Classifications

• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K39/00—Medicinal preparations containing antigens or antibodies
  • A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
  • A61K39/39591—Stabilisation, fragmentation
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K2317/00—Immunoglobulins specific features
  • C07K2317/90—Immunoglobulins specific features characterized by (pharmaco)kinetic aspects or by stability of the immunoglobulin
  • C07K2317/94—Stability, e.g. half-life, pH, temperature or enzyme-resistance

Definitions

• composition of concentrated human immunoglobulins Composition of concentrated human immunoglobulins
• the invention relates to a concentrated human immunoglobulin G composition having improved stability over time.
• the composition according to the invention is particularly suitable for use subcutaneously.
• IgG immunoglobulin G
• examples include primitive immune deficiencies with defective antibody production, Kawasaki disease, immunologic thrombocytopenic purpura in children and adults, secondary immune deficiencies with defective antibody production, in particular chronic lymphocytic leukemia or myeloma associated with recurrent infections, HIV infection of the child with bacterial infections, multifocal motor neuropathies, Guillain-Barré syndrome, severe acute or chronic parvovirus B infections 19, acquired or constitutional immunodeficiency, corticosteroid dermatomyositis, acute myasthenia, idiopathic chronic polyradiculoneuropathy, immune thrombocytopenic purpura, for example associated with HIV infection, stiff man syndrome (Stiffman Syndrome) ), autoimmune neutropenia, resistant autoimmune erythroblastopenia, anti-coa syndrome acquired by auto-antibodies, rheumatoid arthritis, uveitis, etc.
• Stiffman Syndrome stiff man syndrome
• autoimmune neutropenia resistant autoimmune
• IgG compositions suitable for subcutaneous administration may be particularly advantageous.
• This route of administration offers more flexibility and independence to patients, improving their quality of life.
• immunoglobulin compositions comprising high concentrations of IgG have been developed.
• the higher the concentration of IgG the more the problems of stability over time increase.
• oligomers and polymers may activate the complement system with associated risks of anaphylactic reactions.
• These oligomers and polymers are also likely to induce hypotension phenomena in treated patients. This is undesirable and is strictly controlled from the regulatory point of view.
• the formation of protein aggregates due in particular to thermal, mechanical or chemical stress, contributes to these problems of instability.
• the presence of certain detergents conventionally used for intravenous administration can induce a local reaction when administered subcutaneously.
• concentrated IgG compositions that is to say comprising at least 16% IgG, for subcutaneous administration are not entirely satisfactory in terms of stability and local tolerance.
• the Applicant has demonstrated that it is possible to obtain concentrated IgG compositions having a very good tolerance. local after subcutaneous administration and advantageously particularly stable over time. More specifically, the Applicant has developed a formulation combining a high concentration of IgG, with glycine and a nonionic detergent, in which the glycine and the nonionic detergent are in particularly low concentrations, adapted to a use subcutaneously. The combination of nonionic detergent and glycine in such proportions contribute to a high stability of said formulation over time. In addition, the Applicant has demonstrated that such a formulation is particularly well tolerated by patients when administered subcutaneously.
• the concentrated immunoglobulin compositions according to the invention exhibit a substantially physiological osmolality.
• no other excipient than glycine and nonionic detergent is necessary, and in particular no acetate or mannitol or albumin, to ensure greater stability during storage.
• An object of the invention is therefore a pharmaceutical composition
• a pharmaceutical composition comprising:
• immunoglobulin G 200 g / L +/- 5% immunoglobulin G (IgG)
• the composition consists only of water, IgG, glycine and nonionic detergent.
• composition according to the invention advantageously comprises 20% (200 g / l) of IgG.
• IgG concentrations are +/- 5% of the concentration in g / L.
• the IgGs are human IgG, in particular obtained from plasma or plasma fractions.
• the glycine concentration is 215 mM, +/- 5%.
• Such a glycine concentration is particularly suitable for 20% IgG compositions for subcutaneous use.
• the nonionic detergent is chosen from poloxamers, and in particular Pluronic F68®, and polysorbates, preferably polysorbate 20 or polysorbate 80, more preferably polysorbate 80.
• the nonionic detergent is at a concentration of about 20 ppm +/-
• the subject of the invention is also such an IgG composition for use subcutaneously for the treatment of a dysfunction of the immune system, an autoimmune and / or inflammatory disease, an infection or a neurological disease.
• immunoglobulin G or "IgG” in the context of the invention, polyvalent immunoglobulins which are essentially IgG, optionally including IgM. It may be whole immunoglobulins, or fragments such as F (ab ') 2 or F (ab) and any intermediate fraction obtained during the process of manufacturing polyvalent immunoglobulins.
• Stability corresponds to the physical and / or chemical stability of IgG.
• physical stability refers to the reduction or absence of formation of insoluble or soluble aggregates of the dimeric, oligomeric or polymeric forms of Ig, as well as the reduction or absence of any structural denaturation of the molecule. .
• the term "chemical stability” refers to the reduction or absence of any chemical modification of IgG during storage, in the solid state or in dissolved form, under accelerated conditions. For example, the phenomena of hydrolysis, deamidation, and / or oxidation are avoided or delayed. The oxidation of sulfur-containing amino acids is limited.
• the stability of an IgG composition can be evaluated by visual inspection, in particular using an optical fiber device (opalescence, particle formation), by measuring the turbidity using a spectrophotometer measuring the absorbance or optical density at 400 nm, for example, and / or by measurement of the scattered light (dynamic light scattering (DLS)) for measuring the particles in solution of sizes of between 1 nm and 1 pm approximately.
• the IgG concentration is 200 g / L, + 1-5%.
• the concentrations are at the level of the final composition, ready for use.
• the concentrations are determined with respect to the compositions in liquid form, before drying, or after reconstitution as an injectable preparation.
• the Applicant has demonstrated that it is possible to obtain compositions comprising 20% IgG particularly stable over time using a minimum of excipients.
• the 20% IgG compositions advantageously comprise between 200 and 250 mM of glycine, preferably between 200 and 230 mM, preferably between 210 and 220 mM.
• the 20% IgG composition comprises 215 mM glycine, +/- 5%.
• the 20% IgG compositions comprise between 15 and 25 ppm of nonionic detergent.
• the 20% IgG composition comprises 20 ppm of nonionic detergent, +/- 10%.
• the nonionic detergent used in the composition according to the invention is advantageously chosen from polysorbates and in particular from polysorbate 80 (or Tween®80 which is polyoxyethylenesorbitan monooleate), and polysorbate 20 (or Tween Which is polyoyethylene sorbitan monolaurate).
• the nonionic detergent is chosen from poloxamers, and in particular Pluronic® F68 (polyethylenepolypropylene glycol).
• the nonionic detergent is selected from Triton® X 100 (octoxinol 10), polyoxyethylene alkyl ethers and copolymer blocks of ethylene / polypropylene. Nonionic detergents can also be combined with each other.
• the composition is free of mannitol and / or albumin and / or acetate.
• the Applicant has in fact shown that mannitol and / or acetate and / or albumin are not necessary for the stabilization of a 20% IgG composition.
• the composition contains neither mannitol nor albumin. According to another preferred embodiment, the composition does not contain acetate. According to a particularly preferred embodiment, the composition according to the invention does not contain mannitol, acetate or albumin. In an alternative or complementary embodiment, the composition does not contain sugar.
• the Applicant has demonstrated that the compositions according to the invention have an osmolality particularly suitable for administration by injection, especially subcutaneous, and without adding in number and / or amounts of excipients.
• the invention provides 20% IgG compositions having a measured osmolality of about 300 to 400 mOsm / kg, adjusted by glycine.
• the osmolality of the composition refers to the osmolality measured in said composition.
• the osmolality is advantageously measured using an osmometer calibrated with standard solutions, and in particular according to the method recommended by the European Pharmacopoeia, (European Pharmacopoeia 5.0 of 2005 -01/2005: 2.2.35.). Of course, any other method of measuring osmolality can be used.
• the only excipients of the 20% IgG composition according to the invention are glycine and nonionic detergent.
• a formulation allows a good stabilization of immunoglobulin compositions over time and reduction of time and cost of preparation on an industrial scale through the presence of an effective minimum number and amount of excipients.
• such a composition has an osmolality compatible with the administration by injection, in particular by the subcutaneous route.
• the composition consists essentially of IgG, glycine, a nonionic detergent and water, in the sense that any other excipient that may be present would only be in the state of trace.
• the final pH of the composition is advantageously between 4.6 and 5.0.
• the pH is about 4.8 +/- 0.1.
• a pH of 4.8 +/- 0.1 gives indeed particularly satisfactory results in terms of stability over time.
• the final pH refers to the pH of the composition after formulation, i.e., in the ready-to-use composition. Unless otherwise stated, in this specification, the pH of the composition refers to the final pH.
• a preferred IgG composition according to the invention comprises:
• the pH of the composition being between 4.6 and 5.0.
• Another preferred IgG composition according to the invention comprises:
• nonionic detergent preferably polysorbate 80 or Pluronic F68® the pH of the composition being between 4.6 and 5.0.
• a particularly preferred IgG composition according to the invention comprises:
• nonionic detergent 20 ppm of nonionic detergent, +/- 10%, preferably polysorbate 80 or Pluronic F68®, the pH of the composition being 4.8 +/- 0.1.
• Another particularly preferred IgG composition according to the invention comprises:
• nonionic detergent preferably polysorbate 80 or Pluronic F68® the pH of the composition being 4.8.
• the osmolality measured of this 20% IgG composition is advantageously about 300-400 mOsm / kg, +/- 2%, preferably about 340 mOsm / kg, +/- 2%.
• composition according to the invention comprises
• nonionic detergent preferably polysorbate 80 or Pluronic F68®
• the pH of the composition being 4.8
• its osmolality being 340 mOsm / kg, +/- 2%
• said composition being salt-free of acetate, mannitol and albumin.
• a glycine concentration of about 215 mM, +/- 5% combined with a concentration of polysorbate 80 or Pluronic F68® of 20 ppm is sufficient to maintain stability of the 20% immunoglobulin composition over time, while maintaining an osmolality of between 300 and 400 mOsm / kg in the composition, while higher concentrations would have been expected to ensure stability, increasing the osmolality of said compositions.
• a too strong osmolality can be at the origin of a dehydration of cells (exit of intracellular water to the extracellular medium) detrimental to the patient.
• the increase in the amount of the excipients, and in particular the nonionic detergents may cause a decrease in the local tolerance of the composition administered subcutaneously.
• the composition developed by the Applicant in which the number and the amount of the excipients are low, is therefore particularly advantageous for subcutaneous administration.
• compositions according to the invention advantageously comprise human immunoglobulins G.
• Human IgG is usually obtained by fractionation of human blood plasma and presented in an aqueous medium.
• the aqueous medium is composed of water for injection (PPI water) which may contain pharmaceutically acceptable excipients and compatible with IgG.
• the IgG compositions may previously undergo specific virus inactivation / removal steps, such as detergent solvent treatment, pasteurization and / or nanofiltration.
• the composition according to the invention comprises IgG which can be polyclonal or monoclonal. IgGs can be isolated from human or animal blood or produced by other means, for example by molecular biology techniques, for example in cell systems well known to those skilled in the art.
• the composition according to the invention is particularly suitable for highly purified IgG.
• the IgGs of the present invention are obtained by fractionation of human plasma.
• Preferred fractionation methods of human plasma are described by Cohn et al (J. Am Chem Soc., 68, 459, 1946), Kistler et al. (Vox Sang., 7, 1962, 414-424), or Steinbuch et al (French Rev. et.Clin et al., XIV, 1054, 1969).
• a method for preparing an immunoglobulin G composition is also described in the patent application WO2002 / 092632.
• compositions according to the invention are advantageously in liquid form.
• the 20% IgG composition according to the invention in liquid form and after storage for a period of 6 months at 25 ° C., has a level of polymers which is well below the maximum level allowed by the European Pharmacopoeia (3%). advantageously less than about 1%.
• compositions of the invention are pharmaceutical compositions, that is to say adapted for a therapeutic use.
• the pharmaceutical compositions of the invention are thus useful as medicaments, in particular for the treatment of a dysfunction of the immune system, an autoimmune and / or inflammatory disease, an infection or an illness. neurological.
• compositions according to the invention are particularly particularly suitable for the treatment of disorders such as primary immunodeficiency deficiency with lack of antibody production, Kawasaki disease, immunological thrombocytopenic purpura in children and adults, immune deficiency antibodies, particularly chronic lymphocytic leukemia or myeloma associated with repetitive infections, HIV infection of the child with bacterial infections, multifocal motor neuropathies, Guillain- Barré, severe or chronic acute infections with Parvovirus B 19, acquired or constitutional immunodeficiency, corticosteroid dermatomyositis, acute myasthenia, idiopathic chronic polyradiculoneuropathy, immune thrombocytopenic purpura, for example associated with HIV infection , stiff man syndrome (Stiffman syndrome), neutropenia has autoimmune erythroblastopenia, autoantibodies-induced anticoagulation syndrome, rheumatoid arthritis, uveitis.
• disorders such as primary immunodeficiency deficiency with lack of antibody
• composition according to the invention is suitable for the treatment of a human subject, whatever his age, and more particularly an adult subject, child or infant.
• compositions according to the invention may advantageously be subjected to a method for eliminating or inactivating the infectious agents, for example by solvent-detergent treatment or by nanofiltration. Such methods of removing or inactivating infectious agents are well known to those skilled in the art.
• the 20% IgG composition according to the invention is useful in therapy, and especially in injectable form, in particular, subcutaneously.
• the subcutaneous route for the treatment of chronic autoimmune diseases has several advantages, such as improving patient comfort and reducing side effects.
• Subcutaneous administration does not require venous access, which is, in some cases, a decisive advantage when the absence of venous access blocks access to treatment, especially for young children.
• the use of subcutaneous immunoglobulins also reduces some of the side effects associated with intravenous infusions, particularly the risk of systemic reactions. Large changes in circulating levels observed intravenously are avoided, allowing a better regulation of the serum level in the physiological range between infusions.
• SCIGs subcutaneously administered immunoglobulins
• IVIG intravenous immunoglobulins
• IgSC intracranial pressure
• the concentration of the IgSC is a defining characteristic which conditions the injection volume and the number of injection sites and consequently the frequency of administration.
• An immunoglobulin G composition is prepared according to the method as described in application EP1385886.
• the product obtained is then concentrated to 200 g / L by ultrafiltration on Ultracel C cellulose membrane (Millipore®) with a cutoff threshold of 30 kDa in order to obtain the product ready to formulate (PPL).
• compositions The 200 g / L concentrated products ready for formulation are supplemented with glycine, polysorbate 80 or Pluronic L68® and adjusted to the desired pH in order to obtain the following compositions: Table 1: IgG Compositions
• compositions are subjected to stirring stress on a magnetic plate at 440 rpm for 6 hours, the following analyzes are carried out as follows: Turbidity (OD at 400 nm): the turbidity is determined by measuring the absorbance at 400 nm. Water for injection is used as white.
• DLS / S LS This technique tracks the aggregation state of the solution.
• Dynamic Light Scattering measures the size (hydrodynamic diameter) of objects in solution, approximately between 1 nm and 1 mih. Each size population is thus followed.
• SLS static light scattering: follows the phenomenon of aggregation in its together. The measurement is carried out by adding 40 mM NaCl to the solution. The measurement is carried out on a sample diluted at 80 g / l in water for injection.
• Table 3 Intensity of the monomers at 90 ° (%) before and after 2h, 4h and 6h of agitation stress.
• Table 4 Total intensity scattered at 90 ° (u.a) before and after 2h, 4h and 6h stirring stress.
• Sub-Visible Particles Sub visible particles larger than 2 ⁇ m, 10 ⁇ m and 25 ⁇ m are counted using the micro fluidic imaging technique.
• compositions Fl, F2 and F3 with 0 or 10 ppm of surfactant, show unsatisfactory overall stability results, in particular on the measurement of aggregation (DLS) and subvisible particles (MFI), demonstrating a low stability of the composition.
• DLS aggregation
• MFI subvisible particles
• compositions F6 and F7 comprising 30 ppm of surfactant show unsatisfactory results, in particular on the measurement of aggregation (DLS) and subvisible particles (MFI), demonstrating a low stability of the composition.
• DLS aggregation
• MFI subvisible particles
• compositions according to the invention F4 and F5 comprising 20 ppm of surfactant make it possible to obtain a satisfactory stability on all the evaluated criteria.
• An immunoglobulin G composition is prepared according to the method as described in application EP1385886.
• the product obtained is then concentrated to 200 g / L by ultrafiltration on Ultracel C cellulose membrane (Millipore®) with a cutoff threshold of 30 kDa in order to obtain the product ready to formulate (PPF).
• Ultracel C cellulose membrane Millipore®
• compositions for stability are supplemented with glycine, polysorbate 80 or Pluronic F68® and adjusted to the desired pH in order to obtain the following compositions for stability:
• compositions After sterilizing filtration on 0.22pm filter (Sartopore), the compositions are aseptically distributed in glass vials (type I), which are then capped and stored, in a chamber regulated to
• Turbidity (OD at 400 nm): the turbidity is determined by measuring the absorbance at 400 nm. Water for injection is used as white.
• DLS / S LS This technique tracks the aggregation state of the solution.
• DLS dynamic light scattering
• SLS static light scattering
• the measurement is carried out by adding 40 mM NaCl to the solution.
• the measurement is carried out on a sample diluted at 80 g / l in water for injection.
• Sub-Visible Particles Sub visible particles larger than 2 ⁇ m, 10 ⁇ m and 25 ⁇ m are counted using the micro fluidic imaging technique.
• HPSEC High performance size exclusion chromatography is used to assess the level of fragmentation and aggregation of the product. Chromatogram analysis of optical density measurements at 208 nm determines the% of monomers, dimers, polymers and fragments.
• the tests at 40 ° C make it possible to accelerate the phenomena observed during the stability of the compositions because of the high stress applied.
• the methods used show an evolution of the criteria followed, confirming the relevance of the methods selected for the evaluation of the formulations during the stability monitoring.
• the formulations according to the invention F4 and F5 with 20 ppm of surfactant make it possible to maintain the stable compositions over time.

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• 2018
  • 2018-05-24 FR FR1854412A patent/FR3081328B1/fr active Active
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