EP2001454A2 - Use of at least one cytosolic phospholipase a2 inhibitor as a medicine for symptomatic treatment of mucoviscidosis - Google Patents

Abstract

The purpose of this invention is the use of at least one cytosolic phospholipase A2 (cPLA2) inhibitor in the preparation of a medicine intended for preventive and/or remedial symptomatic treatment of mucoviscidosis, and particularly the increased secretion of mucus in mucoviscidosis.

Description

 Use of at least one cytosolic phospholipase A2 inhibitor as a medicament for the symptomatic treatment of cystic fibrosis

The invention is in the field of symptomatic treatment of cystic fibrosis.

Cystic fibrosis (Cystic Fibrosis, CF) is an autosomal recessive genetic disorder caused by a mutation in the CFTR gene, a 250 kb gene that encodes a protein of 1480 amino acids called CFTR protein (Cystic

10 Fibrosis Transmembrane Conductance Regulator). This protein belongs to the family of chloride channels. To date more than 1000 mutations responsible for the disease have been described, but the most common, (discovered in 1989), called ΔF508, is present in about 70% of patients. This mutation consists of the deletion of an amino acid, a phenylalanine at position 508,

Due to a mutation on the tenth exon of the gene. The incidence of mutations in the population is around 1 in 3000 births, but only homozygous patients are sick. Life expectancy is around 35 years in developed countries. Heterozygotes or healthy carriers, phenotypically normal, represent about 4% of the population

20 general.

One of the main consequences of this disease is the modification of the mucus composition (whose viscosity becomes higher than normal) secreted mainly by the secretory epithelia of the respiratory and digestive mucous membranes. Indeed, the alteration of the CFTR channel causes

25. Changes in transepithelial ion fluxes that make the mucus more viscous and thicker, thus preventing its progression to the glottis, in cystic fibrosis patients. This excess of mucus and the absence of mucociliary clearance promote chronic infection with opportunistic bacteria (such as Pseudomonas aeruginosa, Staphilococcus aureus, Haemophilus

Influenzae), which can lead to an exacerbated inflammatory response in the airways, characterized by a massive influx of neutrophils. These pathogens are responsible for most episodes of bronchial infections. The lack of control of this inflammatory process, often associated with bronchial obstruction, causes destruction of lung tissue resulting in a gradual loss of respiratory function until death.

This disease is also characterized by digestive disorders mainly due to exocrine pancreatic insufficiency present in 90% of patients. These respiratory and digestive manifestations dominate the clinical picture, but the respiratory compromise conditions the vital prognosis of the patients.

Chronic infection with P. aeruginosa is the main infectious problem and most often marks a progressive evolutionary course of the disease. Some so-called "mucoid" strains develop within the mucus in the form of microcolonies surrounded by an exopolysaccharide matrix.

This mucoid character is virtually specific to the infection. The incidence of mucoid strains increases with age and the progression of respiratory disease. Bronchopulmonary involvement evolves by relapses which lead in a few months or several decades to chronic respiratory insufficiency. Complications such as recurrent pneumothorax or haemoptysis can make life worse.

Death occurs as a rule following an exacerbation of infectious respiratory signs with signs of right or global heart failure in approximately 1/3 of the cases. At present cystic fibrosis can only be treated symptomatically. Management is mainly based on respiratory physiotherapy and anti-infectious treatment with antibiotics. In addition to its heaviness, this treatment must be followed for life.

Antibiotic therapy must comply with a certain number of binding general principles: appropriate choice of antibiotics, prevention of resistance due to the chronicity of bronchial infection, high doses over long periods of time (at least two weeks), only parenteral administration for antipyocyanic antibiotics.

Molecules make it possible to thin the mucus and in particular rhDNase, catalytic enzyme of hydrolysis of the extracellular DNA which makes it possible to reduce the viscosity of the mucus then easier to evacuate by the physiotherapy. In case of advanced chronic respiratory insufficiency, nocturnal oxygen therapy becomes necessary. But, lung transplantation is the last resort.

There is therefore still a demand for preventive and / or curative treatment of symptoms that would not have the heaviness of current treatments (not to mention transplantation) and which would give patients an improvement in their living conditions.

This is one of the aims of the present invention.

Infection with a pathogen such as Pseudomonas aeruginosa results in activation of inflammatory proteins such as cytosolic phospholipase A2 (cPLA2). The latter moves to the plasma membrane to hydrolyze phospholipids and release arachidonic acid (AA).

AA is then metabolized by two distinct enzymatic pathways involved in inflammation: the cyclooxygenase (COX) pathway that results in the production of prostaglandins, particularly prostaglandin E2 (PGE2); and the lipooxygenase pathway that results in the production of leukotrienes, particularly leukotriene B4 (LTB4).

Previous studies have shown the presence of increased amounts of leukotriene B4 (LTB4) and prostaglandin E2 (PGE2) in bronchoalveolar lavage (BAL) in patients with cystic fibrosis (CF patients). This increase was also found in an animal model of cystic fibrosis in KO (Knock Out) mice whose gene coding for CFTR was deleted, thus not producing CFTR protein (mutant mice called CFTR ^{"; ~} ). has also been established that LTB4 and PGE2 are capable of inducing the secretion of mucus.The inventors have hypothesized a link between the existence of CFTR gene mutation, the causative agent of cystic fibrosis, and increased secretion. mucus in the lung, through increased activity of cPLA2.

The inventors then showed that CFTR mice ^{'' ',} stimulated by instillation with lipopolysaccharide (LPS) of Pseudomonas aeruginosa that causes inflammation of the airways secrete more mucus than wild mice.

By instillation is meant an administration by air, more specifically intratracheally. The inventors have also shown that this secretion of mucus is inhibited by aspirin, a COX inhibitor.

Similarly, the study by the inventors of the secretion of mucus in the respiratory tract of KO mice whose gene encoding cPLA2 has been deleted (mutant mice cPLA2 ^"A ) shows that it is completely abolished. therefore also a role in the regulation of secretion of mucus in the respiratory tract.

Mucus is essentially mucins (glycoproteins) and MUC5AC is one of the most abundant mucins in the lungs of cystic fibrosis patients.

Again, the inventors were able to show by immunohistochemical labeling that MUC5AC is well expressed in CFTR ^"7" mice which show an increase in the amount of mucus in the respiratory tract. The inventors thus confirmed the hypothesis they had emitted by demonstrating for the first time in CFTR ^"7" mice, a probably direct relationship between at least one mutation of the CFTR gene responsible for cystic fibrosis and secretion. of mucus, related to overactivation of cPLA2. The measure of cPLA2 activity in lung homogenates CF patient lung explants having received or not I ¹ ATK confirms this possibility.

Finally, the inventors have shown that the administration intraperitoneally of a specific inhibitor of cPLA2 abolishes the secretion of mucus in the airways, previously secretion induced by Pseudomonas aeruginosa LPS instillation in CFTR mice ^";" , which express cPLA2 at high levels compared to CFTR ^{+ / +} mice.

These results make it possible for the first time to consider the use of a cPLA2 inhibitor in the symptomatic treatment of cystic fibrosis. These surprising and unexpected results suggest a therapeutic, preventive and / or curative role for cPLA2 inhibitors in the treatment of diseases of the respiratory tract leading to increased secretion of mucus, such as cystic fibrosis, but also cystic fibrosis. 'asthma, chronic obstructive pulmonary diseases (COPD) such as laryngitis and pharyngitis, pneumonia, influenza, pneumoconiosis, chronic bronchitis and emphysema or aspergillic asthma.

Thus the subject of the invention is the use of at least one inhibitor of cytosolic phospholipase A2 (cPLA2) in the preparation of a medicament for the symptomatic preventive and / or curative treatment of cystic fibrosis, particularly increased secretion of cystic fibrosis. mucus secretory epithelia of the respiratory and digestive mucous membranes.

By cPLA2 inhibitor is meant any chemical or biological molecule, natural or synthetic, any composition which, whatever its mechanism, induces after administration a decrease, or even a complete inhibition, of the activity of the cPLA2 or the expression of the cPLA2 gene. By way of example, the cakA2 inhibitor ATK (Arachidonyl) may be mentioned as an inhibitor of cPLA2.

Trifluoromethyl Ketone; Ackermann et al., J. Biol. Chem., 1995; 270: 445-50), Pyrrolidine-1 (Seno et al., J. Med Chem 2000; 43: 1041-44), MAFP.

(Methyl arachidonyl fluorophosphonate: Lio et al., Biochim Biophys Acta;

1996; 1302: 55-60), ML3196 (Lehr, J. Med Chem, 1997, 40: 2694-2705), the

4- [4- [2- [2- [bis (4-chlorophenyl) methoxy] ethylsulfonyl] ethoxy] phenyl] -1,1,1-trifluoro-2-butanone, BMS-229724 (Burke et al. J. Pharmacol Exp Ther., 2001, 298: 376-85), 3,3-Dimethyl-6- (3-lauroylureido) -7-oxo-4-thia-1-azabicyclo [3, 2.0] heptane-2-carboxylic acid or the indoles described in international application WO 03/048122, as well as the compounds cited in US Pat. No. 6,630,496 or US Pat. No. 6,350,892.

According to one particular embodiment of the invention, the cPLA2 inhibitor is a siRNA that interferes with the expression of the cPLA2 gene.

Preferably, the inhibitor of cPLA2 used according to the invention is ATK.

According to another embodiment of the invention, it is possible to use a combination of cPLA2 inhibitors (2 or more) in the preparation of a medicament for the treatment of diseases of the respiratory tract leading to a increased secretion of mucus, including the symptomatic treatment of cystic fibrosis.

According to the invention, the cPLA2 inhibitor is used in the preparation a drug for the treatment of cystic fibrosis. It can also be envisaged that the cPLA2 inhibitor is used in the preparation of a medicament for the symptomatic treatment of increased mucus secretion in asthma, chronic obstructive pulmonary disease (COPD) such as laryngitis and pharyngitis, pneumonia, influenza, pneumoconiosis, chronic bronchitis and emphysema or aspergillary asthma.

According to the invention, the cPLA2 inhibitor may be used in the medicament in an amount of between 0.01 mg to 2 g, preferably from 1 mg to 1 g, very preferably from 10 mg to 500 mg.

More particularly, the inhibitor of cPLA2 such as, for example PATK, may especially be administered at doses of between 0.1 mg / kg and 500 mg / kg, preferably 1 mg / kg to 100 mg / kg, very preferably 10 mg / kg to 50 mg / kg. In the context of the invention, the term "symptomatic treatment" refers to the preventive and / or curative treatment of symptoms related to the disease. This treatment also improves the management of patients (reduction of suffering, improvement of the lifespan, slowing of the progression of the disease etc.). The treatment may also be carried out in combination with other ingredients or treatments, such as in particular other active compounds for treating the pathologies or traumas specified in the present application.

For example, it is known that aerial gene therapy trials of cystic fibrosis have given only average results, far from the hopes they may have aroused. In fact, in the aerial gene therapy treatment of instilling into the lungs (for example using aerosol) a nucleic acid vector carrying a non-mutated and functional CFTR gene so that it reaches the epithelial cells of the surface of the lungs, the vector collides with the layer of mucus covering said cells. The mucus thus prevents the vector from reaching its target, at least in sufficient quantity, despite the preliminary treatments using mucus fluidifiers to rid the pulmonary alveolar surface of its mucus barrier. We then understand the importance of having a mucus secretion inhibiting drug which could be administered in addition to gene therapy by air. Thus, according to another aspect, the subject of the invention is the use of at least one inhibitor of cytosolic phospholipase A2 (cPLA2) in the preparation of a medicament for the symptomatic treatment of the increased mucus secretion of cystic fibrosis said medicament to be administered in addition to gene therapy of cystic fibrosis.

The drug according to the invention can take any conceivable form for its administration. For example it may be liquid as a syrup or solution for intramuscular injection or intravenous or solid such as a powder or a tablet.

The administration can be carried out by any method known to those skilled in the art, preferably orally, aerosolized, or by injection, typically intraperitoneal, intra-cerebral, intrathecal, intravenous, intravenous. -arterial or intramuscular. Oral administration is preferred. As a long-term treatment, the preferred route of administration will be sublingual, oral or transcutaneous.

In general, the daily dose of cPLA2 inhibitor used according to the invention will be the minimum dose to obtain the desired therapeutic effect. This dose will depend on various factors such as the weight of the subject to be treated. If necessary, the daily dose may be administered in two, three, four, five, six or more doses taken daily or in multiple sub-doses administered at appropriate intervals during the day.

The amount selected will depend on multiple factors, in particular the route of administration, the duration of administration, the timing of administration, the rate of elimination of the compound, the or various products used in combination with the compound, age, weight and physical condition of the patient, as well as his medical history, and other information known in medicine. The medicaments according to the invention may also comprise at least one other therapeutically active ingredient for simultaneous, separate or spread use over time, in particular during treatment in a subject suffering from a pathology as defined above. . The medicaments according to the invention advantageously comprise one or more excipients or inert carriers, that is to say pharmaceutically inactive and non-toxic vehicles. For example, saline, physiological, isotonic, buffered, etc., solutions compatible with a pharmaceutical use and known to those skilled in the art may be mentioned. The drugs may contain one or more agents or vehicles selected from dispersants, solubilizers, stabilizers, preservatives, etc. Agents or vehicles that can be used in formulations (liquid and / or injectable and / or solid) include methylcellulose, hydroxymethylcellulose, carboxymethylcellulose, cyclodextrins, polysorbate 80, mannitol, gelatin, lactose, vegetable oils or animal, acacia, etc. The compositions may be formulated as injectable suspension, gels, oils, tablets, suppositories, powders, capsules, capsules, etc., optionally using dosage forms or devices providing sustained and / or delayed release. For this type of formulation, an agent such as cellulose, carbonates or starches is advantageously used.

The examples which follow illustrate the invention without limiting it in any way.

Material & Methods ^» Animal Models

The studies were performed on a model of cystic fibrosis established in C57BL / 6J mice (C / ifr ^{mUNC mice} ) maintained on a mixed genetic background by the "Center for Distribution, Typing and Animal Archiving" UPS44 CNRS in Orléans. Wild CFTR ^{+ / +} and mutant CFTR ^{'7 "} mice were weaned at 3-4 weeks and fed a diet supplemented with a commercial laxative, Movicol®, because in the absence of a laxative, CFTR ^'7" mice died. soon after weaning, following intestinal occlusions consecutive to an alteration of the secretion of fluids in the digestive tract.

The experiments were conducted on groups of at least 5 mice per treatment category, 5-7 weeks old.

CPLA2 ^'7' mutant mice and their wild homologs cPLA2 ^{+ / +} , of the same genetic background were obtained from the "Department of Biochemistry and Molecular Biology, Faculty of Medicine, University of Tokyo" (Uozomi et al., Nature, 1997). These mice were 6-8 weeks old at the time of the experiment.

• Products and reagents used Xylazine (Rompum, Bayer-France) Ketamine (Imalgènelooo Merial, Lyon-France)

P. aeruginosa LPS, serotype 10 (Sigma, St. Louis, MO) EDTA, phenylmethylsulphoxide and dithiothreitol (Sigma, St. Louis, MO) Anti-MUC5AC antibody (clone 45 M1, Neomarkers, Fermont, CA, USA) Antibodies mouse IgG (Dako Cytomation Envision System). Amino Ethyl Carbazol (Sigma, St. Louis, MO).

• Experimental protocol

The products used were dissolved in saline solution (or physiological saline) and injected intratracheally (i.t.) or intraperitoneally (i.p.). The usage protocol was as follows:

1) Aspirin (50 mg / kg, i.p.), 30 min before LPS, then daily, once a day;

2) ATK (20 mg / kg, i.p.), 30 min before LPS and 24 h after.

Mice were anesthetized with 2% Xylazine (8 mg / kg) and Ketamine 1000 (40 mg / kg) prior to instillation of LPS (330 μg / kg, i.t.).

Bronchoalveolar lavages (BAL) and lung samples were taken 4 days after instillation of LPS, as indicated below: ^» Bronchoalveolar lavage (BAL)

The mice were anesthetized with Pentobarbital (i.p.) at 200 mg / kg and their incised and cannulated tracheas. The LBAs were performed with successive washings with 1 ml of saline solution containing 5 mM EDTA and protease inhibitors (5 mM phenylmethylsulphoxide, 5 mM dithiothreitol).

The counting of the cells was carried out by an automatic counter Counter ZM (Coultronic, Margency, France). This allows to highlight the state of inflammation of the lungs.

Identification of cell types was obtained by Diff-Quick staining (Baxter Dade AG, Dudingen, Germany) after cytocentrifugation (Hettich-Universal). This makes it possible to count the proportion of polynuclear neutrophilic cells that reflect inflammation.

The results are expressed in number of cells per ml of BAL. • Detection of MUC5AC by ELISA test

50 μl of each wash sample (LBA) were incubated for 1 hour with 50 μl of a bicarbonate-carbonate mixture (50/50) at 40 ^° C. in the wells of 96-well plates (Naked).

After drying, the wells were washed 3 times with PBS and then saturated with 2% BSA, fraction V (Sigma) for 1 h at room temperature.

Wells were again washed 3 times with PBS. 50 μl of primary antibody directed against MU5AC (clone 45 M1) diluted 1/100 in PBS / Tween20 0.05% were then placed in each well and the incubation was continued for 1 h at room temperature. Wells were again washed 3 times with PBS. 100 μl of peroxidase-conjugated anti-mouse IgG secondary antibody (1: 10,000) were then placed in each well and the incubation was continued for 1 hour at room temperature.

After 3 washings of the wells with PBS, the presence of the protein was revealed using 3,3 ', 5,5'-tetramethylbenzidine (TMB) peroxidase, then the reaction was stopped with 1M H ₂ SO ₄ . Absorbance was measured at 450 nm. The results are expressed in milligram of protein per ml of BAL. ^"Fixing the lungs and histology Lungs were perfused with formaldehyde at 4%, to remove blood and immersed in 4% formaldehyde for 48 h at 4 ° C before being embedded in paraffin.

Sections 5 μm thick were stained with Hemotoxylin / Eosin, periodic acid-Schiff (PAS) and Alcian blue (Alcian blue: AB, pH 2.4) according to standard methodologies (Normal and Pathological Histology). P. GANTER & G. Jolies, editions Gauthier-Villars).

After reading under the microscope colored sections with Alcian blue, which stains the mucus, a count of the bronchi and positive cells is realized. The ratio "number of positive bronchi / number of positive cells" makes it possible to establish the score used to evaluate the results of mucus production studies (see below).

• Immunohistochemistry The paraffin sections were treated with the antibody directed against

MLJ5AC (clone 45 M 1) for 2 h at room temperature, at a concentration of 8 μg / ml in 1X PBS.

These sections were then washed with PBS and incubated for 30 min with mouse anti-IgG antibody conjugated with peroxidase. The revelations were made with Amino Ethyl Carbazol.

• Western blot

Mouse lungs were homogenized with lysis buffer (Quiagen RLT buffer). The homogenates were centrifuged, and equivalent amounts of proteins (10-50 μg) were deposited on the 7.5% electrophoresis gel (Tris / Glycine / SDS-polyacrylamide).

The proteins were then transferred to a nitrocellulose membrane (Millipore) previously treated with PBS at 5% milk / Tween 1% o.

The membrane was then incubated for 1 hour at room temperature in the presence of the primary antibody, then washed 3 times with PBS / Tween 1% o.

The membrane was then incubated for 1 hour at room temperature in the presence of the secondary antibody, then washed with PBS / Tween 1% o.

The presence of the proteins on the membrane was then revealed using the ECL system (Amersham 2106) which produces chemiluminescence with peroxidase, according to the supplier's recommendations. The membrane was then exposed in contact with a radiographic film and the latter developed.

Actin control makes it possible to normalize the level of expression of the MU5AC protein. The intensity of the bands is measured with an image analyzer.

^" Measurement of cPLA2 Activity in Bronchial Homogenates of CF Patients Lung Expants. The measurements were made according to Kramer et al. (Biochem J. et al., 248: 779-785 (1987)), with the modifications made by Hidi et al. (J. Immunol 151: 5613-5623 (1993)).

Lung homogenates were prepared according to Filgueiras et al. (Lipids 22: 731-735, (1987)).

50 - 100 μl of lung homogenates were incubated in Tris-HCl buffer containing 1 mM of calcium and 100,000 cpm / ml of the radioactive phosphatidylcholine (1-palmitoyl-214C-arachidonoyl-sn-glycerophosphorylcholine at 52 mCi / mmol , Amersham). After a 30 min incubation, the reactions are stopped by the addition of a chloroform / methanol solution (1: 1).

The lipids were extracted by the method of Bligh and Dyer (Can.

Biochem (1959) 37: 911-918), then evaporated to dryness and subjected to thin layer chromatography (Merck) in the presence of chloroform / methanol / acetic acid / water (65 43, 1, 3) as a solution. migration.

The spots corresponding to phosphatidylcholine (PC) were then recovered and their radioactivity measured using standard scintillation fluid. The cPLA2 activity was expressed in counts per minute (cpm).

Results:

> Mucus production in the airways of CFTR ^{+ / +} and CFTR ^{'7' mice at} day 4 with or without stimulation of P. aeruginosa LPS. The "control" mice received a dose of saline.

Table 1:

These results show the implication of the CFTR gene and the CFTR protein in the production of mucus by the cells of the bronchial epithelium.

> Measurement of the amount of secreted MUC5AC protein in the lungs of CF (- / -) and wild-type (CFTR + / +) mice with or without stimulation of P. aeruginosa LPS (ELISA test)

Table 2

CF mice secrete more MUC5AC in LBAs both in baseline conditions than after LPS treatment compared with WT mice.

* P <0.001 CF mice compared to WT;

# P <0.001 LPS compared to the control.

> Mucus production in the respiratory tract of CFTR ^{+ / +} and CFTR ^{4 " mice at} day 4 with or without stimulation by LPS of P. aeruginosa and with administration of aspirin (asp)

The "control" mice received a dose of saline and a dose of aspirin.

Table 3

The scores obtained are compared with the corresponding scores in Table 1.

These results show that a COX inhibitor causes an inhibition of the mucus production by the cells of the bronchial epithelium, and confirms the implication of the COX in this production.

> Mucus production in the respiratory tract of CFTR ^{+ / +} mice and CTFR ^"7" at 4 ^th day with or without stimulation by P. aeruginosa LPS and ATK administration.

The "control" mice received a dose of saline and a dose of ATK. Table 4

The scores obtained are compared with the corresponding scores in Table 1.

These results show that a cPLA2 inhibitor causes an inhibition of mucus production by the cells of the bronchial epithelium.

> Mucus production in the respiratory pathways mouse cPLA2 ^{+ / +} and cPLA2 ^"/ - to 4 ^ιeme day with or without stimulation with LPS from P. aeruginosa.

The "control" mice received a dose of saline. Table 5

These results demonstrate the involvement of cPLA2 in the production of mucus by the cells of the bronchial epithelium.

> Measurement of the amount of MUC5AC messenger RNA in bronchial homogenates of CFTR + / + and CTFR - / - mice with or without stimulation of P. aeruginosa LPS, by Western blot The primary antibody used is a monoclonal antibody anti-MUC5AC (clone 45 M1, Neomarkers, Fermont, CA, USA)

The secondary antibody used is an anti-mouse IgG antibody (Dako Cytomation Envision System).

For each sample, the measurement of the quantity is related to the measurement of the amount of actin mRNA measured in the same sample for normalization (MUC5AC mRNA / Actin mRNA).

Table 6

These results confirm the increase of MUC5AC mRNAs in CF mice stimulated or not.

* P <0.001 CF compared to WT;

> Measurement of the amount of MUC5AC messenger RNA in bronchial homogenates of CFTR + / + mice with or without stimulation of P. aeruginosa LPS and administration or not of ATK, by

Western blot.

Under conditions identical to the previous experiment, the results show that MUC5AC mRNA expression is completely inhibited in unstimulated CFTR + / + mice and a very small amount of MUC5AC mRNA in stimulated CFTR + / + mice.

> Measurement of the amount of cPLA2 messenger RNAs in mouse bronchial homogenates.

The primary antibody used is an anti-cPLA2 monoclonal antibody (CeII Signaling or Santa Cruz).

The secondary antibody used is an anti-mouse antibody (Dako Cytomation Envision System).

For each sample, the measurement of the quantity is related to the measurement of the amount of actin messenger RNA measured in the same sample for normalization (cPLA2 mRNA / Actin mRNA). Table 7

These results confirm the increase of MUC5AC mRNAs in CF mice stimulated or not.

* P <0.001 CF compared to WT.

> Measurement of cPLA2 activity in bronchial homogenates of lung explants of CF patients. Table 8

These results demonstrate that the use of a cPLA2 inhibitor reports the secretion of mucus in a CF patient to that of a non-CF patient. Histological analyzes and immunohistochemical studies

Histological or immunohistochemical analyzes, performed on sections of the lungs of mice, confirm the results obtained in the measurements of the mucus production by the cells of the bronchial epithelium.

Claims

Use of at least one cytosolic phospholipase A2 inhibitor (cPLA2) in the preparation of a medicament for the symptomatic treatment of cystic fibrosis 2. Use according to claim 1, characterized in that the cPLA2 inhibitor is any chemical or biological molecule, natural or synthetic, or any composition which induces, after administration, a decrease, or even a complete inhibition, of the activity. cPLA2 or the expression of the gene encoding cPLA2. 3. Use according to any one of claims 1 or 2, characterized in that the inhibitor cPLA2 is selected from ATK, Pyrrolidine-1, MAFP, ML3196, BMS-229724, the acid 3,3-Dimethyl-6- (3-lauroylureido) -7-oxo-4-thia-1-azabicyclo [3,2,0] heptane-2-carboxylic acid, siRNA or indoles. 4. Use according to claim 3, characterized in that the inhibitor of cPLA2 is I ¹ ATK. 5. Use according to any one of claims 1 to 4, characterized in that the inhibitor cPLA2 is used in the drug in an amount of 0.01 mg to 2g, preferably 1 mg to 1g, very preferably of 10mg to 500mg. 6. Use according to any one of claims 1 to 5, in a symtomatic preventive and / or curative treatment. 7. Use according to any one of claims 1 to 6, characterized in that the drug is liquid or solid, such as a powder or a tablet. 8. Use according to any one of claims 1 to 7, characterized in that the medicament is intended to be administered orally, by aerosol or by injection, in particular by intraperitoneal, intravenous, intra-arterial route. or intramuscular. 9. Use of at least one cytosolic phospholipase A2 inhibitor (cPLA2) in the preparation of a medicament for the symptomatic treatment of increased secretion of cystic fibrosis mucus, said medicament being intended to be administered in addition to gene therapy of cystic fibrosis.