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
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P7/00—Drugs for disorders of the blood or the extracellular fluid
  • A61P7/02—Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
  • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
  • C07K16/24—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against cytokines, lymphokines or interferons
  • C07K16/241—Tumor Necrosis Factors
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
  • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
  • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
  • C07K16/2839—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the integrin superfamily
  • C07K16/2848—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the integrin superfamily against integrin beta3-subunit-containing molecules, e.g. CD41, CD51, CD61

Definitions

• the present invention relates to the field of pharmaceutical compositions and the manufacture of lyophilized monoclonal antibody and antibody fragment compositions. More particularly, the invention relates to a lyophilized composition of abciximab and infliximab.
• Abciximab is the Fab fragment of the chimeric human-murine monoclonal antibody 7E3 which inhibits platelet aggregation in humans.
• Infliximab is the chimeric human-murine monoclonal antibody against tumor necrosis factor.
• the composition exhibits high glass transition temperatures and improved stability BACKGROUND OF THE INVENTION
• An essential feature in the development of a protein therapeutic is to design a formulation that is stable during shipment and storage before administration to a patient.
• an aqueous formulation is the easiest and most economical to handle for the manufacturer and easiest to administer to the patient.
• most preparations are prepared and stored in a liquid state.
• precise control of storage conditions such as refrigeration and minimization of agitation is necessary to retain stability. Such conditions are often seen as a drawback to commercial application of the product.
• the process has numerous advantages in that the aqueous solution can be processed and filled into dosage containers in a liquid state, dried at low temperatures thereby eliminating adverse thermal effects, and stored in the dried state where it may be more stable.
• the lyophilized product is ordinarily rapidly soluble and is easily reconstituted prior to administration to a patient.
• appropriate care must be taken to choose the conditions and excipients for use in the lyophilization process.
• most protein preparations are at least partially denatured by the freezing and dehydration stresses encountered during lyophilization. Additionally, excipients should be chosen that assure stability during long term storage in the dried solid.
• the glass transition temperature (Tg') of the amorphous phase in the product, which contains the protein must exceed the planned storage temperature. Since water is a plasticizer of the amorphous phase, low residual moisture is needed to insure that Tg' is greater than the highest temperature encountered during shipping and storage.
• the Tg' of the liquid state prior to lyophilization is a critical factor in lyophilization, and is determined by excipients in the liquid state and container configuration. In the case where a dual chamber syringe such as a Lyo-Ject syringe is to be used, a composition with a high Tg' is necessary to be compatible with the uniqueness of the syringe design.
• a monoclonal antibody is a protein molecule produced by fusing a chosen B cell line with an immortal myeloma cell line to produce hybridomas, immortal cells that secrete only the selected antibody type of the selected B cell clone.
• each antibody is formed by the interaction of two identical "heavy" chains and two identical light chains, all of which combine to form a Y shape (the heavy chains span the entire Y, and the light chains the two arms only.
• An antibody according to the present invention includes any protein or peptide containing molecule that comprises at least a portion of an immunoglobulin molecule, such as but not limited to at least one complementarity determining region (CDR) of a heavy or light chain or a ligand binding portion thereof, a heavy chain or light chain variable region, a heavy chain or light chain constant region, a framework region, or any portion thereof, that can be incorporated into an antibody of the present invention.
• An antibody of the invention can include or be derived from any mammal, such as but not limited to a human, a mouse, a rabbit, a rat, a rodent, a primate, or any combination thereof, and the like.
• An antibody fragment as used in the present invention includes the fragment of the immunoglobulin molecule known as the Fab containing the CDR antigen binding site, generated by cleavage of the antibody with the protease papain which cuts at the "hinge" region of the Y shaped antibody molecule producing two Fab fragments.
• monoclonal antibodies are protein molecules, they are generally formulated as aqueous solutions for injection. As such, they have limited shelf life and must be stored under controlled conditions as discussed above.
• the monoclonal antibody abciximab (REOPRO®), is the Fab fragment of the chimeric human-murine monoclonal antibody 7E3.
• Abciximab binds to the glycoprotein (GP) llb/llla receptor of human platelets and inhibits platelet aggregation. Abciximab also binds to the vitronectin ( ⁇ v ⁇ s) receptor found on platelets and vessel wall endothelial and smooth muscle cells. It is indicated as an adjunct to percutaneous coronary intervention for the prevention of cardiac ischemic complications. Abciximab is sold commercially as a clear, colorless, sterile solution for intravenous use and must be stored under refrigerated conditions at 2 to 8 S C. A detailed description of abciximab is disclosed in US patent 5,770,198 hereby incorporated by reference into the present application.
• the monoclonal antibody infliximab (REMICADE®) is a chimeric lgG1 k monoclonal antibody with an approximate molecular weight of 149,100 daltons. It is composed of human constant and murine variable regions. Infliximab binds specifically to human tumor necrosis factor alpha (TNF(alpha)) with an association constant of 10 10 M "1 . Infliximab is produced by a recombinant cell line cultured by continuous perfusion and is purified by a series of steps that includes measures to inactivate and remove viruses. REMICADE is currently supplied as a sterile, white, lyophilized powder for intravenous infusion.
• each single-use vial contains 100 mg infliximab, 500 mg sucrose, 0.5 mg polysorbate 80, 2.2 mg monobasic sodium phosphate, monohydrate, and 6J mg dibasic sodium phosphate, dihydrate. No preservatives are present.
• infliximab is disclosed in US patent 5,656,272 hereby incorporated by reference into the present application.
• a lyophilized product Prior to administration to a patient, a lyophilized product has to be reconstituted with sterile water.
• a convenient way to do this is to deliver the product in a dual chamber syringe such as a Lyo-Ject® syringe
• the Lyo-Ject is a dual-chamber prefilled syringe with in situ lyophilized drug.
• the lyophilized drug and diluent are contained in dual chambers in the same syringe and can be easily reconstituted prior to injection.
• Tg' glass transition temperature
• the object of the present invention is to provide a monoclonal antibody composition with enhanced stability by the application of lyophilization to the aqueous solution. Another object is to provide a lyophilized monoclonal antibody product which exhibits the high Tg' to be compatible with a dual chamber syringe.
• the invention concerns an improved solid pharmaceutical composition comprising a lyophilized monoclonal antibody composition that exhibits enhanced stability in the dry form and a high glass transition temperature.
• a lyophilized monoclonal antibody composition comprising a therapeutically active monoclonal antibody or fragment thereof in admixture with a stabilizing excipient comprised of a combination of a disaccharide and a hydroxyethyl starch
• the monoclonal antibody is abciximab.
• a facile process for the production of the solid composition is also provided by the present invention, comprising the steps of:
• the therapeutically active component of this invention is a monoclonal antibody as described above.
• the monoclonal antibody is abciximab as disclosed in U.S. Pat. Nos. 5,770,198 and 5,976,532, hereby incorporated by reference into the present application.
• the monoclonal antibody protein for use in the invention may be prepared by any techniques that are well known to those skilled in the art, for example by purification from a cell line, a mixed cell line, an immortalized cell or clonal population of immortalized cells, as well known in the art.
• Monoclonal antibodies useful in the present invention include abciximab, discussed above and the anti- TNF ⁇ chimeric monoclonal antibody (infliximab, REMICADE®) Methods for the production of the abciximab monoclonal antibody polypeptides are described in U.S. Pat. No. 5,770,198, hereby incorporated by reference into the present application. Methods for the production of the infliximab monoclonal antibody are described in U.S. Pat. No. 5,656,272 hereby incorporated by reference into the present application.
• the lyophilized monoclonal antibody composition is manufactured from the aqueous solution containing approximately 5 mg/ml polypeptide.
• Appropriately sized vials are filled with the aqueous solution with a fill volume of 2 to 5 ml per vial.
• the vials are then frozen in the lyophilization chamber, preferably at a gradual rate of 0.5°-1 ° C./min, for approximately 2 hours to a temperature of ⁇ -40° C. or until completely frozen.
• the lyophilization chamber pressure is reduced to about 100 microns Hg or less.
• the shelf temperature is then raised to -20° C- +20° C. and held until sublimation of ice is substantially complete.
• the shelf temperature is then gradually raised, preferably at a rate of 0.5° C./min to 30° C and held for about 2 hours or more.
• the lyophilized dry composition prepared by the method of the instant invention exhibits enhanced stability and can be stored at room temperature for 6 months or greater depending on product specifications.
• the sealed vials are intended for use as single dose formulations following reconstitution with appropriate volumes of Sterile Water for Injection. It is intended that the filled vials will allow rapid dispersion of the solid composition upon reconstitution with water in situ giving an appropriate sterile solution of the desired monoclonal antibody concentration for administration.
• the lyophilized product is generally a white powder which undergoes reconstitution in about 1-5 minutes by swirling the vial.
• the vials utilized should be capable of maintaining a sterile environment by being hermetically sealed by means of a stopper and overseal.
• the vials should be of an appropriate size, considering the volume of solution to be held upon reconstitution of the lyophilized composition; and should be made of appropriate material, generally Type I glass.
• the stopper means employed, preferably sterile rubber closures or an equivalent, should provide the appropriate seal but allow entry for the purpose of introducing the diluent for reconstitution.
• the lyophilized monoclonal antibody composition of the invention exhibits enhanced stability in the dry form and a high glass transition temperature and is obtained by lyophilizing the monoclonal antibody in the presence of a stabilizing excipient comprised of a combination of a disaccharide and a hydroxyethyl starch.
• a stabilizing excipient comprised of a combination of a disaccharide and a hydroxyethyl starch.
• the hydroxyethyl starch (HES) used in the invention is obtained from native starch after partial hydrolysis and substitution of the hydroxy groups by hydroxyethyl groups (see E. Nitsch, Chemie der Hydroxyethystarke (HES), Beitr. Anaesth. Intesivmed., 26 (1988) 15-26..
• HES is a known stabilizing agent in the freeze drying of biological material and comes in various commercially available forms depending on the degree of molar substitution.
• the HES has an average molecular weight of 10,000 - 200,000, preferably 20,000-60,000.
• the disaccharide is selected from known disaccharides, such as sucrose, trehalose, maltose and lactose, with the non-reducing disaccharides such as sucrose being preferrred.
• a combination of HES and disaccharide is employed in the lyophilizing process.
• the desired Tg' and stabilizing effect can be customized for the monoclonal antibody preparation employed.
• the combination of HES and disaccharide is employed at an appropriate ratio to provide a Tg' of greater than -26 9 C and sufficient stabilizing effect.
• the ratio of sucrose to HES is 6:4 but a range of 10:1 to 4:6 can be employed.
• the disaccharide and HES should preferably be contained in the aqueous monoclonal antibody solution in an amount of not less than 1 mg, more preferably 3 mg to 65 mg, per milliliter of said solution.
• the combination of disaccharide and HES provides sufficient stability, minimizes aggregation, shortens the lyophilization cycle time and minimizes the formation of visible particles.
• Monoclonal antibody preparation formulated in accordance with the invention exhibits long term stability on storage of 6 months or greater at room temperature.
• the lyophilized monoclonal antibody product of the present invention is generally prepared as follows: to an aqueous solution containing monoclonal antibody in an appropriate concentration, there is added the disaccharide HES in an amount sufficient to make a concentration of not less than 1 mg/ml, preferably 5 to 65 mg/ml. The above mentioned optional excipients may also be added . The therapeutic antibody product is then lyophilized in accordance with the above mentioned procedure.
• ingredients may be included in the formulation of the product of the present invention. These may include buffers to affect the pH of the solution, wetting or emulsifying agents, antimicrobial agents, preservatives, surfactants, isotonizing agents and the like. Also, bulking agents such as sodium bicarbonate, lactose, maltose, mannitol, sorbitol, or dextrose may be included to improve the characteristics of the freeze-dried cake.
• buffers to affect the pH of the solution
• wetting or emulsifying agents such as sodium bicarbonate, lactose, maltose, mannitol, sorbitol, or dextrose
• lactose maltose
• mannitol mannitol
• sorbitol or dextrose
• the lyophilized monoclonal antibody composition of the present invention is preferably formulated for intravenous administration when reconstituted as a solution.
• EXAMPLE 1 Abciximab monoclonal antibody composition containing approximately 2 mg/ml total antibody is prepared in accordance with the procedures outlined in Coller et al., U.S. Pat. No. in US patent 5,770,198.
• sucrose HES in an amount sufficient to make a concentration of not less than 2 mg/ml, preferably 5 to 65 mg/ml and the resultant solution is then sterile filtered.
• the above mentioned optional excipients may also be added .
• the therapeutic antibody product is then lyophilized in accordance with the following procedure: 5 ml vials containing 2 ml drug product are filled and placed into the lyophilization chamber. Shelf temperature is lowered at 0.5 e -1 9 C/min. to -40° C. and held below -40 s C. for 2 hours.
• Chamber pressure is then reduced to 100 microns Hg, the shelf is ramped to 0° C. at 0.5° C./min. and held for 48 hours. Shelf temperature is then raised to approximately +30 Q C. at 0.5 9 C./min. and held for 12 hours. The chamber is then brought to atmospheric pressure with dry nitrogen and the vials are stoppered and removed from the chamber and sealed.
• the vial contents Prior to use in therapy, the vial contents are reconstituted with 5 ml of Sterile Water for Injection, physiological saline or injectable glucose solution to yield the original concentration of solids in the vial.

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• 2002
  • 2002-10-18 EP EP02773798A patent/EP1455822A4/en not_active Withdrawn
  • 2002-10-18 CA CA002466641A patent/CA2466641A1/en not_active Abandoned
  • 2002-10-18 JP JP2003543524A patent/JP2005508992A/ja active Pending
  • 2002-10-18 MX MXPA04004459A patent/MXPA04004459A/es unknown
  • 2002-10-18 WO PCT/US2002/033272 patent/WO2003041637A2/en not_active Application Discontinuation
  • 2002-11-08 TW TW091133319A patent/TW200303213A/zh unknown
  • 2002-11-11 AR ARP020104325A patent/AR037304A1/es not_active Application Discontinuation

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