KR20080104160A - Anti-igf-1r human monoclonal antibody formulation - Google Patents

Abstract

The present invention relates to an anti-IGF-1R human monoclonal antibody formulation, a process for the preparation and uses thereof. ® KIPO & WIPO 2009

Description

Anti-IFF-1R human monoclonal antibody formulation {ANTI-IGF-1R HUMAN MONOCLONAL ANTIBODY FORMULATION}

The present invention relates to anti-IGF-1R human monoclonal antibody formulations, methods of preparation and use thereof.

In one aspect, the invention relates to an IGF-1R formulation comprising the following at a pH of about 5.0 to about 7.0:

About 1 to about 150 mg / mL huMab IGF-1R,

From about 0.001 to about 1% of at least one surfactant, and

About 1 to about 100 mM buffer.

IGF-IR (Type 1 Insulin Growth Factor Receptor) is involved in promoting carcinogenic transformation, growth and survival of cancer cells. High levels of IGF-IR expression have been reported in a wide range of human malignancies. In addition, high levels of IGF-I and IGF-II expression have been noted in tumors and associated cell tissue cells, and can stimulate cancer cell growth in autocrine or paracrine methods. Epidemiological studies indicate that the upper quintile plasma levels of IGF-I correlate with increased risk for prostate cancer, colon cancer, lung cancer and breast cancer. In addition to its role in the proliferation of cancer cells, IGF-IR protects cells from apoptosis due to growth factor loss, adhesion loss or cytotoxic drug treatment.

One promising method of inhibiting the function of IGF-IR in cancer cells is to apply human anti-IGF-IR antibodies that bind to the extracellular domain of IGF-IR and inhibit receptor activity. These antagonistic, fully human monoclonal antibodies, represented by huMab IGF-1R, have been developed that specifically bind to human insulinogenic growth factor I receptor (IGF-IR) and inhibit signal transduction and proliferative function of receptors in cancer cells.

Antibodies included in the formulations of the invention are described for the first time in PCT Patent Application No. WO2005 / 005635, in which the applicant is the owner and in particular the content of the claims is incorporated herein by reference. As described in WO2005 / 005635, the antibody binds to IGF-IR and inhibits the binding of IGF-I and IGF-II to IGF-IR, characterized by the following:

a) is IgG1 isotype,

and 1,: b) IGF-I binding IC ₅₀ value for IGF-II binding to the inhibitory ratio of the IC ₅₀ value of inhibition of IGF-IR to IGF-IR of 1: 3 to 3

c) 80% at 5 nM IGF-IR phosphorylation concentration in a cell phosphorylation assay using HT29 cells in medium containing 0.5% heat inactivated fetal calf serum (FCS) when compared to an assay without the antibody. Above, preferably at least 90%, and

d) Cell phosphorylation assay using 3T3 cells that provide 400,000 to 600,000 molecules IGF-IR per cell in 0.5% heat inactivated fetal calf serum (FCS) containing medium when compared to assays without said antibody. It does not show IGF-IR stimulatory activity measured as pkB phosphorylation at a concentration of 10 μM at.

Antibodies included in the formulations according to the invention show benefits for patients in need of anti-tumor therapy and provide a reduction in tumor growth and large prolongation of the time to progress. Antibodies included in the formulations according to the invention have new and unique properties that bring benefits for patients suffering from diseases associated with IGF control abnormalities, in particular tumor diseases. Antibodies included in the formulations of the present invention are characterized by the characteristics described above. The specific binding properties of the IgG1 isotype, in particular to IGF-IR, inhibit the binding of IGF-I and IGF-II to IGF-IR in the above-mentioned ratios, and IGF- at a concentration 200 times the IC ₅₀ value. It does not activate IGF-IR signals even in IR overexpressing cells. Antibodies that do not have "IGF-I symptomatic activity" offer strong advantages when used as therapeutic agents.

The term “anti-IGF-1R human monoclonal antibody” or “huMAb IGF-IR” especially refers to an antibody described or claimed in WO2005 / 005635, the content of which is hereby incorporated by reference.

The term "antibody" encompasses various forms of antibodies, including, but not limited to, whole antibodies, antibody fragments, human antibodies, humanized antibodies, and genetically modified antibodies, so long as the characteristic properties according to the invention are retained.

An "antibody fragment" comprises a portion of a full length antibody, generally at least an antibody binding portion or variable portion thereof. Examples of antibody fragments include diabodies, single-chain antibody molecules, immunotoxins, and multispecific antibodies formed from antibody fragments. The antibody fragment may also be a single chain having the characteristics of the VH chain, that is, capable of binding together with the VL chain, or of the VL chain that binds to IGF-1R, that is, the characteristic of being able to bind a functional antigen binding pocket with the VH chain. A polypeptide, thereby providing properties that inhibit binding of IGF-I and IGF-II to IGF-IR.

"Antibody fragments" also include fragments which themselves are incapable of providing effector function (ADCC / CDC) but which, in combination with the appropriate antibody constant region (s), provide such function in a manner according to the invention.

As used herein, the term "monoclonal antibody" or "monoclonal antibody composition" refers to the preparation of an antibody molecule of a single amino acid composition. The term “human monoclonal antibody” thus refers to an antibody that exhibits single binding specificities, with variable and constant regions derived from human germline immunoglobulin sequences. In one embodiment, the human monoclonal antibody is prepared by hybridoma, which is a transgenic non-human animal example having a genome comprising a human heavy chain transgene and a human light chain transgene fused to immortalized cells For example, B cells obtained from a transgenic mouse.

The term "chimeric antibody" refers to a monoclonal antibody comprising a variable region, or binding site, from one source or species and at least some of the constant regions derived from different sources or species, usually produced by recombinant DNA technology. . Especially preferred are chimeric antibodies comprising murine variable regions and human constant regions. Such murine / human chimeric antibodies are the product of expressed immunoglobulin genes comprising DNA segments encoding human immunoglobulin constant regions and DNA segments encoding murine immunoglobulin variable regions. Another form of “chimeric antibody” encompassed by the present invention is that in a class or subclass that has been modified or changed from that of the original antibody. Such "chimeric" antibodies also refer to "antibody-changed antibodies". Methods of making chimeric antibodies include conventional recombinant DNA and gene transduction techniques that are now well known in the art. For example, Morrison, S.L., et al., Proc. Natl. Acad Sci. USA 81 (1984) 6851-6855; See US Pat. Nos. 5,202,238 and 5,204,244.

The term “humanized antibody” refers to an antibody having a “complementarity determining site” (CDR) or framework that has been modified to include CDRs of immunoglobulins of different specificity compared to that of the parent immunoglobulin. In a preferred embodiment, murine CDRs are implanted into the backbone of a human antibody to produce a “humanized antibody”. See, for example, Riechmann, L., et al., Nature 332 (1988) 323-327; And Neuberger, M.S., et al., Nature 314 (1985) 268-270. Particularly preferred CDRs correspond to those representing sequences recognizing the antibodies described above for chimeric and bifunctional antibodies.

As used herein, the term “human antibody” is intended to include antibodies having variable and constant regions derived from human germline immunoglobulin sequences. The variable heavy chain is preferably derived from germline sequence DP-50 (GenBank LO6618) and the variable light chain is preferably derived from germline sequence L6 (GenBank X01668). The constant portion of an antibody is a constant portion of human IgG1 type. Such sites may be allogeneic and are described, for example, in Johnson, G., and Wu, T. T., Acids Res. 28 (2000) 214-218 and the databases referenced therein, as long as the introduction properties of the ADCC and preferably the CDC according to the invention are maintained.

As used herein, the term "recombinant human antibody" refers to any human antibody prepared, expressed, produced or isolated by recombinant means such as an antibody isolated from a host cell, such as SP2-0, NS0 or CHO cells, or It is intended to include antibodies isolated from an animal (eg, a mouse) that is transgenic for a human immunoglobulin gene or antibody expressed using a recombinant expression vector transduced into a host cell. Such recombinant human antibodies have variable and constant regions derived from human germline immunoglobulin sequences in rearranged form. Recombinant human antibodies according to the present invention become somatic hypermutations in vivo. Thus, the amino acid sequences of the VH and VL regions of recombinant antibodies are derived from and related to human germline VH and VL sequences, but may not be naturally present in the human antibody germline list in vivo.

As used herein, “binding” refers to antibody binding to IGF-IR, having an affinity of about 10 ⁻¹³ to 10 ⁻⁸ M (K _D ), preferably about 10 ⁻¹³ to 10 ⁻⁹ M. Indicates.

As used herein, the term “nucleic acid molecule” is intended to include DNA molecules and RNA molecules. The nucleic acid molecule may be single helix or double helix, but is preferably double helix DNA.

"Constant regions" are not directly involved in the binding of antibodies to antigens but are included in effector functions (ADCC, complement binding and CDC). The constant region of the antibody according to the invention is a constant region of IgG1 type. Human constant regions having these characteristics are described by Kabat et al., Sequence of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD. (1991) and Bruggemann, M., et al., J. Exp. Med. 166 (1987) 1351-1361; Love, T. W., et al., Methods Enzymol. 178 (1989) 515-527. An example is shown in SEQ ID NO: 5 to 8 in WO 2005/005635. Another useful and preferred constant region is the constant region of an antibody obtainable from hybridoma cells deposited in DSMZ for this invention. The constant regions useful in this invention provide complement binding. The combination of variable and constant regions provides ADCC and optionally CDC.

As used herein, “variable portion” (variable portion of the light chain (VL), variable portion of the heavy chain (VH)) refers to each pair of light and heavy chains that are directly involved in antibody binding to the antigen. The variable human light and heavy chain regions have the same general structure and each region comprises four framework regions (FRs), the sequence of which is widely conserved and in three "hypervariable regions" (or complementarity determining regions, CDRs). Is connected by. The backbone takes the form of β-sheets and the CDRs may form loops connecting the β-sheet structures. CDRs in each chain occur in a three-dimensional structure by the backbone and together with CDRs from the other chain form an antigen binding site. Antibody heavy and light chain CDR3 sites play a particularly important role in the binding specificity / affinity of the antibodies according to the invention, thus providing further objects of the invention.

As used herein, the term "hypervariable" or "antigen binding site of an antibody" refers to an amino acid residue of an antibody that plays a role in antigen binding. Hypervariables include amino acid residues from “complementarity determining sites” or “CDRs”. A "skeleton" or "FR" site is a variable region site except the hypervariables as defined herein. Thus, the light and heavy chains of the antibody include the N- to C-terminal regions FR1, CDR1, FR2, CDR2, FR3, CDR3 and FR4. In particular, CDR3 of the heavy chain is the site most contributing to antigen binding. CDR and FR sites are described by Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD. (1991)] and / or residues from a "hypervariable loop".

As used herein, the term "binding to IGF-IR" refers to an in vitro assay, preferably wherein the antibody binds to the surface and the binding of IGF-IR is measured by Surface Plasmon Resonance (SPR). In binding assays, means binding of the antibody to IGF-IR. Binding means a binding affinity (K _D ) of 10 ⁻⁸ M or less, preferably 10 ⁻¹³ to 10 ⁻⁹ M.

The BIAcore assay (Pharmacia Biosensor AB, Uppsala, Sweden) can test for binding to IGF-IR. The affinity of binding is defined by the terms ka (rate constant for binding of the antibody from the antibody / antigen complex), kd (dissociation constant) and K _D (kd / ka). Antibodies according to the invention exhibit a K _D of 10 ⁻¹⁰ M or less.

In addition, binding of IGF-I and IGF-II to IGF-IR is inhibited by the antibodies according to the invention. Inhibition is measured as IC ₅₀ in the assay for binding of IGF-I / IGF-II to IGF-IR on tumor cells. This assay is described in Example 7. In such assays, the amount of radioisotope labeled IGF-I or IGF-II or its IGF-IR binding fragments bound to IGF-IR, provided on the surface of the tumor cells (eg HT29), is determined by increasing antibody concentration or Measured as no increase. The IC ₅₀ value of the antibody according to the invention for the binding of IGF-I and IGF-II to IGF-IR is 2 nM or less and the ratio of the IC ₅₀ value for binding of IGF-I / IGF-II to IGF-IR is About 1: 3 to 3: 1. The IC ₅₀ value is measured as the average or median of three or more independent measurements. A single IC ₅₀ value can be out of range.

As used herein, the expression “inhibits the binding of IGF-I and IGF-II to IGF-IR” refers to I ¹²⁵ in IGF-IR presented on the surface of HT29 (ATCC HTB-38) tumor cells in an in vitro assay. -Inhibits the binding of labeled IGF-I or IGF-II. Inhibiting means an IC ₅₀ value of 2 nM or less.

As used herein, the term "surfactant" refers to a pharmaceutically acceptable surfactant. In the formulations of the invention, the amount of surfactant is described as a percentage expressed in weight / volume. The most commonly used weight / volume unit is mg / mL. Suitable pharmaceutically acceptable surfactants include, but are not limited to, polyethylene-sorbitan-fatty acid esters, polyethylene-polypropylene glycols, polyoxyethylene-stearates, and sodium dodecyl sulfate. Preferred polyethylene-sorbitan is polyethylene (20) -sorbitan-ester (synonymous with Polysorbate 20 available under the registered trademark of Tween 20 ^™ ) and polyoxyethylene (20) sorbitan monooleate (Tween 80 ^™ Synonymous with Polysorbate 80, which is a registered trademark. Preferred polyethylene-polypropylene glycol is commercially available under the name of Pluronic ^® F68 or Poloxamer 188 ^TM. Preferred polyoxyethylene-stearates are those sold under the registered trademark of Myrj ^™ . Preferred polyoxyethylene monolauryl ethers are those sold under the registered trademark of Brij ^™ . When using polyethylene-sorbitan-polyethylene (20) -sorbitan-ester (Tween 20 ^™ ) and polyoxyethylene (20) sorbitan monooleate (Tween 80 ^™ ), generally from about 0.001 to about 1%, Preferably from about 0.005 to about 0.1% and more preferably from about 0.01% to about 0.02% w / v.

As used herein, the term "buffer" refers to a pharmaceutically acceptable buffer. Suitable pharmaceutically acceptable buffers include, but are not limited to, histidine-buffer, citrate-buffer, succinate-buffer, acetate-buffer and phosphate-buffer. Preferred buffers include L-histidine or a mixture of L-histidine and L-histidine hydrochloride, pH-adjusted with isotonic agents and acids or salts known in the art. The histidine-buffer described above is generally used in an amount of about 1 mM to about 100 mM, preferably about 5 mM to about 50 mM and even more preferably about 20 mM. Independently from the buffer used, the pH is adjusted to a value comprising about 5.0 to about 7.0 and preferably about 5.5 to about 6.5 and more preferably about 6.0.

As used herein, the term "extinguishing agent" refers to a pharmaceutically acceptable tonicity agent. Isotonic agents are used to provide isotonic formulations. Isotonic formulations are liquids or liquids that are reconstituted from a solid form, for example a lyophilized form, and represent some other comparative solution such as physiological saline and a solution having the same tonicity as serum. Suitable isotonic agents include, but are not limited to, sodium chloride, potassium chloride, glucose, glycerin, and any component from the group of amino acids, sugars, and combinations thereof. Isotonic agents are generally used in total amounts from about 5 mM to about 350 mM.

As used herein in connection with a formulation according to the invention, the term "liquid" refers to a formulation that is liquid at temperatures of about 2 to about 8 ° C or more.

As used herein in connection with the formulations according to the invention, the term "freeze-dried" refers to the freezing of the formulation by any freeze-drying method known in the art, for example by a commercially available freeze-drying apparatus, from the frozen contents. Shows the formulation dried by sublimation of ice.

As used herein, the term "amino acid" includes but is not limited to arginine, glycine, ornithine, lysine, histidine, glutamic acid, aspartic acid, isoleucine, leucine, alanine, phenylalanine, tyrosine, tryptophan, methionine, serine, proline Amino acids in an amount from about 1 to about 100 mg / mL.

As used herein, the term "sugar" refers to pharmaceutically acceptable salts used in amounts of about 25 mM to about 500 mM. Suitable sugars include trehalose, sucrose, lactose, glucose, mannose, maltose, galactose, fructose, sorbose, raffinose, glucosamine, N-methylglucosamine (called "meglumine"), galactosamine and nu Including but not limited to laminic acid.

The term "stabilizer" refers to, for example, but is not limited to, the aforementioned amino acids and sugars, as well as commercially available cyclodextrins and dextrans of any kind and molecular weight known in the art. Does not.

The term "antioxidant" refers to a pharmaceutically acceptable antioxidant.

As mentioned above, in one aspect, the present invention relates to an IGF-1R formulation comprising the following at a pH of about 5.0 to about 7.0:

About 1 to about 150 mg / mL huMab IGF-1R,

From about 0.001 to about 1% of at least one surfactant, and

About 1 to about 100 mM buffer.

Formulations according to the invention preferably comprise from about 0.001 to about 1% of one or more surfactants.

In certain embodiments, formulations according to the invention comprise the following having a pH of about 5.0 to about 7.0:

About 1 to about 150 mg / mL huMab IGF-1R,

From about 0.005 to about 0.05% of at least one surfactant, and

About 1 to about 100 mM buffer.

The formulations according to the invention may be in liquid form, in lyophilized form or in liquid form reconstituted from lyophilized form.

In certain embodiments, formulations according to the invention are lyophilized formulations. The lyophilized formulations according to the invention provide the advantage of improved stability with respect to the formation of particulates and high molecular weight aggregates (which are difficult to obtain normally in liquid formulations at the same concentration of the described anti-IGF-1R human monoclonal antibody). Have

The formulations according to the invention can be administered by intravenous injection (i.v.), subcutaneous injection (s.c.) or any other parenteral administration method such as methods known in the pharmaceutical art.

Formulations of the present invention may further comprise one or more isotonic agents in an amount from about 5 mM to about 350 mM. Suitable isotonic agents can be selected from the group consisting of sodium chloride (NaCl), potassium chloride, sugars including glucose, glycerin, amino acids and combinations thereof.

Formulations of the present invention may further comprise a sugar in an amount from about 25 mM to about 500 mM. Suitable sugars are in the group consisting of trehalose, sucrose, lactose, glucose, mannose, maltose, galactose, fructose, sorbose, raffinose, glucosamine, N-methylglucosamine, galactosamine, neuramic acid and combinations thereof. Can be selected.

Formulations of the present invention may further comprise one or more of the following components: antioxidants, ascorbic acid, glutathione, preservatives such as m-cresol, phenol, benzyl alcohol, methylparaben, propylparaben, chlorbutanol, thio Mersal, benzalkonium chloride, cyclodextrins such as hydroxypropyl-β-cyclodextrin, sulfobutylethyl-β-cyclodextrin, β-cyclodextrin, polyethyleneglycol for example PEG 3000, 3350, 4000, 6000, Albumin, human serum albumin (HSA), bobbin serum albumin (BSA), polyhydric alcohol, glycerol, ethanol, mannitol, salts, acetate salts (e.g. sodium acetate), magnesium chloride, calcium chloride, tromethamine, EDTA (e.g. For example Na-EDTA).

The formulations of the present invention may further comprise one or more stabilizers as described above, and components known in the art as "lyoprotectants" such as sugars, sugar alcohols, amino acids and dextran known in the art. have.

In certain embodiments, the formulations of the invention comprise the following formulations as liquids, reconstituted from lyophilized or lyophilized forms:

About 1 to about 150 mg / mL huMab IGF-1R,

0.01% twin 20 w / v,

20 mM L-histidine,

140 mM NaCl,

pH 6.0.

Formulations according to the invention also include the following specific formulations:

25 mg / mLhuMab IGF-1R,

0.01% polysorbate 20,

20 mM L-histidine,

-140 mM NaCl,

pH 6.0.

or

25 mg / mLhuMab IGF-1R,

0.03% polysorbate 20,

20 mM L-histidine,

140 mM NaCl,

pH 6.0.

or

25 mg / mLhuMab IGF-1R,

0.05% polysorbate 20,

20 mM L-histidine,

140 mM NaCl,

pH 6.0.

or

10 mg / mL huMab IGF-1R,

0.01% polysorbate 20,

20 mM L-histidine,

140 mM NaCl,

pH 6.0.

or

10 mg / mL huMab IGF-1R,

0.03% polysorbate 20,

20 mM L-histidine,

140 mM NaCl,

pH 6.0.

or

10 mg / mL huMab IGF-1R,

0.05% polysorbate 20,

20 mM L-histidine,

140 mM NaCl,

pH 6.0.

or

25 mg / mLhuMab IGF-1R,

20 mM L-histidine,

140 mM NaCl,

pH 5.5.

or

25 mg / mLhuMab IGF-1R,

0.01% polysorbate 20,

20 mM L-histidine,

250 mM trehalose dihydrate,

pH 5.5.

or

25 mg / mLhuMab IGF-1R,

20 mM L-histidine,

250 mM trehalose dihydrate,

pH 6.0.

or

25 mg / mLhuMab IGF-1R,

0.01% polysorbate 20,

20 mM L-histidine,

250 mM trehalose dihydrate,

pH 6.0.

or

25 mg / mLhuMab IGF-1R,

20 mM L-histidine,

250 mM trehalose dihydrate,

0.05% polysorbate 20,

pH 6.0.

or

25 mg / mLhuMab IGF-1R,

20 mM L-histidine,

60 mM trehalose dihydrate,

0.01% polysorbate 20,

pH 6.0.

or

25 mg / mLhuMab IGF-1R,

20 mM succinate,

250 mM trehalose dihydrate,

0.01% polysorbate 20,

pH 5.5.

or

25 mg / mLhuMab IGF-1R,

20 mM L-histidine,

60 mM trehalose dihydrate,

0.01% polysorbate 20,

pH 6.0.

In a preferred embodiment according to the invention, the formulation is in lyophilized form and after reconstitution with an appropriate amount of water for injection:

25 mg / mLhuMab IGF-1R,

0.01% polysorbate 20,

20 mM L-histidine,

250 mM trehalose dihydrate,

pH 5.5.

Such formulations show good stability when stored at 2-8 ° C. and 25 ° C. for about 6 months with high molecular weight species of less than 1.5% without the formation of visible and invisible particles. Shaking and multiple freeze-thaw steps were applied to the liquid formulation to stimulate it by physical stress conditions potentially occurring during manufacture, such as pressure filtration and filling, freeze drying and termination processes. This formulation was found to be stable after 1 week shaking at 5 ° C and 25 ° C. The formation of visible and invisible particles could not be detected, and no significant increase in the fraction of high molecular weight species indicating the formation of soluble aggregates was observed.

Intravenous liquid and lyophilized drug formulations according to the invention were developed as follows:

Preparation of Liquid Formulations

About 10 and 25 mg / mL huMab IGF-IR solutions in preparation buffer (about 10 mM histidine buffer with about 150 mM NaCl) were dialyzed with a large amount of water and each buffer salt system for final formulation (exact formulation See table for composition). If necessary, the protein concentration was increased by filtration using a commercially available centrifugal filter device prior to dialysis and then adjusted to the desired protein concentration by dilution with dialysis buffer. If necessary, sugars and salts were added to the dialysis buffer to stabilize proteins and adjust their tonicity. Surfactant was added to the formulation after dialysis as a 2-40 fold stock solution. Alternatively, buffer exchange and concentration were performed using a commercially available tangential flow filtration device such as AKTA CF (manufactured by GE Healthcare) with Sartorius Hydrosart membrane (30,000 Da molecular weight separation). Buffer exchange using an appropriate amount of concentrated stock solution followed by addition of components such as sugars, salts or surfactants. All formulations were sterile filtered through a 0.22 μm low protein binding filter and aseptically aliquoted into sterile 6 mL glass vials sealed with Teflon coated rubber stoppers and AluCrim cap. These formulations were stored at different temperatures at different time intervals and removed for analysis at the time points indicated in the individual paragraphs in the following manner: 1) UV spectroscopy, 2) size exclusion chromatography (SEC) and 3) turbidity of the solution. Light shielding for measurement. In addition, analysis of visible particles was performed on each sample using the Seidenader V90-T instrument. The incidence of invisible particles was assessed with a HIAC Royco apparatus.

Preparation of Lyophilized Formulations

A solution of 25 mg / mL huMab IGF-1R was prepared as described above for the liquid formulation. All formulations were sterile filtered through a 0.22 μm low protein binding filter and aseptically aliquoted into sterile glass vials. The vial was partially sealed with a Teflon coated rubber stopper suitable for use in the freeze drying method and transferred to the drying chamber of the freeze dryer. Any lyophilization method known in the art is intended to be within the scope of the present invention. For example, the lyophilization method used in this study involves cooling the formulation at room temperature to about 5 ° C. (preliminary cooling) and then freezing at −40 ° C. (freezing I) at a rate ramp of about 1 ° C./min to 5 ° C./min. It included. The first drying step may be applied at a rate ramp of 0.3 to 0.5 ° C./min from −40 ° C. to −30 ° C. and then maintained at −30 ° C. (for at least 50 hours at a drying chamber pressure of about 75 to 80 mTorr). The second drying step can be carried out at a rate ramp of 0.1 to 0.3 ° C./min from −30 ° C. to 25 ° C. and maintained at 25 ° C. (for at least 5 hours at a real pressure of about 50 to 80 mTorr). HuMab IGF-IR formulations dried using the lyophilization method described above were found to have a reasonably fast reconstitution time of less than about 5 minutes. Freeze drying was performed in a LyoStar II freeze dryer (FTS System, Stone Ridge, NY, USA and Usifroid Orion, Maurepas, France). All lyophilized cakes in this study had a residual moisture content of about 0.1-5.0% as measured by Karl-Fischer method. Freeze dried vials were stored at different temperatures for different time intervals. The lyophilized formulation was reconstituted with each volume of water for injection (WFI) and analyzed by: 1) UV spectrophotometry, 2) measurement of reconstitution time, 3) size exclusion chromatography (SEC) and 4) solution Light shielding for turbidity measurements. In addition, analysis of visible particles was performed on each sample using a Seidenader V90-T instrument (Seidenader, Marktschwaben, Germany). The incidence of invisible particles was assessed with a HIAC Royco apparatus.

Size exclusion chromatography (SEC) was performed to detect soluble high molecular weight species (aggregates) and low molecular weight hydrolysis products in the formulation. The method used a Waters Alliance 2795 (detection wavelength [lambda] (280 nm)) with a Merck Hitachi 7000 HPLC instrument or UV detector. Both instruments were equipped with TSK G3000 SWXL columns; The method used 0.2MK ₂ HPO ₄ / 0.25M KCL with pH 7.0 as the mobile phase. Flow rate was 0.5 mL / min (isosolvent), run time 30 min at 25 ° C. column temperature. After diluting the samples with a Uvikon 932 (manufactured by Kontron Instruments) at a wavelength of 278 nm and a concentration of 0.5 mg / mL antibody respectively at 280 nm with a Varian Cary Bio UV spectrophotometer, UV spectroscopy was performed to determine protein concentration. To measure turbidity, light shielding was measured in FTU (turbidity units) using a HACH 2100AN turbidimeter at room temperature.

Liquid according to the invention huMAb IGF Composition of -1R drug formulation and stability data after 3 months storage at 2-8 ° C

Formulation A is a liquid formulation with a composition of 25 mg / mL huMab IGF-IR, 20 mM L-histidine, 140 mM NaCl, 0.01% polysorbate 20 at pH 6.0.

Viewpoint Protein concentration High molecular weight species (%)  Turbidity as the FTU (turbidity) Early 24.47 0.82 9.94 4 Weeks 22.35 1.0 10.3 8 Weeks 23.02 1.31 9.83 12 Weeks 23.59 1.46 9.22

Formulation B is a liquid formulation with a composition of 25 mg / mL huMab IGF-IR, 20 mM L-histidine, 140 mM NaCl, 0.03% polysorbate 20 at pH 6.0.

Viewpoint Protein concentration High molecular weight species (%) Turbidity as FTU Early 24.98 0.84 9.79 4 Weeks 23.58 1.01 9.97 8 Weeks 25.43 1.37 10.5 12 Weeks 23.68 1.49 10.3

Formulation C is a liquid formulation with a composition of 25 mg / mL huMab IGF-IR, 20 mM L-histidine, 140 mM NaCl, 0.05% polysorbate 20 at pH 6.0.

Viewpoint Protein concentration High molecular weight species (%) Turbidity as FTU Early 24.42 0.85 10.1 4 Weeks 21.57 1.02 10.5 8 Weeks 25.0 1.36 10.6 12 Weeks 23.97 1.51 10.2

Formulation D is a liquid formulation with a composition of 10 mg / mL huMab IGF-IR, 20 mM L-histidine, 140 mM NaCl, 0.01% polysorbate 20 at pH 6.0.

Viewpoint Protein concentration High molecular weight species (%) Turbidity as FTU Early 9.87 0.63 4.42 4 Weeks 9.61 0.75 4.44 8 Weeks 9.72 0.97 4.53 12 Weeks 9.63 1.11 4.24

Formulation E is a liquid formulation with a composition of 10 mg / mL huMab IGF-IR, 20 mM L-histidine, 140 mM NaCl, 0.03% polysorbate 20 at pH 6.0.

Viewpoint Protein concentration High molecular weight species (%) Turbidity as FTU Early 9.98 0.69 4.26 4 weeks 9.68 0.79 4.72 8 Weeks 9.68 1.05 4.54 12 Weeks 9.62 1.59 4.26

Formulation F is a liquid formulation with a composition of 10 mg / mL huMab IGF-IR, 20 mM L-histidine, 140 mM NaCl, 0.05% polysorbate 20 at pH 6.0.

Viewpoint Protein concentration High molecular weight species (%) Turbidity as FTU Early 9.81 0.67 4.45 4 Weeks 9.69 0.83 4.36 8 Weeks 9.62 1.19 5.01 12 Weeks 9.18 1.45 4.44

Formulation G is a liquid formulation with a composition of 10 mg / mL huMab IGF-IR, 20 mM L-histidine, 140 mM NaCl at pH 5.5.

Viewpoint Protein concentration High molecular weight species (%) Turbidity as FTU Early 24.7 0.8 4.9 4 Weeks 24.6 1.0 3.9 8 Weeks n.d. n.d. n.d. 12 Weeks 24.8 0.8 6.1

Formulation H is a liquid formulation with a composition of 25 mg / mLhuMab IGF-IR, 20 mM L-histidine, 250 mM trehalose dihydrate, 0.01% polysorbate 20 at pH 5.5.

Viewpoint Protein concentration High molecular weight species (%) Turbidity as FTU Early 27.1 1.29 3.58 4 Weeks 26.9 1.27 3.74 8 Weeks 29.33 1.43 5.03 12 Weeks 26.44 1.38 4.10

Formulation I is a liquid formulation having a composition of 25 mg / mL huMab IGF-IR, 20 mM L-histidine, 250 mM trehalose dihydrate at pH 6.0.

Formulation J is a liquid formulation with a composition of 25 mg / mLhuMab IGF-IR, 20 mM L-histidine, 250 mM trehalose dihydrate, 0.01% polysorbate 20 at pH 6.0.

Formulation K is a liquid formulation with a composition of 25 mg / mLhuMab IGF-IR, 20 mM L-histidine, 250 mM trehalose dihydrate, 0.05% polysorbate 20 at pH 6.0.

Formulation L is a liquid formulation with a composition of 25 mg / mLhuMab IGF-IR, 20 mM L-histidine, 60 mM trehalose dihydrate, 0.01% polysorbate 20 at pH 6.0.

Formulation M is a liquid formulation with a composition of 25 mg / mL huMab IGF-IR, 20 mM succinate, 250 mM trehalose dihydrate, 0.01% polysorbate 20 at pH 5.5.

Viewpoint Protein concentration High molecular weight species (%) Turbidity as FTU Early 26.13 1.49 7.06 4 Weeks 27.93 1.46 7.25 8 Weeks 26.59 1.62 8.20 12 Weeks 26.89 1.62 7.10

Lyophilized according to the invention huMAb IGF Composition of -1R drug formulation and stability data after 3 months storage at 2-8 ° C

Formulation N is a lyophilized formulation having a composition of reconstituted solution of 25 mg / mL huMab IGF-IR, 20 mM L-histidine, 60 mM trehalose dihydrate, 0.01% polysorbate 20 at pH 6.0.

Viewpoint Protein concentration High molecular weight species (%) Turbidity as FTU Early 27.40 1.37 3.99 4 Weeks 27.89 1.36 4.20 8 Weeks 27.93 1.43 5.04 12 Weeks 27.34 1.38 4.31

Formulation O is a lyophilized formulation having a composition of reconstituted solution of 25 mg / mL huMab IGF-IR, 20 mM L-histidine, 60 mM sucrose, 0.01% polysorbate 20 at pH 6.0.

Viewpoint Protein concentration High molecular weight species (%) Turbidity as FTU Early 24.77 0.93 5.77 4 Weeks 24.81 0.93 6.79 8 Weeks 24.11 0.88 5.38 12 Weeks 24.32 1.03 3.93

Formulation P is a lyophilized formulation having a composition of reconstituted solution of 25 mg / mL huMab IGF-1R, 20 mM succinate, 250 mM trehalose dihydrate, 0.01% polysorbate 20 at pH 5.5.

Viewpoint Protein concentration High molecular weight species (%) Turbidity as FTU Early 26.9 1.45 7.17 4 Weeks 27.71 1.55 7.44 8 Weeks 26.59 1.43 7.13 12 Weeks 26.89 1.36 7.07

Formulation Q is a lyophilized formulation having a composition of a reconstituted solution of 25 mg / mL huMab IGF-IR, 20 mM L-histidine, 250 mM trehalose dihydrate, 0.01% polysorbate 20 at pH 5.5.

Viewpoint Protein concentration High molecular weight species (%) Turbidity as FTU Early 27.11 1.33 3.59 4 Weeks 28.28 1.22 3.49 8 Weeks 27.81 1.30 4.65 12 Weeks 28.22 1.18 4.04

Claims (14)

Formulations comprising the pH of about 5.0 to about 7.0: About 1 to about 150 mg / mL huMab IGF-1R, From about 0.001 to about 1% of at least one surfactant, and About 1 to about 100 mM buffer. The formulation of claim 1, comprising from about 0.001 to about 1% of at least one surfactant. The formulation according to claim 1, which is in liquid form, lyophilized form or liquid form reconstituted from lyophilized form. The formulation according to claim 1, which can be administered by intravenous injection (i.v.), subcutaneous injection (s.c.) or any other parenteral administration. The formulation of claim 1, further comprising at least one isotonic agent in an amount from about 5 mM to about 350 mM. 6. The formulation according to claim 5, wherein the tonicity agent is selected from the group consisting of sodium chloride (NaCl), potassium chloride, sugars comprising glucose, glycerin, amino acids and combinations thereof. The formulation of claim 1, further comprising a sugar in an amount from about 25 mM to about 500 mM. 8. The method of claim 7, wherein the sugar is trehalose, sucrose, lactose, glucose, mannose, maltose, galactose, fructose, sorbose, raffinose, glucosamine, N-methylglucosamine ("meglumine"), galactose Formulation selected from the group consisting of samine, neuramic acid. The formulation according to claim 1, further comprising at least one component selected from the group consisting of pharmaceutically acceptable: antioxidants, ascorbic acid, glutathione, preservatives in particular m-cresol , Phenol, benzyl alcohol, methyl paraben, propyl paraben, chlorbutanol, thiomersal, benzalkonium chloride, cyclodextrin in particular hydroxypropyl-β-cyclodextrin, sulfobutylethyl-β-cyclodextrin, β-cyclodextrin, Polyethyleneglycol, especially PEG 3000, 3350, 4000 or 6000, albumin, human serum albumin (HSA), bobbin serum albumin (BSA), polyhydric alcohols, glycerol, ethanol, mannitol, salts, acetate salts especially sodium acetate, magnesium chloride, calcium chloride , Tromethamine, EDTA and especially Na-EDTA. The formulation of claim 1, wherein the formulation comprises a pH 6.0: About 1 to about 150 mg / mL huMab IGF-1R, 0.01% twin 20 w / v, 20 mM L-histidine, 140 mM NaCl. The formulation of claim 1, wherein: 25 mg / mLhuMab IGF-1R, 0.01% polysorbate 20, 20 mM L-histidine, 140 mM NaCl, pH 6.0; or 25 mg / mLhuMab IGF-1R, 0.03% polysorbate 20, 20 mM L-histidine, 140 mM NaCl, pH 6.0; or 25 mg / mLhuMab IGF-1R, 0.05% polysorbate 20, 20 mM L-histidine, 140 mM NaCl, pH 6.0; or 10 mg / mL huMab IGF-1R, 0.01% polysorbate 20, 20 mM L-histidine, 140 mM NaCl, pH 6.0; or 10 mg / mL huMab IGF-1R, 0.03% polysorbate 20, 20 mM L-histidine, 140 mM NaCl, pH 6.0; or 10 mg / mL huMab IGF-1R, 0.05% polysorbate 20, 20 mM L-histidine, 140 mM NaCl, pH 6.0; or 25 mg / mLhuMab IGF-1R, 20 mM L-histidine, 140 mM NaCl, pH 5.5; or 25 mg / mLhuMab IGF-1R, 0.01% polysorbate 20, 20 mM L-histidine, 250 mM trehalose dihydrate, pH 5.5; or 25 mg / mLhuMab IGF-1R, 20 mM L-histidine, 250 mM trehalose dihydrate, pH 6.0; or 25 mg / mLhuMab IGF-1R, 0.01% polysorbate 20, 20 mM L-histidine, 250 mM trehalose dihydrate, pH 6.0; or 25 mg / mLhuMab IGF-1R, 20 mM L-histidine, 250 mM trehalose dihydrate, 0.05% polysorbate 20, pH 6.0; or 25 mg / mLhuMab IGF-1R, 20 mM L-histidine, 60 mM trehalose dihydrate, 0.01% polysorbate 20, pH 6.0; or 25 mg / mLhuMab IGF-1R, 20 mM succinate, 250 mM trehalose dihydrate, 0.01% polysorbate 20, pH 5.5; or 25 mg / mLhuMab IGF-1R, 20 mM L-histidine, 60 mM trehalose dihydrate, 0.01% polysorbate 20, pH 6.0. Use of a formulation according to any one of claims 1 to 11 for the manufacture of a medicament useful for treating a disease modulated by an IGF-IR receptor. 13. The use according to claim 12, wherein the disease is selected from the group consisting of breast cancer, colon cancer, non-small cell lung cancer (NSCLC) and prostate cancer. The invention described above.