animal.

The compositions of the invention have an efficacy that may depend on the stage of the anticancer treatment followed by the patient. This efficiency can be optimum when the compositions are used as the sole or the main exogenous source of polyamines. When combined with other foods, cadaverine, putrescine and spermidine intake should be as low as possible. The object of the invention is that the rate of spermine ingested by the patient is much higher than the levels of the other three polyamines. This concentration difference between the spermine and the cadaverine mixture, putrescine and spermidine must be maintained either by the compositions of the invention alone or by the compositions of the invention associated with foods containing little cadaverine, putrescine and spermidine. Since polyamine intake varies from one food to another, there is a nutritional guide for foods (wvvw.guerir.org/faagazine/nutrition-guide-nutrialys.pdf) to maintain a nutrition program with very low polyamine content. concomitantly with the setting of the compositions of the invention.

By "mixture of cadaverine, putrescine and spermidine" is meant the combination of the three natural polyamines corresponding respectively to the formulas NH ₂ - (CH ₂ ) ₅ -NH ₂ , NH ₂ - (CH ₂ ) ₄ -NH ₂ and NH ₂ - (CH ₂ ) ₃ -NH- (CH ₂ ) ₄ -NH ₂ in a composition potentially containing other excipients.

By "spermine" is meant the natural polyamine corresponding to the formula

NH ₂ - (CH ₂ ) ₃ - NH- (CH ₂ ) ₄ - NH- (CH ₂ ) ₃ -NH ₂ .

The concentrations of the various polyamines are expressed in nanomoles (nmol) per gram of composition. The terms nano moles, nmoles and nmol are equivalent and can be used interchangeably in the text of this application. By way of example, this means that in one gram of composition, the concentration of spermine can vary from 150 to 17000 nanomoles. The polyamines of the present invention have the property of being biologically "active" in that they possess at least one biological activity having an effect on:

stabilization, condensation and conformation of DNA (Thomas & Thomas, 2001, CMLS, Cell Mol Life Sci, 58: 244-258),

- the transcription of the RA,

cell growth and proliferation by intervening directly on the cell cycle of cells (Thomas & Thomas 2001),

regulation of the immune response (Soulet D & Rivest S, 2003, J Cell Biol, 162 (2): 257-268),

- modulating the functioning of N-methyl-D-aspartate (NMD A) receptors and are involved in neurodegenerative processes (Soulet & Rivest 2003).

According to a particular embodiment, the spermine concentration of the compositions of the invention varies from 150 to 1650 nmoles per gram of composition.

According to a particular embodiment, the spermine concentration of the compositions of the invention varies from 500 to 1100 nmol per gram of composition.

According to a particular embodiment, the spermine concentration of the compositions of the invention varies from 1100 to 1650 nmol per gram of composition.

According to a more particular embodiment, the spermine concentration of the compositions of the invention ranges from 1650 to 2500 nmol per gram of composition.

According to another embodiment, the concentration of spermine compositions of the invention ranges from 2500 to 5000 nmol per gram of composition.

According to yet another embodiment, the spermine concentration of the compositions of the invention varies from 5,000 to 10,000 nmoles per gram of composition.

According to yet another more particular embodiment, the spermine concentration of the compositions of the invention ranges from 100 to 17000 nmol per gram of composition.

Advantageously, the putrescine concentration of the compositions of the invention varies from 0.19 to 0.32 nmoles per gram of composition.

More preferably, the cadaverine concentration of the compositions of the invention ranges from 0.02 to 0.08 nmol per gram of composition. Even more advantageously, the spermidine concentration of the compositions of the invention ranges from 0.09 to 0.20 nmoles per gram of composition.

When the compositions of the invention constitute the sole or the main food source of the patients, these compositions can be enriched with lipids, proteins, carbohydrates, vitamins, minerals and electrolytes in a quantity allowing the patient not to suffer from malnutrition or deficiencies.

The compositions of the invention contain as a percentage of dry weight relative to the total dry weight: 10% to 35% of lipids, 8% to 30% of proteins, 35% to 80% of carbohydrates, up to 10% of a mixture of vitamins, minerals and electrolytes.

By "food source" is meant all forms of diet, that is to say all foods that may constitute the diet of a human or animal, a diet consisting of meal replacement, or any other food source that keeps the human or animal alive.

By "mixture consisting of vitamins, minerals and electrolytes" is meant vitamins and minerals having a defined role in the body. The vitamins can be selected from a group consisting of vitamin A, vitamin B1, B6, vitamin B, vitamin C, vitamin D3, vitamin K1, riboflavin, pantothenic acid, niacin, folic acid, biotin, choline, inositol. The minerals and electrolytes may be selected from a group consisting of sodium, potassium, calcium, phosphorus, magnesium, iron, zinc, copper, manganese, chlorides, iodine, selenium, chromium, molybdenum. The choice of vitamins, minerals and electrolytes should not be restricted by the lists listed above. Those skilled in the art can adapt the proportions of each of these constituents allowing the patient to receive a balanced diet, meeting his daily nutritional needs.

Polyamines in the body come from three main sources: cell proliferation (physiological and tumoral), diet and intestinal bacteria. In order to control as much as possible the supply of polyamines in the body, it may be necessary to limit not only the exogenous supply via a perfectly controlled diet but also to inhibit the endogenous synthesis of the polyamines by using inhibitors of the enzymes involved in the anabolism of polyamines and / or inhibitors of polyamine transport between the cell and the extracellular medium. "Specific inhibitor" denotes a molecule capable of blocking, totally or partially, directly or indirectly, reversibly or otherwise, the active site of at least one of the enzymes involved in the synthesis of polyamines (ornithine decarboxylase (ODC)). , spermidine-spermine Nl-acetyltransferase or spermine oxidase) in the human body, or animal. The role of the inhibitor of polyamine biosynthesis is to stop or significantly reduce the endogenous production of polyamines in the organism treated with the product according to the present invention. The joint implementation of an inhibitor of polyamine biosynthesis and dietary intake low in polyamines reduces the amount of bioavailable polyamines in the body.

The compositions of the invention optionally contain an inhibitor of the intracellular synthesis of polyamines in a proportion of at most 15% by weight relative to the total dry weight of the composition.

According to a more particular embodiment, the inhibitor of the intracellular synthesis of polyamines of the compositions of the invention is an inhibitor of ornithine decarboxylase, spermidine-spermine Nl-acetyltransferase or spermine oxidase.

Of the ODC inhibitors, alpha-difluoromethylornithine (α-DFMO) is a useful compound, well known to those skilled in the art (Fabian et al., 2002, Clin Cancer Res, 8 (10), 3105). 3,117 / Levin et al., 2003, Clin Cancer Res, 9 (3), 981-990 / Meyskens et al., 2008, Cancer Prev Res, 1 (1), 32-38). This example should in no way restrict the choice of an inhibitor of the endogenous synthesis of polyamines to this compound alone.

Other compounds capable of inhibiting ornithine decarboxylase, spermidine-spermine Nl-acetyltransferase or spermine oxidase may be used. The amounts of inhibitors will be adapted by the skilled person on the basis of the biological activity data of these compounds and his general knowledge.

The transport of the polyamines between the cell and the extracellular medium also allows a fine regulation of the intracellular content of polyamines. Joint implementation of an inhibitor of polyamine transport and dietary intake low in polyamines reduces the amount of bioavailable polyamines in the body. The compositions of the present invention are optionally enriched with at least one polyamine transport inhibitor, in a proportion of at most 15% by weight relative to the total dry weight of the composition.

The polyamine transport inhibitors of the compositions of the present invention are preferably inhibitors capable of acting on the intracellular transport, in particular of putrescine and / or spermidine, but which have no effect on the intracellular transport of spermine.

In order to further reduce the endogenous synthesis of polyamines, it may be envisaged to use antibiotics to limit the intake of polyamines by the bacteria of the intestinal flora.

According to a particular embodiment, the compositions of the invention optionally contain at least one antibiotic. This antibiotic may belong to the group of intestinal antiseptics, such as Ercefuryl ^® . This antibiotic can also, in addition to its bacteriostatic or bactericidal effect, have an antiparasitic effect, such as Flagyl ^® .

The use of antibiotics may lead to reduce the intake of certain vitamins, especially those provided by the intestinal flora of the patient. In this case, it may be necessary to supplement the composition in these vitamins so as not to cause vitamin deficiencies in the patient in case of prolonged administration of the composition.

By "deficiencies" is meant a lack of nutrients that can alter the physical or mental condition of a human or an animal.

According to a particular embodiment, the compositions of the invention may be enriched with vitamins.

Advantageously, the carbohydrates of the compositions of the invention belong to the group comprising glucose polymers, maltodextrins, sucrose, modified starches, glucose monohydrate, dehydrated glucose syrup, glycerol monostearate and mixtures thereof.

Advantageously, the proteins of the compositions of the invention belong to the group comprising soluble milk proteins, soy proteins, serum peptides, egg white powder, potassium caseinate, non-phosphorylated peptides, casein peptides, mixed caseinate, soy isolate and mixtures thereof. Advantageously, the lipids of the compositions of the invention belong to the group comprising butter oil, peanut oil, medium chain triglycerides, grape seed oil, soy evening primrose oil and mixtures thereof.

According to one particular embodiment, the lipids of the compositions of the invention consist of a mixture of at least one oil of animal origin, at least one vegetable oil and glycerol stearate.

In order to control the supply of exogenous sources of polyamines, the compositions used according to the present invention must be able to constitute all or part of the patient's diet. In this sense they must provide an energy ration that can meet the nutritional needs of the patient.

When the patient is a human, the compositions of the invention constitute the daily food ration of a human being and comprise:

- from 75 g to 500 g of carbohydrates,

from 20 g to 185 g of lipids,

from 20 g to 225 g of proteins,

vitamins, minerals and electrolytes in sufficient quantities to meet the daily nutritional needs of a human being,

and optionally an inhibitor of the intracellular synthesis of polyamines at a rate of less than 50 g and preferably at a rate of 0.3 to 10 g per day.

According to a particular embodiment, the compositions of the invention are a submultiple of a daily food ration of a human being and comprise:

from 75 μg to 500 μg of carbohydrates,

from 20 μg to 185 μg of lipids,

from 20 μg to 225 μg of protein,

vitamins, minerals and electrolytes in sufficient quantities to partially meet the daily nutritional requirements of a human being,

and optionally an inhibitor of the intracellular synthesis of polyamines in a proportion of less than 50 μg and preferably in a proportion of 1 / x to 10 μg per day,

and X being an integer from 2 to 8 and corresponding to the number of rations to be ingested by the patient to meet his daily nutritional requirements. When the patient is an animal, the daily food ration is adapted according to the category and mass of the animal, whether it is a pet or a farm animal. The distribution of carbohydrates, lipids and proteins as well as the vitamin, mineral and electrolyte requirements of the daily food ration of an animal are well known to those skilled in the art. Concerning the intracellular synthesis inhibitor of polyamines, the dose is adapted according to the mass of the animal and possibly on the basis of data obtained in humans.

The compositions used according to the present invention may constitute the daily food ration or a sub-multiple of a daily dietary ration of an animal and will have to satisfy the daily nutritional requirements of an animal.

By way of example, when the patient is a mouse, the compositions used according to the present invention constitute the daily dietary ration of a mouse and comprise:

- from 0.6 g to 1.8 g of carbohydrates,

from 0.04 g to 1.2 g of lipids,

from 0.01 g to 0.6 g of proteins,

vitamins, minerals and electrolytes in sufficient quantities to meet the daily nutritional requirements of an animal,

and optionally an inhibitor of the intracellular synthesis of polyamines at a rate of less than 300 mg and preferably at a rate of 40 to 200 mg per day.

As for humans, the compositions used according to the present invention may be a submultiple of a daily food ration of a mouse and include:

from 0.6 / X g to 1.8 μg of carbohydrates,

from 0.04 μg to 1.2 μg of lipids,

from 0.01 / X g to 0.6 X g of proteins,

vitamins, minerals and electrolytes in sufficient quantities to partially meet the daily nutritional requirements of an animal, and optionally an inhibitor of the intracellular synthesis of polyamines in a proportion of less than 300 / X mg and preferably in a proportion of 40 / X to 200 / X mg per day,

and X being an integer from 2 to 8 and corresponding to the number of rations to be ingested by the patient to meet his daily nutritional requirements.

The proportions of the constituents of the compositions of the present invention given in the context of feeding a mouse, are given as an indication and may serve as a basis for the skilled person who can adapt them, thanks to his general knowledge, to other animals.

The compositions of the invention may be in solid, liquid or semi-liquid form. Their formulation is adapted according to the ability of the patient to absorb an oral diet but also enterally or parenterally.

According to a particular embodiment, the compositions of the invention are in a dry form to dissolve extemporaneously in a neutral vehicle.

According to a more particular embodiment, the compositions of the invention include a neutral vehicle making them ready for use.

By "neutral vehicle" is meant an aqueous solution for producing a more or less liquid composition, facilitating the ingestion of the latter by the patient. Thus, the compositions of the invention will have a viscosity range extending from that of water to that of milk drinks for a temperature of 4 ° C to 40 ° C, at normal atmospheric pressure.

The compositions of the invention can be used directly as a medicament.

By "drug" is meant any substance that when ingested by the patient, provides a benefit in terms of welfare, improvement of biomarkers, regression or remission of pathology.

The compositions of the invention may also be included in the formulation of pharmaceutical preparations.

The compositions of the invention can be used in the preparation of a medicament.

The compositions of the invention may be used as a supplement or a food substitute. The compositions of the invention may be used as a nutraceutical supplement.

The metabolism of polyamines plays a key role in cell proliferation. Knowing that a disturbance in the physiology of cell division is almost always harmful, it seems appropriate to analyze the effects of a disruption of the metabolism of polyamines in the context of pathologies involving processes of cellular hyperproliferation, and in particular as part of cancer treatment.

The compositions of the invention can be used in the treatment, in a patient, of pathologies related to cellular hyperproliferation.

The compositions of the invention can be used in the treatment, in a patient, of cancer.

The compositions of the invention may be used in the treatment of pathologies related to cellular hyperproliferation or in the treatment of cancer, said treatment being carried out by administering a unit dose ranging from 6.7 mg to 670 mg of spermine.

In order to control as much as possible the exogenous supply of polyamines, the compositions of the invention may serve as a basis for a diet which may comprise several phases during which the exogenous supply of polyamines is:

- totally provided by the compositions used according to the invention,

- mainly provided by the compositions used according to the invention,

partially provided by the compositions used according to the invention.

By "totally" is meant that the patient's diet is restricted to the compositions of the invention. No food other than the compositions of the invention enter the diet of the patient. During this phase, the control of the polyamine intake is maximal.

By "majority", it is meant the possibility of introducing into the patient's diet a breakfast comprising foods reduced in polyamine content. The rest of the daily food ration is provided by the compositions of the invention.

"Partially" means the possibility of introducing into the patient's diet a breakfast and at least one solid meal including reduced polyamine foods. The rest of the daily food ration is provided by the compositions of the invention.

According to a particular embodiment, the compositions used according to the present invention are administered to the patient according to the following scheme:

(i) administering a first dose of the composition of the invention during a first period of time, and consecutively,

(ii) administering a second dose of the composition of the invention for a second period of time, the second dose is adjusted according to the reaction of the patient to the first, and consecutively,

(iii) administering a third dose of the composition of the invention for a third period of time, the third dose is adjusted according to the reaction of the patient to the second.

By "patient reaction" is meant its physiological ability to benefit from a depleted polyamine diet. The benefit can be appreciated especially from the regression of the tumor mass. The improvement or disappearance of any sign of the disease can be evaluated by a clinical examination, biological examinations or imaging performed in a usual way in the context of cancer. These criteria vary depending on the type of cancer. Three hypotheses are possible:

or the physiological state of the patient improves after administration of a first dose of the composition and in this case, the second dose of the composition will contain at most the same level of polyamines as the first dose,

or the physiological state of the patient shows no improvement after administration of a first dose of the composition and in this case, the second dose of the composition will contain at least the same level of polyamines as the first dose, or Physiological condition of the patient has degraded after administration of a first dose of the composition and in this case, the second dose of the composition will contain a polyamine level either higher or lower than that of the first dose.

The same type of reasoning applies for the administration of the third dose. Thus, the practitioner has a large latitude in the administration scheme of the compositions of the invention.

When the three doses are identical, it amounts to administering a single dose to the patient for a period determined by the practitioner. According to a particular embodiment of the invention, the first period of time varies from 7 to 14 days, in particular 7 days.

According to a particular embodiment of the invention, the second period of time varies from 14 to 21 days, in particular 14 days.

According to a particular embodiment of the invention, the third period of time varies from 28 to 63 days, in particular 63 days.

The first, second, and third time periods constitute a complete cycle of patient treatment. The administration of the compositions of the invention may take place for one, two or more cycles. The renewal of these cycles is left to the discretion of the practitioner.

According to another particular embodiment of the invention:

the first dose of polyamines varies from 6.7 mg to 5.3 g of spermine / day,

the second dose of polyamines varies from 6.7 mg to 5.3 g of spermine / day, and

the third dose of polyamines ranges from 6.7 mg to 5.3 g of spermine / day.

These doses correspond to the doses of polyamines provided by 1 to 8 rations of a diet depleted of cadaverine, putrescine and spermidine and spermine-enriched dose 33.4 mg / kg of food at the dose 3340 mg / kg of food .

The compositions of the present invention provide at least the same daily amount of spermine and up to 800 times more spermine than an average food ration.

According to a particular embodiment, the present invention also relates to compositions comprising:

a mixture of cadaverine, putrescine and spermidine at a concentration varying from 0.3 to 0.6 nmoles per gram of composition,

spermine at a concentration ranging from 150 to 17000 nmol per gram of composition,

for their use in the treatment in a patient of pathologies related to cellular hyperproliferation, the total amount of logically active organic polyamines ingested per day by the patient does not exceed 11334 nanomoles per kcal of ingested composition, in particular 5000 nanomoles per kcal of composition ingested, in particular 1000 nanomoles per kcal of ingested composition, in particular 100 nanomoles per kcal of ingested composition. In order to potentiate the effects of a first anti-cancer treatment, the compositions of the invention may be used as a second therapeutic agent.

The compositions of the invention may be administered to the patient before and / or during and / or after the anti-cancer treatment.

According to a particular embodiment, the present invention also relates to a combination of a composition comprising:

and a chemotherapeutic agent for its simultaneous, separate or successive use in the treatment of pathologies related to cell hyperproliferation, the total amount of bio logically active polyamines ingested per day by the patient not exceeding 11334 nanomoles per kcal of ingested composition , in particular 5000 nanomoles per kcal of ingested composition, in particular 1000 nanomoles per kcal of ingested composition, in particular 100 nanomoles per kcal of ingested composition.

Description of figures

Figure 1: Evolution of tumor volume

This figure represents the variations in tumor volume (in cm ³ , indicated on the ordinate) as a function of the number of days (indicated on the abscissa) elapsed after the grafting of said tumor in male C57BL / 6 mice. The mice are grafted with a solid tumor of Lewis lung carcinoma cells.

Figure 1-A: Four groups of five mice are fed with different diets 6 days after the transplant took place. The start of treatment is indicated by the arrow on the graph.

The curve marked by black circles represents the mice fed a diet containing a normal level of polyamines (125 mg / kg of food or 861 nmol / g of food). The curve represented by empty circles represents the mice fed a diet containing a normal level of polyamines (125 mg / kg of food or 861 nmol / g of food), and a drinking water containing neomycin (2 mg / ml).

The curve represented by black squares represents the mice fed a diet depleted of cadaverine, putrescine and spermidine (51 μg / kg of food or 0.350 nmol / g of food).

The curve represented by empty squares represents the mice fed a diet depleted of cadaverine, putrescine and spermidine (51 μg / kg of food or 0.350 nmol / g of food), and a drinking water containing neomycin (2 mg / ml).

Figure 1-B: Six groups of five mice are fed with different diets 6 days after the transplant took place. The start of treatment is indicated by the arrow on the graph.

The curve marked by black circles represents the mice fed a diet depleted of cadaverine, putrescine and spermidine (51 μg / kg of food or 0.350 nmol / g of food), and enriched in spermine (33.4 mg / kg of food (165 nmole / g of food).

The curve represented by empty circles represents the mice fed with a diet depleted of cadaverine, putrescine and spermidine (51 μg / kg of food or 0.350 nmol / g of food), enriched in spermine (33.4 mg / kg d feed = 165 nmol / g food) and drinking water containing neomycin (2 mg / ml).

The curve represented by black squares represents the mice fed a diet depleted of cadaverine, putrescine and spermidine (51 μg / kg of food or 0.350 nmol / g of food) and enriched in spermine (334 mg / kg of food 1650 nmol / g of food).

The curve represented by empty squares represents the mice fed with a diet depleted of cadaverine, putrescine and spermidine (51 μg / kg of food or 0.350 nmol / g of food), enriched in spermine (334 mg / kg of food 1650 nmol / g of food) and drinking water containing neomycin (2 mg / ml).

The curve represented by black diamonds represents the mice fed with a diet depleted of cadaverine, putrescine and spermidine (51 μg / kg of food 0.350 nmol / g of food) and enriched with spermine (3340 mg / kg of food or 16500 nmol / g of food).

The curve represented by empty squares represents the mice fed a diet depleted of cadaverine, putrescine and spermidine (51 μg / kg of food or 0.350 nmol / g of food), enriched in spermine (3340 mg / kg of food 16500 nmol / g of food) and drinking water containing neomycin (2 mg / ml).

Figure 2: Evolution of pulmonary metastatic spread

This figure represents the percentage of pulmonary metastatic invasion (on the ordinate) of male C57BL / 6 mice, as a function of the diet (on the abscissa) given to the mice that had received a solid tumor transplant of Lewis carcinoma cells. The percentage of pulmonary metastatic invasion is measured 19 days after the transplant took place.

Ten groups of mice are fed different diets. The first group (represented by a black column) is fed with a diet containing a normal level of polyamines (125 mg / kg of food or 861 nmole / g of food). The average value of pulmonary metastatic invasion of mice in this group is 27%.

The second group (represented by a white column) is fed a diet containing a normal level of polyamines (125 mg / kg of food or 861 nmol / g of food), and a drinking water containing neomycin (2 mg / ml). The average value of pulmonary metastatic invasion of the mice of this group is 11%.

The third group (represented by a column containing discontinuous vertical lines) is fed with a diet depleted of cadaverine, putrescine and spermidine (51 μg / kg of food or 0.350 nmol / g of food). The average value of pulmonary metastatic invasion of the mice of this group is 6%.

The fourth group (represented by a column containing horizontal discontinuous lines) is fed a diet depleted of cadaverine, putrescine and spermidine (51 μg / kg feed or 0.350 nmole / g feed), and a drinking water containing neomycin (2 mg / ml). The average value of pulmonary metastatic invasion of the mice of this group is 7%. The fifth group (represented by a column containing vertical continuous lines) is fed with a diet depleted of cadaverine, putrescine and spermidine (51 μg / kg of food or 0.350 nmol / g of food), and enriched with spermine (33 , 4 mg / kg of food or 165 nmol / g of food). The average value of pulmonary metastatic invasion of the mice of this group is 13%.

The sixth group (represented by a column containing horizontal continuous lines) is fed with a diet depleted of cadaverine, putrescine and spermidine (51 μg / kg of food or 0.350 nmol / g of food), enriched in spermine (33, 4 mg / kg of food (165 nmole / g of food) and drinking water containing neomycin (2 mg / ml). The average value of pulmonary metastatic invasion of the mice of this group is 4%.

The seventh group (represented by a column containing continuous lines pointing to the left) is fed with a diet depleted of cadaverine, putrescine and spermidine (51 μg / kg of food or 0.350 nmol / g of food), and enriched in spermine (334 mg / kg of food or 1650 nmol / g of food). The average value of pulmonary metastatic invasion of the mice of this group is 7%.

The eighth group (represented by a column containing continuous lines pointing to the right) is fed with a diet depleted of cadaverine, putrescine and spermidine (51 μg / kg of food or 0.350 nmol / g of food), enriched in spermine ( 334 mg / kg of food (1650 nmol / g of food) and drinking water containing neomycin (2 mg / ml). The average value of pulmonary metastatic invasion of the mice of this group is 7%.

The ninth group (represented by a column containing small dots) is fed with a diet depleted of cadaverine, putrescine and spermidine (51 μg / kg of food or 0.350 nmol / g of food), and enriched in spermine (3340 mg / kg of food is 16500 nmol / g of food). The average value of pulmonary metastatic invasion of the mice of this group is 5%.

The tenth group (represented by a column containing large dots) is fed with a diet depleted of cadaverine, putrescine and spermidine (51 μg / kg of food or 0.350 nmol / g of food), enriched in spermine (3340 mg / kg of food (16500 nmol / g of food) and drinking water containing neomycin (2 mg / ml). The average value of pulmonary metastatic invasion of the mice of this group is 1%. Figure 3: Evolution of tumor levels of polyamines

This figure represents the tumor levels of putrescine, spermidine and spermine (on the ordinate) of male C57BL / 6 mice, as a function of the diet (on the abscissa) given to the mice which have received the transplant of a solid tumor of Lewis carcinoma cells. . The tumor levels of putrescine, spermidine and spermine are measured 19 days after the graft has taken place.

Figure 3-A: Four groups of five mice are fed different diets 6 days after the transplant took place.

The black column represents the mice fed a diet containing a normal level of polyamines (125 mg / kg of food or 861 nmol / g of food).

The white column represents the mice fed a diet containing a normal level of polyamines (125 mg / kg of food or 861 nmol / g of food), and a drinking water containing neomycin (2 mg / ml).

The column containing vertical discontinuous lines represents the mice fed a diet depleted of cadaverine, putrescine and spermidine (51 μg / kg of food or 0.350 nmol / g of food).

The column containing horizontal discontinuous lines represents mice fed a diet depleted of cadaverine, putrescine and spermidine (51 μg / kg of food or 0.350 nmol / g of food), and a drinking water containing neomycin (2 mg / ml).

Figure 3-B: Six groups of five mice are fed with different diets 6 days after the transplant took place.

The column containing vertical continuous lines represents the mice fed a diet depleted of cadaverine, putrescine and spermidine (51 μg / kg of food or 0.350 nmol / g of food), and enriched in spermine (33.4 mg / kg of food (165 nmole / g of food).

The column containing horizontal continuous lines represents the mice fed with a diet depleted of cadaverine, putrescine and spermidine (51 μg / kg of food or 0.350 nmol / g of food), enriched in spermine (33.4 mg / kg d 'food 165 nmole / g of food) and drinking water containing neomycin (2 mg / ml).

The column containing continuous lines pointing to the left represents the mice fed with a diet depleted of cadaverine, putrescine and spermidine (51 μg / kg of food or 0.350 nmol / g of food) and enriched in spermine (334 mg / kg of food is 1650 nmol / g of food).

The column containing straight lines pointing to the right represents the mice fed with a diet depleted of cadaverine, putrescine and spermidine (51 μg / kg of food or 0.350 nmol / g of food), enriched in spermine (334 mg / kg d feed (1650 nmol / g food) and drinking water containing neomycin (2 mg / ml).

The column containing small dots represents the mice fed with a diet depleted of cadaverine, putrescine and spermidine (51 μg / kg of food or 0.350 nmol / g of food) and enriched in spermine (3340 mg / kg of food either 16500 nmol / g of food).

The column containing large dots represents the mice fed a diet depleted of cadaverine, putrescine and spermidine (51 μg / kg of food or 0.350 nmol / g of food), enriched in spermine (3340 mg / kg of food or 16500 nmol / g food) and drinking water containing neomycin (2 mg / ml).

Figure 4: Evolution of the mass of the spleen

This figure represents the mass of the spleen (ordinate) of male C57BL / 6 mice, as a function of the diet (in abscissas) given to the mice which have received or not the transplant of a solid tumor of Lewis carcinoma cells. The mass of the spleen is measured 19 days after the transplant took place.

Ten groups of mice transplanted with a solid tumor of Lewis carcinoma cells are fed different diets. An eleventh group of mice not receiving a graft and placed under a diet containing a normal level of polyamines makes it possible to establish an average reference value of the splenic mass in the mice used in this study.

The first group (represented by a black column) is fed with a diet containing a normal level of polyamines (125 mg / kg of food or 861 nmol / g of food). The average value of the splenic mass of mice in this group is 171 mg.

The second group (represented by a white column) is fed a diet containing a normal level of polyamines (125 mg / kg of food or 861 nmol / g of food), and a drinking water containing neomycin (2 mg / ml). The mean value of the splenic mass of mice in this group is 151 mg.

The third group (represented by a column containing discontinuous vertical lines) is fed with a diet depleted of cadaverine, putrescine and spermidine (51 μg / kg of food or 0.350 nmol / g of food). The mean value of the splenic mass of mice in this group is 88 mg.

The fourth group (represented by a column containing horizontal discontinuous lines) is fed a diet depleted of cadaverine, putrescine and spermidine (51 μg / kg feed or 0.350 nmole / g feed), and a drinking water containing neomycin (2 mg / ml). The mean value of the splenic mass of mice in this group is 63 mg.

The fifth group (represented by a column containing vertical continuous lines) is fed with a diet depleted of cadaverine, putrescine and spermidine (51 μg / kg of food or 0.350 nmol / g of food), and enriched with spermine (33 , 4 mg / kg of food or 165 nmol / g of food). The mean value of the splenic mass of mice in this group is 93 mg.

The sixth group (represented by a column containing horizontal continuous lines) is fed with a diet depleted of cadaverine, putrescine and spermidine (51 μg / kg of food or 0.350 nmol / g of food), enriched in spermine (33, 4 mg / kg of food (165 nmole / g of food) and drinking water containing neomycin (2 mg / ml). The mean value of the splenic mass of mice in this group is 75 mg.

The seventh group (represented by a column containing continuous lines pointing to the left) is fed with a diet depleted of cadaverine, putrescine and spermidine (51 μg / kg of food or 0.350 nmol / g of food), and enriched in spermine (334 mg / kg of food or 1650 nmol / g of food). The mean value of the splenic mass of mice in this group is 98 mg.

The eighth group (represented by a column containing straight lines pointing to the right) is fed with a diet depleted of cadaverine, putrescine and spermidine (51 μg / kg diet or 0.350 nmol / g feed), spermine-enriched (334 mg / kg diet or 1650 nmol / g food) and drinking water containing neomycin (2 mg / ml). The average value of the splenic mass of mice in this group is 100 mg.

The ninth group (represented by a column containing small dots) is fed with a diet depleted of cadaverine, putrescine and spermidine (51 μg / kg of food or 0.350 nmol / g of food), and enriched in spermine (3340 mg / kg of food is 16500 nmol / g of food). The mean value of the splenic mass of mice in this group is 81 mg.

The tenth group (represented by a column containing large dots) is fed with a diet depleted of cadaverine, putrescine and spermidine (51 μg / kg of food or 0.350 nmol / g of food), enriched in spermine (3340 mg / kg of food (16500 nmol / g of food) and drinking water containing neomycin (2 mg / ml). The mean value of the splenic mass of mice in this group is 42 mg.

The eleventh group (represented by a gray column), corresponding to the mice not receiving a graft, is fed with a diet containing a normal level of polyamines (125 mg / kg of food or 861 nmol / g of food) . The mean value of the splenic mass of mice in this group is 61 mg. Examples

Example 1 Effect of Diets Depleted on Cadaverine, Putrescine and Spermidine and Spermine-enriched 1 - Animal Model

1.1 - Murine strain

Male C57BL / 6 mice, 9 weeks old and weighing 20g (January breeding, Genest St Isle, France) are acclimated 1 week before the start of the study. Animal handling is carried out in accordance with the ethical guidelines for experimentation and according to the recommendations of the European Association for Biomedical Research. The animals are enclosed at 5 individuals per cage under standardized conditions (21 ± 1 ° C, 60% relative humidity, cycles of 12 hours of light, 12 hours of darkness), and have access to food and feed. water ad libitum. 1.2 - Tumor model

Lewis lung carcinoma cells (3LL) were obtained from the ECACC (European Collection of Cell Cultures) and grafted by intramuscular injection into the hind paws of male C57BL / 6 mice according to the method previously described (Hergueux J. et al. al., Cell BioL Exp., 1983, 51 (4), 181-191) to produce tumor cells. After 20 days, the tumor cells are collected and dispersed in PBS, counted and diluted to reach 0.5 × 10 ⁶ cells / 100 μl, before being grafted into the right hind paw of the mice. The grafted mice are randomly distributed in the cages in groups of 5 individuals.

The tumor becomes palpable a few days after the transplant has taken place. The animals are sacrificed 19 days after the tumor cell transplant.

2 - Food

The depleted diet of cadaverine, putrescine and spermidine is marketed under the brand CASTASE ™ by Nutrialys (Saint-Grégoire, France). This polyamine-depleted food contains less than 51 μg of polyamines / kg of food.

The rodent feed containing a normal level of polyamines (125 mg of polyamines per kilogram of food or 861 nmol of polyamines / g of feed) corresponds to a standard commercial feed (UAR, standard feed mill).

The products are distributed to the animals according to their profile and their nutritional needs. A mouse consumes about 2 grams per day of feeding.

The treatment is set up 6 days after the transplant, when the tumor is palpable and is administered 7 days out of 7 and includes the ad libitum intake of the normal diet or test solutions as well as the drinking water.

3 - Results

3.1 - Tumor growth The animal model (C57BL / 6 mouse) and the tumor model (Lewis carcinoma) are described previously. The mice are grafted with 0.5 × 10 ⁶ tumor cells by intramuscular injection into one of the hind limbs.

The growth of the tumor is quantified at 6, 8, 11, 14 and 18 days after the graft has taken place by measuring the volume of the tumor, according to the method previously described (Moulinoux et al., Int J. Cancer, 1984, 34 (2), 277-281).

The mice are divided into ten groups and different diets are administered to each group.

The first group is fed a diet containing a normal level of polyamines (125 mg / kg of food or 861 nmol / g of food).

The second group is fed a diet containing a normal level of polyamines and a drinking water containing neomycin (2 mg / ml).

The third group is fed a diet depleted of cadaverine, putrescine and spermidine (51 μg / kg of food or 0.350 nmol / g of food).

The fourth group is fed a diet depleted of cadaverine, putrescine and spermidine, and a drinking water containing neomycin (2 mg / ml).

The fifth group is fed a diet depleted of cadaverine, putrescine and spermidine (51 μg / kg of food or 0.350 nmol / g of food), and enriched in spermine (33.4 mg / kg of food or 165 nmol / g of food).

The sixth group is fed a diet depleted of cadaverine, putrescine and spermidine (51 μg / kg of food or 0.350 nmol / g of food), enriched in spermine (33.4 mg / kg of food or 165 nmol / g of food) and drinking water containing neomycin (2 mg / ml).

The seventh group is fed a diet depleted of cadaverine, putrescine and spermidine (51 μg / kg of food or 0.350 nmol / g of food), and enriched in spermine (334 mg / kg of food or 1650 nmol / g feed).

The eighth group is fed a diet depleted of cadaverine, putrescine and spermidine (51 μg / kg of food or 0.350 nmol / g of food), enriched in spermine (334 mg / kg of food or 1650 nmol / g of food) and drinking water containing neomycin (2 mg / ml).

The ninth group is fed a diet depleted of cadaverine, putrescine and spermidine (51 μg / kg of food or 0.350 nmol / g of food), and enriched in spermine (3340 mg / kg of food or 16500 nmol / g feed). The tenth group is fed a diet depleted of cadaverine, putrescine and spermidine (51 μg / kg of food or 0.350 nmol / g of food), enriched in spermine (3340 mg / kg of food or 16500 nmol / g of food) and drinking water containing neomycin (2 mg / ml).

Neomycin is diluted in drinking water (0.2% w / v) whether the animals are fed with a diet in a solid form of kibble, or with a diet in a liquid or semi-liquid form.

The evolution of tumor volume (in cm ³ ) is shown in Figures 1-A for animals of groups 1 to 4 and in Figure 1-B for animals of groups 5 to 10.

Neomycin has no effect on the evolution of tumor volume when administered in drinking water on the basis of a diet containing a normal level of polyamines.

The depleted diet of cadaverine, putrescine and spermidine causes a 20% decrease in tumor growth on day 19. This decrease is 35% when neomycin is added to this depleted cadaverine, putrescine and spermidine regimen (Figure 1-A).

Spermine supplementation of the cadaverine depleted diet, putrescine and spermidine increases dose-dependently the antitumor effect of this diet by 7%, 17% and 37%, respectively, for spermine doses of 33.4, 334 and 3340 mg / kg of food (Figure 1-B).

3.2 - Pulmonary metastatic invasion

Immediately after the sacrifice of the animals, the lungs are removed, preserved in 10% formalin and lung metastases are visualized using a binocular loupe. Metastatic spread is expressed as a percentage of invasion relative to the total area of the lungs. The percentage of metastatic invasion was measured in the mice of the ten groups previously described, each receiving a distinct diet. The results are shown in Figure 2.

Neomycin supplemented with a diet containing a normal level of polyamines causes a 60% decrease in metastatic invasion compared to that seen in mice that did not receive this antibiotic. The cadaverine depleted diet, putrescine and spermidine caused a 77% decrease in metastatic invasion compared to that observed in mice fed a diet containing a normal level of polyamines. The addition of neomycin to a diet depleted of cadaverine, putrescine and spermidine has no additional benefit.

Spermine supplementation of the depleted cadaverine, putrescine and spermidine regimen does not seem to provide any benefit in terms of the metastatic process. Conversely, for a spermine dose of 33.4 mg / kg of feed, the addition of spermine contributes to an expansion of the metastatic process. This effect disappears for doses of spermine greater than 33.4 mg / kg of food. Pulmonary metastatic invasion is almost nil when neomycin is combined with the depleted cadaverine, putrescine and spermidine regimen and enriched with 3340 mg / kg of composition.

3.3 - Tumor levels of polyamines

The level of polyamines is measured in the tumor formed at the site of inoculation of Lewis carcinoma cells. The tumor was washed and the cells that compose it are dispersed using a Polytron ^® homogenizer in the presence of 0.2 M perchloric acid. After incubation for at least 16 h at 4 ° C, the cells are centrifuged for 15 minutes at 3500 rpm at 4 ° C. The supernatant which contains the polyamines is taken and used for the determination of the polyamines. The polyamines are assayed according to the method previously described (Seiler N et al., 1996, Cancer Research, 56, 5624-5630). The three main polyamines present in mammals (putrescine, spermidine and spermine) were measured for the ten groups of mice each receiving a distinct diet. The results are shown in Figure 3.

Neomycin causes a 17% decrease in putrescine, a 7% increase in spermine level and has no effect on spermidine levels based on a normal diet. In contrast, neomycin causes a 17% increase in putrescine level, a 24% increase in spermidine level and a 5% decrease in spermine levels based on a depleted cadaverine, putrescine and spermidine regimen. .

The depleted cadaverine, putrescine and spermidine regimen induced an increase of 17% and 24%, respectively, in tumor levels of putrescine and spermidine. This diet has no effect on the tumor rate of spermine. Spermine supplementation of the cadaverine depleted diet, putrescine and spermidine induces variations in tumor levels of putrescine, spermidine and spermine.

At the dose of 33.4 mg of spermine / kg of food (ie 165 nmol / g of food), the tumoral rate of putrescine increases by 17%, the tumoral rate of spermidine increases by 25% and the tumoral rate of spermine decreased by 6% compared to the tumor levels of these two polyamines in mice fed a diet containing a normal level of polyamines.

At a dose of 334 mg spermine / kg feed (ie 1650 nmol / g food), the tumor rate of spermidine increases by 32% and the tumoral rate of spermine increases by 12%) relative to the tumor levels of these substances. two polyamines in mice fed a diet containing a normal level of polyamines. The tumoral rate of putrescine remains unchanged.

At a dose of 3340 mg spermine / kg feed (ie 16500 nmol / g feed), the tumoral rate of putrescine decreases by 50%>, the tumoral rate of spermidine decreases by 19% and the tumoral rate of spermine increases. 25% relative to the tumor levels of these two polyamines in mice fed a diet containing a normal level of polyamines. 3.4 - Splenic mass

After euthanizing the animals, the spleen is taken from the mice and weighed using a precision balance.

In addition to the ten groups of mice each having a distinct diet, as previously described, an eleventh group of animals was included for this study. These are male C57BL / 6 mice with the same basic physiological characteristics as the mice in the other 10 groups but who did not undergo a Lewis carcinoma cell transplant. The mice of this eleventh group were fed with the diet containing a normal level of polyamines. The splenic mass of these healthy mice (61 mg) serves as a reference for an animal with no sign of inflammation.

The mice which have been transplanted with Lewis carcinoma cells and placed under a diet containing a normal level of polyamines have a splenic mass. three times higher (171 mg) than healthy mice fed the same diet.

Neomycin causes a decrease in splenic mass irrespective of the nature of the diet ingested by the mice.

The cadaverine depleted diet, putrescine and spermidine induced a decrease of about 50% of the splenic mass in these mice compared to the transplanted mice that received a diet containing a normal level of polyamines.

Up to a concentration of 334 mg / kg of feed (ie 1650 nmol / g feed), spermine enrichment of the cadaverine depleted diet, putrescine and spermidine has no additional effect in terms of decreased splenic mass and therefore inflammation in these animals carrying a tumor.

Neomycin combined with an enrichment of the cadaverine depleted diet, putrescine and spermidine by 3340 mg spermine / kg feed (ie 16500 nmol / g feed) leads to a decrease in splenic mass below that of healthy animals.

EXAMPLE 2 Effect of Diets Depleted on Cadaverine, Putrescine and Spermidine and Spermine-Enriched in Combination with Anticancer Drug 1 - Animal Model

1.1 - Murine strain

Male C57BL / 6 mice, 9 weeks old and weighing 20g (January breeding, Genest St Isle, France) are acclimated 1 week before the start of the study. Animal handling is carried out in accordance with the ethical guidelines for experimentation and according to the recommendations of the European Association for Biomedical Research.

The animals are enclosed at 5 individuals per cage under standardized conditions (21 ± 1 ° C, 60% relative humidity, cycles of 12 hours of light, 12 hours of darkness), and have access to food and feed. water ad libitum.

1.2 - Tumor model

Lewis lung carcinoma cells (3LL) were obtained from ECACC (European Collection of Cell Cultures) and grafted by injection intramuscularly in the hind legs of male C57BL / 6 mice according to the method previously described (Hergueux J. et al., Exp Cell BioL, 1983, 51 (4), 181-191) to produce tumor cells. After 20 days, the tumor cells are collected and dispersed in PBS, counted and diluted to reach 0.5 × 10 ⁶ cells / 100 μl, before being grafted into the right hind paw of the mice. The grafted mice are randomly distributed in the cages in groups of 5 individuals.

The tumor becomes palpable a few days after the transplant has taken place. The animals are sacrificed 19 days after the tumor cell transplant. 2 - Food

3 - Results

3.1 - Tumor growth

The animal model (C57BL / 6 mouse) and the tumor model (Lewis carcinoma) are described previously. The mice are grafted with 0.5 × 10 ⁶ tumor cells by intramuscular injection into one of the hind limbs.

The mice are divided into fifteen groups and different diets are administered to each group. The first group is fed a diet containing a normal level of polyamines (125 mg / kg of food or 861 nmol / g of food).

The second group was fed a diet containing normal levels of polyamines and cyclophosphamide dose of 9 mg.kg ^"1. ^{Week" 1.}

The third group is fed a diet containing a normal level of polyamines and a dose of cyclophosphamide of 90 mg.kg- ¹ week ^-1 .

The fourth group is fed a diet depleted of cadaverine, putrescine and spermidine (51 μg / kg of food or 0.350 nmol / g of food).

The fifth group is fed a diet depleted of cadaverine, putrescine and spermidine (51 μg / kg of food or 0.350 nmol / g of food) and a dose of cyclophosphamide 9 mg.kg ^"1 .week ^{" 1} .

The sixth group is fed a diet depleted of cadaverine, putrescine and spermidine (51 μg / kg of food or 0.350 nmol / g of food) and a cyclophosphamide dose of 90 mg.kg ^"1 .week ^{" 1} .

The seventh group is fed a diet depleted of cadaverine, putrescine and spermidine (51 μg / kg of food or 0.350 nmol / g of food), and enriched in spermine (33.4 mg / kg of food or 165 nmol / g of food).

The eighth group is fed a diet depleted of cadaverine, putrescine and spermidine (51 μg / kg of food or 0.350 nmol / g of food), enriched in spermine (334 mg / kg of food or 1650 nmol / g of 'food).

The ninth group is fed a diet depleted of cadaverine, putrescine and spermidine (51 μg / kg of food or 0.350 nmol / g of food), and enriched in spermine (3340 mg / kg of food or 16500 nmol / g feed).

The tenth group is fed a diet depleted of cadaverine, putrescine and spermidine (51 μg / kg of food or 0.350 nmol / g of food), enriched in spermine (33.4 mg / kg of food or 165 nmol / g of food) and a dose of cyclophosphamide of 9 mg.kg- ¹ week ^-1 .

The eleventh group is fed a diet depleted of cadaverine, putrescine and spermidine (51 μg / kg of food or 0.350 nmol / g of food), enriched in spermine (334 mg / kg of food or 1650 nmol / g of food) and a dose of cyclophosphamide 9 mg.kg ^"1. ^{week" 1.}

The twelfth group is fed a diet depleted of cadaverine, putrescine and spermidine (51 μg / kg of food or 0.350 nmol / g of food), enriched in spermine (3340 mg / kg of feed is 16500 nmol / g of food) and a dose of cyclophosphamide 9 mg.kg ^"1. ^{week" 1.}

The thirteenth group is fed with a diet depleted of cadaverine, putrescine and spermidine (51 μg / kg of food or 0.350 nmol / g of food), enriched in spermine (33.4 mg / kg of food or 165 nmol / g of food) and cyclophosphamide dose of 90 mg.kg ^"1. ^{week" 1.}

The fourth group is fed a diet depleted of cadaverine, putrescine and spermidine (51 μg / kg of food or 0.350 nmol / g of food), enriched in spermine (334 mg / kg of food or 1650 nmol / g of feed) and a dose of cyclophosphamide of 90 mg.kg- ¹ week ^-1 .

The fifteenth group is fed with a diet depleted of cadaverine, putrescine and spermidine (51 μg / kg of food or 0.350 nmol / g of food), enriched in spermine (3340 mg / kg of food or 16500 nmol / g of feed) and a dose of cyclophosphamide of 90 mg.kg- ¹ week ^-1 .

The evolution of tumor volume (in cm ³ ) is shown in Table 1 for animals from groups 1 to 9.

Table 1: Evolution of tumor volume

With regard to the groups 10, 11 and 12, there is an improvement of 5 to 30% relative to the group 5.

With regard to groups 13, 14 and 15, there is an improvement of 1 to 10% compared to group 6.

Cyclophosphamide (Endoxan) administered at 90 mg.kg ^-1 week ^-1 has a significant antiproliferative effect irrespective of the diet provided. The percentage inhibition is of the order of 80 to 95%.

A low dose of cyclophosphamide (Cytoxan ^®) 9 mg.kg ^"1. ^{Week" 1} associated with a depleted diet cadaverine, putrescine and spermidine increases the antitumor efficacy of 25% compared to a diet containing a ratio normal polyamine (125 mg / kg feed, 861 nmol / g feed) combined with 9 mg.kg- ¹ week- ¹ cyclophosphamide.

This depleted diet cadaverine, putrescine and spermidine enriched spermine increases dose-dependently the antitumor effect of cyclophosphamide to 9 mg.kg ^"1. ^{Week" 1} 5 to 30%, depending on the doses of spermine .

This same regime makes it possible to increase, in a dose-dependent manner, the antitumor effect of cyclophosphamide at 90 mg.kg- ¹ week- ¹ from 1 to 10%, depending on the doses of spermine.

These regimens make it possible to potentiate the antitumor effect of cyclophospamide at a low dose thus making it possible to limit its side effects.

Claims

A composition comprising a mixture of cadaverine, putrescine and spermidine at a concentration ranging from 0.3 to 0.6 nmol per gram of composition, said composition also comprising spermine at a concentration ranging from 150 to 17000 nmol per gram of composition.

The composition of claim 1, wherein the concentration of putrescine ranges from 0.19 to 0.32 nmoles per gram of composition, and / or the cadaverine concentration ranges from 0.09 to 0.08 nmoles per gram of composition, and or the concentration of spermidine ranges from 0.09 to 0.20 nmoles per gram of composition.

Composition according to Claim 1 or 2, which contains, as a percentage of dry weight relative to the total dry weight: 10% to 35% of lipids, 8% to 30% of proteins, 35% to 80% of carbohydrates, up to 10% % of a mixture of vitamins, minerals and electrolytes.

Composition according to one of Claims 1 to 3, which optionally contains at least one inhibitor of the intracellular synthesis of polyamines, in particular chosen from the inhibitors of ornithine decarboxylase, spermidine-spermine Nl-acetyltransferase or spermine oxidase, and / or at least one polyamine transport inhibitor, at most 15% by weight relative to the total dry weight of the composition.

Composition according to one of claims 1 to 4, which contains at least one antibiotic, and / or is enriched with vitamins.

Composition according to one of Claims 3 to 5, in which the carbohydrates belong to the group comprising glucose polymers, maltodextrins, sucrose, modified starches, glucose monohydrate, dehydrated glucose syrup, glycerol monostearate and their salts. mixtures, and / or in which the proteins belong to the group comprising the soluble proteins of milk, the soy proteins, serum peptides, egg white powder, potassium caseinate, non-phosphorylated peptides, casein peptides, mixed caseinate, soy isolate and mixtures thereof, and / or wherein the lipids belong to the group consisting of butter oil, peanut oil, medium chain triglycerides, grape seed oil, soybean oil, evening primrose oil and mixtures thereof, and or wherein the lipids consist of a mixture of at least one oil of animal origin, at least one vegetable oil and glycerol stearate.

Composition according to one of Claims 1 to 6, which composition constitutes the daily food ration of a human being and in that it comprises:

- from 75 g to 500 g of carbohydrates,

from 20 g to 185 g of lipids,

from 20 g to 225 g of proteins,

and optionally an inhibitor of the intracellular synthesis of polyamines at a rate of less than 50 g and preferably at a rate of 1 to 10 g per day, or which composition is a submultiple of a daily food ration of a human being and in that it comprises:

- 75 / X g to 500 X g of carbohydrates,

from 20 μg to 185 μg of lipids,

from 20 μg to 225 μg of protein,

and optionally an inhibitor of the intracellular synthesis of polyamines in a proportion of less than 50 μg and preferably in a proportion of 1 / X to 10 × g per day, and X being an integer between 2 and 8 and corresponding to the number of rations to be ingested by the patient to meet his or her daily nutritional needs.

Composition according to one of claims 1 to 7, which composition is in a dry form to dissolve extemporaneously in a neutral vehicle. Composition according to one of claims 1 to 8 for its use as a medicament. 10. Pharmaceutical preparation comprising a composition according to one of claims 1 to 9.

11. Composition according to one of claims 1 to 8 for use as a supplement or food substitute, or as a nutraceutical supplement.

12. Composition according to one of claims 1 to 9 for its use in the treatment, in a patient, pathologies related to cellular hyperproliferation, or for use in the treatment, in a patient, of cancer. 13. The composition for use according to claim 12, wherein the treatment is carried out by administering a unit dose ranging from 6.7 mg to 670 mg spermine.

The composition for use according to claim 12, wherein the composition is administered to the patient according to the following scheme:

(ii) administering a second dose of the composition of the invention for a second period of time, the aforesaid second dose being adjusted according to the reaction of the patient to the first, and consecutively,

(iii) administering a third dose of the composition of the invention for a third period of time, the aforesaid third dose being adjusted according to the reaction of the patient to the second. The composition for use according to claim 12, wherein:

the first period of time varies from 7 to 14 days, in particular 7 days, and or

the second period of time varies from 14 to 21 days, in particular 14 days, and or

the third period of time varies from 28 to 63 days, in particular 63 days.

The composition for use according to claim 12, wherein:

the first dose of polyamines ranging from 6.7 mg to 5.3 g of spermine / day, and

the second dose of polyamines ranging from 6.7 mg to 5.3 g of spermine / day, and

the third dose of polyamines ranging from 6.7 mg to 5.3 g of spermine / day.

17. Composition comprising:

a mixture of cadaverine, putrescine and spermidine at a concentration ranging from 0.3 to 0.6 nmoles per gram of composition,

for its use in the treatment in a patient of pathologies related to cellular hyperproliferation, the total amount of bio logically active polyamines ingested per day by the patient does not exceed 11334 nanomoles per kcal of ingested composition, in particular 5000 nanomoles per kcal of composition ingested, in particular 1000 nanomoles per kcal of ingested composition, in particular 100 nanomoles per kcal of ingested composition.

18. Combination of a composition according to claim 17 and a chemotherapeutic agent for its simultaneous, separate or successive use in the treatment of pathologies related to cellular hyperproliferation.