Nucleic acid vector tablets

Abstract

To provide a method for producing a tablet comprising a nucleic acid vector.SOLUTION: According to a first aspect, the invention relates to a process for the manufacture of a tablet comprising a complex between (i) a nucleic acid, and (ii) a lipid particle comprising a cationic lipid, the lipid particle being preferably a liposome or a micelle, preferably a liposome, the process comprising the following steps of: a) drying, in particular by freeze-drying, an aqueous mixture comprising a complex between (i) a nucleic acid and (ii) a lipid particle comprising a cationic lipid, the lipid particle being preferably a liposome or a micelle, preferably a liposome, and one or more excipient(s) suitable for drying, in particular for freeze-drying; b) mixing the one or more compression excipient(s) with the dry mixture obtained in the step a); and c) compressing the mixture obtained in the step b).SELECTED DRAWING: None

Description

[Technical field]

[0001] The present invention relates to a method for producing a tablet containing a nucleic acid vector. [Background technology]

[0002] Gene therapy uses nucleic acids (especially DNA or RNA) to treat or prevent diseases. Depending on the pathology, the nucleic acid is administered to regulate, repair, replace, add or delete a gene sequence. The therapeutic, preventive or diagnostic effect depends directly on the nucleic acid sequence or the product of the gene expression of that sequence. Therapeutic nucleic acids include, for example, antisense oligonucleotides, siRNA and shRNA. In particular, molecular interfering RNA, or siRNA, combined with small vectors, constitute agents with high therapeutic potential due to their ability to specifically suppress the expression of pathological proteins. Therapeutic nucleic acids can be directly injected into cells as naked nucleic acids, but they are most often transported into the patient's cells using nucleic acid vectors. The nucleic acid vectors can be viral vectors based on polymer or lipid systems, or synthetic vectors.

[0003] Currently, nucleic acid vectors are mainly administered parenterally in the form of colloidal suspensions. This type of formulation and administration route imposes strong limitations, such as the size of the administered vector and the acquisition of a sterile preparation. These limitations are obstacles to the development of nucleic acid vectors as potential new drugs. [Prior art documents] [Patent documents]

[0004] [Patent Document 1] International Publication WO15023775 Summary of the Invention [Problem to be solved by the invention]

[0005] Therefore, there is a need for new formulations of nucleic acid vectors to avoid the above-mentioned limitations while maintaining the biological activity of therapeutic nucleic acids. [Means for solving the problem]

[0006] According to a first aspect, the present invention provides a method for producing a method for treating a cancer cell comprising: (i) a nucleic acid; and (ii) a lipid particle comprising a cationic lipid, preferably a liposome or a micelle, preferably a liposome; The present invention relates to a method for producing a tablet comprising a complex of The method includes: a) - (i) a complex of a nucleic acid and (ii) a lipid particle comprising a cationic lipid, preferably a liposome or a micelle, preferably a liposome; - one or more additives suitable for drying, in particular for freeze-drying; drying the aqueous mixture comprising said compound, in particular by freeze-drying; b) mixing one or more compression additives with the dry mixture obtained in step a); and c) compressing the mixture obtained in step b) Includes.

[0007] In a particular embodiment, the cationic lipid is selected from lipopolyamines, quaternary ammonium, and lipids with cationic head groups of the guanidine or imidazole type, preferably the cationic lipid is selected from dimyristylaminopropylaminopropyl (DMAPAP), N-[1-(2,3-dioleyloxy)propyl]-N,N,N-trimethylammonium chloride (DOTMA), [1,2-bis(oleoyloxy)-3-(trimethylammonio)propane] (DOTAP), 3β[N-(N′,N′-dimethylaminoethane)-carbamoyl]cholesterol (DCChol) and dioctadecylamidoglycylspermine (DOGS), preferably dimyristylaminopropylaminopropyl (DMAPAP). The cationic lipid may be optionally mixed with a neutral lipid, in particular selected from 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), cholesterol, dioleoyl-sn-glycero-3-phosphocholine (DOPC), dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) and N'-(rac-1-[11-(F-octyl)undec-10-enyl]-2-(hexadecyl)glycero-3-phosphoethanoyl)-sperminecarboxamide), preferably the neutral lipid is 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE). In particular, the mixture of cationic lipid and neutral lipid may contain 50 to 99 mol%, preferably about 50 mol%, of cationic lipid in the mixture, and may contain 1 to 50 mol%, preferably about 50 mol%, of neutral lipid in the mixture.

[0008] According to certain embodiments, the complex comprises a therapeutic nucleic acid, in particular an siRNA, miRNA, shRNA, plasmid or mRNA. According to certain embodiments, the charge ratio (+ / -) between the positive charge of the cationic lipid and the negative charge of the nucleic acid is 0.5-10, preferably 4-10, preferably about 8.

[0009] According to the method of the invention, the additive suitable for drying, in particular for freeze-drying, may be a protective agent (cryoprotectant or lyophilization protectant) and / or a bulking agent, in particular selected from trehalose, mannitol, sucrose, sorbitol, lactose, glucose, glycerin, glycine, alanine, lysine, polyethylene glycol, polyvinylpyrrolidone and dextran, optionally in a mixture. In particular, the additive suitable for freeze-drying is trehalose, in a proportion of 10 to 100% by weight of the additive suitable for freeze-drying, optionally mixed with another additive suitable for freeze-drying, preferably mannitol, in a proportion of 0 to 90% by weight of the additive suitable for freeze-drying.

[0010] According to the method of the invention, the compression additive protects the complex of the lipid particles comprising the nucleic acid and the cationic lipid during the compression step c), said compression additive being in particular a lubricant (e.g. magnesium stearate, stearic acid and talc, preferably magnesium stearate), optionally mixed with another compression additive, preferably an excipient (e.g. lactose, starch, calcium phosphate and its derivatives and cellulose and its derivatives, preferably lactose). In particular, the lubricant may represent 0.25 to 5% by weight of the finished tablet, preferably 0.5%.

[0011] In a particular embodiment, the method according to the invention comprises a compression step (step c) carried out at a pressure between 10 and 400 MPa, preferably between 50 and 250 MPa.

[0012] In another aspect, the present invention relates to a tablet obtainable according to the above process. The tablet of the present invention comprises: - liposome or micelle complexes containing nucleic acids and cationic lipids; - one or more suitable lyophilization excipients, preferably trehalose and mannitol, to protect the complex during the lyophilization step a) and / or to obtain a lyophilizate with a suitable texture for the compression step; - one or more suitable compression additives to protect the complex during compression step c) and to maintain the biological activity of the complex. may include.

[0013] According to a particular embodiment, the tablets of the invention are intended for oral, vaginal, rectal, sublingual or transmucosal administration, preferably by the oral route.

[0014] The present invention also relates to a tablet as described above for use in treating a gastrointestinal or vaginal disorder. [Brief description of the drawings]

[0015] [Figure 1] Freeze-drying cycle: Representation of product temperatures during the three stages of the cycle; (a) initiation of freezing, primary drying and secondary drying; (b) focus on freezing in liquid nitrogen. [Diagram 2] Efficacy of tablets containing lipoplexes as a function of the lyophilisate (=lyoph) and the compression stress applied during the compression method (CX at X=50, 100, 150, 200 or 250 MPa): 48 h transfection into B16-Luc cells, triplicate, 0.5 μg luciferase siRNA per well. Measurement of luciferase and indication of the inhibition rate compared to non-transfected cells. Positive control (T+): cells transfected with lipoplexes prepared immediately before use (neither lyophilised nor compressed), negative control (siCtle): cells transfected with lipoplexes containing control siRNA resuspended from the tablets. [Diagram 3] SAXS profile of siRNA lipoplexes (DMAPAP / DOPE+alginate), prepared in 150 mM NaCl buffer at a + / - charge ratio of 8. Peak analysis identifies two phases, a major cubic peak (C1, q=0.084 nm-1) and a minor lamellar peak (L1, q=0.101 nm-1). [Figure 4]SAXS profile of siRNA lipoplexes in dry powder form after lyophilization and subsequent compression steps. siRNA lipoplexes (DMAPAP / DOPE+alginate) prepared at + / - charge ratio 8 in 150 mM NaCl buffer are supplemented with trehalose and maltose and then lyophilized for 30 hours. The lyophilisate is weighed, supplemented with lactose and compressed. SAXS analysis is performed on the lyophilisate or tablet powder placed in a capillary tube. A single large peak (q=0.096 nm-1) is detected. [Diagram 5] SAXS profile of siRNA lipoplexes suspended in NaCl / sugar. Lipoplexes prepared as shown in the legend of Figure 3 are supplemented firstly with a trehalose / mannitol mixture or secondly with a trehalose / mannitol / lactose mixture under the same conditions as the lyophilized or compressed samples, respectively. Peak analysis identifies a single cubic phase, with the position of the first peak slightly shifted between the geometry of the first mixture (C1, q=0.095 nm-1) and that of the second mixture (C1, q=0.097 nm-1). [Figure 6] SAXS profiles of suspended siRNA lipoplexes. The first curve corresponds to a sample that was (A) freeze-dried, then (B) compressed (see legend to Fig. 4) and then resuspended by adding water. The second curve corresponds to a suspended sample that was not freeze-dried or compressed, but supplemented with the same amount of (A) trehalose / mannitol as the freeze-dried resuspended sample or with the same amount of (B) trehalose / mannitol / lactose as the resuspended compressed sample. Peak analysis identifies a single phase in the figure corresponding to the second curve of cubic type (C1, (A) q=0.094 nm-1; (B) q=0.096 nm-1), a cubic phase in the figure of the first curve in (A) (C1, q=0.093 nm-1) and two phases in the figure of the first curve in (B) that are cubic (C1, q=0.086 nm-1) and lamellar (L1, q=0.101 nm-1). DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0016] The present inventors propose to formulate the nucleic acid vector in tablet form for oral, vaginal, rectal, sublingual or transmucosal administration. The "tablet" form facilitates administration while avoiding the formulation and administration constraints mentioned above. In addition, the tablet allows many formulation options to control or regulate the release of the nucleic acid vector based on the therapeutic target. The "tablet" form also represents a breakthrough in the stability of the nucleic acid vector. In particular, it makes it possible to avoid their preparation immediately before use. In addition, the "tablet" form improves both patient comfort and compliance with treatment.

[0017] Prior to the present invention, nucleic acid vector tablets were not available or even conceivable to those skilled in the art due to fears of loss of nucleic acid activity during the steps required to form the tablets. However, contrary to this prejudice, the present inventors propose an innovative means of generating nucleic acid vector tablets.

[0018] The present invention relates to a method for manufacturing a tablet comprising at least one nucleic acid vector. More specifically, the proposed invention makes it possible to formulate nucleic acid vectors in a "tablet" form, while maintaining their therapeutic properties despite the mechanical stresses involved in this forming operation.

[0019] definition "Tablet" means any uniform solid preparation without particular shape or size obtained from compressed powder, which may or may not be agglomerated.

[0020] "Nucleic acid vector" (otherwise referred to below as "complex") refers to any structure known to be suitable for vectorizing nucleic acids; in particular, these terms refer to a complex of (i) a nucleic acid and (ii) a lipid particle comprising a cationic lipid, preferably a liposome or micelle, preferably a liposome.

[0021] "Lipoplex" means a complex of (i) a nucleic acid and (ii) a liposome containing a cationic lipid.

[0022] "Cationic lipid" means a lipid that has an overall positive charge. A cationic lipid contains a cationic polar head and one or more hydrophobic chains.

[0023] "Neutral lipid" means a lipid having a neutral overall charge, more specifically, a zwitterionic lipid.

[0024] "Micelle" means a spherical aggregate of amphiphilic molecules having hydrophilic polar heads and hydrophobic chains.

[0025] "Liposome" means an artificial vesicle formed by concentric lipid bilayers enclosing an aqueous compartment between them.

[0026] For example, the nucleic acid vector can be selected from liposomes, lipids, such as cationic lipids, anionic lipids, amphoteric lipids or uncharged lipids, cationic polymers, polymers, hydrogels, micro- or nanocapsules (biodegradable), microspheres (optionally bioadhesive), cyclodextrins, protein vectors, or any combination thereof. Preferably, the vector can be selected from lipid-based delivery systems, polyimine-based delivery systems, dendrimers, poly(lactide-co-glycolide) (PLGA) particles, systems described in International Publication WO15023775, and similar systems.

[0027] "About" refers to a value + or -10%, preferably + or -5% thereof. For example, about 50 is 45 to 55, preferably 47.5 to 52.5.

[0028] Preparation of nucleic acid vectors The nucleic acids may in particular be deoxyribonucleic acids (DNA) or ribonucleic acids (RNA) of any size, single or double stranded, linear or circular, chemically unmodified or modified (such as phosphorothioate, phosphoramidate and / or 2'-O-methyl type). The size of the nucleic acids may vary widely, for example from 10 base pairs to tens of thousands of base pairs. For example, the size of the nucleic acids may reach about 10,000 base pairs or more for plasmids, about 19-25 nucleotides for double-stranded siRNAs or about 20-30 nucleotides for single-stranded microRNAs. They may be natural, hybrid or synthetic sequences of any origin (prokaryotic, eukaryotic, viral, parasitic, plant, etc.). Preferably, the nucleic acids are therapeutic nucleic acids, i.e. nucleic acids that have a therapeutic effect by regulating, repairing, replacing, adding or deleting gene sequences. Preferably, the nucleic acid is capable of modulating the expression of a protein and can be selected in particular from antisense oligonucleotides, oligonucleotides for exon skipping, oligonucleotides for alternative splicing modification, interfering RNA, messenger RNA or plasmid DNA. Preferably, the nucleic acid is an interfering RNA. In the context of the present invention, an "interfering RNA" is an RNA that inhibits gene expression. In particular, an interfering RNA is a small (in particular 20-25 base pairs, in particular 21-23 base pairs) ribonucleic acid that interferes with a specific messenger RNA, leading to its degradation and the inhibition of its translation into a protein. For example, small interfering RNA (siRNA) and microRNA (miRNA) are interfering RNAs.

[0029] In particular, the nucleic acid is selected from a plasmid, an siRNA, an miRNA, an mRNA and an shRNA, preferably the nucleic acid is an siRNA or an miRNA. Preferably, the nucleic acid is an siRNA.

[0030] The tablet produced according to the invention comprises one or more nucleic acid vectors (or complexes) as defined above. According to a particular embodiment, the tablet comprises a combination of several different nucleic acid vectors, in particular a combination of at least 2, at least 3, at least 4 or even at least 5 different nucleic acid vectors. According to another particular embodiment, the tablet comprises a combination of one or more nucleic acid vectors (or complexes) and at least one probiotic.

[0031] "Probiotic" means a live microorganism that, when administered in sufficient amounts, provides a positive health effect beyond conventional nutritional benefits, such as improved digestion or improved immune defenses.

[0032] According to the present invention, the complex is formed by electrostatic interaction between the negatively charged nucleic acid and the lipid particle comprising the cationic lipid.

[0033] Preferably, the lipid particle comprising the cationic lipid is a micelle or a liposome.

[0034] Even more preferably, the lipid particle is a liposome. According to this embodiment, the complex formed between the nucleic acid and the liposome is called a "lipoplex."

[0035] According to a particular embodiment, the present invention provides: (i) a nucleic acid; and (ii) lipid particles comprising cationic lipids, which are preferably liposomes or micelles, and even more preferably liposomes; The present invention relates to a method for producing a tablet comprising the complex of

[0036] According to certain embodiments, the method of the invention comprises a step of complexing the nucleic acid with a lipid particle comprising a cationic lipid, preferably a liposome or micelle, preferably a liposome, prior to the drying step described in detail below.

[0037] According to a particular embodiment, the nucleic acid can be premixed with an anionic polymer before the step of forming a complex between the nucleic acid and the lipid particle. The anionic polymer is an adjuvant for improving the structure of the complex between the nucleic acid and the lipid particle, and can therefore increase the effectiveness of the complex. The anionic polymer can be selected from anionic polysaccharides and anionic polypeptides, and preferably, the anionic polymer is selected from sodium alginate and sodium polyglutamate. According to a particular embodiment, the anionic polymer is an alginate, in particular sodium alginate. Therefore, the manufacturing method according to the present invention may also include a step of mixing the nucleic acid with the anionic polymer before the step of forming a complex between the nucleic acid and the lipid particle.

[0038] In addition, the method according to the invention may comprise a prior step of forming lipid particles, preferably liposomes or micelles, comprising cationic lipids, preferably liposomes.

[0039] Preferably, the cationic lipid is selected from cationic lipids that can form particles by combining the hydrophobic chains of several lipids. The cationic lipid according to the present invention can be selected from lipopolyamines, quaternary ammonium, and lipids with cationic heads of guanidine or imidazole type. Preferably, the cationic lipid is selected from dimyristylaminopropylaminopropyl (or DMAPAP), N-[1-(2,3-dioleyloxy)propyl]-N,N,N-trimethylammonium chloride (or DOTMA), 1,2-bis(oleoyloxy)-3-(trimethylammonio)propane] (or DOTAP), 3β[N-(N',N'-dimethylaminoethane)-carbamoyl]cholesterol (DCChol), and dioctadecylamidoglycylspermine (or DOGS). Preferably, the cationic lipid is dimyristylaminopropylaminopropyl (or DMAPAP).

[0040] According to a particular embodiment, the cationic lipid is mixed with a neutral lipid.Preferably, the neutral lipid is selected from 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), cholesterol, dioleoyl-sn-glycero-3-phosphocholine (DOPC), dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) and N'-(rac-1-[11-(F-octyl)undec-10-enyl]-2-(hexadecyl)glycero-3-phosphoethanoyl)-sperminecarboxamide).Preferably, the neutral lipid is 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE).

[0041] Thus, a tablet according to the invention may comprise a complex of (i) a nucleic acid and (ii) a lipid particle comprising a cationic lipid optionally mixed with a neutral lipid.

[0042] According to a preferred embodiment, the tablet according to the invention comprises a complex of (i) a nucleic acid and (ii) a lipid particle comprising a mixture of cationic lipids and neutral lipids. Preferably, the lipid particle is a micelle or liposome, preferably a liposome.

[0043] The mixture of cationic lipids and neutral lipids contains cationic lipids and neutral lipids in a specifically defined ratio. Preferably, the ratio of cationic lipids is 50-99 mol% of the lipids in the mixture. In particular, the ratio of cationic lipids may be about 50 mol%, about 60 mol%, about 70 mol%, about 80 mol% or about 90 mol% of the lipids in the mixture. Preferably, the ratio of cationic lipids is about 50 mol% of the lipids in the mixture.

[0044] Preferably, the ratio of neutral fats is 1 to 50 mol% of lipids in the mixture. In particular, the ratio of neutral fats may be about 10 mol%, about 20 mol%, about 30 mol%, about 40 mol%, or about 50 mol% of lipids in the mixture. Preferably, the ratio of neutral fats is about 50 mol% of lipids in the mixture.

[0045] According to a preferred embodiment, the cationic lipid is mixed with the neutral lipid in a 1:1 equimolar ratio.

[0046] The method according to the invention may include a step of mixing the cationic lipids and neutral lipids prior to the step of forming lipid particles, which are then used during the step of forming complexes with nucleic acids.

[0047] According to a particular embodiment, the mixture of cationic lipids and neutral lipids is a mixture of DMAPAP and DOPE.

[0048] According to a particular embodiment, the nucleic acid of the invention is mixed with the lipid particles of the invention according to a defined charge ratio. The "charge ratio" corresponds to the + / - ratio of the number of positive charges of the cationic lipid and the number of negative charges of the nucleic acid. This charge ratio can affect the effectiveness of the complex formed by influencing its physicochemical properties (e.g. size or surface charge). The charge ratio can be different and adapted by the skilled person depending on the lipid and nucleic acid selected and the desired application (target organ or cell type). Preferably, the + / - charge ratio is between 0.5 and 10, for example about 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10. Preferably, the + / - charge ratio is between 4 and 10, for example about 4, 5, 6, 7, 8, 9 or 10. Even more preferably, the charge ratio is about 8.

[0049] According to certain embodiments, the mixture of nucleic acid and lipid particles is an aqueous mixture. Thus, the method according to the invention may comprise the step of preparing an aqueous mixture of nucleic acid and lipid particles comprising cationic lipids.

[0050] Preferably, the aqueous mixture of the nucleic acid and lipid particles is carried out in a buffer solution. In particular, the pH of said buffer solution is about 4 to 8. Preferably, the selected buffer solution is a solution containing NaCl. The buffer solution may contain a NaCl concentration of about 150 mM. For example, the skilled person can use HEPES or Opti-MEM type buffers, or solutions containing glucose.

[0051] Thus, according to a particular embodiment, the method for producing a tablet according to the invention comprises: - forming lipid particles, preferably liposomes, in a buffer; and / or - preparing the nucleic acid in a buffer; and / or - mixing the nucleic acid and the lipid particles in a buffer solution; may include:

[0052] According to a particular embodiment, the mixing time of the nucleic acid and the lipid particles, preferably liposomes, is between 10 seconds and several days, for example between 10 seconds and 10 hours, in particular between 5 minutes and 5 hours, in particular between 10 minutes and 1 hour. Preferably, the mixing time is greater than or equal to 30 minutes, in particular equal to about 30 minutes. The mixing can be carried out over a wide temperature range. However, according to a preferred embodiment, the mixing is carried out at a temperature between 10°C and 30°C, in particular between 15°C and 25°C.

[0053] After the complex has been formed, it may be directly subjected to the following steps of the method according to the invention or may be stored, in particular for at least one day, prior to said following steps.

[0054] "Tablet" formulation The method according to the present invention includes a step of tablet formulation from an aqueous mixture containing nucleic acid and lipid particles as described above. The inventors of the present application have succeeded in making a nucleic acid vector (or complex) into a "tablet" form for potential oral, vaginal, rectal, sublingual or transmucosal administration, while maintaining the biological properties of said nucleic acid vector.

[0055] More specifically, the method according to the invention comprises: - drying the aqueous mixture containing the complex defined above; and - compressing the dry mixture thus obtained Includes.

[0056] Preferably, the method according to the invention comprises the steps of: a)- a complex as defined above; and - one or more additives suitable for drying drying the aqueous mixture comprising: b) mixing one or more compression additives with the dry mixture obtained in step a); and c) compressing the mixture obtained in step b) Includes.

[0057] ·Drying process a): Drying refers to any dehydration process capable of removing water contained in a substance or body. For example, drying can be achieved by spray drying, convection drying or freeze drying. According to a preferred embodiment, drying is performed by freeze drying. According to this embodiment, the additive suitable for drying is an additive suitable for freeze drying. Freeze drying is a low-temperature, reduced pressure drying process that removes water from a previously frozen product by sublimation.

[0058] According to a preferred embodiment, the drying step a) is freeze-drying. According to a particular embodiment, the freeze-drying process carried out comprises the following steps: - freezing an aqueous mixture comprising the complex as defined above and one or more additives suitable for lyophilization; - Primary drying (or sublimation); and - Secondary drying Includes.

[0059] Preferably, the method according to the invention comprises, prior to the drying step, a step of mixing the complex as defined above with an additive suitable for drying.

[0060] An additive suitable for drying is an additive capable of preserving the structure of the nucleic acid vector during the drying step, thereby preserving the biological activity of the nucleic acid. An additive suitable for drying also provides a dry mixture suitable for a compression step. In particular, the additive makes it possible to obtain a dry product whose texture (affected by residual moisture) is suitable for the compression step. The additive also makes it possible to obtain a dry product whose mass and / or mechanical strength is suitable for the compression step, said additive making it possible, for example, to avoid the collapse of the structure of the dry product. Preferably, said additive suitable for drying, preferably for freeze-drying, facilitates the comminution and homogeneous mixing of the compressed additive with the obtained dry product. The optimization of parameters such as texture, mass and / or mechanical strength of the dry product by selecting the nature and amount of additive suitable for drying is conventionally carried out by the skilled person, in particular based on the physicochemical characteristics of the respective additive (presence of eutectic, glass transition temperature, etc.).

[0061] According to a preferred embodiment of the present invention, when freeze-drying is used in step a), the additive suitable for drying is suitable for freeze-drying, thus protecting the complex during the freeze-drying step and / or making it possible to obtain a freeze-dried product with a texture suitable for the compression step.

[0062] According to certain embodiments, additives suitable for lyophilization are protective agents (cryoprotectants or lyoprotectants) and / or bulking agents (or ballasts). Cryoprotective additives are additives that protect the structure of the nucleic acid vector during the freezing process. Lyoprotective additives are additives that protect the structure of the nucleic acid vector during the dehydration process. During lyophilization, these agents replace water molecules and interact with the polar heads of phospholipids (water replacement hypothesis) to form a glassy matrix that helps to maintain the structure of the lipoplexes (vitrification hypothesis). Bulking agents (or ballasts) are additives that maintain the structure and rigidity of the lyophilizate (as opposed to a collapsed state).

[0063] According to a particular embodiment, the additives suitable for lyophilization are selected from trehalose, mannitol, sucrose, sorbitol, lactose, glucose, glycerin, glycine, alanine, lysine, polyethylene glycol, polyvinylpyrrolidone (PVP) and dextran, optionally in a mixture.

[0064] According to a particular embodiment, the additive suitable for lyophilization is trehalose in a proportion of 1-100% by weight, in particular 10-100% by weight, based on the weight of the additive suitable for lyophilization, for example, the proportion of trehalose is about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90% or about 100% by weight of the additive suitable for lyophilization.

[0065] According to certain embodiments, the additive suitable for lyophilization is mannitol in a proportion of 1-90% by weight of the additive suitable for lyophilization, for example, the proportion of mannitol is about 0%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80% or about 90% by weight of the additive suitable for lyophilization.

[0066] Preferably, the excipient suitable for freeze-drying is trehalose optionally mixed with another excipient suitable for freeze-drying selected from mannitol, sucrose, sorbitol, lactose, glucose, glycerin, glycine, alanine, lysine, polyethylene glycol, polyvinylpyrrolidone (PVP) and dextran, preferably mannitol.

[0067] According to a particular embodiment, trehalose is mixed with mannitol in a defined ratio. In particular, the proportion of trehalose may be 10-99% by weight of the trehalose / mannitol mixture. For example, the proportion of trehalose is about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80% or about 90% by weight of the trehalose / mannitol mixture, preferably about 71% by weight of the trehalose / mannitol mixture.

[0068] According to certain embodiments, the proportion of mannitol may be 1-90% by weight of the trehalose / mannitol mixture, for example, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80% or about 90% by weight of the trehalose / mannitol mixture, preferably about 29% by weight of the trehalose / mannitol mixture.

[0069] According to a particular embodiment, the method according to the invention comprises the steps of: - preparing an additive suitable for drying, preferably prepared in an aqueous medium; - mixing an additive suitable for drying with the complex defined above; and - drying the mixture thus obtained, in particular by freeze-drying. may include.

[0070] According to a particular embodiment, the freezing step carried out during lyophilization corresponds to the freezing of a mixture comprising an additive suitable for drying and a complex as defined above. The freezing is carried out in such a way as to preserve the biological activity of the nucleic acid vector, preferably the supramolecular assembly of the nucleic acid vector. The freezing parameters are controlled in order to promote amorphization of the components of the mixture and / or to limit crystal growth.

[0071] Preferably, the freezing kinetics is fast, particularly depending on the freezing temperature. Preferably, the freezing rate is between -40°C / min and -0.5°C / min.

[0072] Preferably, in order to promote amorphization, freezing is carried out in liquid nitrogen, in particular at a temperature of about −190° C. Preferably, the freezing time in liquid nitrogen is from 1 minute to 24 hours, in particular from 1 minute to 1 hour, and said freezing time is in particular about 10 minutes.

[0073] According to certain embodiments, the parameters of the lyophilization process make it possible to preserve the biological activity of said complexes. In particular, the operating parameters controlling the lyophilization cycle (room or storage temperature, pressure in the drying chamber and cycle duration) are appropriately selected in order to maintain the biological activity of the complexes and can be adapted according to the volume of the aqueous mixture containing said complexes (more specifically according to the volume of water to be sublimated).

[0074] According to a particular embodiment, the freeze-drying duration, including primary and secondary drying, is between 10 and 80 hours, preferentially between 15 and 72 hours, more preferentially between 20 and 48 hours, preferably about 30 hours. The freeze-drying time can be adapted by the skilled person based on the volume and formulation of the aqueous mixture to be freeze-dried.

[0075] According to a particular embodiment, drying, in particular by freeze-drying, is carried out at atmospheric pressure.

[0076] ·Mixing process b): The process according to the invention may also include a step of destructuring the product obtained after drying. This may be a step of gentle comminution (low energy) or redispersion of the product. It may be carried out, for example, using vibration or a calibrated grid or in a paddle mixer, for example when mixing with excipients.

[0077] Prior to compression, the optionally milled dry mixture is mixed with one or more compression additives. - to protect the complex of nucleic acid and lipid particles during the compression step c); and / or - to reduce the phenomenon of friction during the compression step c); and / or - delimiting the compartments in which the powder is compressed, allowing a regular supply of the matrix used for compression; and / or - to obtain tablets of sufficient mass and sufficient mechanical strength, and / or - to control or delay the release of the nucleic acid vector, and / or - Allows tablets to disintegrate represents an optional additive.

[0078] According to a particular embodiment, the compression additives are selected from among the additives used in the granular mixture for compression.

[0079] In a preferred embodiment, the compression additive is a lubricant, optionally mixed with at least one other compression additive well known to those skilled in the art, preferably an excipient.

[0080] "Lubricant" refers to any additive that reduces adhesion of powder to the punches and dies of a press and / or reduces friction between the particles and the dies during tablet compression and ejection.

[0081] According to a particular embodiment, the lubricant represents 0.25-5% by weight of the finished tablet. Preferably, the lubricant represents about 0.5% by weight of the finished tablet. According to a preferred embodiment, the lubricant is selected from magnesium stearate, talc, sodium stearyl fumarate and stearic acid, optionally in admixture. Preferably, the lubricant is magnesium stearate.

[0082] "Excipient" refers to any additive that has a filler role when the amount of active ingredient and other additives is insufficient to produce a tablet of suitable size, thus allowing adjustment of the tablet mass and size.

[0083] According to a preferred embodiment, the excipient is selected from lactose, starch and its derivatives, calcium phosphate and its derivatives, in particular dicalcium phosphate, and cellulose and its derivatives, in particular microcrystalline cellulose, optionally in admixture. Preferably, the excipient is lactose.

[0084] The tablet may further contain pharmaceutical additives well known to those skilled in the art, such as flow agents, disintegrants, binders, additives for controlling or delaying the release of the nucleic acid vector, additives that impart mucoadhesive properties to the tablet, and combinations thereof. For example, the tablet may contain one or more of the following additives: sucrose, glucose, mannitol, trehalose, and silica.

[0085] Preferably, the compressed additive is a dry additive, preferably in powder form.

[0086] The process of the invention may also comprise, prior to the mixing step, a step of mixing different compression additives with the dry mixture obtained after drying step a).

[0087] Compression step c): The method according to the invention further comprises a step c) of compression of the mixture obtained at the end of step b). The compression step is carried out under conditions that preserve the biological function of the nucleic acid vector. In particular, the compression step is carried out in such a way as to preserve the therapeutic activity of the nucleic acid vector, for example in the case of siRNA, the expression of the target gene is inhibited.

[0088] The mechanical stress applied during the compression step can be controlled and monitored. More specifically, the pressure applied to the mixture containing the nucleic acid vector according to the present invention is controlled and monitored during the compression step. Preferably, the compression in step c) is performed at a pressure of 10-400 MPa, preferably 50-250 MPa.

[0089] According to a preferred embodiment, the compression in step c) is carried out using a tablet press, such as a rotary press or an alternative press.

[0090] In a particular embodiment, a wet granulation or dry granulation step is carried out before the compression step c). The "wet granulation" or "dry granulation" step consists of increasing the size of the particles to obtain agglomerates to facilitate compression. More specifically, the wet granulation step involves the addition of a binder to obtain granules in the form of agglomerates, which are then dried and optionally sieved before the compression step. Dry granulation consists of strongly compressing the particles to obtain agglomerates, which are then broken down to form particles, which are optionally calibrated before the final shaping step by compression.

[0091] According to a preferred embodiment, the compression step c) is a direct compression, which does not require a prior granulation step. According to this embodiment, the dry mixture obtained at the end of the drying step (step a)), optionally mixed with compression additives (step b)), has properties suitable for direct compression.

[0092] In a particular embodiment, the method according to the invention comprises a step of film-coating (or coating) of the tablets obtained after the compression step. The advantage of this step is the production of better protected tablets, whose stability is improved and / or whose release kinetics are adapted. The nature of the film-coating and / or coating agents and the processes used to carry out this step are well known to those skilled in the art of formulation.

[0093] In a second aspect, the present invention relates to a tablet obtainable by the process described above.

[0094] In particular, the present invention provides: - one or more nucleic acid vectors (or complexes) as above; - one or more additives suitable for drying; - one or more compression additives The present invention relates to a tablet comprising:

[0095] In particular, the tablet of the present invention comprises: - complexes of nucleic acids with liposomes or micelles containing cationic lipids; - one or more suitable lyophilization additives to protect the complex during the lyophilization step a) and / or to obtain a lyophilisate with a suitable texture for the compression step, preferably trehalose and mannitol; - one or more suitable compression additives to protect said complex during compression step c) and to preserve the biological activity of the complex. may also include

[0096] In particular, the finished tablet contains between 1 μg and 1 mg of nucleic acid, preferably about 10 μg.

[0097] The mass of cationic lipid in the finished tablet may be between 18.7 μg and 18.7 mg, preferably about 170 μg.

[0098] The mass of neutral fat in the finished tablet may be 16.5 μg to 16.5 mg, preferably about 150 μg.

[0099] The mass of the polymer in the finished tablet may be between 1 μg and 1 mg, preferably about 10 μg.

[0100] The mass of the additive suitable for drying may be 36 mg to 720 mg, preferably about 126 mg, in the finished tablet. In particular, the mass of trehalose in the finished tablet may be 3.6 mg to 720 mg, preferably about 90 mg. In particular, the mass of mannitol in the finished tablet may be 0 mg to 648 mg, preferably about 36 mg.

[0101] The proportion of additives suitable for compression may be 0.25-95% by weight of the finished tablet. In particular, the proportion of the lubricant, preferably magnesium stearate, may be 0.25-5% by weight, preferably about 0.5% by weight, of the finished tablet. In particular, the proportion of other compression additives may be 0-94.75% by weight of the finished tablet.

[0102] According to a particular embodiment, the tablet according to the invention comprises: - complexes of nucleic acids with liposomes or micelles containing DMAPAP and DOPE, further comprising an alginate, in particular sodium alginate; - trehalose and mannitol; and - Lactose Includes.

[0103] According to a variant of this embodiment, the tablet thus constituted has, after resuspension in water, a synchrotron radiation small angle X-ray scattering (SAXS) profile comprising a cubic phase and a lamellar phase, and optionally a liquid crystalline phase. The SAXS profile can be determined based on the protocol given in the examples. The SAXS profile of a tablet according to the invention after resuspension in water has a peak at q=0.086 nm. -1 and q = 0.101 nm -1 According to a particular embodiment, the lamellar phase is predominant compared to the cubic phase.

[0104] According to a particular embodiment, the tablet according to the invention is intended for oral, vaginal, rectal, sublingual or transmucosal administration. Preferably, the tablet according to the invention is intended for oral administration.

[0105] In another aspect, the present invention relates to a tablet of the present invention for use in the treatment of a local, locoregional or systemic pathology. Preferably, the tablet according to the present invention is used to treat a disease of the gastrointestinal tract. For example, the disease of the gastrointestinal tract may be Crohn's disease (wherein the nucleic acid is, for example, an siRNA that inhibits an inflammatory protein such as TNF-α), ulcerative colitis (wherein the nucleic acid is, for example, an anti-ICAM-1 oligonucleotide, an IL-10 or anti-TNF-α siRNA encoding plasmid), or gastric cancer (wherein the nucleic acid is, for example, an siRNA or an anti-Her2 oligonucleotide). The tablet according to the present invention may also be used to treat a vaginal pathology, in particular vulvovaginal candidiasis, or HSV, HPV or HIV infection (for example, via administration of an antiviral siRNA).

[0106] In addition, the tablets can be used to treat conditions such as spinal muscular atrophy (wherein the nucleic acid is, for example, the antisense oligonucleotide Nusinersen), familial hypercholesterolemia (wherein the nucleic acid is, for example, the antisense oligonucleotide Mipomersen), Duchenne myopathy, or age-related macular degeneration (wherein the nucleic acid is, for example, the aptamer Macugen).

[0107] The tablets according to the invention are stable, ie, substantially retain their biological activity, preferably at room temperature for up to 2 weeks, 1 month, 3 months, 1 year (between formulation and administration of the tablet). EXAMPLES

[0108] Example 1 Preparation of tablets 1. Lipoplex formation The first step consists of the formation of lipoplexes, i.e. complexes of nucleic acids and liposomes. The nucleic acid used in this example is a luciferase-specific siRNA. The liposomes used here contain a cationic lipid (DMAPAP) and a neutral lipid (DOPE). The formation of lipoplexes was carried out using the following components:

[0109] [Table 1]

[0110] The siRNA, liposome and polymer preparations were vortexed. - Liposomes were mixed with NaCl solution to obtain a final volume of 200 μL (10.7 μL liposomes and 189 μL NaCl solution). - Furthermore, siRNA was mixed with polymer in NaCl solution to obtain a final volume of 200 μL (7.5 μL siRNA + 6.5 μL polymer + 186 μL NaCl solution), so that the resulting mixture contains a 1:1 siRNA to polymer ratio by mass. - 200 μL of each of the two preparations obtained were mixed, vortexed and then left at room temperature for 30 minutes to allow the formation of siRNA-liposome complexes.

[0111] The above components were mixed to form lipoplexes with a + / - charge ratio of 8.

[0112] 2. Freeze drying In this example, mannitol and trehalose were used as additives suitable for lyophilization.

[0113] The lipoplex suspension obtained previously was transferred to a 50 mL centrifuge tube. A solution containing 2.5% m / v trehalose and 1% m / v mannitol was prepared. 3.6 mL of this trehalose / mannitol solution was then added to 400 μL of the lipoplex suspension. The centrifuge tube was then immersed in liquid nitrogen for 10 minutes.

[0114] Freeze-drying was performed in a freeze-dryer with a manifold equipped with 12 valves to which the frozen sample tubes were connected. Freeze-drying was carried out for 30 h at a pressure of less than 0.2 mbar (more specifically, 0.1-0.18 mbar) in the freeze-drying chamber. The energy required to compensate for the energy absorbed during sublimation is provided by the surrounding air (room temperature, more specifically, 20-25 °C).

[0115] A temperature sensor was placed in the sample to monitor the temperature of the product during the freezing (see Figure 1b) and freeze-drying (see Figure 1a) processes.

[0116] 3. Compression In this example, magnesium stearate (lubricant) and lactose (filler) were used as additives suitable for compression.

[0117] The previously obtained lyophilisate was first powdered by lightly grinding in a mortar, then 36.2 mg of lactose and 0.8 mg of magnesium stearate were added to the mortar and mixed with the lyophilisate.

[0118] Compression was performed using a STYL'One Evolution compression simulator according to the following parameters: - 6mm diameter chamfer flat punch - Compression Cycle = Cycle 1 compression (default cycle), speed 2%. - Force Control - Dose (die filling height) = 18mm - Manual filling - Compression force = 1.4~7.1kN (50~250MPa)

[0119] Example 2 Example of the composition of tablets containing lipoplexes Table 1 below shows, to illustrate the composition of tablets containing lipoplexes, the different components being expressed in mg.

[0120] [Table 2]

[0121] Example 3 Tablet Efficacy Testing The effectiveness of the tablets is then evaluated in cultured cells expressing the luciferase gene. When these cells are exposed to a vector containing a luciferase-specific siRNA, luciferase activity is inhibited. The test consists of transfecting mouse melanoma cells expressing luciferase (B16-Luc) and determining the residual luciferase activity in the cells after 48 hours. This measurement allows to obtain the percentage of inhibition induced by the siRNA vector studied, relative to the activity of cells not treated or treated with a vector containing a control siRNA (no inhibitory effect).

[0122] To assess the maintenance of this activity after compression, the tablets are solubilized before contacting the cells to be transfected. The lyophilisates (obtained before the compression step) were also rehydrated to analyse the effectiveness of the dry mixture before compression.

[0123] So the test is: - negative control: i.e. lipoplexes containing control siRNA (whose sequence has been selected so as not to inhibit the protein target) resuspended from a compacted form; - positive transfection control: i.e. resuspended lipoplexes (not lyophilized, not compacted); - a rehydrated lyophilisate; or - Rehydrated tablets This consisted of treating B16-Luc cells with

[0124] More precisely, resuspended lipoplexes (corresponding to the positive transfection control) were prepared as described above and then mixed with culture medium: complete medium (DMEM+Glutamax+fetal bovine serum+penicillin / streptomycin) supplemented with geneticin (G418).

[0125] Lyophilisates and tablets were prepared as described above and then dissolved in distilled or Milli-Q water and mixed with the medium.

[0126] The procedure is carried out over 4 days (days 0, 1, 2 and 3): On day 0, cells are prepared on 24-well plates (40,000 cells / well). On day 1, replace the cell culture medium with 1 mL of transfection medium (to add 0.5 μg of siRNA per well), which is the medium containing the negative control, the positive transfection control, the lyophilisate or the tablets. On day 2, the transfection medium is replaced with culture medium (complete medium + G418). On day 3, the cells are lysed and luciferase activity is measured. Tests are performed in triplicate (three repeats per condition).

[0127] result: Tablet Efficacy Testing Cell transfection studies showed superior efficacy of tablets containing lipoplexes (efficacy expressed by the percentage of inhibition of tablets compared to non-transfected cells). In addition, no compressive stress effect was observed on the percentage of inhibition of the luciferase gene over the entire range of applied compressive stresses (Figure 2).

[0128] Example 4 supramolecular structure The small angle X-ray scattering (SAXS) technique was used to identify the supramolecular structure of the siRNA vector, both in suspended particles and in dry forms, such as lyophilisates or tablets. The siRNA used was a control siRNA, each sample containing 100 micrograms of siRNA. This identification is made necessary by the fact that the structure of this type of particles is directly related to their effectiveness in administering nucleic acids. Synchrotron radiation SAXS analysis is the only technique that gives access to this structural identification in the various galenic forms suspended in aqueous media or powders.

[0129] First, we determined the structure of siRNA lipoplexes (DMAPAP / DOPE+alginate) prepared with a + / - charge ratio of 8. Studies performed on lipoplexes suspended in salt buffer (150 mM NaCl) showed that these lipoplexes are in the form of two coexisting structures, a lamellar and a cubic phase (Figure 3).

[0130] Next, we studied the fate of this structure when the suspended vector was subjected to a lyophilization step, a necessary prerequisite before the compression step could be performed afterwards. Figure 4 shows that a signal was detected in both types of samples (lyophilisates and tablets) with q = 0.096 nm -1 It shows a single, somewhat broad peak at 1000 nm, whose position is intermediate between the values ​​of peaks L1 and C1 of suspended lipoplexes (Figure 3), and whose specificity does not allow us to deduce the phase corresponding to this peak. The peak observed in the lyophilized sample is more intense than that of the compressed sample, although the latter remains fully detectable.

[0131] The lyophilization and compression steps require the addition of additives. We therefore evaluated the signals obtained for lipoplexes diluted in NaCl / additive mixtures under the same concentration conditions as the lyophilized and compressed samples (Figure 5). The obtained profile shows the presence of a single cubic phase, whose first peak is slightly offset between the "lyophilized" and "tablet" conditions. The position of this peak is close to the peak detected during the powder analysis, which may indicate that the phase present in the dry form is a cubic phase. The peak corresponding to the lamellar phase is no longer detectable in the presence of additives, either in the dry form or in suspension.

[0132] We then analyzed the SAXS profiles of the resuspended freeze-dried or compressed samples. Previous analysis showed the effect of the presence of additives on the structure of the particles (Figure 5), and we compared each resuspended sample with suspended lipoplexes under conditions of the same additive concentration. The results obtained are shown in Figure 6. It was found that the SAXS profile of the resuspended freeze-dried sample is close to that of the suspended lipoplexes under the same conditions (Figure 6A), suggesting that freeze-drying does not change the structure of the lipoplexes. In the presence of additives, only the cubic phase is detected. On the other hand, in the case of the resuspended compressed sample, the profile obtained allows the clear detection of two phases, lamellar and cubic, whose main peaks have positions similar to those obtained for the initial lipoplex suspension (Figure 3). The profile of this sample is clearly different from that of the lipoplex suspension under conditions of the same additive concentration. It is also different from the initial lipoplex suspension, since the relative proportions of the cubic and lamellar phases are reversed. In the initial suspension, the cubic phase dominates, and in the resuspended compressed sample, the lamellar phase dominates. This result indicates that compression affects the structure by changing the relative proportions of one phase to another, but does not cause new phases to appear or disappear.

[0133] Example 5 In vivo efficacy evaluation Model: Induction of ulcerative colitis in mice by administration of dextran sulfate (DS) in drinking water for 7 days

[0134] Assessment of vector efficacy: Oral administration of tablets containing siRNA lipoplexes. Mice under DS treatment receive tablets on days 1, 3 and 5 (starting on day 0). Each mouse receives a tablet containing 50 μg of siRNA formulated as a lipoplex.

[0135] Three groups of 10 mice were used: Group 1: Drink water and do not receive lipoplex Group 2: DS on days 1, 3, and 5, and tablets containing siRNA lipoplexes against TNF-α Group 3: DS on days 1, 3 and 5, plus tablets containing control siRNA lipoplexes (no inhibitory effect) Group 4: DS and no Lipoplex

[0136] Fecal mass and appearance (presence of blood) are monitored throughout treatment. On day 7, animals are euthanized, colons harvested, colon size measured, colon lysates assayed for inflammatory cytokines, and colon sections analyzed histologically for inflammation and tissue damage.

[0137] Expected Results: A reduction or even disappearance of clinical and biochemical signs of disease after treatment, which is specific to the siRNA sequence, i.e., observed in mice treated with lipoplexes containing anti-TNF-α siRNA but not in mice treated with lipoplexes containing control siRNA.

Claims

1. (i) a nucleic acid; and (ii) a lipid particle comprising a cationic lipid, preferably a liposome or a micelle, preferably a liposome; 1. A method for producing a tablet comprising the steps of: a) - (i) a complex of a nucleic acid and (ii) a lipid particle comprising a cationic lipid, preferably a liposome or a micelle, preferably a liposome; - one or more additives suitable for drying, in particular freeze-drying; drying the aqueous mixture comprising said component, in particular by freeze-drying; b) mixing one or more compression additives with the dry mixture obtained in step a); and c) compressing the mixture obtained in step b) A method comprising:

2. the cationic lipid is selected from lipopolyamines, quaternary ammonium, and lipids with cationic heads of guanidine or imidazole type, preferably the cationic lipid is selected from dimyristylaminopropylaminopropyl (DMAPAP), N-[1-(2,3-dioleyloxy)propyl]-N,N,N-trimethylammonium chloride (DOTMA), [1,2-bis(oleoyloxy)-3-(trimethylammonio)propane] (DOTAP), 3β[N-(N′,N′-dimethylaminoethane)-carbamoyl]cholesterol (DCChol) and dioctadecylamidoglycylspermine (DOGS), preferably dimyristylaminopropylaminopropyl (DMAPAP); 2. The method according to claim 1, characterized in that the cationic lipid is optionally mixed with a neutral lipid, in particular chosen from 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), cholesterol, dioleoyl-sn-glycero-3-phosphocholine (DOPC), dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) and N'-(rac-1-[11-(F-octyl)undec-10-enyl]-2-(hexadecyl)glycero-3-phosphoethanoyl)-sperminecarboxamide), preferably the neutral lipid being 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE).

3. A mixture of cationic lipids and neutral lipids: - 50-99 mol % of cationic lipids in the mixture, preferably about 50 mol % - 1-50 mol % of neutral fats in the mixture, preferably about 50 mol % 3. The method of claim 2, comprising:

4. The method according to any one of claims 1 to 3, wherein the nucleic acid is a therapeutic nucleic acid, in particular a siRNA, miRNA, shRNA, plasmid or mRNA.

5. 5. The method according to claim 1, characterized in that the charge ratio (+ / -) between the positive charges of the cationic lipids and the negative charges of the nucleic acid is between 0.5 and 10, preferably between 4 and 10, preferably about 8.

6. 6. The method according to claim 1 , characterized in that the additive suitable for freeze-drying is a protective agent (cryoprotectant or lyoprotectant) and / or a bulking agent, optionally in the form of a mixture, selected in particular from trehalose, mannitol, sucrose, sorbitol, lactose, glucose, glycerin, glycine, alanine, lysine, polyethylene glycol, polyvinylpyrrolidone and dextran.

7. 7. The method according to claim 6, characterized in that the additive suitable for freeze-drying is trehalose in a proportion of 10 to 100% by weight of the additive suitable for freeze-drying, optionally mixed with another additive suitable for freeze-drying, preferably mannitol, in a proportion of 0 to 90% by weight of the additive suitable for freeze-drying.

8. 8. The method according to claim 1, characterized in that the compression additive protects the complex of the lipid particles comprising the nucleic acid and the cationic lipid during the compression step c), said compression additive being in particular a lubricant (e.g. magnesium stearate, stearic acid and talc, preferably magnesium stearate), optionally mixed with another compression additive, preferably an excipient (e.g. lactose, starch, calcium phosphate and derivatives thereof and cellulose and derivatives thereof, preferably lactose).

9. 9. The process according to claim 8, characterized in that the lubricant represents 0.25 to 5% by weight of the finished tablet, preferably 0.5%.

10. 10. The method according to any one of claims 1 to 9, characterized in that the compression in step c) is carried out at a pressure of 10 to 400 MPa, preferably 50 to 250 MPa.

11. A tablet obtainable by the method according to any one of claims 1 to 10.

12. - liposome or micelle complexes containing nucleic acids and cationic lipids; - one or more suitable lyophilization excipients, preferably trehalose and mannitol, to protect the complex during the lyophilization step a) and / or to obtain a lyophilizate with a suitable texture for the compression step; - one or more suitable compression additives to protect the complex during compression step c) and to maintain the biological activity of the complex. tablets,

13. 13. A tablet according to claim 11 or 12 for oral, vaginal, rectal, sublingual or transmucosal administration, preferably by the oral route.

14. 14. A tablet according to any one of claims 11 to 13 for use in the treatment of a gastrointestinal or vaginal disorder.

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