FR3075603A1 - PHARMACEUTICAL COMPOSITION FOR ORAL ADMINISTRATION OF ANTIBODIES FOR THE TREATMENT OF GASTROINTESTINAL TRACT DISEASES - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for oral administration comprising at least one drug antibody and at least one or more pharmaceutically acceptable excipients, for its use in the treatment of diseases of the gastrointestinal tract, in patients having developed antibodies directed against the drug antibody.

Description

PHARMACEUTICAL COMPOSITION FOR ORAL ADMINISTRATION OF ANTIBODIES FOR THE TREATMENT OF DISEASES OF THE GASTROINTESTINAL TRACT

The present invention relates to compositions for the oral administration of a drug antibody, for use in the treatment of diseases of the gastrointestinal tract, in particular in patients who have developed antibodies against the drug antibody.

TECHNOLOGICAL BACKGROUND

Diseases of the gastrointestinal tract include chronic inflammatory bowel disease, irritable bowel syndrome, Crohn's disease, ulcerative colitis, hemorrhagic rectal ulcer, lesions concomitant with Behçet's disease, pochites and cancer, including small intestine cancer and colorectal cancer.

All these pathologies are characterized by inflammation of the wall of the digestive tract. Thus, in Crohn's disease, this inflammation can be localized at all levels of the digestive system, from the mouth to the anus, although it is most often found in the intestine, whereas in the case of ulcerative colitis, the inflammation is localized in the rectum and colon. These diseases progress by inflammatory attacks of extremely variable duration and frequency according to the patients. These relapses alternate with remission phases.

There is no curative treatment for inflammatory diseases of the gastrointestinal tract, but current anti-inflammatory drugs in the vast majority of cases allow lasting control of the disease, for several years, associated with a satisfactory quality of life.

In patients whose disease is progressive, not regulated by the administration of anti-inflammatory drugs (5-aminosalicylic acid, corticosteroids) or immunosuppressants, doctors use antibody-based biotherapies to stop attacks and heal the lesions of the intestinal mucosa. The most used are the anti-TNFα and anti-IL12 / IL-23 antibodies which specifically block pro-inflammatory cytokines involved in the disease. About 70% of patients respond well to these treatments. However, in half of them, the effectiveness of these drugs deteriorates after one or two years, requiring a change of molecule.

Indeed, it has been found that these monoclonal or polyclonal antibodies, used as medicaments, were for the most part immunogenic, and induced an undesirable immune response in the patient, which is accompanied by the production of specific anti-drug antibodies (ADA - for “Anti Drug Antibody”). This is particularly the case when using adalimumab (an anti-TNFa antibody) marketed under the name Humira®. Indeed, a recent study has shown that 23% to 46% of Crohn's disease patients, treated with infliximab or adalimumab, develop anti-drug antibodies within the first year of treatment (see in particular Ben-Horin, S. and Chowers , Y., Review article: loss of response to anti-TNF treatments in Crohn's disease. Alimentary Pharmacology & Therapeutics, 2011, 33: 987995). The development of these anti-drug antibodies may be due to the patient, especially in the event of repeated immunizations to successive biotherapies, to an active disease or to genetics. This development of anti-drug antibodies may also be due to the treatment itself, and in particular to the dosage (doses and frequency of administration) or to the route of administration. The development of anti-drug antibodies may also be due to the drug antibody itself, in particular by virtue of its origin (murine or human), its structure (fragments of immunoglobulins or whole immunoglobulins) and / or its sequence (precise epitopes which are recognized by T or B lymphocytes). The presence of anti-drug antibodies is harmful to the patient, as they are responsible for a decrease in the effectiveness of treatment with the drug antibody. Indeed, anti-drug antibodies can form immune complexes with the drug antibody, varying in size depending on the patient, and can be quickly eliminated. As the clearance of the drug antibody is increased, its half-life is therefore reduced, resulting in a lower serum concentration and lower efficacy. In addition, certain anti-drug antibody - drug antibody dimers, such as, for example, adalimumab - anti-adalimumab antibody, may be thrombogenic. Finally, certain anti-drug antibodies, known as neutralizers, have the capacity to block the attachment of the drug antibody to its target.

Therefore, it is necessary to develop antibody formulations, in which the efficacy of the drug antibody is maintained and / or improved, to guarantee an effective and lasting treatment of the patient suffering from a disease of the disease. gastrointestinal tract and this even in case of development of anti-drug antibodies.

Formulations of polyclonal or monoclonal antibodies for oral administration are currently in development. But there is still a need to improve the efficacy and / or safety of the drug antibodies administered orally.

The subject of the present invention is therefore a pharmaceutical composition for oral administration comprising at least one drug antibody and pharmaceutically acceptable excipients, for its use in the treatment of diseases of the gastrointestinal tract, in patients having developed antibodies directed against antibody drug.

The pharmaceutical composition according to the invention, by its oral administration, makes it possible to guarantee an effective and lasting treatment of the patient, and this even in the event of the development of anti-drug antibodies.

For the purposes of the present invention, the term “drug antibody” means any antibody which can be administered to a patient and which has therapeutic activity.

Within the meaning of the present invention, the term antibody designates immunoglobulin molecules and immunologically active portions of immunoglobulin molecules, that is to say molecules which contain an antigen binding site which specifically binds an antigen. , whether natural or partially or entirely synthetically produced. The term also covers any polypeptide or protein having a binding domain which is, or is homologous to, an antibody binding domain. These can come from natural sources, or be produced in part or in whole by synthesis.

An antibody is a molecule composed of two heavy polypeptide chains and two light polypeptide chains linked together by disulfide bonds. Each heavy chain is composed of a variable domain (VH domain) and 3 constant domains (CH1, CH2 and CH3 domains), while each light chain is composed of a variable domain (VL domain) and a constant domain (domain CL). In addition, each variable domain (VH or VL domain) is composed of 4 framework regions (FR1, FR2, FR3 and FR4), which have less sequence variability among the antibodies and are involved in the formation of beta sheets, and 3 hypervariable regions, commonly called complementary determining regions 1,2 and 3 (CDR1, CDR2, CDR3), which correspond to 3 loops separating each beta sheet. The 3 CDR regions, juxtaposed in space at the end of the variable domain, are crucial for determining an antibody (or antibody fragment) specificity, since they are the part of the variable domain mainly at contact of the antigen, in particular the CDR3 region of each chain, corresponding to the rearranged region of the heavy and light chains, which is even more variable and more directly in contact with the specific antigen. Examples of antibodies are the immunoglobulin isotypes (eg, IgG, IgE, IgM, IgD and IgA) and their isotypic subclasses. Advantageously, the drug antibody according to the invention can be a polyclonal antibody or a monoclonal antibody. A monoclonal antibody can be referred to herein as mAb.

Since antibodies can be modified in a number of ways, the term antibody should be interpreted to cover any specific binding element or substance having a binding domain with the required specificity. Thus, this term covers fragments of antibodies, derivatives, functional equivalents and homologs of antibodies, humanized antibodies, including any polypeptide comprising an immunoglobulin binding domain, whether natural or totally or partially synthetic. Chimeric molecules comprising an immunoglobulin binding domain, or equivalent, fused to another polypeptide are therefore included.

In one embodiment, the drug antibody of the present invention is a humanized monoclonal antibody. The monoclonal antibody of the present invention can be humanized by modifying the amino acid sequence of the antibody. Methods for reducing the immunogenicity of specific binding molecules of the invention include grafting CDRs onto an appropriate antibody framework structure or remodeling of variable surface residues, e.g. by site-directed mutagenesis or by other commonly used molecular biology techniques. A humanized antibody can be a modified antibody having the variable regions of a non-human antibody, e.g. murine, and the constant region of a human antibody. In one embodiment, the drug antibody of the present invention is a human monoclonal antibody.

In another advantageous embodiment, the drug antibody of the invention can be a fragment of an antibody. Fragments of a whole antibody have been shown to perform the function of binding to the antigen. Examples of binding fragments are:

(i) the Fab fragment consisting of the VL, VH, CL and CH1 domains;

(ii) the Fd fragment consisting of the VH and CH 1 domains;

(iii) the Fv fragment consisting of the VL and VH domains of a single antibody;

(iv) the dAb fragment, which consists of a VH domain;

(v) isolated CDR regions;

(vi) F (ab ') 2 fragments, a bivalent fragment comprising two linked Fab fragments;

(vii) single chain variable fragments (scFv), in which a VH domain and a VL domain are linked by a peptide linker which allows the two domains to combine to form an antigen binding site;

(viii) bispecific single-chain Fv dimers;

(ix) diabodies, multivalent or multispecific fragments constructed by gene fusion;

(x) scFv-Fc consisting of two scFv fragments with an Fc part.

For the purposes of the present invention, the term CH1 domain corresponds to the first constant region domain of the heavy chain of an immunoglobulin which extends, for example, from positions 114-223 in the Kabat numbering system (positions EU 118-215). The CH1 domain is adjacent to the VH domain and the Nterminal region of the hinge region of the immunoglobulin heavy chain, but is not part of the Fc region of the immunoglobulin heavy chain.

For the purposes of the present invention, the term CL domain includes the constant region of an immunoglobulin light chain which extends, e.g. from Kabat position 107A-216 (EU positions 108-214 (kappa)). The CL domain is adjacent to the VL domain and includes the C-terminal region of the immunoglobulin light chain.

In an advantageous embodiment, the drug antibody of the invention can be a single chain variable fragment (scFv). Within the meaning of the present invention, the term variable single-stranded fragment or scFv molecule or scFv fragment or scFv or scFv antibody denotes a single folded polypeptide comprising the VH and VL domains of an antibody linked by a peptide linker. In such a scFv molecule, the VH and VL domains can be in the order VH-linker-VL or VL-linker-VH.

In an advantageous embodiment of the invention, the drug antibody can be chosen without limitation from the group comprising an anti-TNFα antibody, an anti-type 1 interferon receptor antibody, an anti-IL antibody -1, an anti-IL-6 antibody, an anti-IL-8 antibody, an anti-IL-12 antibody, an anti-IL-13 antibody, an anti-IL-17 antibody, an anti-IL-23 antibody, a anti-IL-34 antibody, anti-CD3 antibody, anti-CD4 antibody, anti-CD20 antibody, anti-CD70 antibody, antiIFN-γ antibody, anti-INF-γ receptor antibody, anti-CEA antibody, an anti-integrin antibody, in particular an anti-integrin a or β antibody and advantageously an anti-integrin α4β7 antibody, an anti-b7 antibody, an anti-VEGF antibody and an anti-EGFR antibody,

Examples of anti-TNFα antibodies include adalimumab, infliximab, etanercept, certolizumab, ozoralizumab or golimumab, the list is not exhaustive. In an advantageous embodiment of the invention, the antiTNFα antibody is adalimumab. In an advantageous embodiment of the invention, the drug antibody is adalimumab.

As an example of anti-IL-1 antibodies, mention may in particular be made of canakinumab. Examples of anti-IL-6 antibodies include tocilizumab, sirukumab, olokizumab and elsilimomab. Examples of anti-IL-13 antibodies include lebrikizumab, dectrekumab and anrukinzumab. Examples of anti-IL-17 antibodies include secukinumab, ixekizumab and brodalumab. Examples of anti-IL-23 antibodies include brazikumab, rizankinumab and birakinumab. As an example of anti-IL-12/1 L-23 antibodies, mention may be made in particular of ustekinumab. As an example of anti-CD3 antibodies, mention may in particular be made of visilizumab. Examples of anti-CD4 antibodies include priliximab. Examples of anti-CD20 antibodies include rituximab, ofatumumab, oblituximab, obinutuzumab, hydrocaratuzumab and ocrelizumab, the list is not exhaustive. As an example of anti-IFN-γ antibodies, mention may in particular be made of fontolizumab. As an example of anti-CEA antibodies, mention may in particular be made of labetuzumab. As an example of anti-integrin antibody α4β7, there may be mentioned in particular vedolizumab, etrolizumab and abrilumab. As an example of anti-integrin a antibodies, mention may in particular be made of natalizumab. Examples of anti-VEGF antibodies include bevacizumab and ramucirumab, the list not being exhaustive. Examples of anti-EGFR antibodies include cetuximab and panitumumab. The examples of antibodies mentioned above are not limiting.

The drug antibody used in the present invention can be produced by any technique well known to those skilled in the art. Advantageously, the drug antibody according to the invention is a recombinant drug antibody.

In a particular embodiment, the recombinant drug antibody can be produced by recombination in a host cell, transformed with one or more vector (s) which allow the expression and / or the secretion of the nucleotide sequences coding for the heavy chain and / or the light chain of the antibody. The vector generally includes a promoter, translation initiation and termination signals, as well as suitable regions for transcription regulation. It is stably maintained in the host cell and can possibly have specific signals which specify the secretion of the translated protein. These various elements are chosen and optimized by a person skilled in the art according to the cell host used. Such vectors are prepared by methods commonly used by those skilled in the art, and the resulting clones can be introduced into an appropriate host by standard methods, such as lipofection, electroporation, the use of polycationic agents, thermal shock, or chemical methods. The cell host can be chosen from prokaryotic or eukaryotic systems, for example bacterial cells but also yeast cells or animal cells, in particular non-human mammalian cells. The preferred mammalian cells for the production of the monoclonal antibody are the rat line YB2 / 0, the hamster line CHO, in particular the lines CHO dhfr- and CHO Lec13, PER.C6 ™, EB66, HEK293, K562, NSO, SP2 / 0, BHK or COS. It is also possible to use insect cells such as cells S2, Sf9, Sf21. In a particularly advantageous embodiment, the recombinant drug antibody is produced in mammary non-human mammalian epithelial cells.

Another mode of production is the expression of the recombinant drug antibody in transgenic organisms, for example in plants (Ayala M et al, Methods Mol Biol., 2009; 483, pages 103-134) or in the milk of transgenic animals such as rabbits, goats or pigs (Pollock, DP et al. Journal of Immunological Methods, 1999, 231, pages 147-157). In a particularly advantageous embodiment, the drug antibody is produced in the milk of a non-human transgenic mammal.

According to a preferred embodiment, the recombinant drug antibody is produced in the milk of non-human transgenic mammals, genetically modified to produce this glycoprotein. The mammal can for example be a goat, sheep, bison females, buffalo, camel, llama, mouse, rat, or even a cow, sow, rabbit, or mare. Advantageously, the recombinant drug antibody is produced in transgenic goat milk.

The secretion of the recombinant drug antibody by the mammary glands, allowing its presence in the milk of the transgenic mammal, involves controlling the expression of the recombinant drug antibody in a tissue-dependent manner. Such control methods are well known to those skilled in the art. The expression is controlled by sequences allowing the expression of the glycoprotein in a particular tissue of the animal. These include promoter sequences of the “WAP”, “β-casein”, “β-lactoglobulin” type and possibly sequences of the signal peptide type. Advantageously, the recombinant drug antibody is produced in the mammary glands of a transgenic goat, using an expression vector comprising the sequence of the two chains, under the control of a 5 'β-casein promoter. A process for extracting proteins of interest from the milk of transgenic animals is described in patent EP 0 264 166. Advantageously, more than 4 grams of antibodies are produced per liter of milk, advantageously more than 5, 10, 15, 20, 25, 30, 35 grams per liter of milk, even more advantageously up to 70 grams per liter of milk. Advantageously, the recombinant drug antibody produced by animal transgenesis, in particular in the mammary glands of a transgenic goat.

In a particularly advantageous embodiment, the pharmaceutical composition for oral administration according to the invention comprises at least one or more pharmaceutically acceptable excipients chosen from the group comprising: a carboxymethyl-dextran, a chitosan, a cyclodextrin or a combination thereof .

Advantageously, the pharmaceutical composition of the invention as defined above is devoid of at least one of the following excipients: caprylic acid or a salt of caprylate, glucose, fructose, galactose, mannose, sorbose , sucrose, ribose, deoxyribose, sucrose, maltitol, inulin, lecithin, trehalose, lactose, maltose, raffinose, mannitol, sorbitol, glycerol, arabitol, lactitol , xylitol, polypropylene glycol, polyethylene glycol, polyoxyethylene sorbitan fatty acid ester, polysorbate 20, polysorbate 80, poloxamers, polyoxyethylene alkyl ethers, alkylphenylpolyoxyethylene ethers, of polyoxyethylene-polyoxypropylene copolymer, of sodium dodecylsulphate, of sodium succinate, of sodium chloride, of sodium citrate, of citric acid monohydrate, of sodium phosphate dibasic dihydrate, of phosphate sodium monobasic dihydrate, sodium phosphate monobasic monohydrate or EDTA.

For the purposes of the present invention, the term "devoid" means that the composition of the invention does not contain at least one of the following excipients: caprylic acid or caprylate salt, glucose, fructose, galactose, mannose, sorbose, saccharose, ribose , deoxyribose, sucrose, maltitol, inulin, lecithin, trehalose, lactose, maltose, raffinose, mannitol, sorbitol, glycerol, arabitol, lactitol, xylitol, polypropylene glycol, polyethylene glycol, sorbitan polyoxyethylene fatty acid ester, polysorbate 20 80, poloxamers, polyoxyethylene alkyl ethers, alkylphenylpolyoxyethylene ethers, polyoxyethylene-polyoxypropylene copolymer, sodium dodecylsulphate, sodium succinate, sodium chloride, sodium citrate, citric acid monohydrate, sodium phosphate dibasic dihydrate, phosphate phosphate sodium monobasic dihydrate, sodium phosphate monobasic monohydrate or EDTA, or that they are present in a quantity negligible tee, especially less than 0.001% (m / m).

In another particularly advantageous embodiment, the pharmaceutical composition of the invention as defined above is devoid of molecule with anticoagulant effect, in particular of EDTA.

In another particularly advantageous embodiment, the pharmaceutical composition of the invention as defined above is devoid of surfactant, in particular of polysorbate 20, of polysorbate 80 or of poloxamer.

In another particularly advantageous embodiment, the pharmaceutical composition of the invention as defined above is devoid of laxative molecule, in particular of mannitol, sorbitol, glycerol, lactitol or xylitol.

The excipients mentioned above have in particular been excluded because of their anticoagulant activity such as in particular EDTA, or because they would present a risk of promoting Crohn's disease when they are ingested, such as in particular polysorbate 20, polysorbate 80 or poloxamer, or the fact that these excipients can have a laxative effect, such as mannitol, maltitol, lactitol, xylitol or sorbitol in particular.

In a particularly advantageous embodiment of the invention, the pharmaceutical composition for oral administration comprises:

- a drug antibody, as active ingredient, and

- carboxymethyl-dextran (CMD), as a pharmaceutically acceptable excipient.

In another particularly advantageous embodiment of the invention, the pharmaceutical composition for oral administration comprises:

- a drug antibody, as active ingredient, and

- Chitosan, as a pharmaceutically acceptable excipient.

In another particularly advantageous embodiment of the invention, the pharmaceutical composition for oral administration comprises:

- a drug antibody, as active ingredient, and

- a cyclodextrin, as a pharmaceutically acceptable excipient.

In an advantageous embodiment of the invention, the cyclodextrin can be chosen without limitation from Ι'α-cyclodextrin, β-cyclodextrin, γ-cyclodextrin, one of their derivatives, such as in particular a hydroxypropyl, hydroxyethyl derivative, ethyl or methyl, a beta-sulfobutyl ether cyclodextrin, a branched cyclodextrin, a cyclodextrin-based polymer or a mixture thereof. Advantageously, the cyclodextrin can be I ’hydroxypropyl-p-cyclodextrin (or HPBCD).

In a particularly advantageous embodiment of the invention, the pharmaceutical composition for oral administration comprises:

- a drug antibody as active ingredient, and

- hydroxypropyl-p-cyclodextrin (HPBCD), as a pharmaceutically acceptable excipient.

In another particularly advantageous embodiment of the invention, the pharmaceutical composition for oral administration comprises:

- a drug antibody as active ingredient, and

- a combination of carboxymethyl-dextran (CMD), and of cyclodextrin, in particular hydroxypropyl-p-cyclodextrin (HPBCD), as a pharmaceutically acceptable excipient.

In another particularly advantageous embodiment of the invention, the pharmaceutical composition for oral administration comprises:

- a drug antibody as active ingredient, and

- a combination of carboxymethyl-dextran (CMD) and chitosan, as a pharmaceutically acceptable excipient.

Within the meaning of the present invention, the term “combination” refers to a mixture of pharmaceutically acceptable excipients, advantageously in the form of a single formulation.

In another particularly advantageous embodiment of the invention, the pharmaceutical composition for oral administration comprises:

- a drug antibody as active ingredient, and

- a combination of a cyclodextrin, in particular I ’hydroxypropyl-βcyclodextrin (HPBCD) and chitosan, as a pharmaceutically acceptable excipient.

In another particularly advantageous embodiment of the invention, the pharmaceutical composition for oral administration comprises:

- a drug antibody as active ingredient, and

- a combination of carboxymethyl-dextran (CMD), cyclodextrin, in particular hydroxypropyl-p-cyclodextrin (HPBCD) and chitosan, as a pharmaceutically acceptable excipient.

In a particularly advantageous embodiment of the invention, the pharmaceutical composition for oral administration can also comprise at least one amino acid. Within the meaning of the present invention, the amino acid can be chosen from arginine, glycine, lysine, histidine, glutamate, glutamine, asparagine, isoleucine, leucine, alanine, phenylalanine, threonine, tyrosine, tryptophan, methionine, serine, proline, cysteine, selenocysteine, proline, valine, one of their derived salts, such as in particular a hydrochloride or a phosphate, or a mixture of these. Advantageously, the amino acid is chosen from hydrophilic amino acids. In an advantageous embodiment, the amino acid is glycine or one of these derived salts, such as glycine hydrochloride. In another particularly advantageous embodiment, the amino acid is arginine or one of these derived salts, such as arginine hydrochloride.

In a particularly advantageous embodiment of the invention, the pharmaceutical composition for oral administration comprises:

- a drug antibody as active ingredient,

- carboxymethyl-dextran (CMD), as a pharmaceutically acceptable excipient, and

- an amino acid, in particular glycine, optionally in its hydrochloride form.

In a particularly advantageous embodiment of the invention, the pharmaceutical composition for oral administration comprises:

a drug antibody as active ingredient,

- chitosan, as a pharmaceutically acceptable excipient, and

- an amino acid, in particular glycine, optionally in its hydrochloride form.

In a particularly advantageous embodiment of the invention, the pharmaceutical composition for oral administration comprises:

- a drug antibody as active ingredient,

- a cyclodextrin, in particular hydroxypropyl-p-cyclodextrin (HPBCD), as a pharmaceutically acceptable excipient,

- an amino acid, in particular glycine, optionally in its hydrochloride form.

In a particularly advantageous embodiment of the invention, the pharmaceutical composition for oral administration comprises:

- a drug antibody as active ingredient,

a combination of carboxymethyl-dextran (CMD) and a cyclodextrin, in particular hydroxypropyl-p-cyclodextrin (HPBCD), as a pharmaceutically acceptable excipient, and

- an amino acid, in particular glycine, optionally in its hydrochloride form.

In a particularly advantageous embodiment of the invention, the pharmaceutical composition for oral administration comprises:

- a drug antibody as active ingredient,

a combination of carboxymethyl dextran (CMD) and chitosan, as a pharmaceutically acceptable excipient, and

- an amino acid, in particular glycine, optionally in its hydrochloride form.

In a particularly advantageous embodiment of the invention, the pharmaceutical composition for oral administration comprises:

- a drug antibody as active ingredient,

a combination of a cyclodextrin, in particular I ’hydroxypropyl-βcyclodextrin (HPBCD) and chitosan, as a pharmaceutically acceptable excipient and

- an amino acid, in particular glycine, optionally in its hydrochloride form.

In a particularly advantageous embodiment, the pharmaceutical composition for oral administration according to the invention comprises a drug antibody as active ingredient, and a mixture of carboxymethyl-dextran and cyclodextrin.

In another particularly advantageous embodiment of the invention, the pharmaceutical composition for oral administration consists of:

- a drug antibody as active ingredient, and

- carboxymethyl-dextran (CMD), as a pharmaceutically acceptable excipient.

In another particularly advantageous embodiment of the invention, the pharmaceutical composition for oral administration consists of:

- a drug antibody as active ingredient,

- carboxymethyl-dextran (CMD), and

- a cyclodextrin, in particular hydroxypropyl-p-cyclodextrin (HPBCD).

In another particularly advantageous embodiment of the invention, the pharmaceutical composition for oral administration consists of:

- a drug antibody as active ingredient,

- a combination of carboxymethyl-dextran (CMD) and cyclodextrin, in particular hydroxypropyl-p-cyclodextrin (HPBCD), as a pharmaceutically acceptable excipient,

- an amino acid, in particular glycine, optionally in its hydrochloride form.

In another particularly advantageous embodiment of the invention, the pharmaceutical composition for oral administration consists of:

- a drug antibody as active ingredient, and

- Chitosan, as a pharmaceutically acceptable excipient.

The present invention relates to a pharmaceutical composition for oral administration comprising at least one drug antibody and at least one or more pharmaceutically acceptable excipients, for use in the treatment of diseases of the gastrointestinal tract, in patients having developed antibodies directed against drug antibody.

The term treatment or curative treatment is defined as a treatment leading to a cure or a treatment which alleviates, improves and / or eliminates, reduces and / or stabilizes the symptoms of a disease or the suffering which it causes.

According to the invention, the pharmaceutical composition comprising the drug antibody and at least one or more pharmaceutically acceptable excipients is particularly useful for treating inflammatory bowel disease (IBD), irritable bowel syndrome (IBS), the disease Crohn's disease, ulcerative colitis, hemorrhagic rectal ulcer, lesions concomitant to Behçet's disease, pochitis and cancer, in particular cancer of the gastrointestinal tract and in particular cancer of the small intestine and colorectal cancer.

Another subject of the invention relates to a pharmaceutical composition comprising a pharmaceutically effective amount of drug antibody and at least one or more pharmaceutically acceptable excipients.

Advantageously, the pharmaceutical composition comprising a therapeutically effective amount of drug antibody as defined above as an active ingredient is administered to a patient suffering from or suspected of suffering from a disease of the gastrointestinal tract.

In one embodiment, the patient can be a human, male or female, regardless of age. In another embodiment, the patient can be a non-human animal, for example a dog, a cat, a horse, a cow, a pig, a sheep, a goat or a primate.

Within the meaning of the present invention, the terms therapeutically effective or effective amount to be treated or pharmaceutically effective denote the amount of drug antibody or a composition necessary to inhibit or reverse a disease, and in particular to treat a disease of the gastrointestinal tract. Determination of a therapeutically effective amount depends specifically on factors such as the toxicity and efficacy of the drug. These factors will differ depending on other factors such as activity, relative bioavailability, patient body weight, severity of unwanted side effects and preferred method of administration. Toxicity can be determined using methods well known in the art. The same goes for efficiency. A pharmaceutically effective amount, therefore, is an amount that is considered by the clinician to be toxicologically tolerable, but effective despite the presence of anti-drug antibodies developed by the patient.

The dosage can be adjusted appropriately to achieve the desired levels of drug (e.g. drug antibody), local or systemic, depending on the mode of administration. In cases where the response in a patient is insufficient at such doses, even higher doses (or more effective doses by a different, more localized route of administration) may be used to the extent that the patient's tolerance allows. Multiple doses per day can also be used to reach appropriate levels of antibodies. Dose and dosage are used interchangeably here.

In an advantageous embodiment, the amount of drug antibody administered to the patient is from 0.02 mg / kg to about 100 mg / kg of body weight, i.e. from about 1 mg to about 8 g per day for a person of 55 to 80 kg of body weight. In an even more advantageous embodiment, the dosage of the drug antibody is in the range of from about 0.16 mg / kg to about 32 mg / kg of body weight, or from about 8.75 mg to about 2 , 5 g per day. In an even more advantageous embodiment, the dosage of the drug antibody is in the range of from about 0.16 mg / kg to about 13 mg / kg of body weight, or from about 8.75 mg to about 1 g per day. In an even more advantageous embodiment, the dosage of the drug antibody is in the range of about 0.16 mg / kg to about 6.5 mg / kg of body weight, or about 8.75 mg to about 520 mg. In an even more advantageous embodiment, the dosage of the drug antibody is in the range of about 0.16 mg / kg to about 3.2 mg / kg of body weight, or about 8.75 mg to about 256 mg. In an even more advantageous embodiment, the dosage of the drug antibody is in the range of about 0.16 mg / kg to about 1.3 mg / kg of body weight, or about 8.75 mg to about 104 mg. In a particularly advantageous embodiment, the dosage of the drug antibody is in the range of from about 0.16 mg / kg to about 0.64 mg / kg of body weight, or from about 8.75 mg to about 51 mg.

Generally, advantageous doses of drug antibodies are doses of 8 to 200 mg per day, preferably 8 to 60 mg per day or 15 to 120 mg per day.

In an advantageous embodiment, the pharmaceutical composition according to the invention is administered to the patient one, two, three, four, five, six or seven times a week.

Advantageously, the pharmaceutical composition according to the invention is administered to the patient at the rate of seven times a week, that is to say one administration per day for seven consecutive days. Advantageously, the pharmaceutical composition according to the invention is administered to the patient at the rate of six times a week. Advantageously, the pharmaceutical composition according to the invention is administered to the patient at the rate of five times a week. Advantageously, the pharmaceutical composition according to the invention is administered to the patient at the rate of four times a week. Advantageously, the pharmaceutical composition according to the invention is administered to the patient at the rate of three times a week. Advantageously, the pharmaceutical composition according to the invention is administered to the patient twice a week. Advantageously, the pharmaceutical composition according to the invention is administered to the patient once a week.

In an advantageous embodiment of the invention, the pharmaceutical compositions provided are used for in vivo applications. Advantageously, the compositions used can be in the form of a solid, semi-solid or liquid dosage. In a particularly advantageous embodiment, the pharmaceutical composition according to the invention is in solid form. The pharmaceutical composition in solid form can be obtained by any technique well known to those skilled in the art. Advantageously, the pharmaceutical composition in solid form can be obtained by lyophilization, spray drying, drying (in particular by spraying) or crystallization. The compositions of the present invention can be in all the galenical forms normally used for oral administration, in particular in the form of microgranules, granules, tablets, capsules, capsules, tablets, powder, syrup or any form for solid oral preparation or in any form of oral preparation. In a particularly advantageous manner, the composition of the present invention is administered in the form of capsules. Advantageously, the compositions according to the invention are administered in unit dosage forms suitable for a single administration of precise dosage amounts. In certain embodiments, the compositions according to the invention are sterile.

In a particularly advantageous embodiment, the composition is in the form of a sustained and / or delayed release formulation. For the purposes of the present invention, the term "prolonged and / or delayed release formulation" means a dosage form suitable for targeted release in the gastrointestinal tract, and in particular of the intestine. By "intestine" is meant here all the parts of the intestine, in particular the colon. Such compositions are particularly useful in the treatment of inflammatory bowel diseases, since they allow local action at the site of inflammation (in particular small intestine and / or colon). They also limit the passage of drug antibodies into the bloodstream, limiting the side effects associated with drug antibodies.

In a particularly advantageous embodiment, the pharmaceutical composition according to the invention is administered orally. Several strategies exist for preparing drugs administered orally, the active ingredient (s) of which are only released in the intestine, preferably in the ileum and colon. Some strategies include covalent linking of the drug with a carrier. Excipients and vehicles that are degraded by colon bacteria can also be used. All of these strategies are well known to those of skill in the art.

In a particular embodiment, the pharmaceutical composition in the form of a sustained-release and / or delayed-release formulation comprising the drug antibody and at least one pharmaceutically acceptable excipient as described above can be formulated in solid dosage forms, such as microgranules, granules, tablets, capsules, capsules. Advantageously, the pharmaceutical composition in the form of a sustained-release and / or delayed-release formulation comprising the drug antibody and at least one pharmaceutically acceptable excipient as described above can also comprise at least one coating agent. The coating agent, by its resistance to the acid pH of the stomach (between 1 and 3) makes it possible in particular to protect the composition, comprising the drug antibody and at least one pharmaceutically acceptable excipient, from the acid pH of 1 stomach and their release in the small intestine and the colon where the pH values are close to 7. Advantageously, the coating agent can be a film-forming agent, preferably gastro-resistant or a coating agent. Advantageously, the coating agent is an enteric coating agent. In a particularly advantageous embodiment, the coating agent can be chosen without limitation from the group comprising: derivatives of acrylic acids, derivatives of methacrylic acids, derivatives of ethyl acrylate, derivatives of hydroxypropyl methylcellulose , derivatives of polyvinylacetate phatlates; poly (methyl acrylate -co- methyl methacrylate -co- methacrylic acid, poly (methacrylate acid co-methyl methacrylate), especially marketed under the names Eudragit® S100 Eudragit® FS30D, Eudragit® L100, Eudragit®L12.5, Eudragit® L30D-55, Eudragit®, L10055, Eudragit® S12,5, hydroxypropylmethylcellulose (HPMCP), succinyl acetate of hydroxypropylmethylcellulose, also called HPMCAS and marketed under the name AQOAT®, Shellac or a mixture of these.

In a particular embodiment, the pharmaceutical composition in the form of a sustained and / or delayed release formulation comprising the drug antibody and at least one pharmaceutically acceptable excipient as described above can comprise a monolayer of a single agent coating, a monolayer consisting of a mixture of several coating agents, a multilayer consisting of a single coating agent; a multilayer consisting of a mixture of several different coating agents or a superposition of several monolayers consisting of a single coating agent.

In another embodiment, formulations can be used which are coated with polymers degradable by colon microorganisms (more particularly by bacterial enzymes such as azoreductases and glycosidases), for example azo polymers having a high degree of hydrophilicity. Gels and hydrogels can also be used, in particular hydrogels based on polysaccharides.

The compositions of the invention may comprise or be combined with other therapeutic agents useful in the treatment of inflammatory diseases, autoimmune diseases or cancer.

Another aspect of the invention relates to a process for the preparation of a pharmaceutical composition with sustained and / or delayed release, said process comprising:

a) A step of mixing the drug antibody with one or more pharmaceutically acceptable excipients chosen in a nonlimiting manner from the group comprising: a carboxymethyl-dextran, a chitosan, a cyclodextrin or a combination thereof.

b) A drying step (in the case of pharmaceutical compositions in dry form), in particular by atomization, of the composition obtained in step a),

c) A step of shaping the composition obtained in step b), chosen without limitation from the forms of: microgranules, granules, tablets, capsules, capsules,

d) A coating step.

Advantageously, the coating step d) can be carried out by any technique well known to those skilled in the art. In a particular embodiment, step d) of coating can be carried out by coating, by film-coating, in particular by the addition of film-forming agent, by spraying, by coating in a fluidized bed, by coating dry, by strippable coating. By "dry coating" is meant any solvent-free process using a mechanical action to cover a solid pharmaceutical form with a discrete or continuous layer of inert or active powders. It makes it possible, for example, to obtain composite particles comprising several layers of different compositions.

In a particular embodiment, the coating step d) can be carried out with one or more coating agents by applying:

- a monolayer made up of a single coating agent, or

- a monolayer made up of a mixture of several coating agents, or

- a multilayer consisting of a single coating agent, that is to say a superposition of several monolayers consisting of the same coating agent, or

- a multilayer consisting of a mixture of several coating agents, or

- a superposition of several monolayers made up of a single coating agent, each of the monolayers having a different coating agent.

Another aspect of the invention relates to a pharmaceutical composition with sustained and / or delayed release comprising a drug antibody and at least one or more pharmaceutically acceptable excipients chosen from the group comprising: a carboxymethyl-dextran, a chitosan, a cyclodextrin or a combination of these, for use in the treatment of diseases of the gastrointestinal tract, in patients who have developed antibodies directed against the drug antibody.

Another aspect of the invention relates to a pharmaceutical composition with sustained and / or delayed release comprising a drug antibody and at least one or more pharmaceutically acceptable excipients chosen from the group comprising: a carboxymethyl-dextran, a chitosan, a cyclodextrin or a combination of these, for use in the treatment of a disease of the gastrointestinal tract selected from: inflammatory bowel disease (IBD), irritable bowel syndrome (IBS), Crohn's disease, ulcerative colitis, hemorrhagic rectal ulcer, lesions concomitant to Behçet's disease, pochitis and cancer, in patients who have developed antibodies to the drug antibody. Advantageously, the cancer may be cancer of the gastrointestinal tract and in particular cancer of the small intestine and colorectal cancer.

Another aspect of the invention relates to a sustained release and / or delayed release pharmaceutical composition comprising a drug antibody as an active ingredient and a combination of carboxymethyl dextran and cyclodextrin as pharmaceutically acceptable excipients for use in the treatment of a disease of the gastrointestinal tract in patients who have developed antibodies against the drug antibody, characterized in that:

the drug antibody is chosen without limitation from the group comprising an anti-TNFα antibody, an anti-interferon type 1 receptor antibody, an anti-IL-1 antibody, an anti-IL-6 antibody, anti-IL-8 antibody, anti-IL-12 antibody, anti-IL-13 antibody, anti-IL-17 antibody, anti-IL-23 antibody, anti-IL-34 antibody, antibody anti-CD3, an anti-CD4 antibody, an anti-CD20 antibody, an anti-CD70 antibody, an anti-IFN-y antibody, an anti-INF-γ receptor antibody, an anti-CEA antibody, an antiintegrin antibody, in particular an anti-integrin a or β antibody and advantageously an anti-integrin α4β7 antibody and an anti-b7 antibody, an anti-VEGF antibody, an anti-EGFR antibody, and

- the disease of the gastrointestinal tract is chosen in a nonlimiting way from the group comprising the inflammatory bowel disease (IBD), irritable bowel syndrome (IBS), Crohn's disease, ulcerative colitis, rectal hemorrhagic ulcer, lesions concomitant with Behçet's disease, pochitis and cancer, in particular cancer of the gastrointestinal tract and in particular cancer of the small intestine and colorectal cancer.

Another aspect of the invention is a method of treating diseases of the gastrointestinal tract comprising administering to a patient in need thereof and having developed antibodies directed against the drug antibody of a sustained release pharmaceutical composition comprising a drug antibody and at least one or more pharmaceutically acceptable excipients selected from the group comprising: a carboxymethyl-dextran, a chitosan, a cyclodextrin or a combination thereof.

Another aspect of the invention is a method of treating diseases of the gastrointestinal tract comprising administering to a patient in need thereof and having developed antibodies directed against the drug antibody of a sustained release pharmaceutical composition comprising a drug antibody as active ingredient and a combination of carboxymethyl-dextran and cyclodextrin as pharmaceutically acceptable excipients and, characterized in that:

the drug antibody is chosen without limitation from the group comprising an anti-TNFα antibody, an anti-interferon type 1 receptor antibody, an anti-IL-1 antibody, an anti-IL-6 antibody, anti-IL-8 antibody, anti-IL-12 antibody, anti-IL-13 antibody, anti-IL-17 antibody, anti-IL-23 antibody, anti-IL-34 antibody, antibody anti-CD3, anti-CD4 antibody, anti-CD20 antibody, anti-CD70 antibody, anti-IFN-γ antibody, anti-INF-γ receptor antibody, anti-CEA antibody, anti-integrin antibody, in particular an anti-integrin a or β antibody and advantageously an anti-integrin α4β7 antibody and an anti-β7 antibody, an anti-VEGF antibody, an anti-EGFR antibody, and

- the disease of the gastrointestinal tract is chosen in a nonlimiting way from the group comprising the inflammatory bowel disease (IBD), irritable bowel syndrome (IBS), Crohn's disease, ulcerative colitis, rectal hemorrhagic ulcer, lesions concomitant to Behçet's disease, pochitis and cancer, in particular cancer of the gastrointestinal tract and in particular cancer of the small intestine and colorectal cancer.

FIGURES

Figure 1: Figure 1 relates to the quantification of human IgG in the serum of dogs treated by administration of the anti-TNFa Humira® antibody by the subcutaneous route. The quantification of human IgG was carried out using the FastELYSA kit.

FIG. 2: FIG. 2 relates to the quantification of human IgG in the serum of dogs treated by administration of the composition according to the invention comprising an anti-TNFα antibody by the oral route, at the rate of six repeated administrations. The quantification of human IgG was carried out using the FastELYSA kit. The antiTNFa antibody used has the same amino acid sequence as the antibody marketed as Humira® but is produced in transgenic goat milk.

Figure 3: Figure 3 relates to the binding capacity of the anti-TNFa antibody present in the composition according to the invention to TNFa in the feces of dogs treated by administration of the composition according to the invention comprising an anti-TNFa antibody by orally, at the rate of six repeated administrations. The binding capacity to TNFα was measured by ELISA test. The anti-TNFα antibody used has the same amino acid sequence as the antibody marketed as Humira® but is produced in transgenic goat milk.

Figure 4: Figure 4 relates to the quantification of human IgG in the feces of dogs treated by administration of the composition according to the invention comprising an anti-TNFα antibody by the oral route, in a proportion of six repeated administrations. The quantification of human IgG was carried out using the FastELYSA kit. The anti-TNFa antibody used has the same amino acid sequence as the antibody marketed as Humira® but is produced in transgenic goat milk.

Figure 5: Figure 5 shows the binding capacity of the anti-TNFα antibody at a concentration of 15 ng / ml, present in the serum of dogs (diluted 1/50) treated by administration of the composition according to the invention comprising an anti-TNFα antibody by the oral route. The anti-TNFα antibody used has the same amino acid sequence as the antibody marketed as Humira® but is produced in transgenic goat milk.

EXAMPLES

Example 1: Preparation of an animal model of inflammatory bowel disease (colitis)

Experimental colitis was induced by exposure to TNBS (trinitrobenzene sulfonic acid) in beagle dogs according to the following protocol: on day 0 (DO) of the study, 16 naive dogs underwent laparotomy and a segment of 10 cm long in the proximal part of the ascending colon was isolated. 20 ml of 10% TNBS (Sigma Aldrich) solution was injected into the lumen of the colon and allowed to incubate for 30 minutes. After that, the TNBS was removed by suction and the lumen of the colon was rinsed twice using sterile saline.

In the following examples, the administration of the antibody was carried out from day 2 (D2) after the induction of colitis and two anti-TNFα antibodies were tested:

- the anti-TNFa antibody marketed under the name Humira®, and

- an anti-TNFα antibody having the same amino acid sequence as the antibody marketed under the name Humira® but produced in transgenic goat milk.

Example 2: Routes of administration and dosing regimes tested.

Two routes of administration of an anti-TNFα antibody were tested: the subcutaneous route and the oral route.

The dogs were treated according to the protocol mentioned in Table 1 below.

Table 1: Protocol used

way administration Composition administered Dosing regimen Number of dogs tested Untreated - - 3 Subcutaneous Injectable composition comprising an anti-TNFa antibody marketed under the name Humira® a first dose of 160 mg of Humira® / animal on day 2 after the induction of colitis (D2), then a second dose of 80 mg of Humira® / animal on day 7 after the induction of colitis (D7) ) and a third dose of 40 mg of Humira® / animal on day 21 after the induction of colitis (D21). 3 Oral (6 administrations repeated) * Oral composition according to the invention comprising: - an anti-TNFα antibody having the same amino acid sequence as the antibody marketed under the name Humira® but produced in transgenic goat milk, - in the presence of the mixture of excipients carboxymethyl-dextran (CMD) and cyclodextrin and, - in capsule form (coating agent: Eudragit S100 (3 layers)) Six repeated administrations of 100 mg / animal (in the form of two capsules) in the morning on an empty stomach according to the following therapeutic scheme: a first dose of 100mg / animal on day 2 after induction of colitis (D2). a second dose of 100mg / animal on day 3 after induction of colitis (D3). a third dose of 100mg / animal on day 4 after induction of colitis (D4). a fourth dose of 100 mg / animal on day 5 after the induction of colitis (D5). a fifth dose of 100 mg / animal on day 6 after the induction of colitis (D6). a sixth dose of 100mg / animal on day 7 after induction of colitis (D7). 3

*: In fasting dogs the gastric pH may be high. In order to preserve the gastro-resistant coating of the capsules in treated dogs, a gastric catheter was placed during the surgical intervention on day 0 (DO), in order to be able, by the injection of 75 ml of HCI / KCI, pH 1.0, buffer the gastric medium at acidic pH before administration of the capsules (Polentarutti B et al, J Pharm Pharmacol. 62 (4), 2010).

Example 3 Determination of the Presence of Anti-TNFα Antibodies in the Serum

A blood sample (4 ml) was taken from all the animals from a cephalic vein at DO (before surgery) and at D3, D6, D9, D12, D15, D18, D21, D24, D27 , D30, D35, D42, D49 and D56 after induction of colitis. The dogs were sacrificed on day 56 (D56).

The blood samples were allowed to coagulate at room temperature for 15 to 30 minutes, then centrifuged at 1500 g for 10 minutes at around 4 ° C.

For each blood sample, the resulting serum was separated into 4 aliquots ("250 μl) and transferred, using a disposable plastic pipette, into 1.5 ml polypropylene Eppendorf tubes. The samples were immediately transferred to a freezer (-20 ° C) in an upright position.

The concentrations of human IgG samples were measured using the FastELYSA kit from RD biotech (ref. RDB-3257-10). In a first series of experiments, all the samples were diluted 1/40 in the FastELYSA diluent. Samples with an optical density (OD) outside the standard curve were diluted either to 1/200 (for those with a high OD), or to 1/10 (for those with a low or negative OD).

The results are presented in Figures 1 and 2.

3.1. Subcutaneous administration

It is noted that the dogs having received the composition comprising an anti-TNFa antibody marketed under the name Humira® subcutaneously, at the rate of a first dose of 160 mg of Humira® / animal on day 2 after induction colitis (D2), then a second dose of 80 mg of Humira® / animal on day 7 after induction of colitis (D7) and a third dose of 40 mg of Humira® / animal on day 21 after induction of colitis (D21), have a high level of IgG in the blood. The maximum IgG level is reached on day 9 (D9), with an IgG content of 116,000 ng / ml (see Figure 1).

These results show a significant passage of anti-TNFα antibodies into the bloodstream after subcutaneous administration. It is also observed that the anti-TNFα antibodies are rapidly eliminated.

3.2. Oral administration

In the case of oral administration, two capsules, coated with Eudragit S100 (3 layers), and each containing 56 mg of an anti-TNFa antibody having the same amino acid sequence as the antibody marketed under the name Humira® but produced in transgenic goat milk, formulated in the presence of the mixture of carboxymethyl-dextran excipients (CMD) and cyclodextrin, were administered daily for 6 days, from day 2 to day 7.

It is noted that the dogs having received the composition according to the invention, at the rate of six repeated administrations of two capsules from day 2 to day 7, have an undetectable level of IgG in the blood (see FIG. 2), less than 60 ng / ml, test detection threshold.

These results show the absence of passage of anti-TNFα antibodies into the blood circulation after administration of the composition according to the invention by the oral route, unlike administration by the subcutaneous route.

Example 4 Determination of the Presence of Anti-TNFα Antibodies in the Faeces

Faecal samples (~ 1g) were taken for dogs treated with the oral composition according to the invention at DO (before induction of colitis), and between day 0 (DO) and day 14 (D14) after induction of colitis.

The aliquots were placed in a glass vial with 5 ml of P8340 protease inhibitor cocktail diluted 1: 100. The mixture was vortexed until the sample was homogenized. After centrifuging the sample at 1500 g for 5 min at approximately 4 ° C, 2 aliquots of supernatant were separated in polypropylene tubes and stored, frozen until the TNFα assays of the faeces were developed.

1. Determination of the presence of anti-TNFα antibodies in the faeces

1.1. Assessment of binding to soluble TNFα by ELISA test

The detection in the faeces of antibodies having a binding capacity to TFNa was carried out by ELISA. The plates were coated overnight with mouse anti-human TNFα antibody at 4 pg / ml in PBS. After washing and saturation in PBS + 1% BSA, TNF-α soluble at 100 ng / ml was incubated for 90 minutes at room temperature. After washing, the feces supernatants diluted 1/10 were incubated and then detected with an anti-human IgG antibody (H + L) linked to peroxidase.

The peroxidase activity was then measured by staining of 2,2'-azino-bis (3ethylbenzothiazoline-6-sulfonic acid).

A standard curve was performed with anti-TNFα antibody from 30 ng / ml to 0.47 ng / ml. The sample concentrations obtained with the standard curve were compared with the theoretical concentration to obtain the percentage of binding to TNFα.

1.2. Quantification of human IgG by FastELYSA

Before quantification, all the samples were centrifuged for 5 min at 480 g to remove the faeces still present in the samples. The concentrations of human IgG samples were measured using the FastELYSA kit from RD biotech (ref. RDB-3257-10). All samples were diluted 1/4 in FastELYSA diluent.

1.3. Results obtained

Assessment of binding to soluble TNFα by ELISA test

It is found that anti-TNFα antibodies are indeed present in the faeces of the three dogs treated with the composition according to the invention administered in the amount of six repeated administrations (see Figure 3). The maximum rate observed is 70 ng / ml.

Quantification of human IgG by FastELYSA

To confirm the results of the TFNa ELISA test, the same samples were studied to detect human IgG in faeces by FastELYSA assay (specific for human IgG) independently of their binding to TFNa. The results show that human IgG is well detected in the three dogs in the FastELYSA test (see Figure 4) with a maximum level of 130 ng / ml.

Compared to the anti-TNFa ELISA test which detects only IgG or fragments of IgG binding to TNFa, the levels of total human IgG obtained with the FastELYSA assay remain of the same order of magnitude.

Due to the presence of anti-TNFα antibodies in the feces, it is concluded that the composition according to the invention is indeed delivered to the intestine of dogs treated but without passing into the bloodstream.

Example 5 Evaluation of the Presence of Anti-TFNa Antibody Antibodies (ADA) in Dog Serum

The detection of anti-TFNa antibodies (ADA) in dog serum, capable of neutralizing the binding activity of TFNa, was carried out by ELISA. The plates were coated overnight with mouse anti-human TNFα antibody at 4 pg / ml in PBS. After washing and saturation in PBS + 1% BSA, human TNFα soluble at 100 ng / ml was incubated for 90 minutes at room temperature.

The following method was used to determine the presence of ADA in dog serum samples. Three dogs were treated with oral administration of an anti-TNFα antibody composition. The sera, which in this case do not contain anti-TNFα antibodies, were collected and diluted 1/50, then exogenous anti-TNFα antibody was added to the diluted sera to reach a final concentration of 15 ng / ml, in order to test the inhibition of its binding to TNFα by the serum.

After washing, the samples were incubated and the binding of the anti-TNFα antibody to TNFα was detected with an anti-human IgG antibody (H + L) linked to the peroxidase.

The peroxidase activity was then measured with 2,2'-azino-bis (3ethylbenzothiazoline-6-sulfonic acid).

The standard curve was made with a reference anti-TNFα antibody (TG-Humira, lot 090414 A1) (0.47 to 30 ng / ml).

The sample concentrations obtained were compared with the theoretical concentration of 15 ng / ml and expressed as a percentage of binding to TNFα. The 100% binding percentage corresponds to the anti-TNFα antibody diluted in untreated dog serum and having a concentration of 15 ng / ml. A percentage less than 100% suggests that the dog serum sample contains ADA with neutralizing activity.

The results (see FIG. 5) show that the binding capacity of the anti-TNFα antibody present in the composition according to the invention to TNFα is little or not affected. A TNFα binding capacity of 100% of the anti-TNFα antibody present in the composition according to the invention is observed until day 12 (D12) for dogs 2906997 and 2921244. A moderate decrease in TNFα binding from approximately 25% is observed in a dog (2915112) on the same day (D12). These results indicate that, 12 days after the start of the treatment, the sera of the 3 dogs having received repeated doses of the composition according to the invention comprising an anti-TNFa antibody, did not contain or almost no anti-anti-antibody -TNFa (ADA) neutralizing inhibiting the binding of the anti-TNFa antibody to soluble TNFa.

These results are particularly promising compared with the state of the art data. Indeed, it is known that the intravenous injection of Humira is immunogenic and generates ADA on 7 ^th day (Mabs Myzithras M et al 2016, 8: 1606; Monograph Humira Abbott - Version 18 March 2015).

In total, these results indicate that (1) the composition according to the invention comprising an anti-TNFa antibody can be administered effectively in the dog's intestine (2) without passing into the circulation, even in the presence of lesions of the colon (colitis TNBS-induced experimental study), and (3) without the appearance of neutralizing ADA despite the difference in species, over the 12-day study period.

These results therefore demonstrate that the oral composition according to the invention is particularly suitable for the treatment of patients who have developed antibodies directed against the drug antibody (ADA). Indeed, these drug antibodies, locally active and not passing into the bloodstream, are not inactivated by the patient's ADA. In addition, the absence of passage in the circulation of the drug antibody contributes to limiting the production of ADA by the patient.

Claims (9)

1. Oral pharmaceutical composition comprising at least one drug antibody and at least one pharmaceutically acceptable excipient, for use in the treatment of diseases of the gastrointestinal tract, in patients having developed antibodies directed against the drug antibody. 2. Pharmaceutical composition for use according to claim 1, characterized in that the drug antibody is chosen from the group comprising an anti-TNFα antibody, an anti-type 1 interferon receptor antibody, an anti-IL antibody -1, an anti-IL-6 antibody, an anti-IL-8 antibody, an anti-IL-12 antibody, an anti-IL-13 antibody, an anti-fL-17 antibody, an anti-IL-23 antibody , an anti-IL-34 antibody, an anti-CD3 antibody, an anti-CD4 antibody, an anti-CD20 antibody, an anti-CD70 antibody, an anti-IFN-y antibody, an anti-INF-γ receptor antibody , an anti-CEA antibody, an anti-integrin antibody, an anti-b7 antibody, an anti-VEGF antibody and an anti-EGFR antibody, 3. Pharmaceutical composition for use according to one of claims 1 to 2, characterized in that the drug antibody is produced in mammary non-human mammalian epithelial cells or in the milk of a non-human transgenic mammal. 4. Pharmaceutical composition for its use according to one of claims 1 to 3, characterized in that the pharmaceutically acceptable excipients are chosen from the group comprising: a carboxymethyl-dextran, a chitosan, a cyclodextrin or a combination thereof. 5. Pharmaceutical composition for use according to one of claims 1 to 4, characterized in that the pharmaceutical composition also comprises an amino acid. 6. Pharmaceutical composition for use according to one of claims 1 to 5, characterized in that the pharmaceutical composition is in the form of a formulation with sustained and / or delayed release. 5 7. Pharmaceutical composition for its use according to one of claims 1 to 6, characterized in that the pharmaceutical composition is in the form of a sustained-release and / or delayed-release formulation coated with an enteric coating agent . 10 8. Pharmaceutical composition for use according to claim 1, characterized in that the disease of the gastrointestinal tract can be a chronic inflammatory bowel disease (IBD), irritable bowel syndrome (IBS), Crohn's disease , ulcerative colitis, hemorrhagic rectal ulcer, lesions concomitant to Behçet's disease, pochitis and 15 cancer. 9. Pharmaceutical composition for its use according to claim 1, characterized in that the disease of the gastrointestinal tract can be cancer of the gastrointestinal tract and in particular cancer of the small intestine and cancer 20 colorectal.