BE1031668B1 - PHARMACEUTICAL COMPOSITION FOR INHALATION - Google Patents

Abstract

A pharmaceutical composition for activating macrophages and/or dendritic cells comprising lipid vehicles comprising a pharmaceutical agent, its therapeutic uses and its manufacturing process.

Description

PHARMACEUTICAL COMPOSITION FOR INHALATION

Technical field

The present invention relates to a lipid composition protecting a pharmaceutical agent and intended to be administered into the respiratory tract to a patient.

Prior art

Formulation such as lipid entrapment of pharmaceutical agents is well known.

In particular, the use for this purpose of lipids and in particular cationic lipids selected according to the nature of the pharmaceutical agent to be trapped and the type of vehicles to be obtained is known.

Some of these lipids, however, exhibit clinical toxicity, which limits the options (quantity, nature). In addition, the size of the liposome, including the dispersion of sizes, the stability of the composition in biological media, its ability to trap the pharmaceutical agent, or to release it at the desired location, are all factors that are difficult to optimize, especially jointly.

Ji et al. 2023, describe the use of liposomes for the potentiation of STING (Stimulator of Interferon

Genes): this approach was designed on the basis of cyclic dinucleotides, which are protected by the lipid structure. Different liposome formulations were made based on DOTAP (1,2,- dioleoyl-3-trmethylamonium-propane), cholesterol and a phosphoethanolamine derivatized by polyethylene glycol. Good encapsulation rates were obtained for molar ratios

DOTAP/agonist ratio between 10 and 20 while at a ratio of 2.5, the encapsulation efficiency is poor, which is attributed to too low a density of positive charges. Similarly, at these low concentration ratios, the potentiation of the biological effect by the liposome is lost.

Unfortunately, the low level of encapsulation implies a massive administration of liposomes to a patient, which presents risks of toxicity, in particular due to the large amounts of cationic lipids, abundant in the liposomes of this study.

On the other hand, the modification of the content of a compound is not simple because several physicochemical properties of the liposome must correspond to acceptable values for administration to a patient, such as the size, the dispersion of the size and the electric charge potential; while ensuring the stability of the formulation. In other words, in a formulation, the enrichment in one component is done to the detriment of the others, which risks strongly affecting the balance: the inventors have noticed that this balance is a function of different concentration ratios, which can be strongly affected even following adaptations of the concentrations that could have seemed marginal.

Thus, the inventors noticed that there still remains a need for improved formulations for the benefit of patients.

Brief summary of the invention

A first aspect of the present invention relates to a pharmaceutical composition for activating macrophages and/or dendritic cells comprising lipid vehicles formed from a series of lipids with at least one lipid having a functionalization by a ligand of the mannose receptor (CD206), these lipid vehicles comprising a pharmaceutical agent selected from the group consisting of a nucleic acid, a STING protein agonist or an antagonist of the STING protein, a Toll-like receptor ligand, an immunomodulant, a peptide, or a mixture thereof.

Preferably, this pharmaceutical composition for activating macrophages and/or dendritic cells is administrable by inhalation, advantageously by nebulization and/or by pressurized inhalation and/or by dry powder inhalation and/or by gentle spray inhalation.

A related aspect of the present invention relates to this pharmaceutical composition for activating macrophages and/or dendritic cells for the treatment of cancer, preferably metastatic cancer, preferably said metastasis being outside the lung or the central nervous system and the primary tumor being pulmonary or located at the central nervous system, or said metastasis being pulmonary or located at the central nervous system and the primary tumor being outside the lung or the central nervous system.

A related aspect of the present invention relates to this pharmaceutical composition for activating macrophages and/or dendritic cells for the treatment of an infectious disease at the level of the respiratory and/or systemic tract, or of the central nervous system.

A related aspect of the present invention relates to this pharmaceutical composition for activating macrophages and/or dendritic cells comprising a nucleic acid encoding one or more epitope(s), or a peptide comprising one or more epitopes, said pharmaceutical composition being for vaccination.

A related aspect of the present invention relates to a method for obtaining this pharmaceutical composition for activating macrophages and/or dendritic cells comprising the steps of - solubilizing the lipids in an organic solvent, - evaporating the solvent so as to form a lipid film, - rehydrating with a solution containing the pharmaceutical agent, - extruding and inserting a lipid derivatized with a ligand of the CD206 receptor, - preferably a second step of contacting a solution containing the pharmaceutical agent with the extruded lipids, said lipids comprising a neutral lipid at pH 7.0 and positively charged at pH 5, and said rehydration step being carried out at a pH lower than 7.0.

Brief description of the drawings

Figure 1 compares the stability of different lipid structures.

Figure 2 compares the effect of functionalization on macrophage activation.

Detailed description of an embodiment of the invention

The inventors have succeeded in developing lipid-based formulations which allow significant trapping of one (or more) pharmaceutical agent(s), while retaining advantageous physicochemical properties compatible with clinical use.

Thus, a first aspect of the present invention relates to

A pharmaceutical composition for activating macrophages and/or dendritic cells comprising lipid vehicles formed from a series of lipids with at least one lipid having a functionalization by a ligand of the mannose receptor (CD206) and the lipid vehicles comprise at least one pharmaceutical agent selected from the group consisting of at least one nucleic acid, at least one STING protein agonist or at least one STING protein antagonist, at least one Toll-like receptor ligand, an immunomodulant, a peptide, and mixtures thereof (except the STING agonist with the STING antagonist), this pharmaceutical composition preferably being for inhalation.

The lipid vehicles are advantageously chosen from the group consisting of liposomes, vesicles, micelles, lipoplexes, lipid emulsions, lipid nanocrystals, lipid microspheres, lipid nanoparticles, and mixtures thereof.

In the context of the present invention, the terminology "by inhalation" refers to both nasal administration (e.g., to reach the nasal cavity, central nervous system and/or systemic circulation; e.g., treatment of a tumor, gene therapy, treatment of inflammation or infection) and pulmonary administration (e.g., to treat a disease therein, such as inflammation, infection or tumor), and nasal administration to target both nasal and pulmonary deposition.

In the context of the present invention, the term "peptide" preferably means any chain of at least 2 amino acids linked by peptide bonds. Advantageous peptides are a protein (including an enzyme), an antigen (an epitope), an antibody (or an antibody fragment, a nanobod/y).

Advantageously, the pharmaceutical agent comprises at least one nucleic acid selected from the group consisting of single-stranded DNA, double-stranded DNA (including cDNA), single-stranded RNA (including mRNA), double-stranded RNA, siRNA, shRNA, MIRNA, oligonucleotide (DNA and/or RNA based), and mixtures thereof. The nucleotides, sugars or phosphodiester bonds may advantageously be modified, for example uridine may be substituted or partially substituted by pseudouridine, for example when the nucleic acid is mRNA. The adenosine or cytosine bases may advantageously be methylated, as may the sugars (e.g. in the 2’ position). Peptide nucleic acids, morpholino and/or phosphorothioate bonds may advantageously replace some or all of the phosphodiester bonds. Alternatively, when the nucleic acid is to stimulate an immune response and includes motifs

CpG, cytosine is advantageously not methylated.

Advantageously, the pharmaceutical agent comprises at least one STING protein agonist, preferably selected from the group consisting of CGAMP, 2',3'cGAMP, DMXAA, ADU-S100, MK-1454, MK-2118, GSK-3745417, BMS-986301, SB-11285, IMSA-101, SYN-STING (SYNB1891), BI-1387446, TAK-676, E-7766 ExoSTING, SNX281, HG-381, DN-015089, PC7A, DiABZI, Di-cyclic guanosine monophosphate (cyclic di-GMP; cyclic di-

GMP), MSA-2, ulevostinag, SB 11285, IMSA 101, Dazostinag, BMS-986301, BI 1387446 and their mixtures, preferably 2° 3'CGAMP.

Alternatively, the pharmaceutical agent comprises at least one STING protein antagonist selected from the group consisting of C-176, H-151, Inh-54, RU.521, H-8, A151, SA-1, GSK'932, SN-011 and mixtures thereof.

Preferably, the series of lipids forming the lipid vehicles has a phase transition temperature (MT) of between -30°C and 100°C, preferably between 1°C and 90°C, advantageously between 10°C and 80°C, 20°C and 70°C, for example between 20°C and 60°C.

Preferably (alternatively, or additionally), the series of lipids forming the lipid vehicles has a melting point of between -30°C and 180°C, preferably between 1°C and 160°C, advantageously between 10°C and 155°C, preferentially between 20°C and 150°C, for example between 20°C and 80°C.

These physicochemical properties of the lipid series increase the release of the pharmaceutical agent into the targeted tissues and/or the cytoplasmic release of the pharmaceutical agent and/or its half-life and/or reduce the release of the pharmaceutical agent into non-targeted tissues. In particular, the inventors noted that a high phase transition temperature (Tm) of the lipid series, for example more than 10°C, more than 20°C, is associated with increased rigidity, which is advantageous, at least for certain formulations or modes of administration to the patient.

Advantageously, the at least one lipid having a functionalization by a ligand of the mannose receptor (CD206) is a phospholipid having a functionalization by a ligand of the mannose receptor (CD206),

Advantageously (further) at least one lipid, preferably a phospholipid, is derivatized by polyethylene glycol (PEG).

A PEG of size between 500 Da and 20 kDa is preferred, for example between 1 and 10 kDa, or even between 2 and 5 kDa.

Preferably, a lipid, preferably a phospholipid, derivatized by PEG, advantageously a DSPE-PEG, has a functionalization by a ligand of the mannose receptor (CD206).

Advantageously, between 10% and 100% (mole equivalent), preferably between 20 and 90%, preferably between 30 and 80%, or even between 40 and 50% of the PEG-derivatized lipids are further functionalized with the mannose receptor ligand (CD206). Thus, a portion of the PEG-derivatized lipids is not necessarily functionalized. The inventors have in fact noted that the double optimization, the PEG content and that of the CD206 receptor ligand, was advantageously carried out in this manner. When certain PEG residues are not functionalized by the CD206 receptor ligand, this object of the present invention can be achieved either on the basis of a single lipid, preferably a single phospholipid, derivatized by PEG, of which only a proportion would be functionalized, or on the basis of several lipids, preferably comprising at least ONE phospholipid, which would be derivatized with PEG, with some of these PEG-derivatized molecules which would not be functionalized.

This increases the ability of the ligand to bind to its receptor. In addition, PEG stabilizes lipid vehicles.

Advantageously, the ligand of the mannose receptor (CD206) is selected from the group comprising a mannose, a fucose, an N-acetylglucosamine, an N-acetylgalactosamine, a glycoprotein, a galactocomannan, an a-D-mannopyranoside (eg a-

D-Mannopyranose), a-L-fucopyranose-(1-3)-2-acetamido-2-deoxy-B-D- glucopyranose, Methyl a-D-mannopyranoside, a-L-fucopyranose -{1- 2) -B-D-galactopyranose-(1-4)-B-D-glucopyranose, Methyl 2-

acetamido-2-deoxy-a-D-glucopyranoside, B-D-galactopyranose-({1-3)- (a-L-Fucopyranose-(1-4)) 2-acetamido-d-deoxy-B-D-glycopyranose, a-

D-mannopyranose-{1-2}-a-D-mannopyranose, a polymannose (between 4 and 50, preferably between 5 and 30, or even between 6 and 20 mannose residues), an anti-CD206 antibody, and mixtures thereof.

In the context of the present invention, these ligands may be modified, for example N-acetylglucosamine may be deacetylated. For example, N-acetylglucosamine is incorporated as chitin or chitosan, preferably oligomers of chitin or chitosan (e.g. less than 50 monomers, less than 40 monomers, less than 30 monomers).

Preferably, the lipid vehicles are lipid nanoparticles (LNPs), preferably liposomes or solid lipid nanoparticles (SLNs) or nanostructured lipid carriers (NLCs).

In the context of the present invention, "SLN" preferably means colloidal particles (generally 50 to 500 nm in diameter) prepared from solid lipids (solid at room temperature and body temperature), surfactants and water by high speed or high pressure homogenization methods.

In the context of the present invention, "NLC" preferably means SLNs containing a liquid lipid (oil) fraction that causes structural imperfections of the solid lipids leading to a less ordered crystalline arrangement in order to improve the encapsulation of pharmaceutical agent. They are prepared by methods similar to SLNs.

In the context of the present invention, "liposomes" preferably means spherical lipid vesicles (generally 50 to 500 nm in diameter) composed of one or more lipid bilayers. They result from the emulsification of natural or synthetic lipids in an aqueous medium by methods such as lipid film hydration, the ethanol injection method or by microfluidic methods.

Preferably, the pharmaceutical agent has an overall negative charge.

Indeed, the inventors have managed to incorporate large quantities of this type of agent, even in molar proportion of the cationic (or protonable, see below) lipid potentially present.

Thus, the lipid series preferably comprises a first

A lipid selected from the group consisting of a cationic lipid, a neutral lipid, an anionic lipid, a phospholipid, a triglyceride, a diglyceride, a glycerophospholipid, a sphingomyelin, a fatty acid, a fatty acid salt, and/or a second lipid selected from the group consisting of a cationic lipid, a neutral lipid, an anionic lipid, a phospholipid, a triglyceride, a diglyceride, a glycerophospholipid, a sphingomyelin, a fatty acid, a fatty acid salt, and/or a third lipid selected from the group consisting of a cationic lipid, a neutral lipid, an anionic lipid, a phospholipid, a tigyceride, a = diglyceride a = glycerophospholipid, a sphingomyelin, a fatty acid, a fatty acid salt.

Preferably, the pharmaceutical composition comprises a first lipid being a cationic lipid having at least one quaternary ammonium group, and/or a lipid having at least one protonatable (secondary or tertiary) amine group (and free of negative charge at pH 7.0).

Preferably, the lipid having at least one quaternary ammonium group (and/or first lipid) is selected from the group consisting of DOTAP (18:1 TAP) as well as other TAP derivatives (such as 14:0 TAP, 16:0 TAP, 18:0 TAP), DOTMA, DDAB, DC-Chol,

DODAP, MVL5, DOSPA, GL67, DOBAQ, EPC derivatives (12:0 EPC, 14:0 EPC, 16:0 EPC, 18:0 EPC, 18:1 EPC, 14:1 EPC, 16:0-18.1 EPC), DORI, DC-6-14, and mixtures thereof. DOTAP is preferred.

By protonatable (secondary or tertiary) amine, it is preferably understood in the context of the present invention, an amine which is predominantly in neutral form at pH7.0, (e.g. more than 50%, preferably more than 75%, or even more than 90 or 99% in neutral form at pH7.0) and predominantly in protonated form at pH 5.0 (e.g. more than 50%, preferably more than 75%, or even more than 90 or 99% in protonated form at pH5.0).

An example of a lipid comprising a protonatable amine is 1,2-dioleyloxy-3-dimethylaminopropane (DODMA). It is understood, in the context of the present invention, that other lipid chains than oleic acid can be substituted therefor, for example palmitic acid or stearic acid while retaining the derivatives

Dimethylaminopropane (DMA; such as 18:0 DMA, 16:0 DMA, 14:0

DMA, 18:1 DMA). Conversely (or in addition) substitutions at the aminopropane group are possible, for example at the methyl groups of DMA.

Preferably, the second lipid is a sterol and is advantageously selected from the group consisting of cholesterol and its esters (cholesteryl esters), and its glycosylated derivatives (B-D-glucosyl cholesterol, Galactosyl Cholesterol, BDGL-1), ox-18:2 cholesterol, desmosterol, stigmasterol, lanosterol, 7-dehydrocholesterol,

dihydroxylanosterol, zymosterol, lathosterol, and mixtures thereof. Cholesterol is highly preferred.

Thus, a composition comprising DOTAP and cholesterol is highly preferred.

Preferably, the phospholipid potentially present in this pharmaceutical composition (second lipid in the absence of sterol, third and/or fourth lipid) is chosen from natural, purified or synthetic phospholipids, such as phospholipids extracted from egg or soy, phosphatidic acids, phosphatidylethanolamines (PE), lysophosphatidylethanolamines (LPE) or phosphatidylcholines (PC) or lysophosphatidylcholine (LPC) or phosphatidylserines (PS), or lysophosphatidylserines (LPS) or phosphatidylglycerol (PG) or lysophosphatidylglycerol (LPG) and mixtures thereof, preferably chosen from the group comprising DSPE, DSPE-PEG, DMPE, DPPC,

DSPC, DMPC, DLPC, and mixtures thereof.

In the context of the present invention, preferably, the fatty acid chains constituting the phospholipids, but also the cationic or protonatable lipids can be saturated, unsaturated or polyunsaturated, provided that the parameters below of melting point and/or phase transition temperature (measured on all the lipids used) are within the ranges described. Thus, the composition according to the invention can advantageously contain unsaturated fatty acids and saturated fatty acids.

The fatty acid chains constituting the above phospholipids have a size advantageously between 14 and 20 carbon atoms, such as 16 or 18 carbon atoms. Shorter or longer sizes are possible (in low contents, e.g. less than 20 mol% of all the lipids) provided that the parameters below of melting point and/or phase transition temperature (measured on all the lipids used) remain within the ranges described.

Advantageously, the anionic lipid, when present, is chosen from phosphatidylinositol (Pls) and their phosphates (PIPs), phosphatidylserines (PS; e.g. DPPS), phosphatidic acid (PA; e.g.

DPPA) and phosphatidylglycerols, (eg DPPG or DSPG) and mixtures thereof.

Preferably, the composition comprises a phosphatidylcholine, such as DPPC (1,2-dipalmitoylphosphatidylcholine).

The inventors have noticed that this lipid having a polar head and two unsaturated hydrocarbon chains (di-palmtoyl) increases the rigidity of lipid structures and/or allows a reduction in the (relative) DOTAP content. An incorporation of DPPC in a molar ratio of between 0.1 and 0.95, preferably between 0.2 and 0.90, preferably between 0.5 and 0.8, or between 0.6 and 0.7 (mole phosphatidylcholine, e.g. DPPC: total moles of lipids) relative to all lipids is preferred.

Preferably, the lipid carriers of the pharmaceutical composition have a molar ratio of the first lipid to the second lipid (or to the third lipid if the second lipid is absent) of between 0.3 and 20, preferably between 0.5 and 10, such as between 1 and 5.

Preferably, the lipid vehicles of the pharmaceutical composition have a molar ratio of said at least one lipid having functionalization by a ligand of the mannose receptor (CD206) to the total lipids of between 0.01 to 02, preferably between 0.02 and 0.1, preferably between 0.03 and 0.05.

Advantageously, the molar proportion of said at least one pharmaceutical agent relative to said at least one lipid having functionalization by a ligand of the mannose receptor (CD206) is between 5 (or 10) and 150, preferably between 20 and 100, or even between 30 and 80.

Advantageously, the lipid vehicles have a molar proportion of said cationic and/or ionizable lipid relative to the pharmaceutical agent of between 30 and 0.1, preferably between 10 and 0.2, advantageously between 3 and 0.5, preferably between 2.5 and 0.8, preferentially between 2.4 and 1.

Preferably, the pharmaceutical composition has a loading rate of the pharmaceutical agent of between 0.1 and 80% relative to the mass of the lipid vehicles, preferably between 1 and 70%, preferably between 10 and 60%, preferably between 15 and 50%, such as between 1 and 40% or even between 25 and 30%.

Advantageously, the average size of the liodic vehicles is between 50 and 200 nm, preferably between 60 and 190 nm, advantageously between 70 and 180 nm, preferentially between 80 and 180 nm, preferably between 90 and 20 180 nm, more preferably between 100 and 180 nm, preferably between 110 and 180 nm, advantageously between 120 and 180 nm, preferentially between 120 and 170 nm, preferably between 120 and 160 nm.

Alternatively, the average size of the lipid vehicles is between 350 and 1500 nm, preferably between 350 and 1400 nm, advantageously between 350 and 1300 nm, preferentially between 350 and 1200 nm, advantageously between 350 and 1100 nm, preferentially between 350 and 1000 nm, preferably between 350 and 900 nm, preferably between 350 and 800 nm, more particularly between 350 and 700 nm,

advantageously between 350 and 600 NM, for example between 400 and 600 nm.

Alternatively the average size of lipid vehicles is between 200 and 350 nm.

Preferably the size (mean) and mean diameter (Dh) of lipid vehicles are measured by dynamic light scattering (DLS).

Advantageously, the lipid vehicles have a polydispersity index of between 0.05 and 0.9, preferably between 01 and 08, advantageously between 02 and 0.75, preferentially between 0.3 and 0.7, preferably between 0.4 and 0.6; preferably the polydispersity index being measured by dynamic light scattering (DLS). A preferred apparatus, both for measuring particle size, Pdi (see below) or zeta potential (see below; even if other apparatus are also used) is the Malvern Zetasizer nano ZS (Malvern Instruments SA,

Worcestershire, UK).

Advantageously, in the context of the present invention, the polydispersity index is defined by the formula: 5 AQp (1x5 — VERS)

PI = 2745 The ee pe

VAC,

Bst where = DLS is the average particle diameter, ti is the quantity of particles with size Xi and xi is the diameter of the spherical particle.

Alternatively, the polydispersity index is advantageously between 0.05 and 0.9, preferably between

005 and 08 advantageously between 0.05 and 0.7, more advantageously between 0.05 and 0.6, even more advantageously between 0.05 and 0.5, preferentially between 0.05 and 0.4, advantageously between 0.05 and 0.3, more particularly between 0.05 and 0.25, preferably between 0.05 and 0.2, for example between 0.06 and 0.2, preferentially between 0.07 and 0.2, advantageously between 0.08 and 0.2, measured by dynamic light scattering (DLS).

Indeed, depending on the applications, lipid vehicle particles of the same faults are preferred, while for other applications, a bimodal or even multimodal distribution is preferred.

Preferably, the lipid vehicles have a zeta potential of between -60 mV and 100 mV, preferably between -40 mV and 80 mV, advantageously between -20 mV and 60 mV, preferentially between 0 mV and 50 mV, preferably between 10 mV and 40 mV, advantageously between 10 mV and 30 mV, advantageously measured by “laser doppler electrophoresis”. Preferably, this measurement is carried out in an aqueous solution of NaCl at 0.009% (mass:volume).

Advantageously, said pharmaceutical agent is present in a mass proportion of between 0.1 and 80%, preferably between 1 and 70%, preferably between 5 and 60%, preferably between 10 and 50% or between 20 and 40%, relative to the mass of the lipid vehicles.

Advantageously, the composition contains at least one metal ion such as Mn2?+, Co2+ or Zn?* in order to potentiate the activity of the pharmaceutical agent. Preferably, this or these metal ions are incorporated in a content greater than 10 PPM (parts per million; content by weight; mass of the metal ion(s): total mass (dry) of the lipid composition), preferably at a content of between 100 PPM and 50% by mass (Mass of the metal ion(s): total mass (dry) of the lipid composition), preferably between 300 PPM and 10%, preferably between 800 PPM and 1% (by mass).

A related aspect of the present invention relates to a pharmaceutical composition for activating macrophages and/or dendritic cells in liquid form such as a solution, dispersion or suspension, or in the form of a powder to be redissolved and redispersed before administration, or in the form of a dry powder for inhalation.

Advantageously, this pharmaceutical composition for activating macrophages and/or dendritic cells in liquid form may be frozen to ensure good stability during storage or distribution of the drug. This composition will then be thawed before administration and will allow the lipid vehicles to be reconstituted in an advantageous manner. This further ensures that the formulation retains maximum activity.

Advantageously, this pharmaceutical composition for activating macrophages and/or dendritic cells comprises at least one excipient chosen from the group consisting of a buffer chosen from the group comprising phosphate, sulfonate buffers (MES, TES, HEPES, MOPS, PIPES, TAPS, TAPSO), acetate, bicine, and/or at least one salt chosen from the group of inorganic salts and organic salts, and/or at least one surfactant chosen from the group comprising cholic acids and their salts,

phospholipids (e.g. phosphatidylcholines or lecithin, phosphatidylglycerols), lipids or triglycerides, sorbitan esters, polyethoxylated sorbitans, fatty acids, preferably lauric, palmitic, stearic, erucic or behenic acid, esters of these fatty acids, or derivatives of these fatty acids, such as salts, preferably chosen from magnesium stearate, sodium stearyl fumarate and sodium stearyl lactylate, sodium lauryl sulfate, magnesium lauryl sulfate, natural components of pulmonary surfactant such as phospholipids or cholesterol; and sucroesters (sugar esters, for example esters between sucrose or glucose and fatty acids) and/or at least one amino acid selected from the group consisting of histidine, leucine, isoleucine, threonine, lysine, valine, methionine, phenylalanine, mixtures thereof and derivatives thereof, such as acesulfame K or aspartame and/or a sugar, preferably selected from the group consisting of monosaccharides (such as glucose or arabinose), disaccharides (such as lactose, maltose, sucrose, dexirose, irehalose, maltitol and mixtures thereof or a bulking agent selected from polyols such as sorbitol, mannitol and xylitol), polysaccharides (such as dextran, chitosan, starch, cellulose, and derivatives thereof), oligosaccharides (such as cyclodexirin and dexirins).

An advantageous option for the pharmaceutical composition for activating macrophages and/or dendritic cells is drying in the form of a powder to ensure good stability during storage, to be dissolved or redispersed for example just before administration to a patient (e.g. less than 1 hour). This composition will then be redissolved or redispersed before administration in an aqueous solvent such as water or a physiological solution or buffer such as a 0.9% NaCl solution (mass:volume) or

PBS, and will allow the lipid vehicles to be reconstituted in an advantageous manner. This further ensures that the formulation retains maximum activity.

Alternatively OR additionally, the pharmaceutical composition for activating macrophages and/or dendritic cells according to the invention is in the form of a powder to ensure good stability during storage or distribution of the drug, to be dissolved or redispersed and comprises at least one bulking agent such as a polyol and/or a sugar alcohol such as sorbitol, mannitol, maltitol (sometimes considered a disaccharide) and xylitol, sugars (crystalline) including monosaccharides (glucose, arabinose), disaccharides (lactose, maltose, sucrose, dextrose, trehalose), and polysaccharides (dextran, chitosan, starch, cellulose and its derivatives), or oligosaccharides (dextrins, cyclodextrins) and/or fatty acid salts (e.g. magnesium stearate) or fatty acids or their derivatives (e.g. esters), such as lauric acid, palmitic acid, stearic acid, erucic acid, behenic acid, or a phospholipid (e.g. lecithin, phosphatidylcholine phosphatidylglycerol; preferably not incorporated in the lipid vehicle), triglycerides (preferably not incorporated in the lipid vehicle), sugar esters, and/or an amino acid (see above for the liquid composition) and mixtures thereof. Dextran, trehalose, mannitol and lactose are preferred.

Advantageously, a bulking agent excipient will be used, such as lactose, trehalose, sucrose or mannitol. In this case, the composition, even in powder form, will advantageously comprise a buffer and in addition one or more of the compounds listed above, for example a surfactant.

Preferably, the liquid form to be administered comprising the pharmaceutical composition according to the invention comprises from 50 to 99% water (mass of water: total mass of the composition), preferably from 70 to 98% water, or even from 80 to 95 (or 90%) % water; in other words, the solution comprises from 1 to 50% of the pharmaceutical composition, preferably from 2 to 30%, or even from 5 or 10 to 20%.

The liquid form to be administered comprising the pharmaceutical composition for activating macrophages and/or dendritic cells further comprises at least one buffer selected from the group consisting of phosphate, sulfonate buffers (MES, TES, HEPES, MOPS, PIPES, TAPS, TAPSO), acetate, bicine, and/or at least one salt selected from the group consisting of inorganic salts (sodium chloride, calcium carbonate, sodium or potassium phosphate), organic salts (e.g. sodium lactate, potassium citrate), and/or at least one surfactant selected from the group consisting of cholic acids and their salts (faurocholate, glycocholate), phospholipids (not incorporated into the lipid vehicle), lipids (not incorporated into the lipid vehicle), sorbitan esters (e.g. SPAN 85), polyethoxylated sorbitan (e.g. Tween®80) and/or at least one amino acid selected from the group consisting of histidine, leucine, isoleucine, threonine, lysine, valine, methionine, phenylalanine, their mixtures and their derivatives, such as acesulfame K or aspartame and their mixtures.

Another advantageous option for the pharmaceutical composition for activating macrophages and/or dendritic cells is to be in the form of a dry powder for inhalation and/or intended to be administered using a dry powder inhaler. This composition will have advantageous properties of dispersion and aerosolization of the powder in the air, allowing the delivery of adapted doses of powder into the patient's respiratory tract (inhalable powder). These properties include an aerodynamic size adapted to the route of administration. This composition will allow the lipid vehicles to be reconstituted advantageously in physiological fluids once the powder has been deposited in the respiratory tract. This further ensures that the formulation retains maximum activity.

Preferably the geometric diameter (particle size) and/or the aerodynamic size for lung administration is less than 5 um, preferably between 0.5 um and 5 um, preferably between 1 um and 3 um.

Preferably the geometric diameter (particle size) and/or the aerodynamic size for nasal administration is greater than 5 um, preferably between 5 um and 120 um, preferably between 10 um and 60 um, preferably between 20 and 40 um, preferably between 20 and 30 um.

Thus, a bimodal distribution of particle size and/or aerodynamic size of the pharmaceutical composition in the form of powder for inhalation is advantageous and allows for nasal deposition and pulmonary deposition by nasal administration.

In the context of the present invention, "aerodynamic size" is preferably understood to mean a parameter describing the aerodynamic behavior of the particles; thus, this parameter takes into account the geometric diameter (therefore the particle size), but also the density and shape of the particles.

The particle size (and geometric diameter) is advantageously determined by laser diffraction. The aerodynamic size (and aerodynamic diameter) is advantageously determined using an impaction test (using cascade impactors) described in the pharmacopoeias (such as the European Pharmacopoeia for example).

Alternatively OR additionally, the pharmaceutical composition for activating macrophages and/or dendritic cells according to the invention is administered in the form of a dry powder and comprises at least one bulking agent such as a polyol and/or a sugar alcohol such as sorbitol, mannitol, maltitol (sometimes considered a disaccharide) and xylitol, sugars (crystalline) including monosaccharides (glucose, arabinose), disaccharides (lactose, maltose, sucrose, dextrose, trehalose), and polysaccharides (dextran, chitosan, starch, cellulose and its derivatives), or oligosaccharides (dextrins, cyclodextrins) and/or fatty acid salts (e.g. magnesium stearate) or fatty acids or their derivatives (e.g. esters), such as lauric acid, palmitic acid, stearic acid, erucic acid, behenic acid, or a phospholipid (e.g. lecithin, phosphatidylcholine phosphatidylglycerol; preferably not incorporated in the lipid vehicle), triglycerides (preferably not incorporated in the lipid vehicle), sugar esters, and/or an amino acid (see above for liquid composition) and mixtures thereof. Dextran, trehalose, mannitol and lactose are preferred.

Advantageously, an excipient (bulking agent) will be used, such as lactose, trehalose, sucrose or mannitol.

Advantageously, when the pharmaceutical composition is intended for administration to the patient in the form of dry powder for inhalation, the above excipients comprise at least one lipid or fatty acid-based substance (e.g. magnesium stearate, phospholipid, sucroester). This promotes good mixing homogeneity and/or advantageous aerosolization of the solid particles.

Another excipient is also advantageous for good aerodynamic properties, such as leucine. It should be noted that some surfactants include fatty acids, or even phospholipids or triglycerides, which are also potential components of the lipid vehicles of the present invention. However, these surfactants are not incorporated within the structure of the lipid vehicles and are advantageously incorporated into a dry formulation of these lipid vehicles already formed. In this case, the composition, even in powder form, will advantageously comprise a buffer and in addition one or more of the compounds listed above, for example a surfactant.

Examples of drying methods are freeze-drying, spray-drying, spray-congealing or spray-chilling, spray-freeze-drying, supercritical fluid drying.

Advantageously, this pharmaceutical composition is administrable into the respiratory tract, preferably by nebulization and/or by pressurized inhalation and/or by dry powder inhalation and/or by gentle spray inhalation (for example carried out using the Respimat® SofMist system).

A related aspect of the present invention is a pharmaceutical composition for activating macrophages and/or dendritic cells present in the respiratory tract (as described above), being in a lung tumor or in a central nervous system tumor.

Furthermore, according to another associated aspect of the present invention, this pharmaceutical composition for activating macrophages and/or dendritic cells is particularly advantageous in the case of metastatic cancer: - either the primary tumor is located in the lung or the central nervous system and the composition according to the present invention stimulates the activity of the immune system at the level of metastases outside the lung or the central nervous system (aloscopal effect), - or the primary tumor is located outside the lung or the central nervous system and a metastasis is found at the level of the lung or the central nervous system: the macrophages are activated there, and the immune system is then stimulated to attack the primary tumor and/or metastases outside the lung or the central nervous system.

Furthermore, according to another related aspect of the present invention, this pharmaceutical composition for activating macrophages and/or dendritic cells is particularly advantageous in combination with other therapies such as immunotherapies, preferably immune brake inhibitors, preferably an anti-PDI, an anti-PD-L1, an anti-CTLA-4 and their combinations, in cancer.

Other features and advantages of the present invention will be drawn from the non-limiting description which follows, and with reference to the drawings and examples.

Examples.-

It is understood that the present invention is in no way limited to the embodiments described above and that many modifications may be made thereto without departing from the scope of the appended claims.

Example 1. Lipid vehicles based on soy lecithin

The inventors first tested a lipid mixture of soy lecithin, cholesterol and DSPE-PEG, in a soy lecithin lipid: cholesterol: DSPE-PEG molar ratio of 8:1:0.05. Briefly, the lipids were first weighed in a container, then dissolved, here in a dichloromethane:methanol mixture (50:50; V:V) with stirring for 30 minutes. When solubilization is complete, the solvent is evaporated (rotovapor) at 30 ° C under a pressure of 16000 Pa for 1 hour, leading to the formation of the lipid film. The film is then subjected to an air flow to remove traces of residual solvent. An aqueous solution (water or buffer) is then added to the film and the mixture is stirred for about 30 minutes at a temperature above the highest phase transition temperature characterizing the lipids in the mixture. The suspension is then extruded, here using the

Liposofast LP-50 (4 extrusions through 3 filters - 1, 0.4 and 0.1 um, at 600 psi; therefore approximately 4.1 106 Pa) to finally obtain the lipid vehicles.

The lipid vehicles obtained had a mean diameter (Dh) of the order of 110 to 150 nm and polydispersity indices (Pdl)

of approximately 0.1 to 0.2; the zeta potential was neutral to slightly negative in the absence of DSPE-PEG and strongly negative in its presence (-20 to -35

MV). For a zeta potential measurement, the sample was diluted in a 0.009% NaCl solution (mass/volume) so as to obtain attenuator and conductivity values suitable for a good measurement.

This formulation was taken up for the entrapment of a pharmaceutical agent, here, 2'3'-CGAMP (CGAMP) with entrapment efficiencies (EE%) of 30 to 45% obtained. The best results were obtained for compositions involving a higher cGAMP/lipid ratio (0.08 mole/mole of lipid). These formulations, in addition to presenting low EE% values, did not show any improvement in the activation of THP1-dual macrophages (THP1-

Dual"M Cells, invivogen #thpd-nfis) in vitro.

Example 2 — Lipid vehicles based on

DOTAP/cholesterol

The inventors used a cationic lipid DOTAP (lipid 1 according to the present invention) with cholesterol (lipid 2) at a concentration of 3.9 mg/mL and 1.4 mg/mL respectively and by incorporating 100 ug/ML of 2'3'-CGAMP (CGAMP). These vehicles were prepared in the absence (composition 1) or in the presence of DSPE-PEG at 0.05 mole equivalents (relative to cholesterol).

The physical characteristics are shown in the table below: 2| 128.2+0.76 [0.104+0015| 6832055

In addition, the inventors obtained EE% close to 100%.

In vitro THP1-dual macrophage activation assays (THP1-DualTM

Cells, invivogen #thpd-nfis) showed a 4-fold increased efficacy for both compositions, compared to the same concentration of free cGAMP. Similarly, the inventors verified in vitro that the DOTAP/cholesterol-based formulations better protect CGAMP from enzymatic degradation by observing a better preservation of THP1-dual macrophage activation after in vitro incubation of lipid vehicles (vs free CGAMP) with the enzyme

Recombinant human ENPPI (#6136-EN-010, biotechne, USA).

Although the addition of DSPE negatively affected the polydispersity index pdi, the inventors note that this addition is not too problematic.

Example 3 - Effect of the addition of DSPE-PEG

The inventors then tested different proportions in

DSPE-PEG (Figure 1). Composition 2 above is retained. Other compositions (3-5) are developed with 0.1, 0.5 and 1 mole equivalents of DSPE-PEG. The cGAMP loading rate is then reduced from 1.88% (composition 2), then 1.55%, then 0.94% and finally 0.60%.

Composition 3 shows properties (diameter, pdi, % encapsulation) very close to composition 2. The particle diameter is reduced to 115 then 55 nm at compositions 4-5, while the pdi increases to about 0.26. The zeta potential drops to almost 0 for composition 5. EE% drops to about 89% then 62% at compositions 4-5. Composition 3 still allows very good activation of macrophages

THP 1-dual in vitro, reduced for 4, and the effect is almost completely lost for 5.

On the other hand, compositions 3 and 4 offer better protection of cGAMP against enzymatic degradation by observing a better preservation of the activation of THP1-dual macrophages after in vitro incubation of lipid vehicles (vs free cGAMP) with the recombinant human ENPPT enzyme.

Example 4 — functionalization

Compositions 2 and 4 (0.05 and 0.5 mole equivalents in DSPE-

PEG) served as a basis, with replacement of either the entire DSPE-

PEG by DSPE-PEG-mannose (compositions 6 and 7), i.e. half

IO (compositions 8 and 9).

The diameter and pdi index increase sharply for composition 7. The pdi index also increases for composition 9.

The EE% remains close to 100% each time.

The inventors then carried out an activation test of immunosuppressive macrophages (type M2) or tumor-associated macrophages (TAMs) LLC1 (Lewis lung carcinoma, ATCC CRL1642) obtained according to a protocol similar to that described in Kwart et al. (Cell

Reports 41 (2022) 111769), from bone marrow-derived macrophages (BMDMs). Composition 6 shows a much higher activation than composition 2 on both types of macrophages (M2 and TAMs), as illustrated in Figure 2 for M2. The inventors also observed that composition 6 administered endotracheally to healthy mice induced activation of alveolar macrophages and dendritic cells.

Example 5 - increasing the load rate

In order to increase the cGAMP loading rate in the lipid vehicles, the amount of DOTAP was reduced to 1.5 mole equivalents relative to cholesterol and the amount of both lipids was reduced to 1/10 relative to composition 1 for compositions 10-16.

Different concentrations of cGAMP were tested from 100 (composition 10) to 150, 200, 250, 300, 360, and 720 ug/ML. The loading rate increased from 16.86 (composition 10) to 23.33; 28.86; 33.65; 37.83; 42.20, and 59.35%. The DOTAP/CGAMP ratio decreased from 3.5/1 (composition 10) to 2.35/1; 1.76/1; 1.4/1; 1.2/1, 1/1, and 0.5/1. The particle diameter increased slightly up to composition 12 (loading rate 28.86%), then sharply when the cGAMP concentration was increased. The pdi was constant up to composition 12, then increased sharply.

Conversely, the zeta potential falls beyond composition 12.

EE% remains close to 100% up to this composition. THP1 dual macrophage activation remains significantly higher than for free CGAMP.

Starting from composition 12, the addition of DPPC to DOTAP (here, in a molar ratio 2:1) is possible, but the loading rate in

CGAMP is reduced. Dual THP] macrophage activation remains significantly higher than for free cGAMP, but reduced by half at low cGAMP concentrations. However, this type of formulation is interesting because it is associated with increased stiffness (also reflected by an increased Tm of the lipid series), which is useful in some modes of administration.

Example 6: Improving the stiffness properties of formulations

The inventors observed an instantaneous release of cGAMP trapped in compositions when these formulations were diluted in a large volume of PBS. In order to overcome this limitation, the inventors prepared new formulations by adding an excipient with a higher TM (DPPC). Starting from composition 12, DPPC was added at 3, 5, 8 and 10 mole equivalents relative to cholesterol for compositions 13, 14, 15 and 16, respectively. The results demonstrated that increasing the amount of DPPC decreases the fraction of CGAMP instantly released after a 200X dilution in PBS. Compositions 12, 13, 14, 15 and 16: 100%; 59.48; 69.57; 47.83: 32.18%.

Example 7- use of an ionizable lipid

Starting from composition 14, the inventors replaced the

DOTAP by DODMA in the same molar proportions. The lipid vehicle preparation protocol described above was adapted so that the lipid film rehydration buffer was a 0.01M acetate buffer at pH 5.0 in order to ionize DODMA and thus promote cGAMP trapping. The EE% was similar to that obtained for condition 14.

Example 8- combining a composition with a standard treatment

A composition of the present invention was administered endotracheally in combination with an anti-PD1 monoclonal antibody (clone RPM1-14 #BP0146, BioXcell) to mice bearing lung tumors. The treatment combination resulted in superior activity to the corresponding monotherapies.

Claims (27)

1. Composition = pharmaceutical = for activating macrophages and/or dendritic cells comprising lipid vehicles formed from a series of lipids with at least one lipid having a functionalization by a ligand of the mannose receptor (CD206), said lipid vehicles being selected from the group consisting of liposomes, vesicles, micelles, lipoplexes, lipid emulsions, lipid nanocrystals, lipid microspheres, lipid nanoparticles, and mixtures thereof, said lipid vehicles comprising a pharmaceutical agent selected from the group consisting of - a nucleic acid, - a STING protein agonist or an antagonist of the STING protein, - a ligand of Toll-like receptors, - an immunomodulant, a peptide, OR - a mixture thereof. 2. = Pharmaceutical = composition for activating macrophages and/or dendritic cells according to claim 1, in which the series of lipids forming the lipid vehicles has a phase transition temperature (TM) of between 10°C and 80°C, preferably between 20°C and 60°C. 3. Composition = pharmaceutical = for activating macrophages and/or dendritic cells according to claim 1 or 2 being for inhalation. 4. Pharmaceutical composition = for activating macrophages and/or dendritic cells according to any one of the preceding claims, in which the lipid vehicles are lipid nanoparticles (LNPs), preferably liposomes or solid lipid nanoparticles (SLN) or nanostructured lipid carriers (NLC). 5. Composition = pharmaceutical = for activating macrophages and/or dendritic cells according to any one of the preceding claims, in which the pharmaceutical agent has an overall negative charge. 6. = Pharmaceutical = composition for activating macrophages and/or dendritic cells according to any one of the preceding claims, in which the series of lipids comprises a first lipid being cationic and/or ionizable. 7. = Pharmaceutical = composition for activating macrophages and/or dendritic cells according to claim 6, in which the lipid vehicles have a molar proportion of said cationic and/or ionizable lipid relative to the pharmaceutical agent of between 30 and 0.1, preferably between 10 and 0.2, advantageously between 3 and 0.5, preferably between 2.5 and 0.8, preferentially between 2.4 and 1. 8. = Pharmaceutical = composition for activating macrophages and/or dendritic cells according to claim 6 or 7, in which the series of lipids comprises a second lipid selected from the group of sterols, a phospholipid, a triglyceride, a diglyceride, a glycerophospholipid and a sphingomyelin, preferably a sterol or a phospholipid, very preferably a sterol. 9. = Pharmaceutical = composition for activating macrophages and/or dendritic cells according to claim 8, in which the lipid vehicles have a molar ratio of the first lipid to the second lipid of between 0.3 and 20, preferably between 0.5 and 10, preferably between 1 and 5 (moles of the first lipid: moles of the second lipid), said second lipid preferably being a sterol, such as cholesterol and/or a phospholipid such as DPPC and/or DSPC. 10. = Pharmaceutical = composition for activating macrophages and/or dendritic cells according to any one of the preceding claims, in which said at least one lipid having a functionalization by a ligand of the mannose receptor (CD206) is a phospholipid having a functionalization by a ligand of the mannose receptor (CD206), preferably a phospholipid-PEG having a functionalization by a ligand of the mannose receptor (CD206), advantageously a DSPE-PEG having a functionalization by a ligand of the mannose receptor (CD206). 11. Pharmaceutical composition = for activating macrophages and/or dendritic cells according to any one of the preceding claims, in which said ligand of the mannose receptor (CD206) is selected from the group comprising a mannose, a fucose, an N-acetylglucosamine, an N-acetylgalactosamine, a glycoprotein, a galactocomannan, an a-D-mannopyranoside, a polymannose, an anti-CD206 antibody, and mixtures thereof. 12. = Pharmaceutical = composition for activating macrophages and/or dendritic cells according to any one of the preceding claims, in which the lipid vehicles have a molar proportion of said at least one lipid having a functionalization by a ligand of the mannose receptor (CD206) to the total lipids of between 0.01 and 0.2, preferably of between 0.02 and 0.1, preferably between 0.03 and 0.05. 13. Composition = pharmaceutical = for activating macrophages and/or dendritic cells according to any one of the preceding claims, in which the lipid vehicles have a molar proportion of said at least one pharmaceutical agent relative to said at least one lipid having a functionalization by a ligand of the mannose receptor (CD206) of between 5 and 150. 14. = Pharmaceutical = composition for activating macrophages and/or dendritic cells according to any one of the preceding claims, in which said pharmaceutical agent is present in a loading rate of between 0.1 and 80% relative to the mass of the lipid vehicles, preferably between 1 and 70%, preferably between 10 and 60%, preferably between 15 and 50%, such as between 20 and 40% or even between 25 and 30%. 15. = Pharmaceutical = composition for activating macrophages and/or dendritic cells according to any one of the preceding claims, in which the lipid vehicles have an average size, measured by dynamic light scattering (DLS), of between 50 and 200 nm, preferably between 60 and 190 nm, advantageously between 70 and 180 nm, preferentially between 80 and 180 nm, preferably between 90 and 180 nm, more preferably between 100 and 180 nm, preferably between 110 and 180 nm, advantageously between 120 and 180 nm, preferentially between 120 and 170 nm, preferably between 120 and 160 nm. 16. = Pharmaceutical = composition for activating macrophages and/or dendritic cells according to any one of the preceding claims 1 to 14, in which the lipid vehicles have an average size, measured by dynamic light scattering (DLS), of between 350 and 1500 nm, preferably between 350 and 1400 nm, advantageously between 350 and 1300 nm, preferentially between 350 and 1200 nm, advantageously between 350 and 1100 nm, preferentially between 350 and 1000 nm, preferably between 350 and 900 nm, preferably between 350 and 800 nm, more particularly between 350 and 700 nm, advantageously between 350 and 600 nm, for example between 400 and 600 nm. 17. Pharmaceutical composition = for activating macrophages and/or dendritic cells according to any one of the preceding claims further comprising a metal ion, preferably chosen from Mn, Co2+ or Zn?*, and/or at a content of at least 10 PPM. 18. Pharmaceutical composition = for activating macrophages and/or dendritic cells according to any one of the preceding claims, in which the lipid vehicles have a polydispersity index of between 0.05 and 0.9, preferably between 0.1 and 0.8, advantageously between 0.2 and 0.75, preferentially between 0.3 and 0.7, preferably between 0.4 and 0.6 measured by dynamic light scattering (DLS), preferably this polydispersity index being defined by the formula: y VAO (Va? Vits) PI=2T5 0 dt a VAO, i=t where = DS is the average of the particle diameter, Oet: the quantity of particles with a size Xi and xi being the diameter of the spherical particle. 19. = Pharmaceutical = macrophage and/or dendritic cell activation composition according to any one of the preceding claims, in which the lipid vehicles have a zeta potential of between -60 mV and 100 mV, preferably between -40 mV and 80 MV, advantageously between -20 mV and 60 MV, preferentially between 0 mV and 50 mV, preferably between 10 mV and 40 mV, advantageously between 10 mV and 30 mV, preferably measured by laser doppler electrophoresis, preferably in an aqueous solution of NaCl at 0.009% (mass:volume). 20. Composition = pharmaceutical = for activating macrophages and/or dendritic cells according to any one of the preceding claims being in dry form, preferably intended to be dissolved or redispersed in a physiological aqueous solvent, or to be administered using a dry powder inhaler. 21. Composition = pharmaceutical = for activating macrophages and/or dendritic cells according to any one of the preceding claims 1 to 19 comprising from 50 to 98% water (mass of water: total mass of the composition), preferably from 70 to 95% water, or even from 80 to 90% water. 22. Composition = pharmaceutical = for activating macrophages and/or dendritic cells according to any one of the preceding claims further comprising a buffer selected from the group comprising phosphate, sulfonate buffers (MES, TES, HEPES, MOPS, PIPES, TAPS, TAPSO), acetate, bicine, and/or at least one salt selected from the group consisting of inorganic salts and organic salts, and/or at least one surfactant selected from the group consisting of cholic acids and their salts, phospholipids (e.g. phosphatidylcholines or lecithin, phosphatidylglycerols), lipids or triglycerides, sorbitan esters, polyethoxylated sorbitans, fatty acids, preferably lauric, palmitic, stearic, erucic or behenic acid, esters of these fatty acids, or derivatives of these fatty acids, such as salts, preferably chosen from magnesium stearate, sodium stearyl fumarate and sodium stearyl lactylate, sodium lauryl sulfate, magnesium lauryl sulfate, natural pulmonary surfactants; and sucroesters (sugar esters, for example esters between sucrose OR glucose and fatty acids) and/or a sugar, preferably selected from the group consisting of monosaccharides (such as glucose or arabinose), disaccharides (such as lactose, maltose, sucrose, dextrose, trehalose, maltitol and mixtures thereof or a bulking agent selected from polyols such as sorbitol, mannitol and xylitol), polysaccharides (such as dexiran, chitosan, starch, cellulose, and derivatives thereof), oligosaccharides (such as cyclodextrin and dexirins), and/or at least one amino acid selected from the group consisting of histidine, leucine, isoleucine, threonine, lysine, valine, methionine, phenylalanine, mixtures thereof and their derivatives, such as acesulfame K or aspartame. 23. Composition = pharmaceutical = for activating macrophages and/or dendritic cells according to any one of the preceding claims being administrable by nebulization and/or by pressurized inhalation and/or by dry powder inhalation and/or by gentle spray inhalation. 24, Composition = pharmaceutical = for activating macrophages and/or dendritic cells according to any one of the preceding claims being for the treatment of a cancer, preferably a metastatic cancer, preferably said metastasis being outside the lung or the central nervous system and the primary tumor being pulmonary or located at the level of the central nervous system, or said metastasis being pulmonary or located at the level of the central nervous system and the primary tumor being outside the lung or the central nervous system. 25. Composition = pharmaceutical = for activating macrophages and/or dendritic cells according to any one of the preceding claims 1 to 23 being for the treatment of an infectious disease at the level of the respiratory and/or systemic tract, or of the central nervous system. 26. Pharmaceutical composition according to any one of the preceding claims 1 to 23 comprising a nucleic acid encoding one or more epitope(s), or a peptide comprising one or more epitopes, said pharmaceutical composition being for vaccination. 27. A method for obtaining the pharmaceutical composition for activating macrophages and/or dendritic cells according to any one of the preceding claims, comprising the steps of - solubilizing the lipids in an organic solvent or in a mixture of organic solvents, - evaporating the solvent so as to form a lipid film, - rehydrating with a solution containing the pharmaceutical agent, - extruding and inserting a lipid derivatized with a ligand of the CD206 receptor, - preferably a second step of bringing a solution containing the pharmaceutical agent into contact with the extruded lipids, said lipids comprising a neutral lipid at pH 70 and positively charged at pH 5, and said rehydration step being carried out at a pH lower than 7.0.